Reno-Hepatoprotective effects of Bryophyllum pinnatum against Hepatic Ischemia- Reperfusion Injury in male Wistar rats

Reno-Hepatoprotective effects of Bryophyllum pinnatum against Hepatic Ischemia- Reperfusion Injury in male Wistar rats | Research Square window.SnipcartSettings = { analytics: { enabled: false } }; (function() { var accessVector = localStorage.getItem('access_vector') || ''; window.dataLayer = window.dataLayer || []; if (accessVector) { window.dataLayer.push({ user: { profile: { profileInfo: { snid: accessVector } } } }); } })(); (function(w,d,s,l,i){w[l]=w[l]||[];w[l].push({'gtm.start':new Date().getTime(),event:'gtm.js'});var f=d.getElementsByTagName(s)[0],j=d.createElement(s),dl=l!='dataLayer'?'&l='+l:'';j.async=true;j.src='https://www.googletagmanager.com/gtm.js?id='+i+dl;f.parentNode.insertBefore(j,f);})(window,document,'script','dataLayer','GTM-K279D39R'); Browse Preprints In Review Journals COVID-19 Preprints AJE Video Bytes Research Tools Research Promotion AJE Professional Editing AJE Rubriq About Preprint Platform In Review Editorial Policies Our Team Advisory Board Help Center Sign In Submit a Preprint Cite Share Download PDF Research Article Reno-Hepatoprotective effects of Bryophyllum pinnatum against Hepatic Ischemia- Reperfusion Injury in male Wistar rats Oluwakemi V. Olayinka, Oladele A. Afolabi, Richard A. Ajike, Opeyemi S. Hammed, and 4 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-6205039/v1 This work is licensed under a CC BY 4.0 License Status: Posted Version 1 posted You are reading this latest preprint version Abstract Hepatic ischemia-reperfusion injury (HIRI) is indicated in postoperative complications in hepatic transplantation and surgeries. HIRI can lead to remote renal injury through systemic inflammatory and oxidative cascades. While current therapeutic options are effective, their potential toxic effects can pose additional challenges, thereby hindering hepatic and renal recovery, and highlighting the need for alternative therapies. Bryophyllum pinnatum (BP), popularly called “life plant” is widely used in traditional medicine, and is rich in flavonoids, triterpenoids, and bufadienolides, which have shown remarkable antioxidant and anti-inflammatory activities in various animal models. Studies have demonstrated the ability of BP to scavenge free radicals, suppress acute inflammation, and inhibit pro-inflammatory cytokines. Its nephroprotective effects in renal injury models, as well as its hepatoprotective effects in chemically-induced hepatocarcinogenesis have also been documented. Despite these promising findings, its effect against HIRI-induced kidney damage remains unexplored. This study investigates the reno-hepatoprotective potential of BP methanolic leaf extract in male Wistar rats subjected to HIRI. Forty male Wistar rats were divided into four groups (n=10); group 1- Sham operated, group 2-HIRI, group 3-LDH (low-dose B. pinnatum (100 mg/kg), group 4-HDH (high-dose B. pinnatum (200 mg/kg). BP was administered orally via gavage for 14 days before HIRI induction. Following treatment, animals were anesthetized with ketamine (50 mg/kg) and xylazine (10 mg/kg), and HIRI was induced by clamping the hepatic artery, portal vein and bile duct for 60 minutes to induce ischemia after which the clamp was removed for hepatic reperfusion which lasted for 24-hours. Animals were euthanized under deep anesthesia (ketamine 50 mg/kg and xylazine 10 mg/kg, ip) before sacrifice. Then, blood, kidney and liver tissues were harvested for biochemical and histological analysis. Data were analyzed using one way ANOVA with Graph pad prism. Tukey’s post-hoc test was used for multiple comparison. P<0.05 was considered statistically significant. Biochemical analyses showed a significant restoration in SOD, catalase, and GSH levels and reductions in MDA and inflammatory markers (TNF-α, MPO) in the B. pinnatum groups. In addition, altered ALT, ALP, and GGT levels were restored, and urea and creatine levels raised as a secondary effect of HIRI were reduced in the B. pinnatum -treated groups. Reduced caspase-3 activity was also observed in B. pinnatum -treated groups, indicating lower apoptosis levels. Histological analyses showed improved cytoarchitecture, with preservation of renal corpuscles and reduced inflammation. Conclusively, B. pinnatum exhibits reno-hepatoprotective effects on HIRI-induced kidney damage, potentially modulating oxidative stress, inflammation, and apoptotic pathways. Bryophyllum pinnatum hepatic ischemia-reperfusion injury reno-hepatoprotection oxidative stress Figures Figure 1 Figure 2 Figure 3 Figure 4 Figure 5 Figure 6 Figure 7 Figure 8 Figure 9 Figure 10 Figure 11 Figure 12 Figure 13 Figure 14 Figure 15 Figure 16 Figure 17 Figure 18 Figure 19 1 Introduction Ischemia-reperfusion injury (IRI) is characterized by a complex pathophysiological cascade that aggravates tissue damage during blood flow restoration after a period of ischemia. This phenomenon significantly impacts morbidity and mortality rates in clinical practice, particularly in organ transplantation and major surgical procedures ( 1 ). The liver exhibits particular vulnerability to IRI, making hepatic ischemia-reperfusion injury (HIRI) a major concern in hepatic surgery and liver transplantation ( 2 ). In the pathogenesis of HIRI, oxygen deprivation, during ischemia, triggers ATP depletion, which in turn disrupts ionic homeostasis and causes calcium overload. The reperfusion phase on the other hand, exacerbates injury through reactive oxygen species (ROS) generation, including superoxide anions and hydroxyl radicals, leading to cellular damage through lipid peroxidation and protein oxidation ( 3 ). Furthermore, oxidized phospholipids can act as damage-associated molecular patterns (DAMPs), which then go on to initiate inflammatory cascades that exacerbate the injury ( 4 ). The systemic effect of HIRI can also affect renal function, with reported incidences of acute kidney injury following orthotopic liver transplantation ranging from 12–94% ( 5 ). This hepatorenal interaction is traceable to multiple pathways, including Kupffer cell activation and subsequent pro-inflammatory cytokine release, such as, tumour necrosis factor-alpha (TNF-α) and interleukin-6 (IL-6), which promote systemic inflammation and remote organ injury ( 6 ). Additionally, sinusoidal endothelial cell damage promotes microthrombosis and vasoconstriction through von Willebrand factor release and reduced nitric oxide production ( 6 ). The damage to the kidneys in this context is mediated by oxidative stress, mitochondrial dysfunction, and heightened susceptibility to apoptosis; a combination that significantly impairs renal function ( 5 , 7 ). Current therapeutic strategies to mitigate HIRI-induced renal injury primarily focus on supportive measures and specific pharmacological interventions targeting oxidative stress and inflammation ( 8 ). Additionally, ischemic preconditioning and pharmacological approaches to reduce mitochondrial permeability transition pore (MPTP) formation remain limited by variability in individual response and operational complexities in surgical contexts ( 9 ). The limitations of these existing treatments, including timing constraints and potential side effects, necessitate the exploration of alternative therapeutic approaches, such as natural compounds with multi-targeted actions capable of modulating both oxidative and inflammatory pathways without the drawbacks of synthetic interventions ( 10 ). Among natural compounds, Bryophyllum pinnatum , is well known to possess therapeutic potential suitable for this study. In traditional medicine, Bryophyllum pinnatum has long been utilized for various ailments, including liver and kidney disorders ( 11 , 12 ). Its phytochemical analyses reveal it to be rich in bioactive compounds, including flavonoids, triterpenoids, and bufadienolides ( 13 ). These constituents demonstrate significant antioxidant and anti-inflammatory properties, particularly in suppressing pro-inflammatory cytokine production and nuclear factor-kappa B activation ( 14 , 15 ). In a landmark study by Ojewole ( 11 ) to scientifically evaluate the ethnomedicinal applications of B. pinnatum leaves, the author concluded that aqueous extract of B. pinnatum leaves possessed anti-inflammatory properties, evidenced by the significant inhibition of fresh egg albumin-induced acute inflammation, as well as antinociceptive and anti-diabetic properties. Chibli et al . ( 16 ) also demonstrated B. pinnatum ’s anti-inflammatory property in acute and chronic mice ear edema models induced by different irritant agents. Recent studies have further shown B. pinnatum 's capacity to scavenge free radicals and inhibit lipid peroxidation, due to active components such as quercetin, kaempferol, and bryophyllins ( 17 ). In a study by Tatsimo et al . ( 18 ), methanolic extracts of B. pinnatum showed strong antioxidative and antimicrobial activities, confirming their traditional use in the treatment of infectious and free radical damages. A recent review by Sharma et al. ( 19 ) indicated that B. pinnatum is effective in the treatment of kidney and liver diseases, and possesses anti-leishmanial, neuropharmacological, antibacterial, immunosuppressive, anti-tumour, and cytotoxic activities. Its nephroprotective effect is confirmed in the study by Yadav et al . ( 20 ) which showed in their study on the effect of B. pinnatum on ethylene glycol (EG)-induced renal calculi in rats, that B. pinnatum effectively reversed EG-induced damage comparable to the standard drug Cystone. Likewise, Anadozie et al. ( 21 ) concluded that B. pinnatum is able to inhibit arginase II activity and consequently prevent carbon tetrachloride (CCl4)-induced renal oxidative damage in rats. Other studies to examine the renoprotective effects of B. pinnatum, such as those by Shukla et al. ( 22 ), Shinde et al. ( 23 ), Uahomo and Isirima ( 24 ), and Pal et al. ( 25 ), all show that B. pinnatum is able to attenuate renal damage in experimental studies. Apart from having renoprotective properties, B. pinnatum has also been reported to possess hepatoprotective properties. Afzal et al . ( 26 ) concluded in their study that B. pinnatum is able to mitigate chemically-induced hepatocarcinogenesis due to its ability to upregulate antioxidant activities and scavenge free radicals that damage cells. Kombathula and Chernapalli ( 27 ) confirmed this in their study and attested that the plant’s components—bryophyllin A and β-sitosterol—strongly interact with the Pi3K and C-met pathways respectively to mediate their effect. In another model of butylglycol-induced hepatotoxicity, the researchers observed that groups treated with B. pinnatum showed reduced liver injury markers (ALP and AST) compared to the positive control ( 28 ). In addition, Ajah et al. ( 29 ) examined the efficacy of B. pinnatum in attenuating damage to the kidneys and liver, as well as the lipid profile of rats fed with high-salt diet. They concluded that the ethanolic extract of B. pinnatum was highly potent in preventing cell damage caused by high salt intake. However, its potential role in mitigating HIRI-induced kidney injury remains unexplored. Since Bryophyllum pinnatum ’s pleiotropic effects might provide comprehensive protection against both direct and remote organ injury through upregulation of antioxidant enzymes, modulation of oxidative stress, inflammation, and apoptotic pathways, this study aims to evaluate the reno-hepatoprotective effects of Bryophyllum pinnatum methanolic extract on kidney injury following hepatic ischemia-reperfusion in male Wistar rats. 2 Materials and Methods 2.1 Chemicals and Reagents All chemicals and reagents used in this study were of analytical grade. Ketamine and xylazine for anaesthesia were obtained from the Veterinary Department, University of Ibadan. Streptomycin injection, 0.9% normal saline, 10% formalin, and distilled water were procured from the Physiology Laboratory, LAUTECH. Biochemical assay kits for liver function markers (ALT, AST, GGT) and kidney function markers (urea, creatinine) were purchased from Randox Laboratories Ltd., UK. Stains for histological analysis, including Hematoxylin and Eosin, Masson's trichrome, and Periodic acid-Schiff (PAS), were also utilized. 2.2 Experimental Animals and Design Forty male Wistar rats weighing 120-150g were obtained from animal house of physiology department, LAUTECH, Ogbomoso, Oyo State, Nigeria. The animals were housed under standard laboratory conditions at the animal facility of Faculty of Basic Medical Sciences (FBMS), LAUTECH and were acclimatized for 2 weeks, fed with standard rat chow and watered ad libitum . Ethical approval was obtained from Ethics Committee in Animal Care and Use in Research of FBMS with reference number: ERCFBMSLAUTECH: 058/08/2024. The rats were randomly divided into four groups (n = 10 per group): Sham group : Sham-operated, no treatment HIRI group : Hepatic ischemia-reperfusion injury, no treatment BP Low Dose + HIRI : 100 mg/kg Bryophyllum pinnatum extract + Hepatic ischemia-reperfusion injury BP High Dose + HIRI : 200 mg/kg Bryophyllum pinnatum extract + Hepatic ischemia-reperfusion injury The Bryophyllum pinnatum extract was administered orally via gavage for 14 days prior to the induction of HIRI, as described by Anadozie et al . ( 21 ). 2.3 Plant Material Preparation Fresh Bryophyllum pinnatum leaves were collected from farm house of Faculty of Agricultural Sciences, Ladoke Akintola University of Technology (LAUTECH), Ogbomoso, Oyo State, Nigeria. The plant was identified and authenticated by Dr. Mrs. A.F. Ogundola, a botanist at the Department of Pure and Applied Biology, LAUTECH. The plant was assigned the voucher number LHO 747 , and kept in the Herbarium at the Department of Botany, LAUTECH, Ogbomoso, Nigeria. The leaves were washed, air-dried, and pulverized into a fine powder using an electric blender. The powdered leaves were macerated in 80% methanol for 72 hours, and the extract was concentrated using a rotary evaporator and further dried in a vacuum oven. 2.3.1 Phytochemical Profiling Qualitative screening of the Bryophyllum pinnatum extract was performed to detect the presence of major phytochemical groups, including alkaloids, flavonoids, tannins, saponins, terpenoids, glycosides, steroids, and anthraquinones, using standard chemical tests (Mayer's, Wagner's, alkaline reagent, ferric chloride, foam, Salkowski, Keller-Killiani, Liebermann-Burchard, and Borntrager's tests, respectively). For quantitative analysis, spectrophotometric methods were employed to determine the total flavonoid content (aluminium chloride colorimetric method), total tannin content (Folin-Ciocalteu method), and total phenolic content (Folin-Ciocalteu method) (Table 1 ). Table 1 Qualitative and selected Quantitative analyses of B. pinnatum Phytochemical Group Qualitative Result Quantitative Result (where applicable) Alkaloids +++ Not quantified Flavonoids +++ 24.3 ± 1.8 mg QE/g dry weight Tannins ++ 18.7 ± 1.2 mg TAE/g dry weight Saponins ++ Not quantified Terpenoids +++ Not quantified Phenolic compounds +++ 37.2 ± 2.5 mg GAE/g dry weight Glycosides ++ Not quantified Steroids + Not quantified Anthraquinones + Not quantified Key : +++ (strongly present), ++ (moderately present), + (weakly present), - (absent) QE: Quercetin Equivalent, TAE: Tannic Acid Equivalent, GAE: Gallic Acid Equivalent 2.4 HIRI Induction Procedure Hepatic ischemia-reperfusion injury (HIRI) was induced following the method described by Şener et al . ( 30 ) with slight modifications. Briefly, after overnight fasting, rats were anaesthetised with intraperitoneal injections of ketamine (50 mg/kg) and xylazine (10 mg/kg) ( 31 ). The abdominal area was shaved and disinfected, and a midline laparotomy was performed. The hepatic artery, portal vein supplying the median and left lateral lobes and the bile duct were clamped using microvascular clamp to induce 70% hepatic ischemia. After 60 minutes of ischemia, the clamp was removed to allow reperfusion which lasted for 24 hours. Sham-operated animals underwent the same procedure without vascular occlusion. 2.5 Sample Collection Animals were fasted overnight and twenty-four hours after reperfusion, the animals were anaesthetized using intraperitoneal injection of ketamine (50 mg/kg) and xylazine (10 mg/kg) ( 31 ). Confirmation of anaesthesia was determined by absence of corneal reflex and pedal withdrawal. Blood samples were collected via the retro-orbital puncture in plain bottles. Following this, the liver and kidneys were excised, washed in ice-cold saline, and weighed. Also, the collected blood was allowed to clot and centrifuged at 5000rpm for 10 minutes to obtain serum. Portions of the liver and kidney tissues were fixed in 10% neutral buffered formalin for histological examination. 2.6 Biochemical Analyses Serum and tissue homogenates were used for various biochemical assays. Liver function markers (ALT, AST, GGT) and kidney function markers (urea, creatinine) were determined using commercial kits on an automated analyzer (Biosystem A25 Random Access Analyzer). Oxidative stress markers , including superoxide dismutase (SOD) activity, malondialdehyde (MDA) levels, glutathione (GSH) content, and catalase activity, were measured in liver and kidney tissue homogenates using established spectrophotometric methods ( 32 , 33 , 34 , 35 ). Total protein content in the tissue homogenates was determined by the Bradford method ( 36 ). Inflammatory markers , including tumour necrosis factor-alpha (TNF-α) in serum and myeloperoxidase (MPO) activity in liver and kidney tissues, were assessed using ELISA and spectrophotometric techniques, respectively (RandD Systems; 37). Serum nitrite levels, as an indicator of nitric oxide production, were measured using the Griess reaction ( 38 ). Caspase-3 activity, a marker of apoptosis, was determined in liver and kidney tissue homogenates using a colorimetric assay kit (Sigma-Aldrich, USA). 2.7 Histological Examination Fixed liver and kidney tissues were processed using standard histological techniques and stained with Hematoxylin and Eosin (HandE) for general morphological assessment. Additional stains, including Masson's trichrome for liver fibrosis and Periodic acid-Schiff (PAS) for renal basement membrane and tubular brush border changes, were also performed. The stained sections were examined under a light microscope (Olympus BX51, Tokyo, Japan) and photographed. 2.8 Statistical Analysis Data were analysed using GraphPad Prism software (version 9.0, GraphPad Software Inc., San Diego, CA, USA). Results were expressed as mean ± standard error of the mean (SEM). Differences between groups were analysed using one-way analysis of variance (ANOVA) followed by Tukey's post hoc test for multiple comparisons. 3 Results 3.1 Liver Function Markers Alanine aminotransferase (ALT) There was significant increase in ALT concentration in the HIRI group compared with the sham group. Aspartate aminotransferase (AST) AST concentration was significantly increased in the HIRI group when compared with the sham operated group. Significant decrease in AST concentration was also observed in both the low-dose BP and high-dose BP groups. Gamma-glutamyl transferase (GGT) There was a statistically significant increase in the GGT concentration of the HIRI group when compared with the sham operated group. GGT was however significantly reduced in the high-dose BP group when compared with the sham and low-dose BP groups. 3.2 Kidney Function Markers Creatinine There was a statistically significant increase in creatinine levels of the HIRI group when compared with the sham operated group. The low-dose and high dose groups both significantly reduced creatinine levels when compared with the HIRI group. Urea There was a statistically significant increase in the urea concentration of the HIRI group when compared with the sham operated group. Urea concentration was however significantly reduced by both the low-dose BP and high-dose BP groups. Furthermore, the high-dose group significantly decreased urea concentration when compared with the low-dose group. 3.3 Hepatic Oxidative Stress Markers Hepatic Glutathione (GSH) Hepatic GSH was significantly reduced in the HIRI group when compared with the sham group. Also, both the low-dose BP and high-dose BP showed a significant increase in GSH concentration when compared to the HIRI group. Hepatic Total Protein Hepatic total protein was significantly reduced in the HIRI group when compared with the sham group. Also, both the low-dose BP and high-dose BP groups showed a significant increase in total protein concentration when compared to the HIRI group. Significant decrease was further observed in the high-dose BP group when compared with the low-dose BP group. Hepatic Malondialdehyde (MDA) MDA concentration significantly increased in the livers of the rats of the HIRI group when compared with the sham group. Both the low-dose BP and high-dose BP groups, however, were able to significantly reduce MDA concentration when compared to the HIRI group. Furthermore, there was a significant decrease in the high-dose BP group when compared with the low-dose BP group. Hepatic Superoxide dismutase (SOD) Hepatic SOD concentration was significantly decreased in HIRI group when compared with the sham group. In addition to this, both low-dose BP and high-dose BP groups showed a significant increase in SOD concentration when compared with the HIRI group. Hepatic Catalase There was a statistically significant decrease in hepatic catalase concentration in the HIRI group when compared with the sham group. There was also a statistically significant increase in catalase concentration in the low-dose BP and high-dose BP groups when compared with the HIRI group. 3.4 Renal Oxidative Stress Markers Renal Glutathione (GSH) Renal GSH was significantly reduced in the HIRI group when compared with the sham group. Also, both the low-dose BP and high-dose BP showed a significant increase in GSH concentration when compared to the HIRI group. Renal Total Protein Total protein was significantly reduced in the HIRI group when compared with the sham group. Also, both the low-dose BP and high-dose BP showed a significant increase in total protein concentration when compared to the HIRI group. R enal Malondialdehyde (MDA) Renal MDA concentration was significantly increased in the HIRI group when compared to the HIRI group. Furthermore, both low-dose BP and high-dose BP groups had a significantly reduced MDA concentration in comparison with the HIRI group. Renal Superoxide dismutase (SOD) The renal concentration of SOD was significantly reduced in the HIRI group when compared with the sham group. Only the high-dose BP group, however, showed a significant increase in SOD concentration when compared with the HIRI group. R enal Catalase Catalase concentration was significantly reduced in the kidneys of the animals in the HIRI group when compared with the sham group. Also, both low-dose BP and high-dose BP groups showed significant increase in catalase concentration when compared with the HIRI group. 3.5 Inflammatory Markers Tumour necrosis factor (TNF-α) There was a statistically significant increase in TNF-α concentration in the HIRI group when compared with the sham group. Significant decrease was also observed in the low-dose BP and high-dose BP groups when compared with the HIRI group. Furthermore, the high-dose BP group had a statistically significant reduction in TNF-α concentration when compared with the low-dose BP group. Myeloperoxidase (MPO) There was a statistically significant increase in MPO concentration in the HIRI group compared with the sham group. Also, the high-dose BP group had its MPO concentration significantly reduced when compared with the HIRI group. Serum nitrite There was no statistically significant difference in nitrite concentration among the groups. 3.6 Apoptotic Marker Caspase-3 There was a statistically significant increase in caspase-3 concentration in the HIRI group when compared with the sham group. The low-dose BP and high-does BP groups significantly reduced caspase-3 concentration with respect to the HIRI group. In addition, there was a significant decrease in caspase-3 concentration in the high-dose BP group when compared with the low-dose BP group. 3.7 Histology of the Liver Sham group (SH) The hepatic tissue appears normal. The hepatocytes (H) appear polygonal with well-outlined vesicular nucleus (N). The hepatocytes are separated by the sinusoids (S) with thin endothelial lining, free from collections and inflammatory cells. The central vein (CV) and bile duct (BD) also appear normal HIRI group (HH) The hepatic tissue appears distorted. The hepatocytes (H) appear thickened with well-outlined vesicular nucleus (N). The hepatocytes are separated by the sinusoids (S) with hemorrhagic lesions (circle). There are focal areas of inflammatory cell infiltration (box) and focal vascular congestion (star) in the central vein (CV). The bile duct (BD) appear normal. Low-dose BP group (LDH) The hepatic tissue appears normal. The hepatocytes (H) appear polygonal with well-outlined vesicular nucleus (N). The hepatocytes are separated by the sinusoids (S) with thin endothelial lining, free from collections and inflammatory cells. The central vein (CV) and bile duct (BD) also appear normal. High-dose BP (HDH) The hepatic tissue appears distorted. The hepatocytes (H) appear polygonal with well-outlined vesicular nucleus (N). The hepatocytes are separated by the sinusoids (S) and it is free of lesion. There is focal vascular congestion (star) in the central vein (CV). 3.8 Histology of the Kidney Sham group (SH) The histology of the renal tissue appears normal. The renal corpuscle (arrow) consists of the glomerulus (G) containing podocytes and separated by a well-defined Bowman’s space (BS). The renal tubules (RT) are lined by columnar-cubiodal epithelium and separated by interstitium that is free from congestions and collections. HIRI group (HH) The histology of the renal tissue appears distorted. The renal corpuscle (arrow) consists of the glomerulus (G) containing podocytes and separated by an ill-defined Bowman’s space (BS). The renal tubules (RT) are lined by columnar-cubiodal epithelium and separated by interstitium with focal areas of hemorrhagic lesions (black circle). There are also focal areas of vascular congestions (star) and inflammatory cell infiltration (box). Low-dose BP (LDH) The histology of the renal tissue appears normal. The renal corpuscle (arrow) consists of the glomerulus (G) containing podocytes and separated by a well-defined Bowman’s space (BS). The renal tubules (RT) are lined by columnar-cubiodal epithelium and separated by interstitium that is free from congestions and collections. High-dose BP (HDH) The histology of the renal tissue appears distorted. The renal corpuscle (arrow) consists of the glomerulus (G) containing podocytes and separated by an ill-defined Bowman’s space (BS). The renal tubules (RT) are lined by columnar-cubiodal epithelium and separated by interstitium with focal areas of hemorrhagic lesions (black circle). 4 Discussion The present study demonstrates that HIRI induces significant remote organ dysfunction, particularly affecting renal integrity. The relationship between hepatic damage and renal dysfunction (hepatorenal syndrome), highlights the need for protective interventions that can mitigate the systemic impact of HIRI ( 39 ). B. pinnatum is rich in flavonoids, alkaloids, and terpenes, which are known for their potent antioxidant and anti-inflammatory properties. Of particular interest is the presence of quercetin and kaempferol derivatives, which have shown renoprotective effects in other models of acute kidney injury ( 12 ). The observed elevation in serum ALT, AST, and GGT levels in the HIRI group confirms substantial hepatocellular damage, consistent with previous findings ( 40 ). As demonstrated by Ginès and Schrier ( 41 ), severe liver injury can induce systemic vasodilation, leading to reduced effective arterial blood volume and subsequent activation of the renin-angiotensin-aldosterone system (RAAS). This activation results in renal vasoconstriction and compromised renal perfusion. Also, the release of damage-associated molecular patterns (DAMPs) from injured hepatocytes can trigger a systemic inflammatory response. Released pro-inflammatory cytokines and chemokines can directly impact renal function through leukocyte infiltration and tubular cell injury ( 42 ). Consequently, oxidative stress initiated in the liver, evidenced by elevated protein degradation, GGT and MDA levels, as well as reduced GSH, SOD, and catalase levels, can propagate systemically, affecting distant organs including the kidneys. Oxidative stress is one of the primary mechanisms implicated in IRI ( 3 , 43 ). In the current study, B. pinnatum significantly ameliorated oxidative stress in both liver and kidney tissues, as evidenced by reductions in MDA levels and increases in endogenous antioxidant enzymes including superoxide dismutase SOD and catalase. These results are in alignment with earlier studies, which highlight the antioxidant properties of B. pinnatum as a contributor to its therapeutic effects ( 12 , 17 ). The antioxidative effects observed may be attributed to the plant’s phytochemical constituents, including flavonoids such as quercetin and kaempferol, known for their ROS-scavenging capabilities ( 44 , 45 ). These compounds enhance the activity of nuclear factor erythroid 2-related factor 2 (Nrf2), a major regulator of the cellular antioxidant response. Activation of the Nrf2 pathway also upregulates the expression of genes encoding antioxidant enzymes, which mitigate oxidative stress by neutralising ROS and preserving mitochondrial integrity ( 46 ). The increased activity of SOD and catalase observed in this study supports the hypothesis that B. pinnatum activates Nrf2, aligning with previous findings that document the role of flavonoids in Nrf2-mediated antioxidative pathways ( 47 , 48 ). Moreover, HIRI-induced kidney injury is strongly associated with inflammation, involving the release of pro-inflammatory cytokines ( 6 , 7 ). Inflammatory cascade analysis revealed that B. pinnatum administration led to significant reductions in TNF-α and MPO levels, suggesting an attenuation of the inflammatory cascade. This anti-inflammatory effect may result from the inhibition of nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB), which has been identified as a central mediator of inflammation in ischemia-reperfusion injury ( 1 , 49 ). The phytoconstituents in B. pinnatum , including triterpenoids and flavonoids, likely play a role in modulating NF-κB activation. These compounds have been shown to suppress NF-κB activity by preventing the phosphorylation and degradation of its inhibitor, IκBα, thus impeding the transcription of pro-inflammatory cytokines ( 44 ). This NF-κB inhibition reduces the recruitment of neutrophils and macrophages to the site of injury, thereby decreasing MPO levels, as observed in the present study. Furthermore, the downregulation of TNF-α may have limited secondary damage induced by ROS, creating a positive feedback loop that mitigates both inflammation and oxidative stress. Such multi-faceted anti-inflammatory effects underscore the therapeutic potential of B. pinnatum in ameliorating HIRI-induced renal injury ( 11 , 12 ). Furthermore, our biochemical analyses revealed significant elevations in serum creatinine and urea levels in the HIRI group, indicating compromised glomerular filtration and renal dysfunction. These findings align with previous studies demonstrating the development of acute kidney injury following hepatic ischemia-reperfusion ( 39 , 50 ). However, the nephroprotective effects of B. pinnatum were evidenced by improved levels of creatinine and urea. These markers were significantly reduced in B. pinnatum -treated groups compared to untreated HIRI controls, indicating preserved glomerular filtration rate (GFR) and reduced renal damage. Notably, this nephroprotective property was demonstrated in a dose-dependent manner, with the higher dose showing superior efficacy in preserving renal function. This preservation of renal function is consistent with prior findings suggesting that B. pinnatum has protective effects on renal tissue under conditions of oxidative stress and inflammation ( 5 , 45 ). The reduction in creatinine and urea levels observed in this study may stem from multiple mechanisms, including the antioxidant and anti-inflammatory effects of B. pinnatum , as well as its role in inhibiting apoptosis. Apoptosis in ischemia-reperfusion injury is mediated by both intrinsic (mitochondrial) and extrinsic (death receptor) pathways, with caspase-3 serving as an important executor enzyme ( 4 ). In line with this, our study revealed a significant elevation in caspase-3 levels in the HIRI group, indicating enhanced apoptotic activity. The attenuation of caspase-3 activity observed here suggests that B. pinnatum may inhibit apoptosis, potentially through modulation of mitochondrial pathways and inhibition of the mitochondrial permeability transition pore (mPTP) opening, as well as the modulation of Bcl-2 family proteins ( 51 , 52 ). Reduced apoptosis not only preserves renal tubular structure but also minimises functional impairment, which aligns with the observed maintenance of kidney function markers. Histopathological examination provided visual confirmation of these biochemical findings. The HIRI group showed characteristic renal tissue damage, including ill-defined Bowman's spaces and inflammatory cell infiltration. B. pinnatum treatment preserved normal renal architecture, supporting its nephroprotective effects at the tissue level. 5 Conclusion The nephroprotective effects of B. pinnatum appear to be mediated through multiple mechanisms: ( 1 ) antioxidant properties, evidenced by restoration of GSH, SOD, and catalase levels; ( 2 ) anti-inflammatory effects, demonstrated by reduced TNF-α and MPO levels; and ( 3 ) anti-apoptotic activity, shown by normalised caspase-3 levels. These findings suggest that B. pinnatum could serve as a promising therapeutic agent for preventing HIRI-induced kidney injury, though further clinical studies are warranted to establish its efficacy in human subjects. Abbreviations ALT Alanine Aminotransferase AST Aspartate Aminotransferase GGT Gamma-glutamyl transferase BP Bryophyllum pinnatum HIRI Hepatic Ischemia-Reperfusion Injury IRI Ischemia-Reperfusion Injury MDA Malondialdehyde ROS Reactive Oxygen Species SOD Superoxide Dismutase GSH Reduced Glutathione Declarations Ethics Approval and Consent to Participate: Ethical approval was obtained from Ethics Committee in Animal Care and Use in Research of FBMS with reference number: ERCFBMSLAUTECH: 058/08/2024. Consent for Publication: Not applicable Competing Interests: The authors declare that there is no conflict of interest in this manuscript Funding Declaration: No funding was received for this study Clinical trial number : Not applicable Authors' contributions: OVO, OAA, and OSH designed the study; OVO, OAA, RAA, and OSH performed the experiments; RAA, ABA, AWS, and SOO provided new reagents and analytical tools; OVO, ABA, OSH, and OSH analysed the research data; OVO, RAA, OSH, OSH, AWS, and SOO contributed to the writing of the manuscript; all authors read and approved the final manuscript. Acknowledgements: The authors acknowledge the support of the Department of Physiology, LAUTECH, and technical staff of the animal facility. References Yellon, D.M., and Hausenloy, D.J. (2013). Myocardial reperfusion injury. New England Journal of Medicine , 370, 112–119. Elias-Miró, M., Jiménez-Castro, M. B., Rodés, J., and Peralta, C. (2013). Current knowledge on oxidative stress in hepatic ischemia/reperfusion. Free radical research, 47(8), 555-568. Murphy, M. P., and Hartley, R. C. (2018). Mitochondria as a therapeutic target for common pathologies. Nature Reviews Drug Discovery , 17(12), 865-886. Kalogeris, T., Baines, C. P., Krenz, M., and Korthuis, R. J. (2012). Cell biology of ischemia/reperfusion injury. International review of cell and molecular biology , 298, 229-317. Trinh, E., Alam, A., Tchervenkov, J., and Cantarovich, M. (2017). Impact of acute kidney injury following liver transplantation on long‐term outcomes. Clinical Transplantation , 31(1), e12863. Lai, I. R., Chang, K. J., Tsai, H. W., and Chen, C. F. (2008). Pharmacological preconditioning with simvastatin protects liver from ischemia-reperfusion injury by heme oxygenase-1 induction. Transplantation , 85(5), 732-738. Awad, A.S., Rouse, M., Huang, L., Vergis, A.L., Reutershan, J., Cathro, H.P., Linden, J. and Okusa, M.D., (2009). Compartmentalization of neutrophils in the kidney and lung following acute ischemic kidney injury. Kidney international , 75(7), pp.689-698. Casillas-Ramírez, A., Zaouali, A., Padrissa-Altés, S., Ben Mosbah, I., Pertosa, A., Alfany-Fernández, I., Bintanel-Morcillo, M., Xaus, C., Rimola, A., Rodés, J. and Rosello-Catafau, J., (2009). Insulin-like growth factor and epidermal growth factor treatment: new approaches to protecting steatotic livers against ischemia-reperfusion injury. Endocrinology , 150(7), pp.3153-3161. Buchholz, B., Donato, M., D’Annunzio, V., and Gelpi, R. J. (2014). Ischemic postconditioning: mechanisms, comorbidities, and clinical application. Molecular and cellular biochemistry , 392, 1-12. Radomska-Leśniewska, D. M., and Skopiński, P. (2012). Review paper N-acetylcysteine as an anti-oxidant and anti-inflammatory drug and its some clinical applications. Central European journal of immunology, 37(1), 57-66. Ojewole, J. A. (2005). Antinociceptive, anti-inflammatory and antidiabetic effects of Bryophyllum pinnatum (Crassulaceae) leaf aqueous extract. Journal of ethnopharmacology , 99(1), 13-19. Fürer, K., Simões-Wüst, A. P., von Mandach, U., Hamburger, M., and Potterat, O. (2016). Bryophyllum pinnatum and related species used in anthroposophic medicine: constituents, pharmacological activities, and clinical efficacy. Planta medica , 82(11/12), 930-941. Fernandes, J. M., Cunha, L. M., Azevedo, E. P., Lourenço, E. M., Fernandes-Pedrosa, M. F., and Zucolotto, S. M. (2019). Kalanchoe laciniata and Bryophyllum pinnatum: an updated review about ethnopharmacology, phytochemistry, pharmacology and toxicology. Revista Brasileira de Farmacognosia, 29(4), 529-558. Shahrajabian, M. H., Sun, W., and Cheng, Q. (2022). The importance of flavonoids and phytochemicals of medicinal plants with antiviral activities. Mini-Reviews in Organic Chemistry, 19(3), 293-318. Akinwumi, I. A., and Ambali, O. A. (2024). Cardiac glycosides from african medicinal plants as promising therapeutics. Tropical Journal of Phytochemistry and Pharmaceutical Sciences , 3(2), 158-167. Chibli, L.A., Rodrigues, K.C., Gasparetto, C.M., Pinto, N.C., Fabri, R.L., Scio, E., Alves, M.S., Del-Vechio-Vieira, G. and Sousa, O.V., 2014. Anti-inflammatory effects of Bryophyllum pinnatum (Lam.) Oken ethanol extract in acute and chronic cutaneous inflammation. Journal of ethnopharmacology , 154 (2), pp.330-338. Selvakumar, P. (2022). Phytochemical and pharmacological profile review of Bryophyllum pinnatum . Biomedical And Biotechnology Research Journal (Bbrj) , 6(3), 295-301. Tatsimo, S.J.N., Tamokou, J.D.D., Havyarimana, L., Csupor, D., Forgo, P., Hohmann, J., Kuiate, J.R. and Tane, P., (2012). Antimicrobial and antioxidant activity of kaempferol rhamnoside derivatives from Bryophyllum pinnatum. BMC Research notes, 5, pp.1-6. Sharma, G., Jangra, A., Sihag, S., Chaturvedi, S., Yadav, S., and Chhokar, V. (2024). Bryophyllum pinnatum (Lam.) Oken: unravelling therapeutic potential and navigating toxicity. Physiology and Molecular Biology of Plants , 30 (9), 1413-1427. Yadav, M., Gulkari, V. D., and Wanjari, M. M. (2016). Bryophyllum pinnatum leaf extracts prevent formation of renal calculi in lithiatic rats. Ancient science of life, 36(2), 90-97. Anadozie, S. O., Akinyemi, J. A., Agunbiade, S., Ajiboye, B. O., and Adewale, O. B. (2018). Bryophyllum pinnatum inhibits arginase II activity and prevents oxidative damage occasioned by carbon tetrachloride (CCl4) in rats. Biomedicine and Pharmacotherapy , 101, 8-13. Shukla, A. B., Mandavia, D. R., Barvaliya, M. J., Baxi, S. N., and Tripathi, C. R. (2014). Evaluation of anti-urolithiatic effect of aqueous extract of Bryophyllum pinnatum (Lam.) leaves using ethylene glycol-induced renal calculi. Avicenna journal of phytomedicine , 4 (3), 151. Shinde, S. K., Chaware, V. J., and Redasani, V. K. (2024). Anti-urolithiatic effect of Ethanolic extract of Bryophyllum pinnatum (Lam) against chemicals induced Renal calculi in rats. Research Journal of Science and Technology , 16 (4), 283-290. Uahomo, P. O., and Isirima, J. C. Attenuating Ketamine-Induced Nephrotoxicity with Bryophyllum pinnatum Extract: Biochemical and Histological Investigation. Pal, S. K., Lahon, L. C., Sarkar, B. K., Paul, S. R., and Debroy, B. (2020). Effect of methanol extract of Bryophyllum pinnatum leaves on ethylene glycol-induced urolithiasis in adult male albino rats. Journal of Entomology and Zoology Studies , 8 (4), 782-786. Afzal, M., Kazmi, I., and Anwar, F. (2013). Antineoplastic potential of Bryophyllum pinnatum Lam. on chemically induced hepatocarcinogenesis in rats. Pharmacognosy research , 5 (4), 247. Kombathula, L., and Chernapalli, V. (2024). Evaluation of efficacy of selected Bryophyllum pinnatum phytochemicals in hepatocarcinogenic therapy through in silico approach: Efficacy of B. Pinnatum phytochemicals on hepatocarcinoma. Indian Journal of Experimental Biology (IJEB) , 62 (09), 752-759. Osifeso, O. O., Umoren, O. D., Bamidele, A. A., Obika, C. G., and Adeogun, O. A. (2025). Protective Effect of Bryophyllum pinnatum (Lam.) Oken (Miracle Leaf) Extract on Butylglycol-Induced Hepatotoxicity. World News of Natural Sciences , 59 , 239-249. Ajah, O., Unegbu, C. C., Onwusonye, C. J., Nnorom, C. M., and Duru, C. A. (2023). Effect of ethanol extract of Bryophyllum pinnatum leaf on lipid profile, renal and hepatic function biomarkers of high salt fed Albino rats. Journal of Medicinal Herbs, , 13 (4), 75-81. Şener, G., Sehirli, Ö., Ipçi, Y., Ercan, F., şirvanci, S., Gedik, N., and Yeĝen, B. Ç. (2005). Aqueous garlic extract alleviates ischaemia-reperfusion-induced oxidative hepatic injury in rats. Journal of Pharmacy and Pharmacology , 57(1), 145-150. Lorenzini, F., Tambara Filho, R., Gomes, R. P. X., Martino-Andrade, A. J., Erdmann, T. R., and Matias, J. E. F. (2012). Long-term effects of the testicular torsion on the spermatogenesis of the contralateral testis and the preventive value of the twisted testis orchiepididymectomy. Acta cirurgica brasileira, 27, 388-395. Nandi, A., and Chatterjee, I. B. (1988). Assay of superoxide dismutase activity in animal tissues. Journal of biosciences , 13, 305-315. Beuge, J. A., Aust, S. D. (1978) Microsomal lipid peroxidation. Methods Enzymol . 52: 302–311. Beutler, E. (1975) Glutathione. In: Beutler, E. (ed.) Red blood cell metabolism: a manual of biochemical methods. Grune and Stratton, New York, pp 112–114. Aebi, H. (1974). Catalase. In Methods of enzymatic analysis (pp. 673-684). Academic press. Inchiosa, M. A. (1964). Direct biuret determination of total protein in tissue homogenates. The Journal of Laboratory and Clinical Medicine , 63(2), 319-324. Hillegass, L. M., Griswold, D. E., Brickson, B., AlbrightsonWinslow, C. (1990) Assessment of myeloperoxidase activity in whole rat kidney. J. Pharmacol. Methods 24: 285–295. Giovannoni, G., Land, J. M., Keir, G., Thompson, E. J., and Heales, S. J. R. (1997). Adaptation of the nitrate reductase and Griess reaction methods for the measurement of serum nitrate plus nitrite levels. Annals of Clinical Biochemistry , 34(2), 193-198. Nastos, C., Kalimeris, K., Papoutsidakis, N., Tasoulis, M.K., Lykoudis, P.M., Theodoraki, K., Nastou, D., Smyrniotis, V. and Arkadopoulos, N., (2014). Global consequences of liver ischemia/reperfusion injury. Oxidative medicine and cellular longevity , 2014(1), p.906965. Jaeschke, H., and Woolbright, B. L. (2012). Current strategies to minimize hepatic ischemia–reperfusion injury by targeting reactive oxygen species. Transplantation reviews , 26(2), 103-114. Ginès, P., and Schrier, R. W. (2009). Renal failure in cirrhosis. New England Journal of Medicine , 361(13), 1279-1290. Bernardi, M., Moreau, R., Angeli, P., Schnabl, B., and Arroyo, V. (2015). Mechanisms of decompensation and organ failure in cirrhosis: from peripheral arterial vasodilation to systemic inflammation hypothesis. Journal of hepatology , 63(5), 1272-1284. Eltzschig, H. K., and Eckle, T. (2011). Ischemia and reperfusion—from mechanism to translation. Nature medicine , 17(11), 1391-1401. Kamboj, A., and Saluja, A. (2009). Bryophyllum pinnatum (Lam.) Kurz.: phytochemical and pharmacological profile: a review. Pharmacognosy Reviews , 3(6), 364. Akbari, B., Baghaei‐Yazdi, N., Bahmaie, M., and Mahdavi Abhari, F. (2022). The role of plant‐derived natural antioxidants in reduction of oxidative stress. BioFactors , 48(3), 611-633. Ma, Y. F., Wu, Z. H., Gao, M., and Loor, J. J. (2018). Nuclear factor erythroid 2-related factor 2 antioxidant response element pathways protect bovine mammary epithelial cells against H2O2-induced oxidative damage in vitro. Journal of dairy science , 101(6), 5329-5344. Kobayashi, A., Kang, M. I., Watai, Y., Tong, K. I., Shibata, T., Uchida, K., and Yamamoto, M. (2006). Oxidative and electrophilic stresses activate Nrf2 through inhibition of ubiquitination activity of Keap1. Molecular and cellular biology , 26(1), 221-229. Ashrafizadeh, M., Ahmadi, Z., Mohammadinejad, R., Farkhondeh, T., and Samarghandian, S. (2020). Curcumin activates the Nrf2 pathway and induces cellular protection against oxidative injury. Current Molecular Medicine , 20(2), 116-133. Perkins, N. D. (2007). Integrating cell-signalling pathways with NF-κB and IKK function. Nature reviews Molecular cell biology , 8(1), 49-62. Levey, A. S., Inker, L. A., and Coresh, J. (2014). GFR estimation: from physiology to public health. American Journal of Kidney Diseases , 63(5), 820-834. Chouchani, E.T., Pell, V.R., Gaude, E., Aksentijević, D., Sundier, S.Y., Robb, E.L., Logan, A., Nadtochiy, S.M., Ord, E.N., Smith, A.C. and Eyassu, F., 2014. Ischaemic accumulation of succinate controls reperfusion injury through mitochondrial ROS. Nature , 515(7527), pp.431-435. Czabotar, P. E., Lessene, G., Strasser, A., and Adams, J. M. (2014). Control of apoptosis by the BCL-2 protein family: implications for physiology and therapy. Nature reviews Molecular cell biology , 15(1), 49-63. Plates Plates 1 and 2 are available in the Supplementary Files section. Additional Declarations No competing interests reported. Supplementary Files Plates.docx Cite Share Download PDF Status: Posted Version 1 posted You are reading this latest preprint version Research Square lets you share your work early, gain feedback from the community, and start making changes to your manuscript prior to peer review in a journal. As a division of Research Square Company, we’re committed to making research communication faster, fairer, and more useful. We do this by developing innovative software and high quality services for the global research community. Our growing team is made up of researchers and industry professionals working together to solve the most critical problems facing scientific publishing. Also discoverable on Platform About Our Team In Review Editorial Policies Advisory Board Help Center Resources Author Services Accessibility API Access RSS feed Manage Cookie Preferences © Research Square 2026 | ISSN 2693-5015 (online) Privacy Policy Terms of Service Do Not Sell My Personal Information {"props":{"pageProps":{"initialData":{"identity":"rs-6205039","acceptedTermsAndConditions":true,"allowDirectSubmit":true,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":458172472,"identity":"733f4e51-e056-44de-8772-1809e80adca7","order_by":0,"name":"Oluwakemi V. 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represents significance at (P\u0026lt;0.01 and P\u0026lt;0.001) respectively vs HIRI group.\u003c/p\u003e","description":"","filename":"image2.png","url":"https://assets-eu.researchsquare.com/files/rs-6205039/v1/e2625f8c7ffbc7cb0ccf00c2.png"},{"id":83216396,"identity":"d3b355cc-6251-4e84-a4e6-d6dc9c7b3dfb","added_by":"auto","created_at":"2025-05-21 09:08:45","extension":"png","order_by":3,"title":"Figure 3","display":"","copyAsset":false,"role":"figure","size":20215,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eEffect of \u003c/strong\u003e\u003cem\u003e\u003cstrong\u003eBryophyllum pinnatum \u003c/strong\u003e\u003c/em\u003e\u003cstrong\u003eon Gamma-glutamyl transferase (GGT) concentration following HIRI in Male Wistar Rats\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eData were presented as mean ± SEM, n=10 (number of rat per group). * represents significance at (P\u0026lt;0.001) vs sham group. # represents significance at (P\u0026lt;0.001) vs HIRI group. ^ represents significance at (P\u0026lt;0.01) vs low-dose group.\u003c/p\u003e","description":"","filename":"image3.png","url":"https://assets-eu.researchsquare.com/files/rs-6205039/v1/f8aaa926278a68f75797d83c.png"},{"id":83213376,"identity":"0e9cea7a-9e78-4793-b89b-fe34c1444aa4","added_by":"auto","created_at":"2025-05-21 08:44:45","extension":"png","order_by":4,"title":"Figure 4","display":"","copyAsset":false,"role":"figure","size":20781,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eEffect of \u003c/strong\u003e\u003cem\u003e\u003cstrong\u003eBryophyllum pinnatum \u003c/strong\u003e\u003c/em\u003e\u003cstrong\u003eon Creatinine concentration following HIRI in Male Wistar Rats\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eData were presented as mean ± SEM, n=10 (number of rat per group). * represents significance at (P\u0026lt;0.05) vs sham group. # represents significance at (P\u0026lt;0.01 and P\u0026lt;0.001) vs HIRI group.\u003c/p\u003e","description":"","filename":"image4.png","url":"https://assets-eu.researchsquare.com/files/rs-6205039/v1/df00d188f815914c9f717c74.png"},{"id":83211643,"identity":"4bfbc36d-b445-4ca7-b436-2e958ca170c1","added_by":"auto","created_at":"2025-05-21 08:36:45","extension":"png","order_by":5,"title":"Figure 5","display":"","copyAsset":false,"role":"figure","size":20255,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eEffect of \u003c/strong\u003e\u003cem\u003e\u003cstrong\u003eBryophyllum pinnatum \u003c/strong\u003e\u003c/em\u003e\u003cstrong\u003eon Urea concentration following HIRI in Male Wistar Rats\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eData were presented as mean ± SEM, n=10 (number of rat per group). * represents significance at (P\u0026lt;0.001) vs sham group. # represents significance at (P\u0026lt;0.001 and P\u0026lt;0.001) vs HIRI group. ^ represents significance at (P\u0026lt;0.001) vs low-dose BP group.\u003c/p\u003e","description":"","filename":"image5.png","url":"https://assets-eu.researchsquare.com/files/rs-6205039/v1/f133449b3f5037e0d2e280c3.png"},{"id":83211650,"identity":"48b289cd-1a3e-46d2-9dbb-d028758c0864","added_by":"auto","created_at":"2025-05-21 08:36:45","extension":"png","order_by":6,"title":"Figure 6","display":"","copyAsset":false,"role":"figure","size":25117,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eEffect of \u003c/strong\u003e\u003cem\u003e\u003cstrong\u003eBryophyllum pinnatum \u003c/strong\u003e\u003c/em\u003e\u003cstrong\u003eon hepatic GSH concentration following HIRI in Male Wistar Rats\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eData were presented as mean ± SEM, n=10 (number of rat per group). * represents significance at (P\u0026lt;0.001) vs sham group. # represents significance at (P\u0026lt;0.01 and P\u0026lt;0.001) vs HIRI group.\u003c/p\u003e","description":"","filename":"image6.png","url":"https://assets-eu.researchsquare.com/files/rs-6205039/v1/e3123c07794c8be4f4f63d7a.png"},{"id":83215420,"identity":"54bb257b-55ac-4a9a-9a2c-2928752258bd","added_by":"auto","created_at":"2025-05-21 09:00:45","extension":"png","order_by":7,"title":"Figure 7","display":"","copyAsset":false,"role":"figure","size":24057,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eEffect of \u003c/strong\u003e\u003cem\u003e\u003cstrong\u003eBryophyllum pinnatum \u003c/strong\u003e\u003c/em\u003e\u003cstrong\u003eon hepatic total protein concentration following HIRI in Male Wistar Rats\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eData were presented as mean ± SEM, n=10 (number of rat per group). * represents significance at (P\u0026lt;0.001) vs sham group. # represents significance at (P\u0026lt;0.001 and P\u0026lt;0.05) vs HIRI group. ^ represent significance at (P\u0026lt;0.01) vs low-dose BP group.\u003c/p\u003e","description":"","filename":"image7.png","url":"https://assets-eu.researchsquare.com/files/rs-6205039/v1/627ed64ca6fdaab273aa1623.png"},{"id":83211651,"identity":"60804237-9c89-428c-a50c-1adf6366885b","added_by":"auto","created_at":"2025-05-21 08:36:45","extension":"png","order_by":8,"title":"Figure 8","display":"","copyAsset":false,"role":"figure","size":23836,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eEffect of \u003c/strong\u003e\u003cem\u003e\u003cstrong\u003eBryophyllum pinnatum \u003c/strong\u003e\u003c/em\u003e\u003cstrong\u003eon hepatic MDA concentration following HIRI in Male Wistar Rats\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eData were presented as mean ± SEM, n=10 (number of rat per group). * represents significance at (P\u0026lt;0.001) vs sham group. # represents significance at (P\u0026lt;0.001 and P\u0026lt;0.001) vs HIRI group. ^ represent significance at (P\u0026lt;0.001) vs low-dose BP group.\u003c/p\u003e","description":"","filename":"image8.png","url":"https://assets-eu.researchsquare.com/files/rs-6205039/v1/cb1f4aba474000c9e505dfe2.png"},{"id":83213377,"identity":"1122753f-236e-4e83-bbf3-ec8018da3a24","added_by":"auto","created_at":"2025-05-21 08:44:45","extension":"png","order_by":9,"title":"Figure 9","display":"","copyAsset":false,"role":"figure","size":23126,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eEffect of \u003c/strong\u003e\u003cem\u003e\u003cstrong\u003eBryophyllum pinnatum \u003c/strong\u003e\u003c/em\u003e\u003cstrong\u003eon hepatic SOD concentration following HIRI in Male Wistar Rats\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eData were presented as mean ± SEM, n=10 (number of rat per group). * represents significance at (P\u0026lt;0.001) vs sham group. # represents significance at (P\u0026lt;0.001 and P\u0026lt;0.001) vs HIRI group.\u003c/p\u003e","description":"","filename":"image9.png","url":"https://assets-eu.researchsquare.com/files/rs-6205039/v1/ddeda1c3d9cbd1dabedc3504.png"},{"id":83213381,"identity":"b5b02c5d-8b2d-4062-bd8c-c2298a84c55f","added_by":"auto","created_at":"2025-05-21 08:44:45","extension":"png","order_by":10,"title":"Figure 10","display":"","copyAsset":false,"role":"figure","size":23286,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eEffect of \u003c/strong\u003e\u003cem\u003e\u003cstrong\u003eBryophyllum pinnatum \u003c/strong\u003e\u003c/em\u003e\u003cstrong\u003eon hepatic Catalase concentration following HIRI in Male Wistar Rats\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eData were presented as mean ± SEM, n=10 (number of rat per group). * represents significance at (P\u0026lt;0.001) vs sham group. # represents significance at (P\u0026lt;0.05 and P\u0026lt;0.01) vs HIRI group.\u003c/p\u003e","description":"","filename":"image10.png","url":"https://assets-eu.researchsquare.com/files/rs-6205039/v1/18253cc520681a84a2931052.png"},{"id":83213378,"identity":"a13090bb-cc53-4000-96ea-fa5997c2506e","added_by":"auto","created_at":"2025-05-21 08:44:45","extension":"png","order_by":11,"title":"Figure 11","display":"","copyAsset":false,"role":"figure","size":24205,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eEffect of \u003c/strong\u003e\u003cem\u003e\u003cstrong\u003eBryophyllum pinnatum \u003c/strong\u003e\u003c/em\u003e\u003cstrong\u003eon renal GSH concentration following HIRI in Male Wistar Rats\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eData were presented as mean ± SEM, n=10 (number of rat per group). * represents significance at (P\u0026lt;0.001) vs sham group. # represents significance at (P\u0026lt;0.05) vs HIRI group. ^ represents significance at P\u0026lt;0.05) vs low-dose BP group.\u003c/p\u003e","description":"","filename":"image11.png","url":"https://assets-eu.researchsquare.com/files/rs-6205039/v1/fb0eb0714c2f1f290c9c1d7e.png"},{"id":83213383,"identity":"91d24a9b-50ae-4e99-aa1d-a774e2d5b53a","added_by":"auto","created_at":"2025-05-21 08:44:45","extension":"png","order_by":12,"title":"Figure 12","display":"","copyAsset":false,"role":"figure","size":24823,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eEffect of \u003c/strong\u003e\u003cem\u003e\u003cstrong\u003eBryophyllum pinnatum \u003c/strong\u003e\u003c/em\u003e\u003cstrong\u003eon renal total protein concentration following HIRI in Male Wistar Rats\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eData were presented as mean ± SEM, n=10 (number of rat per group). * represents significance at (P\u0026lt;0.05) vs sham group. # represents significance at (P\u0026lt;0.05 and P\u0026lt;0.001) vs HIRI group.\u003c/p\u003e","description":"","filename":"image12.png","url":"https://assets-eu.researchsquare.com/files/rs-6205039/v1/d45567f43e17786b4bbe0761.png"},{"id":83211654,"identity":"2630e36b-b0a0-4960-80d2-3d5cd6473cb5","added_by":"auto","created_at":"2025-05-21 08:36:45","extension":"png","order_by":13,"title":"Figure 13","display":"","copyAsset":false,"role":"figure","size":22376,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eEffect of \u003c/strong\u003e\u003cem\u003e\u003cstrong\u003eBryophyllum pinnatum \u003c/strong\u003e\u003c/em\u003e\u003cstrong\u003eon renal MDA concentration following HIRI in Male Wistar Rats\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eData were presented as mean ± SEM, n=10 (number of rat per group). * represents significance at (P\u0026lt;0.01) vs sham group. # represents significance at (P\u0026lt;0.01 and P\u0026lt;0.01) vs HIRI group.\u003c/p\u003e","description":"","filename":"image13.png","url":"https://assets-eu.researchsquare.com/files/rs-6205039/v1/5083bcec347421afa5d17ded.png"},{"id":83214400,"identity":"09a8088f-a679-4f59-81ff-bbc5eb8550f1","added_by":"auto","created_at":"2025-05-21 08:52:45","extension":"png","order_by":14,"title":"Figure 14","display":"","copyAsset":false,"role":"figure","size":22382,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eEffect of \u003c/strong\u003e\u003cem\u003e\u003cstrong\u003eBryophyllum pinnatum \u003c/strong\u003e\u003c/em\u003e\u003cstrong\u003eon renal SOD concentration following HIRI in Male Wistar Rats\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eData were presented as mean ± SEM, n=10 (number of rat per group). * represents significance at (P\u0026lt;0.001) vs sham group. # represents significance at (P\u0026lt;0.01) vs HIRI group.\u003c/p\u003e","description":"","filename":"image14.png","url":"https://assets-eu.researchsquare.com/files/rs-6205039/v1/c61c63177215d0c56a812d4c.png"},{"id":83211657,"identity":"d612485d-c472-472c-bb16-804b1328dac2","added_by":"auto","created_at":"2025-05-21 08:36:45","extension":"png","order_by":15,"title":"Figure 15","display":"","copyAsset":false,"role":"figure","size":22819,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eEffect of \u003c/strong\u003e\u003cem\u003e\u003cstrong\u003eBryophyllum pinnatum \u003c/strong\u003e\u003c/em\u003e\u003cstrong\u003eon renal catalase concentration following HIRI in Male Wistar Rats\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eData were presented as mean ± SEM, n=10 (number of rat per group). * represents significance at (P\u0026lt;0.001) vs sham group. # represents significance at (P\u0026lt;0.001 and P\u0026lt;0.001) vs HIRI group.\u003c/p\u003e","description":"","filename":"image15.png","url":"https://assets-eu.researchsquare.com/files/rs-6205039/v1/a5168ccc1385478567bc7913.png"},{"id":83211693,"identity":"3933c512-9fff-4b3c-b1b6-cc285bef0463","added_by":"auto","created_at":"2025-05-21 08:36:46","extension":"png","order_by":16,"title":"Figure 16","display":"","copyAsset":false,"role":"figure","size":19956,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eEffect of \u003c/strong\u003e\u003cem\u003e\u003cstrong\u003eBryophyllum pinnatum \u003c/strong\u003e\u003c/em\u003e\u003cstrong\u003eon TNF-α concentration following HIRI in Male Wistar Rats\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eData were presented as mean ± SEM, n=10 (number of rat per group). * represents significance at (P\u0026lt;0.001) vs sham group. # represents significance at (P\u0026lt;0.001 and P\u0026lt;0.001) vs HIRI group. ^ represents significance at (P\u0026lt;0.01) vs low-dose BP group.\u003c/p\u003e","description":"","filename":"image16.png","url":"https://assets-eu.researchsquare.com/files/rs-6205039/v1/ac695d1f4cd3f2e11f9e96fc.png"},{"id":83211660,"identity":"b74c7a97-932b-40ca-a2f8-534364bde0a9","added_by":"auto","created_at":"2025-05-21 08:36:45","extension":"png","order_by":17,"title":"Figure 17","display":"","copyAsset":false,"role":"figure","size":9476,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eEffect of \u003c/strong\u003e\u003cem\u003e\u003cstrong\u003eBryophyllum pinnatum \u003c/strong\u003e\u003c/em\u003e\u003cstrong\u003eon MPO concentration following HIRI in Male Wistar Rats\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eData were presented as mean ± SEM, n=10 (number of rat per group). * represents significance at (P\u0026lt;0.01) vs sham group. # represents significance at (P\u0026lt;0.05) vs HIRI group.\u003c/p\u003e","description":"","filename":"image17.png","url":"https://assets-eu.researchsquare.com/files/rs-6205039/v1/33605fcadb24d0bdc158006e.png"},{"id":83211664,"identity":"278c8bb9-22b4-4655-a02c-eb60973b143d","added_by":"auto","created_at":"2025-05-21 08:36:45","extension":"png","order_by":18,"title":"Figure 18","display":"","copyAsset":false,"role":"figure","size":19971,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eEffect of \u003c/strong\u003e\u003cem\u003e\u003cstrong\u003eBryophyllum pinnatum \u003c/strong\u003e\u003c/em\u003e\u003cstrong\u003eon nitrite concentration following HIRI in Male Wistar Rats\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eData were presented as mean ± SEM, n=10 (number of rat per group).\u003c/p\u003e","description":"","filename":"image18.png","url":"https://assets-eu.researchsquare.com/files/rs-6205039/v1/2ceb43556658368758d46698.png"},{"id":83213390,"identity":"c49d6bb7-aee9-4dcd-b994-21ac06ffd198","added_by":"auto","created_at":"2025-05-21 08:44:46","extension":"png","order_by":19,"title":"Figure 19","display":"","copyAsset":false,"role":"figure","size":19266,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eEffect of \u003c/strong\u003e\u003cem\u003e\u003cstrong\u003eBryophyllum pinnatum \u003c/strong\u003e\u003c/em\u003e\u003cstrong\u003eon caspase-3 concentration following HIRI in Male Wistar Rats\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eData were presented as mean ± SEM, n=10 (number of rat per group). * represents significance at (P\u0026lt;0.001) vs sham group. # represents significance at (P\u0026lt;0.001 and P\u0026lt;0.001) vs HIRI group. ^ represents significance at (P\u0026lt;0.01) vs low-dose BP group.\u003c/p\u003e","description":"","filename":"image19.png","url":"https://assets-eu.researchsquare.com/files/rs-6205039/v1/d9bd579c98914d1774315d62.png"},{"id":89484346,"identity":"83c387a0-374d-40a2-8031-f65d4c75d5f7","added_by":"auto","created_at":"2025-08-20 12:32:05","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":2383175,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-6205039/v1/8b3f2d1d-f49b-4704-a7ab-6be58325a27e.pdf"},{"id":83213372,"identity":"70918c47-c077-4568-9dfb-d2db867a2814","added_by":"auto","created_at":"2025-05-21 08:44:45","extension":"docx","order_by":1,"title":"","display":"","copyAsset":false,"role":"supplement","size":522611,"visible":true,"origin":"","legend":"","description":"","filename":"Plates.docx","url":"https://assets-eu.researchsquare.com/files/rs-6205039/v1/7ee7070df5eae260d9ac8854.docx"}],"financialInterests":"No competing interests reported.","formattedTitle":"Reno-Hepatoprotective effects of Bryophyllum pinnatum against Hepatic Ischemia- Reperfusion Injury in male Wistar rats","fulltext":[{"header":"1 Introduction","content":"\u003cp\u003eIschemia-reperfusion injury (IRI) is characterized by a complex pathophysiological cascade that aggravates tissue damage during blood flow restoration after a period of ischemia. This phenomenon significantly impacts morbidity and mortality rates in clinical practice, particularly in organ transplantation and major surgical procedures (\u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e1\u003c/span\u003e). The liver exhibits particular vulnerability to IRI, making hepatic ischemia-reperfusion injury (HIRI) a major concern in hepatic surgery and liver transplantation (\u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eIn the pathogenesis of HIRI, oxygen deprivation, during ischemia, triggers ATP depletion, which in turn disrupts ionic homeostasis and causes calcium overload. The reperfusion phase on the other hand, exacerbates injury through reactive oxygen species (ROS) generation, including superoxide anions and hydroxyl radicals, leading to cellular damage through lipid peroxidation and protein oxidation (\u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e3\u003c/span\u003e). Furthermore, oxidized phospholipids can act as damage-associated molecular patterns (DAMPs), which then go on to initiate inflammatory cascades that exacerbate the injury (\u003cspan citationid=\"CR4\" class=\"CitationRef\"\u003e4\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eThe systemic effect of HIRI can also affect renal function, with reported incidences of acute kidney injury following orthotopic liver transplantation ranging from 12\u0026ndash;94% (\u003cspan citationid=\"CR5\" class=\"CitationRef\"\u003e5\u003c/span\u003e). This hepatorenal interaction is traceable to multiple pathways, including Kupffer cell activation and subsequent pro-inflammatory cytokine release, such as, tumour necrosis factor-alpha (TNF-α) and interleukin-6 (IL-6), which promote systemic inflammation and remote organ injury (\u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e6\u003c/span\u003e). Additionally, sinusoidal endothelial cell damage promotes microthrombosis and vasoconstriction through von Willebrand factor release and reduced nitric oxide production (\u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e6\u003c/span\u003e). The damage to the kidneys in this context is mediated by oxidative stress, mitochondrial dysfunction, and heightened susceptibility to apoptosis; a combination that significantly impairs renal function (\u003cspan citationid=\"CR5\" class=\"CitationRef\"\u003e5\u003c/span\u003e, \u003cspan citationid=\"CR7\" class=\"CitationRef\"\u003e7\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eCurrent therapeutic strategies to mitigate HIRI-induced renal injury primarily focus on supportive measures and specific pharmacological interventions targeting oxidative stress and inflammation (\u003cspan citationid=\"CR8\" class=\"CitationRef\"\u003e8\u003c/span\u003e). Additionally, ischemic preconditioning and pharmacological approaches to reduce mitochondrial permeability transition pore (MPTP) formation remain limited by variability in individual response and operational complexities in surgical contexts (\u003cspan citationid=\"CR9\" class=\"CitationRef\"\u003e9\u003c/span\u003e). The limitations of these existing treatments, including timing constraints and potential side effects, necessitate the exploration of alternative therapeutic approaches, such as natural compounds with multi-targeted actions capable of modulating both oxidative and inflammatory pathways without the drawbacks of synthetic interventions (\u003cspan citationid=\"CR10\" class=\"CitationRef\"\u003e10\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eAmong natural compounds, \u003cem\u003eBryophyllum pinnatum\u003c/em\u003e, is well known to possess therapeutic potential suitable for this study. In traditional medicine, \u003cem\u003eBryophyllum pinnatum\u003c/em\u003e has long been utilized for various ailments, including liver and kidney disorders (\u003cspan citationid=\"CR11\" class=\"CitationRef\"\u003e11\u003c/span\u003e, \u003cspan citationid=\"CR12\" class=\"CitationRef\"\u003e12\u003c/span\u003e). Its phytochemical analyses reveal it to be rich in bioactive compounds, including flavonoids, triterpenoids, and bufadienolides (\u003cspan citationid=\"CR13\" class=\"CitationRef\"\u003e13\u003c/span\u003e). These constituents demonstrate significant antioxidant and anti-inflammatory properties, particularly in suppressing pro-inflammatory cytokine production and nuclear factor-kappa B activation (\u003cspan citationid=\"CR14\" class=\"CitationRef\"\u003e14\u003c/span\u003e, \u003cspan citationid=\"CR15\" class=\"CitationRef\"\u003e15\u003c/span\u003e). In a landmark study by Ojewole (\u003cspan citationid=\"CR11\" class=\"CitationRef\"\u003e11\u003c/span\u003e) to scientifically evaluate the ethnomedicinal applications of \u003cem\u003eB. pinnatum\u003c/em\u003e leaves, the author concluded that aqueous extract of \u003cem\u003eB. pinnatum\u003c/em\u003e leaves possessed anti-inflammatory properties, evidenced by the significant inhibition of fresh egg albumin-induced acute inflammation, as well as antinociceptive and anti-diabetic properties. Chibli \u003cem\u003eet al\u003c/em\u003e. (\u003cspan citationid=\"CR16\" class=\"CitationRef\"\u003e16\u003c/span\u003e) also demonstrated \u003cem\u003eB. pinnatum\u003c/em\u003e\u0026rsquo;s anti-inflammatory property in acute and chronic mice ear edema models induced by different irritant agents.\u003c/p\u003e \u003cp\u003eRecent studies have further shown \u003cem\u003eB. pinnatum\u003c/em\u003e's capacity to scavenge free radicals and inhibit lipid peroxidation, due to active components such as quercetin, kaempferol, and bryophyllins (\u003cspan citationid=\"CR17\" class=\"CitationRef\"\u003e17\u003c/span\u003e). In a study by Tatsimo \u003cem\u003eet al\u003c/em\u003e. (\u003cspan citationid=\"CR18\" class=\"CitationRef\"\u003e18\u003c/span\u003e), methanolic extracts of \u003cem\u003eB. pinnatum\u003c/em\u003e showed strong antioxidative and antimicrobial activities, confirming their traditional use in the treatment of infectious and free radical damages. A recent review by Sharma et al. (\u003cspan citationid=\"CR19\" class=\"CitationRef\"\u003e19\u003c/span\u003e) indicated that \u003cem\u003eB. pinnatum\u003c/em\u003e is effective in the treatment of kidney and liver diseases, and possesses anti-leishmanial, neuropharmacological, antibacterial, immunosuppressive, anti-tumour, and cytotoxic activities.\u003c/p\u003e \u003cp\u003eIts nephroprotective effect is confirmed in the study by Yadav \u003cem\u003eet al\u003c/em\u003e. (\u003cspan citationid=\"CR20\" class=\"CitationRef\"\u003e20\u003c/span\u003e) which showed in their study on the effect of \u003cem\u003eB. pinnatum\u003c/em\u003e on ethylene glycol (EG)-induced renal calculi in rats, that \u003cem\u003eB. pinnatum\u003c/em\u003e effectively reversed EG-induced damage comparable to the standard drug Cystone. Likewise, Anadozie et al. (\u003cspan citationid=\"CR21\" class=\"CitationRef\"\u003e21\u003c/span\u003e) concluded that \u003cem\u003eB. pinnatum\u003c/em\u003e is able to inhibit arginase II activity and consequently prevent carbon tetrachloride (CCl4)-induced renal oxidative damage in rats. Other studies to examine the renoprotective effects of \u003cem\u003eB.\u003c/em\u003e pinnatum, such as those by Shukla et al. (\u003cspan citationid=\"CR22\" class=\"CitationRef\"\u003e22\u003c/span\u003e), Shinde et al. (\u003cspan citationid=\"CR23\" class=\"CitationRef\"\u003e23\u003c/span\u003e), Uahomo and Isirima (\u003cspan citationid=\"CR24\" class=\"CitationRef\"\u003e24\u003c/span\u003e), and Pal et al. (\u003cspan citationid=\"CR25\" class=\"CitationRef\"\u003e25\u003c/span\u003e), all show that \u003cem\u003eB. pinnatum\u003c/em\u003e is able to attenuate renal damage in experimental studies.\u003c/p\u003e \u003cp\u003eApart from having renoprotective properties, \u003cem\u003eB. pinnatum\u003c/em\u003e has also been reported to possess hepatoprotective properties. Afzal \u003cem\u003eet al\u003c/em\u003e. (\u003cspan citationid=\"CR26\" class=\"CitationRef\"\u003e26\u003c/span\u003e) concluded in their study that \u003cem\u003eB. pinnatum\u003c/em\u003e is able to mitigate chemically-induced hepatocarcinogenesis due to its ability to upregulate antioxidant activities and scavenge free radicals that damage cells. Kombathula and Chernapalli (\u003cspan citationid=\"CR27\" class=\"CitationRef\"\u003e27\u003c/span\u003e) confirmed this in their study and attested that the plant\u0026rsquo;s components\u0026mdash;bryophyllin A and β-sitosterol\u0026mdash;strongly interact with the Pi3K and C-met pathways respectively to mediate their effect. In another model of butylglycol-induced hepatotoxicity, the researchers observed that groups treated with \u003cem\u003eB. pinnatum\u003c/em\u003e showed reduced liver injury markers (ALP and AST) compared to the positive control (\u003cspan citationid=\"CR28\" class=\"CitationRef\"\u003e28\u003c/span\u003e). In addition, Ajah et al. (\u003cspan citationid=\"CR29\" class=\"CitationRef\"\u003e29\u003c/span\u003e) examined the efficacy of \u003cem\u003eB. pinnatum\u003c/em\u003e in attenuating damage to the kidneys and liver, as well as the lipid profile of rats fed with high-salt diet. They concluded that the ethanolic extract of \u003cem\u003eB. pinnatum\u003c/em\u003e was highly potent in preventing cell damage caused by high salt intake. However, its potential role in mitigating HIRI-induced kidney injury remains unexplored.\u003c/p\u003e \u003cp\u003eSince \u003cem\u003eBryophyllum pinnatum\u003c/em\u003e\u0026rsquo;s pleiotropic effects might provide comprehensive protection against both direct and remote organ injury through upregulation of antioxidant enzymes, modulation of oxidative stress, inflammation, and apoptotic pathways, this study aims to evaluate the reno-hepatoprotective effects of \u003cem\u003eBryophyllum pinnatum\u003c/em\u003e methanolic extract on kidney injury following hepatic ischemia-reperfusion in male Wistar rats.\u003c/p\u003e"},{"header":"2 Materials and Methods","content":"\u003cdiv id=\"Sec3\" class=\"Section2\"\u003e \u003ch2\u003e2.1 Chemicals and Reagents\u003c/h2\u003e \u003cp\u003eAll chemicals and reagents used in this study were of analytical grade. Ketamine and xylazine for anaesthesia were obtained from the Veterinary Department, University of Ibadan. Streptomycin injection, 0.9% normal saline, 10% formalin, and distilled water were procured from the Physiology Laboratory, LAUTECH. Biochemical assay kits for liver function markers (ALT, AST, GGT) and kidney function markers (urea, creatinine) were purchased from Randox Laboratories Ltd., UK. Stains for histological analysis, including Hematoxylin and Eosin, Masson's trichrome, and Periodic acid-Schiff (PAS), were also utilized.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec4\" class=\"Section2\"\u003e \u003ch2\u003e2.2 Experimental Animals and Design\u003c/h2\u003e \u003cp\u003eForty male Wistar rats weighing 120-150g were obtained from animal house of physiology department, LAUTECH, Ogbomoso, Oyo State, Nigeria. The animals were housed under standard laboratory conditions at the animal facility of Faculty of Basic Medical Sciences (FBMS), LAUTECH and were acclimatized for 2 weeks, fed with standard rat chow and watered \u003cem\u003ead libitum\u003c/em\u003e. Ethical approval was obtained from Ethics Committee in Animal Care and Use in Research of FBMS with reference number: \u003cb\u003eERCFBMSLAUTECH: 058/08/2024.\u003c/b\u003e\u003c/p\u003e \u003cp\u003eThe rats were randomly divided into four groups (n\u0026thinsp;=\u0026thinsp;10 per group):\u003c/p\u003e \u003cp\u003e \u003col\u003e \u003cspan\u003e \u003cli\u003e \u003cp\u003e \u003cb\u003eSham group\u003c/b\u003e: Sham-operated, no treatment\u003c/p\u003e \u003c/li\u003e \u003c/span\u003e \u003cspan\u003e \u003cli\u003e \u003cp\u003e \u003cb\u003eHIRI group\u003c/b\u003e: Hepatic ischemia-reperfusion injury, no treatment\u003c/p\u003e \u003c/li\u003e \u003c/span\u003e \u003cspan\u003e \u003cli\u003e \u003cp\u003e \u003cb\u003eBP Low Dose\u0026thinsp;+\u0026thinsp;HIRI\u003c/b\u003e: 100 mg/kg \u003cem\u003eBryophyllum pinnatum\u003c/em\u003e extract\u0026thinsp;+\u0026thinsp;Hepatic ischemia-reperfusion injury\u003c/p\u003e \u003c/li\u003e \u003c/span\u003e \u003cspan\u003e \u003cli\u003e \u003cp\u003e \u003cb\u003eBP High Dose\u0026thinsp;+\u0026thinsp;HIRI\u003c/b\u003e: 200 mg/kg \u003cem\u003eBryophyllum pinnatum\u003c/em\u003e extract\u0026thinsp;+\u0026thinsp;Hepatic ischemia-reperfusion injury\u003c/p\u003e \u003c/li\u003e \u003c/span\u003e \u003c/ol\u003e \u003c/p\u003e \u003cp\u003eThe \u003cem\u003eBryophyllum pinnatum\u003c/em\u003e extract was administered orally via gavage for 14 days prior to the induction of HIRI, as described by Anadozie \u003cem\u003eet al\u003c/em\u003e. (\u003cspan citationid=\"CR21\" class=\"CitationRef\"\u003e21\u003c/span\u003e).\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec5\" class=\"Section2\"\u003e \u003ch2\u003e2.3 Plant Material Preparation\u003c/h2\u003e \u003cp\u003eFresh \u003cem\u003eBryophyllum pinnatum\u003c/em\u003e leaves were collected from farm house of Faculty of Agricultural Sciences, Ladoke Akintola University of Technology (LAUTECH), Ogbomoso, Oyo State, Nigeria. The plant was identified and authenticated by Dr. Mrs. A.F. Ogundola, a botanist at the Department of Pure and Applied Biology, LAUTECH. The plant was assigned the voucher number \u003cb\u003eLHO 747\u003c/b\u003e, and kept in the Herbarium at the Department of Botany, LAUTECH, Ogbomoso, Nigeria.\u003c/p\u003e \u003cp\u003eThe leaves were washed, air-dried, and pulverized into a fine powder using an electric blender. The powdered leaves were macerated in 80% methanol for 72 hours, and the extract was concentrated using a rotary evaporator and further dried in a vacuum oven.\u003c/p\u003e \u003cdiv id=\"Sec6\" class=\"Section3\"\u003e \u003ch2\u003e2.3.1 Phytochemical Profiling\u003c/h2\u003e \u003cp\u003eQualitative screening of the \u003cem\u003eBryophyllum pinnatum\u003c/em\u003e extract was performed to detect the presence of major phytochemical groups, including alkaloids, flavonoids, tannins, saponins, terpenoids, glycosides, steroids, and anthraquinones, using standard chemical tests (Mayer's, Wagner's, alkaline reagent, ferric chloride, foam, Salkowski, Keller-Killiani, Liebermann-Burchard, and Borntrager's tests, respectively).\u003c/p\u003e \u003cp\u003eFor quantitative analysis, spectrophotometric methods were employed to determine the total flavonoid content (aluminium chloride colorimetric method), total tannin content (Folin-Ciocalteu method), and total phenolic content (Folin-Ciocalteu method) (Table\u0026nbsp;\u003cspan refid=\"Tab1\" class=\"InternalRef\"\u003e1\u003c/span\u003e).\u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab1\" border=\"1\"\u003e \u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 1\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003eQualitative and selected Quantitative analyses of \u003cem\u003eB. pinnatum\u003c/em\u003e\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"3\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\"\u003e \u003cp\u003ePhytochemical Group\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c2\"\u003e \u003cp\u003eQualitative Result\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c3\"\u003e \u003cp\u003eQuantitative Result (where applicable)\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eAlkaloids\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e+++\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eNot quantified\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eFlavonoids\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e+++\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e24.3\u0026thinsp;\u0026plusmn;\u0026thinsp;1.8 mg QE/g dry weight\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eTannins\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e++\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e18.7\u0026thinsp;\u0026plusmn;\u0026thinsp;1.2 mg TAE/g dry weight\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eSaponins\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e++\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eNot quantified\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eTerpenoids\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e+++\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eNot quantified\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003ePhenolic compounds\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e+++\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e37.2\u0026thinsp;\u0026plusmn;\u0026thinsp;2.5 mg GAE/g dry weight\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eGlycosides\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e++\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eNot quantified\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eSteroids\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eNot quantified\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eAnthraquinones\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eNot quantified\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003ctfoot\u003e \u003ctr\u003e\u003ctd colspan=\"3\"\u003e\u003cb\u003eKey\u003c/b\u003e: +++ (strongly present), ++ (moderately present), + (weakly present), - (absent) QE: Quercetin Equivalent, TAE: Tannic Acid Equivalent, GAE: Gallic Acid Equivalent\u003c/td\u003e\u003c/tr\u003e \u003c/tfoot\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003c/div\u003e \u003c/div\u003e \u003cdiv id=\"Sec7\" class=\"Section2\"\u003e \u003ch2\u003e2.4 HIRI Induction Procedure\u003c/h2\u003e \u003cp\u003eHepatic ischemia-reperfusion injury (HIRI) was induced following the method described by Şener \u003cem\u003eet al\u003c/em\u003e. (\u003cspan citationid=\"CR30\" class=\"CitationRef\"\u003e30\u003c/span\u003e) with slight modifications. Briefly, after overnight fasting, rats were anaesthetised with intraperitoneal injections of ketamine (50 mg/kg) and xylazine (10 mg/kg) (\u003cspan citationid=\"CR31\" class=\"CitationRef\"\u003e31\u003c/span\u003e). The abdominal area was shaved and disinfected, and a midline laparotomy was performed. The hepatic artery, portal vein supplying the median and left lateral lobes and the bile duct were clamped using microvascular clamp to induce 70% hepatic ischemia. After 60 minutes of ischemia, the clamp was removed to allow reperfusion which lasted for 24 hours. Sham-operated animals underwent the same procedure without vascular occlusion.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec8\" class=\"Section2\"\u003e \u003ch2\u003e2.5 Sample Collection\u003c/h2\u003e \u003cp\u003eAnimals were fasted overnight and twenty-four hours after reperfusion, the animals were anaesthetized using intraperitoneal injection of ketamine (50 mg/kg) and xylazine (10 mg/kg) (\u003cspan citationid=\"CR31\" class=\"CitationRef\"\u003e31\u003c/span\u003e). Confirmation of anaesthesia was determined by absence of corneal reflex and pedal withdrawal. Blood samples were collected via the retro-orbital puncture in plain bottles. Following this, the liver and kidneys were excised, washed in ice-cold saline, and weighed. Also, the collected blood was allowed to clot and centrifuged at 5000rpm for 10 minutes to obtain serum. Portions of the liver and kidney tissues were fixed in 10% neutral buffered formalin for histological examination.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec9\" class=\"Section2\"\u003e \u003ch2\u003e2.6 Biochemical Analyses\u003c/h2\u003e \u003cp\u003eSerum and tissue homogenates were used for various biochemical assays. Liver function markers (ALT, AST, GGT) and kidney function markers (urea, creatinine) were determined using commercial kits on an automated analyzer (Biosystem A25 Random Access Analyzer).\u003c/p\u003e \u003cp\u003e \u003cb\u003eOxidative stress markers\u003c/b\u003e, including superoxide dismutase (SOD) activity, malondialdehyde (MDA) levels, glutathione (GSH) content, and catalase activity, were measured in liver and kidney tissue homogenates using established spectrophotometric methods (\u003cspan citationid=\"CR32\" class=\"CitationRef\"\u003e32\u003c/span\u003e, \u003cspan citationid=\"CR33\" class=\"CitationRef\"\u003e33\u003c/span\u003e, \u003cspan citationid=\"CR34\" class=\"CitationRef\"\u003e34\u003c/span\u003e, \u003cspan citationid=\"CR35\" class=\"CitationRef\"\u003e35\u003c/span\u003e). Total protein content in the tissue homogenates was determined by the Bradford method (\u003cspan citationid=\"CR36\" class=\"CitationRef\"\u003e36\u003c/span\u003e).\u003c/p\u003e \u003cp\u003e \u003cb\u003eInflammatory markers\u003c/b\u003e, including tumour necrosis factor-alpha (TNF-α) in serum and myeloperoxidase (MPO) activity in liver and kidney tissues, were assessed using ELISA and spectrophotometric techniques, respectively (RandD Systems; 37). Serum nitrite levels, as an indicator of nitric oxide production, were measured using the Griess reaction (\u003cspan citationid=\"CR38\" class=\"CitationRef\"\u003e38\u003c/span\u003e). Caspase-3 activity, a marker of apoptosis, was determined in liver and kidney tissue homogenates using a colorimetric assay kit (Sigma-Aldrich, USA).\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec10\" class=\"Section2\"\u003e \u003ch2\u003e2.7 Histological Examination\u003c/h2\u003e \u003cp\u003eFixed liver and kidney tissues were processed using standard histological techniques and stained with Hematoxylin and Eosin (HandE) for general morphological assessment. Additional stains, including Masson's trichrome for liver fibrosis and Periodic acid-Schiff (PAS) for renal basement membrane and tubular brush border changes, were also performed. The stained sections were examined under a light microscope (Olympus BX51, Tokyo, Japan) and photographed.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec11\" class=\"Section2\"\u003e \u003ch2\u003e2.8 Statistical Analysis\u003c/h2\u003e \u003cp\u003eData were analysed using GraphPad Prism software (version 9.0, GraphPad Software Inc., San Diego, CA, USA). Results were expressed as mean\u0026thinsp;\u0026plusmn;\u0026thinsp;standard error of the mean (SEM). Differences between groups were analysed using one-way analysis of variance (ANOVA) followed by Tukey's post hoc test for multiple comparisons.\u003c/p\u003e \u003c/div\u003e"},{"header":"3 Results","content":"\u003cdiv id=\"Sec13\" class=\"Section2\"\u003e\n \u003ch2\u003e3.1 Liver Function Markers\u003c/h2\u003e\n \u003cp\u003e\u003cstrong\u003eAlanine aminotransferase (ALT)\u003c/strong\u003e\u003c/p\u003e\n \u003cp\u003eThere was significant increase in ALT concentration in the HIRI group compared with the sham group.\u003c/p\u003e\n \u003cp\u003e\u003cstrong\u003eAspartate aminotransferase (AST)\u003c/strong\u003e\u003c/p\u003e\n \u003cp\u003eAST concentration was significantly increased in the HIRI group when compared with the sham operated group. Significant decrease in AST concentration was also observed in both the low-dose BP and high-dose BP groups.\u003c/p\u003e\n \u003cp\u003e\u003cstrong\u003eGamma-glutamyl transferase (GGT)\u003c/strong\u003e\u003c/p\u003e\n \u003cp\u003eThere was a statistically significant increase in the GGT concentration of the HIRI group when compared with the sham operated group. GGT was however significantly reduced in the high-dose BP group when compared with the sham and low-dose BP groups.\u003c/p\u003e\n\u003c/div\u003e\n\u003cdiv id=\"Sec14\" class=\"Section2\"\u003e\n \u003ch2\u003e3.2 Kidney Function Markers\u003c/h2\u003e\n \u003cp\u003e\u003cstrong\u003eCreatinine\u003c/strong\u003e\u003c/p\u003e\n \u003cp\u003eThere was a statistically significant increase in creatinine levels of the HIRI group when compared with the sham operated group. The low-dose and high dose groups both significantly reduced creatinine levels when compared with the HIRI group.\u003c/p\u003e\n \u003cp\u003e\u003cstrong\u003eUrea\u003c/strong\u003e\u003c/p\u003e\n \u003cp\u003eThere was a statistically significant increase in the urea concentration of the HIRI group when compared with the sham operated group. Urea concentration was however significantly reduced by both the low-dose BP and high-dose BP groups. Furthermore, the high-dose group significantly decreased urea concentration when compared with the low-dose group.\u003c/p\u003e\n\u003c/div\u003e\n\u003cdiv id=\"Sec15\" class=\"Section2\"\u003e\n \u003ch2\u003e3.3 Hepatic Oxidative Stress Markers\u003c/h2\u003e\n \u003cp\u003e\u003cstrong\u003eHepatic Glutathione (GSH)\u003c/strong\u003e\u003c/p\u003e\n \u003cp\u003eHepatic GSH was significantly reduced in the HIRI group when compared with the sham group. Also, both the low-dose BP and high-dose BP showed a significant increase in GSH concentration when compared to the HIRI group.\u003c/p\u003e\n \u003cp\u003e\u003cstrong\u003eHepatic Total Protein\u003c/strong\u003e\u003c/p\u003e\n \u003cp\u003eHepatic total protein was significantly reduced in the HIRI group when compared with the sham group. Also, both the low-dose BP and high-dose BP groups showed a significant increase in total protein concentration when compared to the HIRI group. Significant decrease was further observed in the high-dose BP group when compared with the low-dose BP group.\u003c/p\u003e\n \u003cp\u003e\u003cstrong\u003eHepatic Malondialdehyde (MDA)\u003c/strong\u003e\u003c/p\u003e\n \u003cp\u003eMDA concentration significantly increased in the livers of the rats of the HIRI group when compared with the sham group. Both the low-dose BP and high-dose BP groups, however, were able to significantly reduce MDA concentration when compared to the HIRI group. Furthermore, there was a significant decrease in the high-dose BP group when compared with the low-dose BP group.\u003c/p\u003e\n \u003cp\u003e\u003cstrong\u003eHepatic Superoxide dismutase (SOD)\u003c/strong\u003e\u003c/p\u003e\n \u003cp\u003eHepatic SOD concentration was significantly decreased in HIRI group when compared with the sham group. In addition to this, both low-dose BP and high-dose BP groups showed a significant increase in SOD concentration when compared with the HIRI group.\u003c/p\u003e\n \u003cp\u003e\u003cstrong\u003eHepatic Catalase\u003c/strong\u003e\u003c/p\u003e\n \u003cp\u003eThere was a statistically significant decrease in hepatic catalase concentration in the HIRI group when compared with the sham group. There was also a statistically significant increase in catalase concentration in the low-dose BP and high-dose BP groups when compared with the HIRI group.\u003c/p\u003e\n\u003c/div\u003e\n\u003cdiv id=\"Sec16\" class=\"Section2\"\u003e\n \u003ch2\u003e3.4 Renal Oxidative Stress Markers\u003c/h2\u003e\n \u003cp\u003e\u003cstrong\u003eRenal Glutathione (GSH)\u003c/strong\u003e\u003c/p\u003e\n \u003cp\u003eRenal GSH was significantly reduced in the HIRI group when compared with the sham group. Also, both the low-dose BP and high-dose BP showed a significant increase in GSH concentration when compared to the HIRI group.\u003c/p\u003e\n \u003cp\u003e\u003cstrong\u003eRenal Total Protein\u003c/strong\u003e\u003c/p\u003e\n \u003cp\u003eTotal protein was significantly reduced in the HIRI group when compared with the sham group. Also, both the low-dose BP and high-dose BP showed a significant increase in total protein concentration when compared to the HIRI group.\u003c/p\u003e\n \u003cp\u003eR\u003cstrong\u003eenal Malondialdehyde (MDA)\u003c/strong\u003e\u003c/p\u003e\n \u003cp\u003eRenal MDA concentration was significantly increased in the HIRI group when compared to the HIRI group. Furthermore, both low-dose BP and high-dose BP groups had a significantly reduced MDA concentration in comparison with the HIRI group.\u003c/p\u003e\n \u003cp\u003e\u003cstrong\u003eRenal Superoxide dismutase (SOD)\u003c/strong\u003e\u003c/p\u003e\n \u003cp\u003eThe renal concentration of SOD was significantly reduced in the HIRI group when compared with the sham group. Only the high-dose BP group, however, showed a significant increase in SOD concentration when compared with the HIRI group.\u003c/p\u003e\n \u003cp\u003eR\u003cstrong\u003eenal Catalase\u003c/strong\u003e\u003c/p\u003e\n \u003cp\u003eCatalase concentration was significantly reduced in the kidneys of the animals in the HIRI group when compared with the sham group. Also, both low-dose BP and high-dose BP groups showed significant increase in catalase concentration when compared with the HIRI group.\u003c/p\u003e\n\u003c/div\u003e\n\u003cdiv id=\"Sec17\" class=\"Section2\"\u003e\n \u003ch2\u003e3.5 Inflammatory Markers\u003c/h2\u003e\n \u003cp\u003e\u003cstrong\u003eTumour necrosis factor (TNF-\u0026alpha;)\u003c/strong\u003e\u003c/p\u003e\n \u003cp\u003eThere was a statistically significant increase in TNF-\u0026alpha; concentration in the HIRI group when compared with the sham group. Significant decrease was also observed in the low-dose BP and high-dose BP groups when compared with the HIRI group. Furthermore, the high-dose BP group had a statistically significant reduction in TNF-\u0026alpha; concentration when compared with the low-dose BP group.\u003c/p\u003e\n \u003cp\u003e\u003cstrong\u003eMyeloperoxidase (MPO)\u003c/strong\u003e\u003c/p\u003e\n \u003cp\u003eThere was a statistically significant increase in MPO concentration in the HIRI group compared with the sham group. Also, the high-dose BP group had its MPO concentration significantly reduced when compared with the HIRI group.\u003c/p\u003e\n \u003cp\u003e\u003cstrong\u003eSerum nitrite\u003c/strong\u003e\u003c/p\u003e\n \u003cp\u003eThere was no statistically significant difference in nitrite concentration among the groups.\u003c/p\u003e\n\u003c/div\u003e\n\u003cdiv id=\"Sec18\" class=\"Section2\"\u003e\n \u003ch2\u003e3.6 Apoptotic Marker\u003c/h2\u003e\n \u003cp\u003e\u003cstrong\u003eCaspase-3\u003c/strong\u003e\u003c/p\u003e\n \u003cp\u003eThere was a statistically significant increase in caspase-3 concentration in the HIRI group when compared with the sham group. The low-dose BP and high-does BP groups significantly reduced caspase-3 concentration with respect to the HIRI group. In addition, there was a significant decrease in caspase-3 concentration in the high-dose BP group when compared with the low-dose BP group.\u003c/p\u003e\n\u003c/div\u003e\n\u003cdiv id=\"Sec19\" class=\"Section2\"\u003e\n \u003ch2\u003e3.7 Histology of the Liver\u003c/h2\u003e\n \u003cp\u003e\u003cstrong\u003eSham group (SH)\u003c/strong\u003e\u003c/p\u003e\n \u003cp\u003eThe hepatic tissue appears normal. The hepatocytes (H) appear polygonal with well-outlined vesicular nucleus (N). The hepatocytes are separated by the sinusoids (S) with thin endothelial lining, free from collections and inflammatory cells. The central vein (CV) and bile duct (BD) also appear normal\u003c/p\u003e\n \u003cp\u003e\u003cstrong\u003eHIRI group (HH)\u003c/strong\u003e\u003c/p\u003e\n \u003cp\u003eThe hepatic tissue appears distorted. The hepatocytes (H) appear thickened with well-outlined vesicular nucleus (N). The hepatocytes are separated by the sinusoids (S) with hemorrhagic lesions (circle). There are focal areas of inflammatory cell infiltration (box) and focal vascular congestion (star) in the central vein (CV). The bile duct (BD) appear normal.\u003c/p\u003e\n \u003cp\u003e\u003cstrong\u003eLow-dose BP group (LDH)\u003c/strong\u003e\u003c/p\u003e\n \u003cp\u003eThe hepatic tissue appears normal. The hepatocytes (H) appear polygonal with well-outlined vesicular nucleus (N). The hepatocytes are separated by the sinusoids (S) with thin endothelial lining, free from collections and inflammatory cells. The central vein (CV) and bile duct (BD) also appear normal.\u003c/p\u003e\n \u003cp\u003e\u003cstrong\u003eHigh-dose BP (HDH)\u003c/strong\u003e\u003c/p\u003e\n \u003cp\u003eThe hepatic tissue appears distorted. The hepatocytes (H) appear polygonal with well-outlined vesicular nucleus (N). The hepatocytes are separated by the sinusoids (S) and it is free of lesion. There is focal vascular congestion (star) in the central vein (CV).\u003c/p\u003e\n\u003c/div\u003e\n\u003cdiv id=\"Sec20\" class=\"Section2\"\u003e\n \u003ch2\u003e3.8 Histology of the Kidney\u003c/h2\u003e\n \u003cp\u003e\u003cstrong\u003eSham group (SH)\u003c/strong\u003e\u003c/p\u003e\n \u003cp\u003eThe histology of the renal tissue appears normal. The renal corpuscle (arrow) consists of the glomerulus (G) containing podocytes and separated by a well-defined Bowman\u0026rsquo;s space (BS). The renal tubules (RT) are lined by columnar-cubiodal epithelium and separated by interstitium that is free from congestions and collections.\u003c/p\u003e\n \u003cp\u003e\u003cstrong\u003eHIRI group (HH)\u003c/strong\u003e\u003c/p\u003e\n \u003cp\u003eThe histology of the renal tissue appears distorted. The renal corpuscle (arrow) consists of the glomerulus (G) containing podocytes and separated by an ill-defined Bowman\u0026rsquo;s space (BS). The renal tubules (RT) are lined by columnar-cubiodal epithelium and separated by interstitium with focal areas of hemorrhagic lesions (black circle). There are also focal areas of vascular congestions (star) and inflammatory cell infiltration (box).\u003c/p\u003e\n \u003cp\u003e\u003cstrong\u003eLow-dose BP (LDH)\u003c/strong\u003e\u003c/p\u003e\n \u003cp\u003eThe histology of the renal tissue appears normal. The renal corpuscle (arrow) consists of the glomerulus (G) containing podocytes and separated by a well-defined Bowman\u0026rsquo;s space (BS). The renal tubules (RT) are lined by columnar-cubiodal epithelium and separated by interstitium that is free from congestions and collections.\u003c/p\u003e\n \u003cp\u003e\u003cstrong\u003eHigh-dose BP (HDH)\u003c/strong\u003e\u003c/p\u003e\n \u003cp\u003eThe histology of the renal tissue appears distorted. The renal corpuscle (arrow) consists of the glomerulus (G) containing podocytes and separated by an ill-defined Bowman\u0026rsquo;s space (BS). The renal tubules (RT) are lined by columnar-cubiodal epithelium and separated by interstitium with focal areas of hemorrhagic lesions (black circle).\u003c/p\u003e\n\u003c/div\u003e"},{"header":"4 Discussion","content":"\u003cp\u003eThe present study demonstrates that HIRI induces significant remote organ dysfunction, particularly affecting renal integrity. The relationship between hepatic damage and renal dysfunction (hepatorenal syndrome), highlights the need for protective interventions that can mitigate the systemic impact of HIRI (\u003cspan citationid=\"CR39\" class=\"CitationRef\"\u003e39\u003c/span\u003e). \u003cem\u003eB. pinnatum\u003c/em\u003e is rich in flavonoids, alkaloids, and terpenes, which are known for their potent antioxidant and anti-inflammatory properties. Of particular interest is the presence of quercetin and kaempferol derivatives, which have shown renoprotective effects in other models of acute kidney injury (\u003cspan citationid=\"CR12\" class=\"CitationRef\"\u003e12\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eThe observed elevation in serum ALT, AST, and GGT levels in the HIRI group confirms substantial hepatocellular damage, consistent with previous findings (\u003cspan citationid=\"CR40\" class=\"CitationRef\"\u003e40\u003c/span\u003e). As demonstrated by Gin\u0026egrave;s and Schrier (\u003cspan citationid=\"CR41\" class=\"CitationRef\"\u003e41\u003c/span\u003e), severe liver injury can induce systemic vasodilation, leading to reduced effective arterial blood volume and subsequent activation of the renin-angiotensin-aldosterone system (RAAS). This activation results in renal vasoconstriction and compromised renal perfusion. Also, the release of damage-associated molecular patterns (DAMPs) from injured hepatocytes can trigger a systemic inflammatory response. Released pro-inflammatory cytokines and chemokines can directly impact renal function through leukocyte infiltration and tubular cell injury (\u003cspan citationid=\"CR42\" class=\"CitationRef\"\u003e42\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eConsequently, oxidative stress initiated in the liver, evidenced by elevated protein degradation, GGT and MDA levels, as well as reduced GSH, SOD, and catalase levels, can propagate systemically, affecting distant organs including the kidneys. Oxidative stress is one of the primary mechanisms implicated in IRI (\u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e3\u003c/span\u003e, \u003cspan citationid=\"CR43\" class=\"CitationRef\"\u003e43\u003c/span\u003e). In the current study, \u003cem\u003eB. pinnatum\u003c/em\u003e significantly ameliorated oxidative stress in both liver and kidney tissues, as evidenced by reductions in MDA levels and increases in endogenous antioxidant enzymes including superoxide dismutase SOD and catalase. These results are in alignment with earlier studies, which highlight the antioxidant properties of \u003cem\u003eB. pinnatum\u003c/em\u003e as a contributor to its therapeutic effects (\u003cspan citationid=\"CR12\" class=\"CitationRef\"\u003e12\u003c/span\u003e, \u003cspan citationid=\"CR17\" class=\"CitationRef\"\u003e17\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eThe antioxidative effects observed may be attributed to the plant\u0026rsquo;s phytochemical constituents, including flavonoids such as quercetin and kaempferol, known for their ROS-scavenging capabilities (\u003cspan citationid=\"CR44\" class=\"CitationRef\"\u003e44\u003c/span\u003e, \u003cspan citationid=\"CR45\" class=\"CitationRef\"\u003e45\u003c/span\u003e). These compounds enhance the activity of nuclear factor erythroid 2-related factor 2 (Nrf2), a major regulator of the cellular antioxidant response. Activation of the Nrf2 pathway also upregulates the expression of genes encoding antioxidant enzymes, which mitigate oxidative stress by neutralising ROS and preserving mitochondrial integrity (\u003cspan citationid=\"CR46\" class=\"CitationRef\"\u003e46\u003c/span\u003e). The increased activity of SOD and catalase observed in this study supports the hypothesis that \u003cem\u003eB. pinnatum\u003c/em\u003e activates Nrf2, aligning with previous findings that document the role of flavonoids in Nrf2-mediated antioxidative pathways (\u003cspan citationid=\"CR47\" class=\"CitationRef\"\u003e47\u003c/span\u003e, \u003cspan citationid=\"CR48\" class=\"CitationRef\"\u003e48\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eMoreover, HIRI-induced kidney injury is strongly associated with inflammation, involving the release of pro-inflammatory cytokines (\u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e6\u003c/span\u003e, \u003cspan citationid=\"CR7\" class=\"CitationRef\"\u003e7\u003c/span\u003e). Inflammatory cascade analysis revealed that \u003cem\u003eB. pinnatum\u003c/em\u003e administration led to significant reductions in TNF-α and MPO levels, suggesting an attenuation of the inflammatory cascade. This anti-inflammatory effect may result from the inhibition of nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB), which has been identified as a central mediator of inflammation in ischemia-reperfusion injury (\u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e1\u003c/span\u003e, \u003cspan citationid=\"CR49\" class=\"CitationRef\"\u003e49\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eThe phytoconstituents in \u003cem\u003eB. pinnatum\u003c/em\u003e, including triterpenoids and flavonoids, likely play a role in modulating NF-κB activation. These compounds have been shown to suppress NF-κB activity by preventing the phosphorylation and degradation of its inhibitor, IκBα, thus impeding the transcription of pro-inflammatory cytokines (\u003cspan citationid=\"CR44\" class=\"CitationRef\"\u003e44\u003c/span\u003e). This NF-κB inhibition reduces the recruitment of neutrophils and macrophages to the site of injury, thereby decreasing MPO levels, as observed in the present study. Furthermore, the downregulation of TNF-α may have limited secondary damage induced by ROS, creating a positive feedback loop that mitigates both inflammation and oxidative stress. Such multi-faceted anti-inflammatory effects underscore the therapeutic potential of \u003cem\u003eB. pinnatum\u003c/em\u003e in ameliorating HIRI-induced renal injury (\u003cspan citationid=\"CR11\" class=\"CitationRef\"\u003e11\u003c/span\u003e, \u003cspan citationid=\"CR12\" class=\"CitationRef\"\u003e12\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eFurthermore, our biochemical analyses revealed significant elevations in serum creatinine and urea levels in the HIRI group, indicating compromised glomerular filtration and renal dysfunction. These findings align with previous studies demonstrating the development of acute kidney injury following hepatic ischemia-reperfusion (\u003cspan citationid=\"CR39\" class=\"CitationRef\"\u003e39\u003c/span\u003e, \u003cspan citationid=\"CR50\" class=\"CitationRef\"\u003e50\u003c/span\u003e). However, the nephroprotective effects of \u003cem\u003eB. pinnatum\u003c/em\u003e were evidenced by improved levels of creatinine and urea. These markers were significantly reduced in \u003cem\u003eB. pinnatum\u003c/em\u003e-treated groups compared to untreated HIRI controls, indicating preserved glomerular filtration rate (GFR) and reduced renal damage. Notably, this nephroprotective property was demonstrated in a dose-dependent manner, with the higher dose showing superior efficacy in preserving renal function. This preservation of renal function is consistent with prior findings suggesting that \u003cem\u003eB. pinnatum\u003c/em\u003e has protective effects on renal tissue under conditions of oxidative stress and inflammation (\u003cspan citationid=\"CR5\" class=\"CitationRef\"\u003e5\u003c/span\u003e, \u003cspan citationid=\"CR45\" class=\"CitationRef\"\u003e45\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eThe reduction in creatinine and urea levels observed in this study may stem from multiple mechanisms, including the antioxidant and anti-inflammatory effects of \u003cem\u003eB. pinnatum\u003c/em\u003e, as well as its role in inhibiting apoptosis. Apoptosis in ischemia-reperfusion injury is mediated by both intrinsic (mitochondrial) and extrinsic (death receptor) pathways, with caspase-3 serving as an important executor enzyme (\u003cspan citationid=\"CR4\" class=\"CitationRef\"\u003e4\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eIn line with this, our study revealed a significant elevation in caspase-3 levels in the HIRI group, indicating enhanced apoptotic activity. The attenuation of caspase-3 activity observed here suggests that \u003cem\u003eB. pinnatum\u003c/em\u003e may inhibit apoptosis, potentially through modulation of mitochondrial pathways and inhibition of the mitochondrial permeability transition pore (mPTP) opening, as well as the modulation of Bcl-2 family proteins (\u003cspan citationid=\"CR51\" class=\"CitationRef\"\u003e51\u003c/span\u003e, \u003cspan citationid=\"CR52\" class=\"CitationRef\"\u003e52\u003c/span\u003e). Reduced apoptosis not only preserves renal tubular structure but also minimises functional impairment, which aligns with the observed maintenance of kidney function markers. Histopathological examination provided visual confirmation of these biochemical findings. The HIRI group showed characteristic renal tissue damage, including ill-defined Bowman's spaces and inflammatory cell infiltration. \u003cem\u003eB. pinnatum\u003c/em\u003e treatment preserved normal renal architecture, supporting its nephroprotective effects at the tissue level.\u003c/p\u003e"},{"header":"5 Conclusion","content":"\u003cp\u003eThe nephroprotective effects of \u003cem\u003eB. pinnatum\u003c/em\u003e appear to be mediated through multiple mechanisms: (\u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e1\u003c/span\u003e) antioxidant properties, evidenced by restoration of GSH, SOD, and catalase levels; (\u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2\u003c/span\u003e) anti-inflammatory effects, demonstrated by reduced TNF-α and MPO levels; and (\u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e3\u003c/span\u003e) anti-apoptotic activity, shown by normalised caspase-3 levels. These findings suggest that \u003cem\u003eB. pinnatum\u003c/em\u003e could serve as a promising therapeutic agent for preventing HIRI-induced kidney injury, though further clinical studies are warranted to establish its efficacy in human subjects.\u003c/p\u003e"},{"header":"Abbreviations","content":"\u003cp\u003eALT\u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp;\u0026nbsp;Alanine Aminotransferase\u003c/p\u003e\n\u003cp\u003eAST\u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp;Aspartate Aminotransferase\u003c/p\u003e\n\u003cp\u003eGGT\u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp;\u0026nbsp;Gamma-glutamyl transferase\u003c/p\u003e\n\u003cp\u003eBP \u003cem\u003eBryophyllum pinnatum\u003c/em\u003e\u003c/p\u003e\n\u003cp\u003eHIRI\u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp;Hepatic Ischemia-Reperfusion Injury\u003c/p\u003e\n\u003cp\u003eIRI\u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp;\u0026nbsp;Ischemia-Reperfusion Injury\u003c/p\u003e\n\u003cp\u003eMDA\u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp;Malondialdehyde\u003c/p\u003e\n\u003cp\u003eROS\u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp;\u0026nbsp;Reactive Oxygen Species\u003c/p\u003e\n\u003cp\u003eSOD\u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp;\u0026nbsp;Superoxide Dismutase\u003c/p\u003e\n\u003cp\u003eGSH \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp;Reduced Glutathione\u003c/p\u003e"},{"header":"Declarations","content":"\u003cp\u003e\u003cstrong\u003eEthics Approval and Consent to Participate:\u0026nbsp;\u003c/strong\u003eEthical approval was obtained from Ethics Committee in Animal Care and Use in Research of FBMS with reference number: \u003cstrong\u003eERCFBMSLAUTECH: 058/08/2024.\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eConsent for Publication:\u0026nbsp;\u003c/strong\u003eNot applicable\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eCompeting Interests:\u0026nbsp;\u003c/strong\u003eThe authors declare that there is no conflict of interest in this manuscript\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eFunding Declaration:\u0026nbsp;\u003c/strong\u003eNo funding was received for this study\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eClinical trial number\u003c/strong\u003e: Not applicable\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAuthors\u0026apos; contributions:\u0026nbsp;\u003c/strong\u003eOVO, OAA, and OSH designed the study; OVO, OAA, RAA, and OSH performed the experiments; RAA, ABA, AWS, and SOO provided new reagents and analytical tools; OVO, ABA, OSH, and OSH analysed the research data; OVO, RAA, OSH, OSH, AWS, and SOO contributed to the writing of the manuscript; all authors read and approved the final manuscript.\u003cstrong\u003e\u003cbr\u003e\u0026nbsp;Acknowledgements:\u0026nbsp;\u003c/strong\u003eThe authors acknowledge the support of the Department of Physiology, LAUTECH, and technical staff of the animal facility.\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\n\u003cli\u003eYellon, D.M., and Hausenloy, D.J. (2013). Myocardial reperfusion injury. \u003cem\u003eNew England Journal of Medicine\u003c/em\u003e, 370, 112\u0026ndash;119.\u003c/li\u003e\n\u003cli\u003eElias-Mir\u0026oacute;, M., Jim\u0026eacute;nez-Castro, M. B., Rod\u0026eacute;s, J., and Peralta, C. (2013). Current knowledge on oxidative stress in hepatic ischemia/reperfusion. Free radical research, 47(8), 555-568.\u003c/li\u003e\n\u003cli\u003eMurphy, M. P., and Hartley, R. C. (2018). Mitochondria as a therapeutic target for common pathologies. \u003cem\u003eNature Reviews Drug Discovery\u003c/em\u003e, 17(12), 865-886.\u003c/li\u003e\n\u003cli\u003eKalogeris, T., Baines, C. P., Krenz, M., and Korthuis, R. J. (2012). Cell biology of ischemia/reperfusion injury. \u003cem\u003eInternational review of cell and molecular biology\u003c/em\u003e, 298, 229-317.\u003c/li\u003e\n\u003cli\u003eTrinh, E., Alam, A., Tchervenkov, J., and Cantarovich, M. (2017). Impact of acute kidney injury following liver transplantation on long‐term outcomes. \u003cem\u003eClinical Transplantation\u003c/em\u003e, 31(1), e12863.\u003c/li\u003e\n\u003cli\u003eLai, I. R., Chang, K. J., Tsai, H. W., and Chen, C. F. (2008). Pharmacological preconditioning with simvastatin protects liver from ischemia-reperfusion injury by heme oxygenase-1 induction. \u003cem\u003eTransplantation\u003c/em\u003e, 85(5), 732-738.\u003c/li\u003e\n\u003cli\u003eAwad, A.S., Rouse, M., Huang, L., Vergis, A.L., Reutershan, J., Cathro, H.P., Linden, J. and Okusa, M.D., (2009). Compartmentalization of neutrophils in the kidney and lung following acute ischemic kidney injury. \u003cem\u003eKidney international\u003c/em\u003e, 75(7), pp.689-698.\u003c/li\u003e\n\u003cli\u003eCasillas-Ram\u0026iacute;rez, A., Zaouali, A., Padrissa-Alt\u0026eacute;s, S., Ben Mosbah, I., Pertosa, A., Alfany-Fern\u0026aacute;ndez, I., Bintanel-Morcillo, M., Xaus, C., Rimola, A., Rod\u0026eacute;s, J. and Rosello-Catafau, J., (2009). Insulin-like growth factor and epidermal growth factor treatment: new approaches to protecting steatotic livers against ischemia-reperfusion injury. \u003cem\u003eEndocrinology\u003c/em\u003e, 150(7), pp.3153-3161.\u003c/li\u003e\n\u003cli\u003eBuchholz, B., Donato, M., D\u0026rsquo;Annunzio, V., and Gelpi, R. J. (2014). Ischemic postconditioning: mechanisms, comorbidities, and clinical application. \u003cem\u003eMolecular and cellular biochemistry\u003c/em\u003e, 392, 1-12.\u003c/li\u003e\n\u003cli\u003eRadomska-Leśniewska, D. M., and Skopiński, P. (2012). Review paper N-acetylcysteine as an anti-oxidant and anti-inflammatory drug and its some clinical applications. Central European journal of immunology, 37(1), 57-66.\u003c/li\u003e\n\u003cli\u003eOjewole, J. A. (2005). Antinociceptive, anti-inflammatory and antidiabetic effects of \u003cem\u003eBryophyllum pinnatum\u003c/em\u003e (Crassulaceae) leaf aqueous extract. \u003cem\u003eJournal of ethnopharmacology\u003c/em\u003e, 99(1), 13-19.\u003c/li\u003e\n\u003cli\u003eF\u0026uuml;rer, K., Sim\u0026otilde;es-W\u0026uuml;st, A. P., von Mandach, U., Hamburger, M., and Potterat, O. (2016). \u003cem\u003eBryophyllum pinnatum\u003c/em\u003e and related species used in anthroposophic medicine: constituents, pharmacological activities, and clinical efficacy. \u003cem\u003ePlanta medica\u003c/em\u003e, 82(11/12), 930-941.\u003c/li\u003e\n\u003cli\u003eFernandes, J. M., Cunha, L. M., Azevedo, E. P., Louren\u0026ccedil;o, E. M., Fernandes-Pedrosa, M. F., and Zucolotto, S. M. (2019). Kalanchoe laciniata and Bryophyllum pinnatum: an updated review about ethnopharmacology, phytochemistry, pharmacology and toxicology. Revista Brasileira de Farmacognosia, 29(4), 529-558.\u003c/li\u003e\n\u003cli\u003eShahrajabian, M. H., Sun, W., and Cheng, Q. (2022). The importance of flavonoids and phytochemicals of medicinal plants with antiviral activities. Mini-Reviews in Organic Chemistry, 19(3), 293-318.\u003c/li\u003e\n\u003cli\u003eAkinwumi, I. A., and Ambali, O. A. (2024). Cardiac glycosides from african medicinal plants as promising therapeutics. \u003cem\u003eTropical Journal of Phytochemistry and Pharmaceutical Sciences\u003c/em\u003e, 3(2), 158-167.\u003c/li\u003e\n\u003cli\u003eChibli, L.A., Rodrigues, K.C., Gasparetto, C.M., Pinto, N.C., Fabri, R.L., Scio, E., Alves, M.S., Del-Vechio-Vieira, G. and Sousa, O.V., 2014. Anti-inflammatory effects of Bryophyllum pinnatum (Lam.) Oken ethanol extract in acute and chronic cutaneous inflammation. \u003cem\u003eJournal of ethnopharmacology\u003c/em\u003e, \u003cem\u003e154\u003c/em\u003e(2), pp.330-338.\u003c/li\u003e\n\u003cli\u003eSelvakumar, P. (2022). Phytochemical and pharmacological profile review of \u003cem\u003eBryophyllum pinnatum\u003c/em\u003e. \u003cem\u003eBiomedical And Biotechnology Research Journal (Bbrj)\u003c/em\u003e, 6(3), 295-301.\u003c/li\u003e\n\u003cli\u003eTatsimo, S.J.N., Tamokou, J.D.D., Havyarimana, L., Csupor, D., Forgo, P., Hohmann, J., Kuiate, J.R. and Tane, P., (2012). Antimicrobial and antioxidant activity of kaempferol rhamnoside derivatives from Bryophyllum pinnatum. BMC Research notes, 5, pp.1-6.\u003c/li\u003e\n\u003cli\u003eSharma, G., Jangra, A., Sihag, S., Chaturvedi, S., Yadav, S., and Chhokar, V. (2024). Bryophyllum pinnatum (Lam.) Oken: unravelling therapeutic potential and navigating toxicity. \u003cem\u003ePhysiology and Molecular Biology of Plants\u003c/em\u003e, \u003cem\u003e30\u003c/em\u003e(9), 1413-1427.\u003c/li\u003e\n\u003cli\u003eYadav, M., Gulkari, V. D., and Wanjari, M. M. (2016). Bryophyllum pinnatum leaf extracts prevent formation of renal calculi in lithiatic rats. Ancient science of life, 36(2), 90-97.\u003c/li\u003e\n\u003cli\u003eAnadozie, S. O., Akinyemi, J. A., Agunbiade, S., Ajiboye, B. O., and Adewale, O. B. (2018). \u003cem\u003eBryophyllum pinnatum\u003c/em\u003e inhibits arginase II activity and prevents oxidative damage occasioned by carbon tetrachloride (CCl4) in rats. \u003cem\u003eBiomedicine and Pharmacotherapy\u003c/em\u003e, 101, 8-13.\u003c/li\u003e\n\u003cli\u003eShukla, A. B., Mandavia, D. R., Barvaliya, M. J., Baxi, S. N., and Tripathi, C. R. (2014). Evaluation of anti-urolithiatic effect of aqueous extract of Bryophyllum pinnatum (Lam.) leaves using ethylene glycol-induced renal calculi. \u003cem\u003eAvicenna journal of phytomedicine\u003c/em\u003e, \u003cem\u003e4\u003c/em\u003e(3), 151.\u003c/li\u003e\n\u003cli\u003eShinde, S. K., Chaware, V. J., and Redasani, V. K. (2024). Anti-urolithiatic effect of Ethanolic extract of Bryophyllum pinnatum (Lam) against chemicals induced Renal calculi in rats. \u003cem\u003eResearch Journal of Science and Technology\u003c/em\u003e, \u003cem\u003e16\u003c/em\u003e(4), 283-290.\u003c/li\u003e\n\u003cli\u003eUahomo, P. O., and Isirima, J. C. Attenuating Ketamine-Induced Nephrotoxicity with Bryophyllum pinnatum Extract: Biochemical and Histological Investigation.\u003c/li\u003e\n\u003cli\u003ePal, S. K., Lahon, L. C., Sarkar, B. K., Paul, S. R., and Debroy, B. (2020). Effect of methanol extract of Bryophyllum pinnatum leaves on ethylene glycol-induced urolithiasis in adult male albino rats. \u003cem\u003eJournal of Entomology and Zoology Studies\u003c/em\u003e, \u003cem\u003e8\u003c/em\u003e(4), 782-786.\u003c/li\u003e\n\u003cli\u003eAfzal, M., Kazmi, I., and Anwar, F. (2013). Antineoplastic potential of Bryophyllum pinnatum Lam. on chemically induced hepatocarcinogenesis in rats. \u003cem\u003ePharmacognosy research\u003c/em\u003e, \u003cem\u003e5\u003c/em\u003e(4), 247.\u003c/li\u003e\n\u003cli\u003eKombathula, L., and Chernapalli, V. (2024). Evaluation of efficacy of selected Bryophyllum pinnatum phytochemicals in hepatocarcinogenic therapy through in silico approach: Efficacy of B. Pinnatum phytochemicals on hepatocarcinoma. \u003cem\u003eIndian Journal of Experimental Biology (IJEB)\u003c/em\u003e, \u003cem\u003e62\u003c/em\u003e(09), 752-759.\u003c/li\u003e\n\u003cli\u003eOsifeso, O. O., Umoren, O. D., Bamidele, A. A., Obika, C. G., and Adeogun, O. A. (2025). Protective Effect of Bryophyllum pinnatum (Lam.) Oken (Miracle Leaf) Extract on Butylglycol-Induced Hepatotoxicity. \u003cem\u003eWorld News of Natural Sciences\u003c/em\u003e, \u003cem\u003e59\u003c/em\u003e, 239-249.\u003c/li\u003e\n\u003cli\u003eAjah, O., Unegbu, C. C., Onwusonye, C. J., Nnorom, C. M., and Duru, C. A. (2023). Effect of ethanol extract of Bryophyllum pinnatum leaf on lipid profile, renal and hepatic function biomarkers of high salt fed Albino rats. \u003cem\u003eJournal of Medicinal Herbs,\u003c/em\u003e, \u003cem\u003e13\u003c/em\u003e(4), 75-81.\u003c/li\u003e\n\u003cli\u003eŞener, G., Sehirli, \u0026Ouml;., Ip\u0026ccedil;i, Y., Ercan, F., şirvanci, S., Gedik, N., and Yeĝen, B. \u0026Ccedil;. (2005). Aqueous garlic extract alleviates ischaemia-reperfusion-induced oxidative hepatic injury in rats. \u003cem\u003eJournal of Pharmacy and Pharmacology\u003c/em\u003e, 57(1), 145-150.\u003c/li\u003e\n\u003cli\u003eLorenzini, F., Tambara Filho, R., Gomes, R. P. X., Martino-Andrade, A. J., Erdmann, T. R., and Matias, J. E. F. (2012). Long-term effects of the testicular torsion on the spermatogenesis of the contralateral testis and the preventive value of the twisted testis orchiepididymectomy. Acta cirurgica brasileira, 27, 388-395.\u003c/li\u003e\n\u003cli\u003eNandi, A., and Chatterjee, I. B. (1988). Assay of superoxide dismutase activity in animal tissues. \u003cem\u003eJournal of biosciences\u003c/em\u003e, 13, 305-315.\u003c/li\u003e\n\u003cli\u003eBeuge, J. A., Aust, S. D. (1978) Microsomal lipid peroxidation. \u003cem\u003eMethods Enzymol\u003c/em\u003e. 52: 302\u0026ndash;311.\u003c/li\u003e\n\u003cli\u003eBeutler, E. (1975) Glutathione. In: Beutler, E. (ed.) Red blood cell metabolism: a manual of biochemical methods. Grune and Stratton, New York, pp 112\u0026ndash;114.\u003c/li\u003e\n\u003cli\u003eAebi, H. (1974). Catalase. In Methods of enzymatic analysis (pp. 673-684). Academic press.\u003c/li\u003e\n\u003cli\u003eInchiosa, M. A. (1964). Direct biuret determination of total protein in tissue homogenates. \u003cem\u003eThe Journal of Laboratory and Clinical Medicine\u003c/em\u003e, 63(2), 319-324.\u003c/li\u003e\n\u003cli\u003eHillegass, L. M., Griswold, D. E., Brickson, B., AlbrightsonWinslow, C. (1990) Assessment of myeloperoxidase activity in whole rat kidney. \u003cem\u003eJ. Pharmacol. Methods\u003c/em\u003e 24: 285\u0026ndash;295.\u003c/li\u003e\n\u003cli\u003eGiovannoni, G., Land, J. M., Keir, G., Thompson, E. J., and Heales, S. J. R. (1997). Adaptation of the nitrate reductase and Griess reaction methods for the measurement of serum nitrate plus nitrite levels. \u003cem\u003eAnnals of Clinical Biochemistry\u003c/em\u003e, 34(2), 193-198.\u003c/li\u003e\n\u003cli\u003eNastos, C., Kalimeris, K., Papoutsidakis, N., Tasoulis, M.K., Lykoudis, P.M., Theodoraki, K., Nastou, D., Smyrniotis, V. and Arkadopoulos, N., (2014). Global consequences of liver ischemia/reperfusion injury. \u003cem\u003eOxidative medicine and cellular longevity\u003c/em\u003e, 2014(1), p.906965.\u003c/li\u003e\n\u003cli\u003eJaeschke, H., and Woolbright, B. L. (2012). Current strategies to minimize hepatic ischemia\u0026ndash;reperfusion injury by targeting reactive oxygen species. \u003cem\u003eTransplantation reviews\u003c/em\u003e, 26(2), 103-114.\u003c/li\u003e\n\u003cli\u003eGin\u0026egrave;s, P., and Schrier, R. W. (2009). Renal failure in cirrhosis. \u003cem\u003eNew England Journal of Medicine\u003c/em\u003e, 361(13), 1279-1290.\u003c/li\u003e\n\u003cli\u003eBernardi, M., Moreau, R., Angeli, P., Schnabl, B., and Arroyo, V. (2015). Mechanisms of decompensation and organ failure in cirrhosis: from peripheral arterial vasodilation to systemic inflammation hypothesis. \u003cem\u003eJournal of hepatology\u003c/em\u003e, 63(5), 1272-1284.\u003c/li\u003e\n\u003cli\u003eEltzschig, H. K., and Eckle, T. (2011). Ischemia and reperfusion\u0026mdash;from mechanism to translation. \u003cem\u003eNature medicine\u003c/em\u003e, 17(11), 1391-1401.\u003c/li\u003e\n\u003cli\u003eKamboj, A., and Saluja, A. (2009). \u003cem\u003eBryophyllum pinnatum\u003c/em\u003e (Lam.) Kurz.: phytochemical and pharmacological profile: a review. \u003cem\u003ePharmacognosy Reviews\u003c/em\u003e, 3(6), 364.\u003c/li\u003e\n\u003cli\u003eAkbari, B., Baghaei‐Yazdi, N., Bahmaie, M., and Mahdavi Abhari, F. (2022). The role of plant‐derived natural antioxidants in reduction of oxidative stress. \u003cem\u003eBioFactors\u003c/em\u003e, 48(3), 611-633.\u003c/li\u003e\n\u003cli\u003eMa, Y. F., Wu, Z. H., Gao, M., and Loor, J. J. (2018). Nuclear factor erythroid 2-related factor 2 antioxidant response element pathways protect bovine mammary epithelial cells against H2O2-induced oxidative damage in vitro. \u003cem\u003eJournal of dairy science\u003c/em\u003e, 101(6), 5329-5344.\u003c/li\u003e\n\u003cli\u003eKobayashi, A., Kang, M. I., Watai, Y., Tong, K. I., Shibata, T., Uchida, K., and Yamamoto, M. (2006). Oxidative and electrophilic stresses activate Nrf2 through inhibition of ubiquitination activity of Keap1. \u003cem\u003eMolecular and cellular biology\u003c/em\u003e, 26(1), 221-229.\u003c/li\u003e\n\u003cli\u003eAshrafizadeh, M., Ahmadi, Z., Mohammadinejad, R., Farkhondeh, T., and Samarghandian, S. (2020). Curcumin activates the Nrf2 pathway and induces cellular protection against oxidative injury. \u003cem\u003eCurrent Molecular Medicine\u003c/em\u003e, 20(2), 116-133.\u003c/li\u003e\n\u003cli\u003ePerkins, N. D. (2007). Integrating cell-signalling pathways with NF-\u0026kappa;B and IKK function. \u003cem\u003eNature reviews Molecular cell biology\u003c/em\u003e, 8(1), 49-62.\u003c/li\u003e\n\u003cli\u003eLevey, A. S., Inker, L. A., and Coresh, J. (2014). GFR estimation: from physiology to public health. \u003cem\u003eAmerican Journal of Kidney Diseases\u003c/em\u003e, 63(5), 820-834.\u003c/li\u003e\n\u003cli\u003eChouchani, E.T., Pell, V.R., Gaude, E., Aksentijević, D., Sundier, S.Y., Robb, E.L., Logan, A., Nadtochiy, S.M., Ord, E.N., Smith, A.C. and Eyassu, F., 2014. Ischaemic accumulation of succinate controls reperfusion injury through mitochondrial ROS. \u003cem\u003eNature\u003c/em\u003e, 515(7527), pp.431-435.\u003c/li\u003e\n\u003cli\u003eCzabotar, P. E., Lessene, G., Strasser, A., and Adams, J. M. (2014). Control of apoptosis by the BCL-2 protein family: implications for physiology and therapy. \u003cem\u003eNature reviews Molecular cell biology\u003c/em\u003e, 15(1), 49-63.\u003c/li\u003e\n\u003c/ol\u003e"},{"header":"Plates","content":"\u003cp\u003ePlates 1 and 2 are available in the Supplementary Files section.\u003c/p\u003e"}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":true,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":false,"hideJournal":true,"highlight":"","institution":"","isAcceptedByJournal":false,"isAuthorSuppliedPdf":false,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":false,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true},"keywords":"Bryophyllum pinnatum, hepatic ischemia-reperfusion injury, reno-hepatoprotection, oxidative stress","lastPublishedDoi":"10.21203/rs.3.rs-6205039/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-6205039/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003eHepatic ischemia-reperfusion injury (HIRI) is indicated in postoperative complications in hepatic transplantation and surgeries. HIRI can lead to remote renal injury through systemic inflammatory and oxidative cascades. While current therapeutic options are effective, their potential toxic effects can pose additional challenges, thereby hindering hepatic and renal recovery, and highlighting the need for alternative therapies. \u003cem\u003eBryophyllum pinnatum \u003c/em\u003e(BP), popularly called “life plant” is widely used in traditional medicine, and is rich in flavonoids, triterpenoids, and bufadienolides, which have shown remarkable antioxidant and anti-inflammatory activities in various animal models. Studies have demonstrated the ability of BP to scavenge free radicals, suppress acute inflammation, and inhibit pro-inflammatory cytokines. Its nephroprotective effects in renal injury models, as well as its hepatoprotective effects in chemically-induced hepatocarcinogenesis have also been documented. Despite these promising findings, its effect against HIRI-induced kidney damage remains unexplored. This study investigates the reno-hepatoprotective potential of BP methanolic leaf extract in male Wistar rats subjected to HIRI.\u003c/p\u003e\n\u003cp\u003eForty male Wistar rats were divided into four groups (n=10); group 1- Sham operated, group 2-HIRI, group 3-LDH (low-dose \u003cem\u003eB. pinnatum\u003c/em\u003e (100 mg/kg), group 4-HDH (high-dose \u003cem\u003eB. pinnatum\u003c/em\u003e (200 mg/kg). BP was administered orally via gavage for 14 days before HIRI induction. Following treatment, animals were anesthetized with ketamine (50 mg/kg) and xylazine (10 mg/kg), and HIRI was induced by clamping the hepatic artery, portal vein and bile duct for 60 minutes to induce ischemia after which the clamp was removed for hepatic reperfusion which lasted for 24-hours. Animals were euthanized under deep anesthesia (ketamine 50 mg/kg and xylazine 10 mg/kg, ip) before sacrifice. Then, blood, kidney and liver tissues were harvested for biochemical and histological analysis. Data were analyzed using one way ANOVA with Graph pad prism. Tukey’s post-hoc test was used for multiple comparison. P\u0026lt;0.05 was considered statistically significant.\u003c/p\u003e\n\u003cp\u003eBiochemical analyses showed a significant restoration in SOD, catalase, and GSH levels and reductions in MDA and inflammatory markers (TNF-α, MPO) in the \u003cem\u003eB. pinnatum\u003c/em\u003e groups. In addition, altered ALT, ALP, and GGT levels were restored, and urea and creatine levels raised as a secondary effect of HIRI were reduced in the \u003cem\u003eB. pinnatum\u003c/em\u003e-treated groups. Reduced caspase-3 activity was also observed in \u003cem\u003eB. pinnatum\u003c/em\u003e-treated groups, indicating lower apoptosis levels. Histological analyses showed improved cytoarchitecture, with preservation of renal corpuscles and reduced inflammation.\u003c/p\u003e\n\u003cp\u003eConclusively, \u003cem\u003eB. pinnatum\u003c/em\u003e exhibits reno-hepatoprotective effects on HIRI-induced kidney damage, potentially modulating oxidative stress, inflammation, and apoptotic pathways.\u003c/p\u003e","manuscriptTitle":"Reno-Hepatoprotective effects of Bryophyllum pinnatum against Hepatic Ischemia- Reperfusion Injury in male Wistar rats","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2025-05-21 08:36:40","doi":"10.21203/rs.3.rs-6205039/v1","editorialEvents":[{"type":"communityComments","content":0}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true}}],"origin":"","ownerIdentity":"0ac4bf07-9a3e-4e2f-af3a-4b7452506c0c","owner":[],"postedDate":"May 21st, 2025","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"posted","subjectAreas":[],"tags":[],"updatedAt":"2025-08-20T12:23:53+00:00","versionOfRecord":[],"versionCreatedAt":"2025-05-21 08:36:40","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-6205039","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-6205039","identity":"rs-6205039","version":["v1"]},"buildId":"8U1c8b4HqxoKbykW_rLl7","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}