Effect of Canarium Schweinfurthii (Africa Black Olive) Oil on Restraint Stressed Induced Renal Dysfunction in Wistar Rat

Effect of Canarium Schweinfurthii (Africa Black Olive) Oil on Restraint Stressed Induced Renal Dysfunction in Wistar Rat | 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 Effect of Canarium Schweinfurthii (Africa Black Olive) Oil on Restraint Stressed Induced Renal Dysfunction in Wistar Rat Ezekiel Babalola, Eneche Joseph Gabriel, Egbunu Rabbi Enyojo, and 1 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-6874801/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 Background Chronic stress regardless of its origin has server repercussion on health, notably impacting the renal system. the kidney are vulnerable to prolonged stress, during stress the renal tissue are exposed to oxidative damage. This study aim to evaluate the protective effect of canarium schweinfurhii oil on renal dysfunction of Wistar rats subjected to restraint stress on the following biomarkers Creatinine, Urea, Blood urea nitrogen and Total protein respectively. Result A total of twenty-eight (28) apparently healthy adult male Wistar rat weighing between 150−170g were used for this study. They were allocated into four groups of seven rat (n = 7 rat/group). Group I: control group and administered with 10ml/kg/bw of water, Group II: stress group + 10ml/kg/bw of water, Group III: this group under goes stress and treated with 10ml/kg (1ml/100g) of canarium schweinfurhii oil, Group IV: this group under goes stress and treated with 20ml/kg (2ml/100g) of canarium schweinfurhii oil. All group were treated for 21 days. All data were expressed as mean ± SEM. The data obtained were statistically analyzed using analysis of variance (ANOVA) with Tukey’s multiple comparison post hoc tests to compare the level of significance between control and experimental groups. All statistical analysis were evaluated using SPSS version 17.0 software. The values of P < 0.05 were considered as significant. Conclusion The findings concluded canarium schweinfurhii oil at a dose of 10ml/kg (1ml/100g) provided some protection against stress-induced renal dysfunction General Cell Biology & Physiology Stress renal dysfunction canarium schweinfurhii Figures Figure 1 Figure 2 Figure 3 Figure 4 Introduction Day-to-day activities in human life often involve various forms of stress. Student are frequently subjected to academic stress from exams, lectures, and assignments, while adults navigate the complexities of work-related and relationship stress (Akinpelu et al., 2019 ). Chronic stress, regardless of its origin, can have severe repercussions on health, notably impacting the renal system. The kidneys which are responsible fir filtering blood, regulating fluid balance, and maintaining electrolyte levels, are especially vulnerable to prolonged stress. Elevated stress hormones like cortisol can lead to hypertension and other risk factors for kidney disease. During stress renal tissues are vulnerable to oxidative damage (Ayodele et al., 2016 ). The excessive production of ROS can lead to mitochondrial dysfunction, cellular injury and dysfunction leading to renal failure (Dantzer et al., 2014 ). Stress is a biological response that occurs in response to any external or internal stimulus (Dantzer et al., 2014 ). The body's reaction to these stressors, or stimuli, is known as the stress response. Stress response is a set of physiological changes that help the body adapt to the stressor and return back to homeostasis (Shenghua et al., 2014 ). Restraint stress refers to the application of physical force or restraints on experimental animals, which leads to a state of stress and agitation. It is often used as a model to induce anxiety and depression like behaviors in experimental animals for studying the physiological and behavioral responses associated with stress (Fenga et al., 2012 ). One of the physiological responses observed during stress is the activation of the hypothalamicpituitary-adrenal (HPA) axis. The HPA axis is responsible for regulating the body's response to stress through the release of stress hormones such as corticotrophin-releasing hormone (CRH) by the hypothalamus, adrenocorticotropic hormone (ACTH) by the pituitary gland, and cortisol by the adrenal glands (Deussing et al ., 2018). Restraint stress rapidly increases the release of CRH and ACTH, which in turn triggers the release of cortisol which is the primary stress hormone (Ana et al., 2015 ). Canarium scweinfurthii (Africa black olive) an African olive tree species, is known for its rich phytochemical composition, which includes antioxidant compounds such as flavonoids and phenolic compounds (Ayoade et al ., 2015). These bioactive compounds have been reported to possess various health-promoting properties, including anti-inflammatory and antioxidant effects (Olivier et al., 2016 ). With these properties Canarium scweinfurthii have the potency to mitigate the effect of stress (Adachukwu et al., 2019 ). Material and methods Plant materials Fresh “ Canarium scweinfurthii (Africa black olive)” fruit was obtained from Pankshin in Plateau State, Nigeria. The identification of the plant was done at the department of plant and environmental biology, Prince Abubaker Audu University, Anyigba, Kogi State, Nigeria given the Voucher number: KSU-PT-B−066. Animals materials and management A total of 28 apparently healthy adult male Wistar rats weighing between 150g−170g were used for this study. The animals were purchased from the Animal House of College of Health Sciences, Benue State University, Benue State, Nigeria. The animals were kept in cages under normal environmental temperature and were fed with standard pellet diet and water given ad-libitum. The rats were allowed to acclimatize to the laboratory environment for two weeks before the commencement of the experiment. Chemicals and reagents All chemicals and reagents used for this study were obtained commercially which includes; ketamine was purchased from Nigorite, Anyingba Kogi State, Urease reagent, Jaffe reagent, biuret reagent and buffer solution was purchased from Medex supply Nigeria Ltd, Ilorin Kwara state. Methods Animal grouping The animals were grouped into 4 groups containing 7 rats each and drug dosage was determined using the method as discribed by Oghenesuvwe et al., 2014 . Group I (n = 7): this group serves as the Control group which will not undergo restraint stress, all animal in this group will be administered with 10ml/kg of water Group II (n = 7): this group serves as the stress group without administration of Canarium scweinfurthii (Africa black olive) oil, all animal in this group will be administered with 10ml/kg of water. Group III (n = 7): this group under goes restraint stress and treated with 10 ml/kg (1ml/100g) of Canarium scweinfurthii (Africa black olive) oil. Group IV (n = 7) this group under goes restraint stress and treated with 20ml/kg (2ml/100g) of Canarium scweinfurthii (Africa black olive) oil. Restraint stress induction The Wistar rats were restraint using a locally constructed wooden restraint cage with dimensions of 30 cm (L) × 3 cm (B) × 3 cm (H). The restraint stress induction was carried out following the method as described by Ezekiel et al., ( 2016 ) with little modification. Each Wistar rat were housed individually in a multi-compartment cage, The Wistar rats were exposed to chronic restraint stress for 6 hours (8am−2pm) per day for 21 days by keeping them in the restraint cages without food and water during the time of the experiment. The un-restrained rats were left undisturbed in their home cages but without access to food or water during the time of the experiment (Ezekiel et al., 2016 ). Sample collection At the end of the three weeks study the animals were anaesthetized with ketamine inter-peritoneal. And was humanely sacrificed. 5ml syringe were used to collect blood samples via cardiac puncture method into and EDTA sample bottles. The bloods sample were centrifuged at 2012.4 xg for 10 minutes in the EDTA tube to obtain plasma. Renal function analysis Creatinine, urea, blood urea nitrogen, and total protein were evaluated with commercially available kits from Medex supply Nigeria Ltd, Ilorin Kwara state. via spectrophotometry Statistical Analysis All data were expressed as mean ± SEM. The data obtained were statistically analyzed using analysis of variance (ANOVA) with Tukey’s multiple comparison post hoc tests to compare the level of significance between control and experimental groups. All statistical analysis were evaluated using SPSS version 17.0 software. The values of P < 0.05 were considered as significant Result Effect of canarium schweinfurthii (Africa black olive) oil on restraint stress - induced renal dysfunction in Wistar rat on creatinine level. The effect of canarium schweinfurthii (Africa black olive) oil on restraint stress-induced renal dysfunction in Wistar rat on creatinine level is shown in Fig. 1 . Creatinine level for the four groups was as follows: positive control, 62.69 ± 16.18; Stressed + DW, 50.71 ± 14.95; 10 ml/kg of C.S (Group C), 67.75 ± 14.75; 20ml/kg of C.S (Group D), 75.12 ± 19.51, respectively. There was no significant difference between the groups (p˃0.05). Effect of canarium schweinfurthii (Africa black olive) oil on restraint stress - induced renal dysfunction in Wistar rat on urea level. The effect of canarium schweinfurthii (Africa black olive) oil on restraint stress-induced renal dysfunction in Wistar rat on urea level is shown in Fig. 2 . Urea level for the four groups was as follows: positive control (Group A), 26.94 ± 6.36; Negative control (Group B), 15.84 ± 3.57; 10 ml/kg of C.S (Group C), 70.91 ± 5.15; 20ml/kg of C.S (Group D), 51.50 ± 9.65, respectively. There was no significant difference between the groups (p˃0.05). Effect of canarium schweinfurthii (Africa black olive) oil on restraint stress - induced renal dysfunction in Wistar rat on blood urea nitrogen level. The effect of canarium schweinfurthii (Africa black olive) oil on restraint stress-induced renal dysfunction in Wistar rat on blood urea nitrogen level is shown in Fig. 3 . blood Urea nitrogen level for the four groups was as follows: positive control (Group A), 12.58 ± 3.70; Negative control (Group B), 7.40 ± 1.66; 10 ml/kg of C.S (Group C), 33.11 ± 2.40 20ml/kg of C.S (Group D), 24.05 ± 4.31, respectively. There was no significant difference between the groups (p˃0.05). Effect of canarium schweinfurthii (Africa black olive) oil on restraint stress - induced renal dysfunction in Wistar rat on total protein level. The effect of canarium schweinfurthii (Africa black olive) oil on restraint stress-induced renal dysfunction in Wistar rat on total protein level is shown in Fig. 4 . total protein level for the four groups was as follows: positive control (Group A), 5.09 ± 0.71; Negative control (Group B), 4.60 ± 0.48; 10 ml/kg of C.S (Group C), 4.99 ± 0.35; 20ml/kg of C.S (Group D), 4.31 ± 0.49, respectively. There was no significant difference between the groups (p˃0.05). Discussion The effect of plant extract of canarium schweinfurthii oil on serum creatinine level produce a non-significant increase in serum creatinine levels in stressed Wistar rats on comparison with the negative control (P ˃ 0.05). Negative Control (stressed + DW) showed a reduction in creatinine levels despite stress. This might indicate that restraint stress did not have a strong impact on skeletal muscles due to immobilization resulting in decrease creatinine levels. Administration of canarium schweinfurthii oil at a dose of 10 ml/kg shows elevated creatinine levels compared to negative control, indicating that CSO at 10 mg/kg does not fully protect against stress-induced increases in creatinine, though it still provides some benefit compared to 20 ml/kg of CSO. The fact that creatinine levels in this group are lower than in Group 4 suggests that the low dose might be somewhat effective but is not optimal. canarium schweinfurthii oil at a dose of 20 ml/kg has the highest creatinine levels, suggesting that a higher dose of CSO might have toxic effects to the kidneys, leading to impaired function and increased creatinine levels. This result disagrees with the findings of Okwuosa et al. , (2010) who report that the stem bark extracts of canarium schweinfurthii significantly reduced plasma creatinine levels and preserving renal histoarchitecture. They concluded that stem bark extracts of canarium schweinfurthii produce a fall in plasma creatinine and urea in present of acetaminophen-induced nephroprotective in rat. The result of the current study shows that the administration of canarium schweinfurthii oil given at a dose of 10 ml/kg and 20 ml/kg produce a non-significant increase in serum urea and BUN levels in stressed Wistar rats on comparison with the negative control (P ˃ 0.05). negative control shows the lowest urea and BUN levels among the stressed groups, possibly because stress with water intervention might initially lower protein catabolism allowing the animals to adapt to the stress condition over time. Administration of CSO at a dose of 10 ml/kg shows increased urea and BUN levels compared to negative control. This suggests that at a low dose (10 ml/kg), CSO might not provide sufficient protection against stress-induced renal dysfunction and might exacerbate stress effects due to mild toxicity or insufficient adaptive response. Administration of CSO at a dose of 20 ml/kg shows increased urea and BUN levels compared to negative control, but lower than CSO administration at 10 ml/kg. This indicates that a higher dose of CSO (20 ml/kg) might begin to offer some protective effects, although it is still not enough to completely mitigate the stress-induced increase in urea and BUN levels. At higher dose CSO (20 ml/kg) likely influence metabolic pathways affecting protein catabolism and urea production. The higher dose might better support metabolic balance under stress when compared to the lower dose. The result is consistent with earlier study done by Ansari et al., ( 2021 ). Who report that Thymus serrulatus essential oil has a protective effect on Cadmium-Induced Nephrotoxicity in Rats, through Suppression of Oxidative Stress and Downregulation of NF-κB, iNOS, and Smad2 mRNA Expression The result of the current study shows that the administration of canarium schweinfurthii oil given at a dose of 10 ml/kg and 20 ml/kg produce a non-significant increase in serum total protein levels in stressed Wistar rats on comparison with the negative control (P ˃ 0.05). Negative Control shows decreased total protein levels due to stress. Stress triggers the release of stress hormones like cortisol, which promotes protein breakdown (catabolism) to provide amino acids for energy and gluconeogenesis. This leads to reduced total protein levels in the blood. CSO at a dose of 10 ml/kg likely mitigate stress-induced protein catabolism, potentially by modulating the release of stress hormones and by enhancing the body’s antioxidant defenses. This would help maintain higher protein levels by reducing catabolism, while the high dose of CSO 20ml/kg might still offer some protective effects against stress-induced protein degradation, it might not be as effective as the lower dose. Higher doses could introduce mild toxicity, potentially by affecting liver function and overall metabolism. This could reduce its effectiveness in promoting protein synthesis or protecting against protein degradation. This agrees with the research carried out by Gurevich, (2013). Who study the effect of blood protein concentration on drug-dosing regimes: practical guidance? Conclusion In this study Group B (stressed + DW) showed the lowest levels of blood urea nitrogen (BUN) and total protein, indicating significant stress impact leading to increased protein catabolism and reduced protein synthesis. Group C (stressed + 10ml/kg of CSO) exhibited the highest BUN levels and elevated creatinine levels, suggesting increased protein breakdown, renal stress, and mild kidney impairment due to the low dose of CSO. Group D (stressed + 20 ml/kg of CSO) showed elevated but lower BUN and creatinine levels compared to Group C (stressed + 10ml/kg of CSO), implying that the higher dose of CSO was more effective in mitigating stress effects and supporting kidney function. Total protein levels increased in Group C (stressed + 10ml/kg of CSO) compared to Group B (stressed + DW), indicating partial protective effects of the low CSO dose, whereas Group D (stressed + 10ml/kg of CSO) displayed even higher total protein levels, suggesting a dose-dependent enhancement of protein synthesis and stress adaptation. Urea levels followed a similar trend, with Group B (stressed + DW) having the lowest levels, Group C (stressed + 10ml/kg of CSO) the highest, and Group D (stressed + 10ml/kg of CSO) showing elevated levels but lower than Group C (stressed + 10ml/kg of CSO). Overall, the findings suggest that while both doses of CSO provided some protection against stress-induced biochemical changes, the higher dose (20 ml/kg) was more effective in reducing stress impacts, supporting protein metabolism, and maintaining renal function. Recommendations Future similar study on the effect of canarium schweinfurthii on stress induced renal dysfunction should include measurement of uric acid levels and electrolyte such as chloride, biacarbonate etc following administration of different doses. Other future similar study on the effect of canarium schweinfurthii on stress induced renal dysfunction should be done on Wistar rat with metabolic disorder such as diabetes, obesity etc. Similar future study should include the effect of canarium schweinfurthii on stress induced Wistar rat should be done on important organs such as the lung, brain and heart that are capable of damaging during stress. Abbreviations CSO canarium schweinfurhii oil BUN blood urea nitrogen EDTA ethylenediaminetetraacetic acid Declarations a. Ethics approval and consent to participate All experimental procedures were approved by the Institutional Animal Care and Use Committee (IACUC) of [Prince Abubaker Audu University, in accordance with international guidelines for the care and use of laboratory b Consent for publication Not applicable. c. Availability of data and material The datasets generated and/or analyzed during the current study are available from the corresponding author on reasonable request. d. Competing interests The authors declare that they have no competing interests. e. Funding No external funding was received for this study f. Authors' contributions Ezekiel, B. conceived and designed the experiments. Ezekiel, B and Egbunu, R.E. performed the experiments. Eneche, J.G. analyzed the data. Ejeh, A.O. contributed reagents/materials/analysis tools. [Initials] wrote the manuscript. All authors read and approved the final manuscript. g. Acknowledgements The authors wish to thank Dr. O.A Daikwo for his technical support and valuable input throughout this study. References Adachukwu NN, Pius OU, William RW, Njemuwa NN, Karl F (2019) Fatty Acid Profile and Production of Fatliquor from Canarium schweinfurthii Mesocarp Oil. Pertanika J Sci Technol 27(4):2221–2243 Akinnuga AM, Jeje SO, Bamidele O, Amaku EE, Otogo FO, and Sunday V. E (2014) Virgin Coconut Oil: Remedial Effects on Renal Dysfunction in Diabetic Rats. Hindawi Publishing Corporation Physiol J 10:1–5 Akinpelu LA, Aiyelero OM, Olayiwola G (2019) Ethanol leaf extract of Milicia excelsa mitigates anxiety and depressive-like behaviours induced by acute restraint stress in mice. GSC Biol Pharm Sci 6(2):30–39 Akram E, Sara M, Jalal ZM, Eidi M, Rezazadeh S (2012) Antinociceptive and anti-inflammatory effects of olive oil (Olea europeae L.) in mice. Pharm Biol 50(3):332–337 Ana PP, Carda KC, Marchi ER, André SM, Antunes-Rodrigues J, Claudia MP, Carlos RT (2015) Acute restraint stress induces endothelial dysfunction: role of vasoconstrictor prostanoids and oxidative stress. Stress 18(2):233–243 Annerstedt M, Jönsson P, Wallergård M, Johansson G, Karlson B, Grahn P, Åse MH, Währborg P (2014) Inducing physiological stress recovery with sounds of nature in a virtual reality forest — Results from a pilot study. Physiol Behav 118:240–250 Ansari MN, Rehman UN, Karim A, Imam F, Hamad AM (2021) Protective Effect of Thymus serrulatus Essential Oil on Cadmium-Induced Nephrotoxicity in Rats, through Suppression of Oxidative Stress and Downregulation of NF-κB, iNOS, and Smad2 mRNA Expression. Molecules 26(5):12–52 Ayodele OM, Kolawole IA, Gabriel OO, Titilola AS (2016) Restraint Stress Impairs Glucose Homeostasis Through Altered Insulin Signalling in Sprague-Dawley Rat. Nigeria J Physiological Sci 31:23–29 Chinaka EJ (2018) Viscosity and biodiesel characteristics of wild Canarium schweinfurthii Engl. fruit oil. Reserved Agricultural Eng 64:169–175 Chomini MS, Francis MJ, Ishaya M, Chomini AE, Peter MK (2021) Assessment of Proximate and Physico-Chemical Potentials of Crude Kernel Oil Extracts of Canarium schweinfurthii Engl. Nigeria J Pure Appl Sci 34(1):3941–3950 Christine N, Stefan B, Thomas R (2015) Chemical composition and anti-termitic activity of essential oil from Canarium schweinfurthii Engl. Ind Crops Prod 71:75–79 Dantzer B, Fletcher QE, Boonstra R, Sheriff MJ (2014) Measures of physiological stress: a transparent or opaque window into the status, management and conservation of species? Conserv Physiol 2:1–18 Deussing MJ, Chen A (2018) The corticotropin-releasing factor family: physiology of the stress response. Physiol Rev 98:2225–2286 El-Saber G, Batiha J, Oluwafemi T, Hazem MS, Opeyemi AA, Aya AA, Titilade K, Ayandeyi T, Athanasios A, Marios P (2023) Bioactive compounds, pharmacological actions and pharmacokinetics of Cupressus sempervirens. Naunyn-Schmiedeberg's. Archives Pharmacol 396(3):389–403 Ezekiel I, Ayo JO, Magaji RA, Tanko Y (2016) Effect of Taurine on Chronic Restraint Stress Induced Behavioural Deficits in Rodents. J Nat Sci Res 6(11):2224–3186 Faizah MN, Mohd AA, Haziq HN, Nurul AMY, Mohamad SIM (2024) Protective Effects of Tualang Honey on the Testes of Rats Subjected to Chronic Combined Stress. Malaysian J Med Health Sci 20(2):34–41 Fenga Z, Liub L, Zhanga C, Zhengc T, Wangc J, Lina M, Zhaoc Y, Wanga X, Arnold JL, Wenwei H (2012) Chronic restraint stress attenuates p53 function and promotes tumorigenesis. GenetiCSO 18(109):7013–7018 Gonçalves M, María RP, Anna C, Emma BR, Giulia A, Paula S (2024) A Narrative Review of Metabolomic Insights into Olive Oil’s Nutritional Value. Appl Sci 14(10):23–45 Gong S, Miao YL, Jiao GZ, Sun MJ, Li H, Lin J, Luo JM, Tan JH (2015) Dynamics and Correlation of Serum Cortisol and Corticosterone under Different Physiological or Stressful Conditions in Mice. Public Libr Sci 10(2):1–14 Haihua G, Wibke BW (2023) The Interplay of Autophagy and Oxidative Stress in the Kidney: What Do We Know? Nephron , 147 (10): 627–642 Hassanzadeh-Taheri M, Jahani F, Erfanian Z, Moodi H, Hosseini M (2019) The impact of long-term consumption of diets enriched with olive, cottonseed or sesame oils on kidney morphology: A stereological study. An Acad Bras Cienc 9(1):1–11 Ijeoma AD, Tochukwu DM (2020) Profiling and comparison of fatty acids in the oils from the fruits of Dacryodes edulis and Canarium schweinfurthii. J Med Plants Stud 8(5):213–217 Kiin-Kabari DB, Umunna PS, Giami SY (2020) Physicochemical properties and fatty acid profile of African elemi fruit pulp oil compared with kernel oil. Eur J Agric Food Sci 2(6):1–5 Mishnev M, Korolev A, Zadorin A, Astashkin V (2024) Cyclic Thermomechanical Loading of Epoxy Polymer: Modeling with Consideration of Stress Accumulation and Experimental Verification. Polymers , 16: 910–939 Nyam MA, Makut MD, Itelima UJ, Daniel AM (2014) Nutritional Potential of the Fruits of Black Olive (Canarium schweinfurthii Linn) from Plateau State. Nigeria Pakistan J Nutrition 13(6):335–339 Oghenesuvwe EE, Ekene NE, Ajaghaku DL (2014) Guidelines on dosage calculation and stock solution preparation in experimental animals’ studies. J Nat Sci Res 4(18):100–106 Olivier TT, Armel JS, Francis NT (2016) A Review on Traditional Uses, Phytochemical Composition and Pharmacological Profile of Canarium scweinfurthii Eng. Nat Sci 14(11):17–22 Shenghua Z, Ruoyang S, Junhui W, Jun-Feng W, Xin-Min L (2014) Unpredictable chronic mild stress not chronic restraint stress induces depressive behaviours in mice. NeuroReport 25:1151–1155 Tang N, Liu XT, Lin LX, Yang WX, Li LQ, Guo-En W, Yan-Hua W (2024) Erzhiwan Ameliorates Restraint Stress- and Monobenzone-Induced Depigmentation in Mice by Inhibiting Macrophage Migration Inhibitory Factor and 8-Hydroxy-2-Deoxyguanosine. Clin Cosmet Invest Dermatology 17:147–158 Teixeira-Santos L, Albino- Teixeira A, Pinho D (2021) An alternative method for oral drug administration by voluntary intake in male and female mice. Lab Anim 55(1):76–80 Additional Declarations The authors declare no competing interests. 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-6874801","acceptedTermsAndConditions":true,"allowDirectSubmit":true,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":470006476,"identity":"a0d7ced3-d57b-4d1e-b053-6e91ad392365","order_by":0,"name":"Ezekiel Babalola","email":"data:image/png;base64,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","orcid":"https://orcid.org/0009-0009-0520-1809","institution":"Kogi state University, anyigba , kogi state","correspondingAuthor":true,"prefix":"","firstName":"Ezekiel","middleName":"","lastName":"Babalola","suffix":""},{"id":470006477,"identity":"a07c187b-eb54-4dbd-a049-ac0925f86767","order_by":1,"name":"Eneche Joseph Gabriel","email":"","orcid":"","institution":"Kogi state University anyigba kogi state","correspondingAuthor":false,"prefix":"","firstName":"Eneche","middleName":"Joseph","lastName":"Gabriel","suffix":""},{"id":470006478,"identity":"bb9e5175-351b-45c3-a677-283b25bc8b76","order_by":2,"name":"Egbunu Rabbi Enyojo","email":"","orcid":"","institution":"Kogi state University anyigba kogi state","correspondingAuthor":false,"prefix":"","firstName":"Egbunu","middleName":"Rabbi","lastName":"Enyojo","suffix":""},{"id":470006479,"identity":"d5663a90-3847-4ec5-8f5b-0d045cff797c","order_by":3,"name":"Ejeh Ojone Aminat","email":"","orcid":"","institution":"Kogi state University anyigba kogi state","correspondingAuthor":false,"prefix":"","firstName":"Ejeh","middleName":"Ojone","lastName":"Aminat","suffix":""}],"badges":[],"createdAt":"2025-06-11 20:41:57","currentVersionCode":1,"declarations":{"humanSubjects":false,"vertebrateSubjects":true,"conflictsOfInterestStatement":false,"humanSubjectEthicalGuidelines":false,"humanSubjectConsent":false,"humanSubjectClinicalTrial":false,"humanSubjectCaseReport":false,"vertebrateSubjectEthicalGuidelines":true},"doi":"10.21203/rs.3.rs-6874801/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-6874801/v1","draftVersion":[],"editorialEvents":[],"editorialNote":"","failedWorkflow":false,"files":[{"id":84571326,"identity":"4cc42359-bdbc-42d0-826b-d02ccdbe64f2","added_by":"auto","created_at":"2025-06-13 15:32:16","extension":"png","order_by":1,"title":"Figure 1","display":"","copyAsset":false,"role":"figure","size":21538,"visible":true,"origin":"","legend":"\u003cp\u003eCreatinine level in the blood following 21 days of oral administration of \u003cem\u003ecanarium schweinfurthii\u003c/em\u003e (Africa black olive) oil in different dosage (10ml/kg and 20ml/kg). All the error bars are SEMs. DW (distilled water), CSO (\u003cem\u003ecanarium schweinfurthii \u003c/em\u003eoil).\u003c/p\u003e","description":"","filename":"1.png","url":"https://assets-eu.researchsquare.com/files/rs-6874801/v1/b8d0f3317ddf1b1fe89b6b2d.png"},{"id":84571327,"identity":"dfbef59c-6a36-47b9-96e0-7fa9532aa1e9","added_by":"auto","created_at":"2025-06-13 15:32:16","extension":"png","order_by":2,"title":"Figure 2","display":"","copyAsset":false,"role":"figure","size":17972,"visible":true,"origin":"","legend":"\u003cp\u003eUrea level in the blood following 21 days of oral administration of \u003cem\u003ecanarium schweinfurthii\u003c/em\u003e (Africa black olive) oil in different dosage (10ml/kg and 20ml/kg). All the error bars are SEMs. DW (distilled water), CSO (\u003cem\u003ecanarium schweinfurthii \u003c/em\u003eoil).\u003c/p\u003e","description":"","filename":"2.png","url":"https://assets-eu.researchsquare.com/files/rs-6874801/v1/5b4675e9dfdf903fd9b7b46a.png"},{"id":84572048,"identity":"a541b622-2dc5-4eeb-8801-e84da62ec145","added_by":"auto","created_at":"2025-06-13 15:40:17","extension":"png","order_by":3,"title":"Figure 3","display":"","copyAsset":false,"role":"figure","size":17991,"visible":true,"origin":"","legend":"\u003cp\u003eBlood urea nitrogen level following 21 days of oral administration of \u003cem\u003ecanarium schweinfurthii\u003c/em\u003e (Africa black olive) oil in different dosage (10ml/kg and 20ml/kg). All the error bars are SEMs. DW (distilled water), CSO (\u003cem\u003ecanarium schweinfurthii \u003c/em\u003eoil).\u003c/p\u003e","description":"","filename":"3.png","url":"https://assets-eu.researchsquare.com/files/rs-6874801/v1/bf8b1325e82f8a767167e7c7.png"},{"id":84571328,"identity":"7cf18427-64d0-4ae0-be81-f9f97b01341f","added_by":"auto","created_at":"2025-06-13 15:32:17","extension":"png","order_by":4,"title":"Figure 4","display":"","copyAsset":false,"role":"figure","size":16381,"visible":true,"origin":"","legend":"\u003cp\u003eTotal protein level in the blood following 21 days of oral administration of \u003cem\u003ecanarium schweinfurthii\u003c/em\u003e (Africa black olive) oil in different dosage (10ml/kg and 20ml/kg). All the error bars are SEMs. DW (distilled water), CSO (\u003cem\u003ecanarium schweinfurthii \u003c/em\u003eoil).\u003c/p\u003e","description":"","filename":"4.png","url":"https://assets-eu.researchsquare.com/files/rs-6874801/v1/9403b642bb4809c5a90df213.png"},{"id":84572279,"identity":"b62bb4d1-7bc9-454e-b3b1-12e8fe610356","added_by":"auto","created_at":"2025-06-13 15:48:17","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":777886,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-6874801/v1/47e698e3-b6d9-4e34-a5dd-f1c78572e03c.pdf"}],"financialInterests":"The authors declare no competing interests.","formattedTitle":"\u003cp\u003eEffect of Canarium Schweinfurthii (Africa Black Olive) Oil on Restraint Stressed Induced Renal Dysfunction in Wistar Rat\u003c/p\u003e","fulltext":[{"header":"Introduction","content":"\u003cp\u003eDay-to-day activities in human life often involve various forms of stress. Student are frequently subjected to academic stress from exams, lectures, and assignments, while adults navigate the complexities of work-related and relationship stress (Akinpelu et al., \u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e2019\u003c/span\u003e). Chronic stress, regardless of its origin, can have severe repercussions on health, notably impacting the renal system. The kidneys which are responsible fir filtering blood, regulating fluid balance, and maintaining electrolyte levels, are especially vulnerable to prolonged stress. Elevated stress hormones like cortisol can lead to hypertension and other risk factors for kidney disease. During stress renal tissues are vulnerable to oxidative damage (Ayodele et al., \u003cspan citationid=\"CR8\" class=\"CitationRef\"\u003e2016\u003c/span\u003e). The excessive production of ROS can lead to mitochondrial dysfunction, cellular injury and dysfunction leading to renal failure (Dantzer et al., \u003cspan citationid=\"CR12\" class=\"CitationRef\"\u003e2014\u003c/span\u003e). Stress is a biological response that occurs in response to any external or internal stimulus (Dantzer et al., \u003cspan citationid=\"CR12\" class=\"CitationRef\"\u003e2014\u003c/span\u003e). The body's reaction to these stressors, or stimuli, is known as the stress response. Stress response is a set of physiological changes that help the body adapt to the stressor and return back to homeostasis (Shenghua et al., \u003cspan citationid=\"CR28\" class=\"CitationRef\"\u003e2014\u003c/span\u003e). Restraint stress refers to the application of physical force or restraints on experimental animals, which leads to a state of stress and agitation. It is often used as a model to induce anxiety and depression like behaviors in experimental animals for studying the physiological and behavioral responses associated with stress (Fenga et al., \u003cspan citationid=\"CR17\" class=\"CitationRef\"\u003e2012\u003c/span\u003e). One of the physiological responses observed during stress is the activation of the hypothalamicpituitary-adrenal (HPA) axis. The HPA axis is responsible for regulating the body's response to stress through the release of stress hormones such as corticotrophin-releasing hormone (CRH) by the hypothalamus, adrenocorticotropic hormone (ACTH) by the pituitary gland, and cortisol by the adrenal glands (Deussing \u003cem\u003eet al\u003c/em\u003e., 2018). Restraint stress rapidly increases the release of CRH and ACTH, which in turn triggers the release of cortisol which is the primary stress hormone (Ana et al., \u003cspan citationid=\"CR5\" class=\"CitationRef\"\u003e2015\u003c/span\u003e).\u003cem\u003eCanarium\u003c/em\u003e scweinfurthii (Africa black olive) an African olive tree species, is known for its rich phytochemical composition, which includes antioxidant compounds such as flavonoids and phenolic compounds (Ayoade \u003cem\u003eet al\u003c/em\u003e., 2015). These bioactive compounds have been reported to possess various health-promoting properties, including anti-inflammatory and antioxidant effects (Olivier et al., \u003cspan citationid=\"CR27\" class=\"CitationRef\"\u003e2016\u003c/span\u003e). With these properties \u003cem\u003eCanarium\u003c/em\u003e scweinfurthii have the potency to mitigate the effect of stress (Adachukwu et al., \u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e2019\u003c/span\u003e).\u003c/p\u003e"},{"header":"Material and methods","content":"\u003cp\u003e\u003cb\u003ePlant materials\u003c/b\u003e\u003c/p\u003e\u003cp\u003eFresh “\u003cem\u003eCanarium\u003c/em\u003e scweinfurthii (Africa black olive)” fruit was obtained from Pankshin in Plateau State, Nigeria. The identification of the plant was done at the department of plant and environmental biology, Prince Abubaker Audu University, Anyigba, Kogi State, Nigeria given the Voucher number: KSU-PT-B−066.\u003c/p\u003e\u003cp\u003e\u003cb\u003eAnimals materials and management\u003c/b\u003e\u003c/p\u003e\u003cp\u003eA total of 28 apparently healthy adult male Wistar rats weighing between 150g−170g were used for this study. The animals were purchased from the Animal House of College of Health Sciences, Benue State University, Benue State, Nigeria. The animals were kept in cages under normal environmental temperature and were fed with standard pellet diet and water given ad-libitum. The rats were allowed to acclimatize to the laboratory environment for two weeks before the commencement of the experiment.\u003c/p\u003e\u003cp\u003e\u003cb\u003eChemicals and reagents\u003c/b\u003e\u003c/p\u003e\u003cp\u003eAll chemicals and reagents used for this study were obtained commercially which includes; ketamine was purchased from Nigorite, Anyingba Kogi State, Urease reagent, Jaffe reagent, biuret reagent and buffer solution was purchased from Medex supply Nigeria Ltd, Ilorin Kwara state.\u003c/p\u003e\u003cp\u003e\u003cb\u003eMethods Animal grouping\u003c/b\u003e\u003c/p\u003e\u003cp\u003eThe animals were grouped into 4 groups containing 7 rats each and drug dosage was determined using the method as discribed by Oghenesuvwe et al., \u003cspan citationid=\"CR26\" class=\"CitationRef\"\u003e2014\u003c/span\u003e.\u003c/p\u003e\u003cp\u003eGroup I (n = 7): this group serves as the Control group which will not undergo restraint stress, all animal in this group will be administered with 10ml/kg of water\u003c/p\u003e\u003cp\u003eGroup II (n = 7): this group serves as the stress group without administration of \u003cem\u003eCanarium\u003c/em\u003e scweinfurthii (Africa black olive) oil, all animal in this group will be administered with 10ml/kg of water.\u003c/p\u003e\u003cp\u003eGroup III (n = 7): this group under goes restraint stress and treated with 10 ml/kg (1ml/100g) of \u003cem\u003eCanarium\u003c/em\u003e scweinfurthii (Africa black olive) oil.\u003c/p\u003e\u003cp\u003eGroup IV (n = 7) this group under goes restraint stress and treated with 20ml/kg (2ml/100g) of \u003cem\u003eCanarium\u003c/em\u003e scweinfurthii (Africa black olive) oil.\u003c/p\u003e\u003cp\u003e\u003cb\u003eRestraint stress induction\u003c/b\u003e\u003c/p\u003e\u003cp\u003eThe Wistar rats were restraint using a locally constructed wooden restraint cage with dimensions of 30 cm (L) × 3 cm (B) × 3 cm (H). The restraint stress induction was carried out following the method as described by Ezekiel et al., (\u003cspan citationid=\"CR15\" class=\"CitationRef\"\u003e2016\u003c/span\u003e) with little modification. Each Wistar rat were housed individually in a multi-compartment cage, The Wistar rats were exposed to chronic restraint stress for 6 hours (8am−2pm) per day for 21 days by keeping them in the restraint cages without food and water during the time of the experiment. The un-restrained rats were left undisturbed in their home cages but without access to food or water during the time of the experiment (Ezekiel et al., \u003cspan citationid=\"CR15\" class=\"CitationRef\"\u003e2016\u003c/span\u003e).\u003c/p\u003e\u003cp\u003e\u003cb\u003eSample collection\u003c/b\u003e\u003c/p\u003e\u003cp\u003eAt the end of the three weeks study the animals were anaesthetized with ketamine inter-peritoneal. And was humanely sacrificed. 5ml syringe were used to collect blood samples via cardiac puncture method into and EDTA sample bottles. The bloods sample were centrifuged at 2012.4 xg for 10 minutes in the EDTA tube to obtain plasma.\u003c/p\u003e\u003cp\u003e\u003cb\u003eRenal function analysis\u003c/b\u003e\u003c/p\u003e\u003cp\u003eCreatinine, urea, blood urea nitrogen, and total protein were evaluated with commercially available kits from Medex supply Nigeria Ltd, Ilorin Kwara state. via spectrophotometry\u003c/p\u003e\u003cp\u003e\u003cb\u003eStatistical Analysis\u003c/b\u003e\u003c/p\u003e\u003cp\u003eAll data were expressed as mean ± SEM. The data obtained were statistically analyzed using analysis of variance (ANOVA) with Tukey’s multiple comparison post hoc tests to compare the level of significance between control and experimental groups. All statistical analysis were evaluated using SPSS version 17.0 software. The values of P \u0026lt; 0.05 were considered as significant\u003c/p\u003e"},{"header":"Result","content":"\u003cp\u003e\u003cb\u003eEffect of\u003c/b\u003e \u003cb\u003ecanarium schweinfurthii\u003c/b\u003e \u003cb\u003e(Africa black olive) oil on restraint stress\u003c/b\u003e-\u003cb\u003einduced renal dysfunction in Wistar rat on creatinine level.\u003c/b\u003e\u003c/p\u003e\u003cp\u003eThe effect of \u003cem\u003ecanarium schweinfurthii\u003c/em\u003e (Africa black olive) oil on restraint stress-induced renal dysfunction in Wistar rat on creatinine level is shown in Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003e. Creatinine level for the four groups was as follows: positive control, 62.69 ± 16.18; Stressed + DW, 50.71 ± 14.95; 10 ml/kg of C.S (Group C), 67.75 ± 14.75; 20ml/kg of C.S (Group D), 75.12 ± 19.51, respectively. There was no significant difference between the groups (p˃0.05).\u003c/p\u003e\u003cp\u003e\u003c/p\u003e\u003cp\u003e\u003cb\u003eEffect of\u003c/b\u003e \u003cb\u003ecanarium schweinfurthii\u003c/b\u003e \u003cb\u003e(Africa black olive) oil on restraint stress\u003c/b\u003e-\u003cb\u003einduced renal dysfunction in Wistar rat on urea level.\u003c/b\u003e\u003c/p\u003e\u003cp\u003eThe effect of \u003cem\u003ecanarium schweinfurthii\u003c/em\u003e (Africa black olive) oil on restraint stress-induced renal dysfunction in Wistar rat on urea level is shown in Fig.\u0026nbsp;\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e2\u003c/span\u003e. Urea level for the four groups was as follows: positive control (Group A), 26.94 ± 6.36; Negative control (Group B), 15.84 ± 3.57; 10 ml/kg of C.S (Group C), 70.91 ± 5.15; 20ml/kg of C.S (Group D), 51.50 ± 9.65, respectively. There was no significant difference between the groups (p˃0.05).\u003c/p\u003e\u003cp\u003e\u003c/p\u003e\u003cp\u003e\u003cb\u003eEffect of\u003c/b\u003e \u003cb\u003ecanarium schweinfurthii\u003c/b\u003e \u003cb\u003e(Africa black olive) oil on restraint stress\u003c/b\u003e-\u003cb\u003einduced renal dysfunction in Wistar rat on blood urea nitrogen level.\u003c/b\u003e\u003c/p\u003e\u003cp\u003eThe effect of \u003cem\u003ecanarium schweinfurthii\u003c/em\u003e (Africa black olive) oil on restraint stress-induced renal dysfunction in Wistar rat on blood urea nitrogen level is shown in Fig.\u0026nbsp;\u003cspan refid=\"Fig3\" class=\"InternalRef\"\u003e3\u003c/span\u003e. blood Urea nitrogen level for the four groups was as follows: positive control (Group A), 12.58 ± 3.70; Negative control (Group B), 7.40 ± 1.66; 10 ml/kg of C.S (Group C), 33.11 ± 2.40 20ml/kg of C.S (Group D), 24.05 ± 4.31, respectively. There was no significant difference between the groups (p˃0.05).\u003c/p\u003e\u003cp\u003e\u003c/p\u003e\u003cp\u003e\u003cb\u003eEffect of\u003c/b\u003e \u003cb\u003ecanarium schweinfurthii\u003c/b\u003e \u003cb\u003e(Africa black olive) oil on restraint stress\u003c/b\u003e-\u003cb\u003einduced renal dysfunction in Wistar rat on total protein level.\u003c/b\u003e\u003c/p\u003e\u003cp\u003eThe effect of \u003cem\u003ecanarium schweinfurthii\u003c/em\u003e (Africa black olive) oil on restraint stress-induced renal dysfunction in Wistar rat on total protein level is shown in Fig.\u0026nbsp;\u003cspan refid=\"Fig4\" class=\"InternalRef\"\u003e4\u003c/span\u003e. total protein level for the four groups was as follows: positive control (Group A), 5.09 ± 0.71; Negative control (Group B), 4.60 ± 0.48; 10 ml/kg of C.S (Group C), 4.99 ± 0.35; 20ml/kg of C.S (Group D), 4.31 ± 0.49, respectively. There was no significant difference between the groups (p˃0.05).\u003c/p\u003e"},{"header":"Discussion","content":"\u003cp\u003eThe effect of plant extract of \u003cem\u003ecanarium schweinfurthii\u003c/em\u003e oil on serum creatinine level produce a non-significant increase in serum creatinine levels in stressed Wistar rats on comparison with the negative control (P ˃ 0.05). Negative Control (stressed\u0026thinsp;+\u0026thinsp;DW) showed a reduction in creatinine levels despite stress. This might indicate that restraint stress did not have a strong impact on skeletal muscles due to immobilization resulting in decrease creatinine levels. Administration of \u003cem\u003ecanarium schweinfurthii\u003c/em\u003e oil at a dose of 10 ml/kg shows elevated creatinine levels compared to negative control, indicating that CSO at 10 mg/kg does not fully protect against stress-induced increases in creatinine, though it still provides some benefit compared to 20 ml/kg of CSO. The fact that creatinine levels in this group are lower than in Group 4 suggests that the low dose might be somewhat effective but is not optimal. \u003cem\u003ecanarium schweinfurthii\u003c/em\u003e oil at a dose of 20 ml/kg has the highest creatinine levels, suggesting that a higher dose of CSO might have toxic effects to the kidneys, leading to impaired function and increased creatinine levels. This result disagrees with the findings of Okwuosa \u003cem\u003eet al.\u003c/em\u003e, (2010) who report that the stem bark extracts of \u003cem\u003ecanarium schweinfurthii\u003c/em\u003e significantly reduced plasma creatinine levels and preserving renal histoarchitecture. They concluded that stem bark extracts of \u003cem\u003ecanarium schweinfurthii\u003c/em\u003e produce a fall in plasma creatinine and urea in present of acetaminophen-induced nephroprotective in rat.\u003c/p\u003e \u003cp\u003eThe result of the current study shows that the administration of \u003cem\u003ecanarium schweinfurthii\u003c/em\u003e oil given at a dose of 10 ml/kg and 20 ml/kg produce a non-significant increase in serum urea and BUN levels in stressed Wistar rats on comparison with the negative control (P ˃ 0.05). negative control shows the lowest urea and BUN levels among the stressed groups, possibly because stress with water intervention might initially lower protein catabolism allowing the animals to adapt to the stress condition over time. Administration of CSO at a dose of 10 ml/kg shows increased urea and BUN levels compared to negative control. This suggests that at a low dose (10 ml/kg), CSO might not provide sufficient protection against stress-induced renal dysfunction and might exacerbate stress effects due to mild toxicity or insufficient adaptive response. Administration of CSO at a dose of 20 ml/kg shows increased urea and BUN levels compared to negative control, but lower than CSO administration at 10 ml/kg. This indicates that a higher dose of CSO (20 ml/kg) might begin to offer some protective effects, although it is still not enough to completely mitigate the stress-induced increase in urea and BUN levels. At higher dose CSO (20 ml/kg) likely influence metabolic pathways affecting protein catabolism and urea production. The higher dose might better support metabolic balance under stress when compared to the lower dose. The result is consistent with earlier study done by Ansari et al., (\u003cspan citationid=\"CR7\" class=\"CitationRef\"\u003e2021\u003c/span\u003e). Who report that Thymus serrulatus essential oil has a protective effect on Cadmium-Induced Nephrotoxicity in Rats, through Suppression of Oxidative Stress and Downregulation of NF-κB, iNOS, and Smad2 mRNA Expression\u003c/p\u003e \u003cp\u003eThe result of the current study shows that the administration of \u003cem\u003ecanarium schweinfurthii\u003c/em\u003e oil given at a dose of 10 ml/kg and 20 ml/kg produce a non-significant increase in serum total protein levels in stressed Wistar rats on comparison with the negative control (P ˃ 0.05). Negative Control shows decreased total protein levels due to stress. Stress triggers the release of stress hormones like cortisol, which promotes protein breakdown (catabolism) to provide amino acids for energy and gluconeogenesis. This leads to reduced total protein levels in the blood. CSO at a dose of 10 ml/kg likely mitigate stress-induced protein catabolism, potentially by modulating the release of stress hormones and by enhancing the body\u0026rsquo;s antioxidant defenses. This would help maintain higher protein levels by reducing catabolism, while the high dose of CSO 20ml/kg might still offer some protective effects against stress-induced protein degradation, it might not be as effective as the lower dose. Higher doses could introduce mild toxicity, potentially by affecting liver function and overall metabolism. This could reduce its effectiveness in promoting protein synthesis or protecting against protein degradation. This agrees with the research carried out by Gurevich, (2013). Who study the effect of blood protein concentration on drug-dosing regimes: practical guidance?\u003c/p\u003e"},{"header":"Conclusion","content":"\u003cp\u003eIn this study Group B (stressed\u0026thinsp;+\u0026thinsp;DW) showed the lowest levels of blood urea nitrogen (BUN) and total protein, indicating significant stress impact leading to increased protein catabolism and reduced protein synthesis. Group C (stressed\u0026thinsp;+\u0026thinsp;10ml/kg of CSO) exhibited the highest BUN levels and elevated creatinine levels, suggesting increased protein breakdown, renal stress, and mild kidney impairment due to the low dose of CSO. Group D (stressed\u0026thinsp;+\u0026thinsp;20 ml/kg of CSO) showed elevated but lower BUN and creatinine levels compared to Group C (stressed\u0026thinsp;+\u0026thinsp;10ml/kg of CSO), implying that the higher dose of CSO was more effective in mitigating stress effects and supporting kidney function. Total protein levels increased in Group C (stressed\u0026thinsp;+\u0026thinsp;10ml/kg of CSO) compared to Group B (stressed\u0026thinsp;+\u0026thinsp;DW), indicating partial protective effects of the low CSO dose, whereas Group D (stressed\u0026thinsp;+\u0026thinsp;10ml/kg of CSO) displayed even higher total protein levels, suggesting a dose-dependent enhancement of protein synthesis and stress adaptation. Urea levels followed a similar trend, with Group B (stressed\u0026thinsp;+\u0026thinsp;DW) having the lowest levels, Group C (stressed\u0026thinsp;+\u0026thinsp;10ml/kg of CSO) the highest, and Group D (stressed\u0026thinsp;+\u0026thinsp;10ml/kg of CSO) showing elevated levels but lower than Group C (stressed\u0026thinsp;+\u0026thinsp;10ml/kg of CSO). Overall, the findings suggest that while both doses of CSO provided some protection against stress-induced biochemical changes, the higher dose (20 ml/kg) was more effective in reducing stress impacts, supporting protein metabolism, and maintaining renal function.\u003c/p\u003e \u003cdiv id=\"Sec3\" class=\"Section2\"\u003e \u003ch2\u003eRecommendations\u003c/h2\u003e \u003cp\u003e \u003cul\u003e \u003cli\u003e \u003cp\u003eFuture similar study on the effect of \u003cem\u003ecanarium schweinfurthii\u003c/em\u003e on stress induced renal dysfunction should include measurement of uric acid levels and electrolyte such as chloride, biacarbonate etc following administration of different doses.\u003c/p\u003e \u003c/li\u003e \u003cli\u003e \u003cp\u003eOther future similar study on the effect of \u003cem\u003ecanarium schweinfurthii\u003c/em\u003e on stress induced renal dysfunction should be done on Wistar rat with metabolic disorder such as diabetes, obesity etc.\u003c/p\u003e \u003c/li\u003e \u003cli\u003e \u003cp\u003eSimilar future study should include the effect of \u003cem\u003ecanarium schweinfurthii\u003c/em\u003e on stress induced Wistar rat should be done on important organs such as the lung, brain and heart that are capable of damaging during stress.\u003c/p\u003e \u003c/li\u003e \u003c/ul\u003e \u003c/p\u003e \u003c/div\u003e"},{"header":"Abbreviations","content":"\u003cdiv class=\"DefinitionList\"\u003e \u003cdiv class=\"DefinitionListEntry\"\u003e \u003cdiv class=\"Term\"\u003eCSO\u003c/div\u003e \u003cdiv class=\"Description\"\u003e \u003cp\u003ecanarium schweinfurhii oil\u003c/p\u003e \u003c/div\u003e \u003c/div\u003e \u003cdiv class=\"DefinitionListEntry\"\u003e \u003cdiv class=\"Term\"\u003eBUN\u003c/div\u003e \u003cdiv class=\"Description\"\u003e \u003cp\u003eblood urea nitrogen\u003c/p\u003e \u003c/div\u003e \u003c/div\u003e \u003cdiv class=\"DefinitionListEntry\"\u003e \u003cdiv class=\"Term\"\u003eEDTA\u003c/div\u003e \u003cdiv class=\"Description\"\u003e \u003cp\u003eethylenediaminetetraacetic acid\u003c/p\u003e \u003c/div\u003e \u003c/div\u003e \u003c/div\u003e"},{"header":"Declarations","content":"\u003cp\u003ea. \u003cstrong\u003eEthics approval and consent to participate\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eAll experimental procedures were approved by the Institutional Animal Care and Use Committee (IACUC) of [Prince Abubaker Audu University, in accordance with international guidelines for the care and use of laboratory\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eb Consent for publication\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eNot applicable.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003ec. Availability of data and material\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe datasets generated and/or analyzed during the current study are available from the corresponding author on reasonable request.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003ed. Competing interests\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe authors declare that they have no competing interests.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003ee. Funding\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eNo external funding was received for this study\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003ef. Authors\u0026apos; contributions\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eEzekiel, B. conceived and designed the experiments.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eEzekiel, B and Egbunu, R.E.\u0026nbsp;performed the experiments.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eEneche, J.G. analyzed the data.\u003c/p\u003e\n\u003cp\u003eEjeh, A.O. \u0026nbsp;contributed reagents/materials/analysis tools. [Initials] wrote the manuscript. All authors read and approved the final manuscript.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eg. Acknowledgements\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe authors wish to thank Dr. O.A Daikwo \u0026nbsp;for his technical support and valuable input throughout this study.\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\u003cli\u003e\u003cspan\u003eAdachukwu NN, Pius OU, William RW, Njemuwa NN, Karl F (2019) Fatty Acid Profile and Production of Fatliquor from Canarium schweinfurthii Mesocarp Oil. Pertanika J Sci Technol 27(4):2221\u0026ndash;2243\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eAkinnuga AM, Jeje SO, Bamidele O, Amaku EE, Otogo FO, and Sunday V. E (2014) Virgin Coconut Oil: Remedial Effects on Renal Dysfunction in Diabetic Rats. Hindawi Publishing Corporation Physiol J 10:1\u0026ndash;5\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eAkinpelu LA, Aiyelero OM, Olayiwola G (2019) Ethanol leaf extract of Milicia excelsa mitigates anxiety and depressive-like behaviours induced by acute restraint stress in mice. GSC Biol Pharm Sci 6(2):30\u0026ndash;39\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eAkram E, Sara M, Jalal ZM, Eidi M, Rezazadeh S (2012) Antinociceptive and anti-inflammatory effects of olive oil (Olea europeae L.) in mice. Pharm Biol 50(3):332\u0026ndash;337\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eAna PP, Carda KC, Marchi ER, Andr\u0026eacute; SM, Antunes-Rodrigues J, Claudia MP, Carlos RT (2015) Acute restraint stress induces endothelial dysfunction: role of vasoconstrictor prostanoids and oxidative stress. Stress 18(2):233\u0026ndash;243\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eAnnerstedt M, J\u0026ouml;nsson P, Wallerg\u0026aring;rd M, Johansson G, Karlson B, Grahn P, \u0026Aring;se MH, W\u0026auml;hrborg P (2014) Inducing physiological stress recovery with sounds of nature in a virtual reality forest \u0026mdash; Results from a pilot study. Physiol Behav 118:240\u0026ndash;250\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eAnsari MN, Rehman UN, Karim A, Imam F, Hamad AM (2021) Protective Effect of Thymus serrulatus Essential Oil on Cadmium-Induced Nephrotoxicity in Rats, through Suppression of Oxidative Stress and Downregulation of NF-κB, iNOS, and Smad2 mRNA Expression. Molecules 26(5):12\u0026ndash;52\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eAyodele OM, Kolawole IA, Gabriel OO, Titilola AS (2016) Restraint Stress Impairs Glucose Homeostasis Through Altered Insulin Signalling in Sprague-Dawley Rat. Nigeria J Physiological Sci 31:23\u0026ndash;29\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eChinaka EJ (2018) Viscosity and biodiesel characteristics of wild Canarium schweinfurthii Engl. fruit oil. Reserved Agricultural Eng 64:169\u0026ndash;175\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eChomini MS, Francis MJ, Ishaya M, Chomini AE, Peter MK (2021) Assessment of Proximate and Physico-Chemical Potentials of Crude Kernel Oil Extracts of Canarium schweinfurthii Engl. Nigeria J Pure Appl Sci 34(1):3941\u0026ndash;3950\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eChristine N, Stefan B, Thomas R (2015) Chemical composition and anti-termitic activity of essential oil from Canarium schweinfurthii Engl. Ind Crops Prod 71:75\u0026ndash;79\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eDantzer B, Fletcher QE, Boonstra R, Sheriff MJ (2014) Measures of physiological stress: a transparent or opaque window into the status, management and conservation of species? Conserv Physiol 2:1\u0026ndash;18\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eDeussing MJ, Chen A (2018) The corticotropin-releasing factor family: physiology of the stress response. Physiol Rev 98:2225\u0026ndash;2286\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eEl-Saber G, Batiha J, Oluwafemi T, Hazem MS, Opeyemi AA, Aya AA, Titilade K, Ayandeyi T, Athanasios A, Marios P (2023) Bioactive compounds, pharmacological actions and pharmacokinetics of Cupressus sempervirens. Naunyn-Schmiedeberg's. Archives Pharmacol 396(3):389\u0026ndash;403\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eEzekiel I, Ayo JO, Magaji RA, Tanko Y (2016) Effect of Taurine on Chronic Restraint Stress Induced Behavioural Deficits in Rodents. J Nat Sci Res 6(11):2224\u0026ndash;3186\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eFaizah MN, Mohd AA, Haziq HN, Nurul AMY, Mohamad SIM (2024) Protective Effects of Tualang Honey on the Testes of Rats Subjected to Chronic Combined Stress. Malaysian J Med Health Sci 20(2):34\u0026ndash;41\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eFenga Z, Liub L, Zhanga C, Zhengc T, Wangc J, Lina M, Zhaoc Y, Wanga X, Arnold JL, Wenwei H (2012) Chronic restraint stress attenuates p53 function and promotes tumorigenesis. GenetiCSO 18(109):7013\u0026ndash;7018\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eGon\u0026ccedil;alves M, Mar\u0026iacute;a RP, Anna C, Emma BR, Giulia A, Paula S (2024) A Narrative Review of Metabolomic Insights into Olive Oil\u0026rsquo;s Nutritional Value. Appl Sci 14(10):23\u0026ndash;45\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eGong S, Miao YL, Jiao GZ, Sun MJ, Li H, Lin J, Luo JM, Tan JH (2015) Dynamics and Correlation of Serum Cortisol and Corticosterone under Different Physiological or Stressful Conditions in Mice. Public Libr Sci 10(2):1\u0026ndash;14\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eHaihua G, Wibke BW (2023) The Interplay of Autophagy and Oxidative Stress in the Kidney: What Do We Know? \u003cem\u003eNephron\u003c/em\u003e, 147 (10): 627\u0026ndash;642\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eHassanzadeh-Taheri M, Jahani F, Erfanian Z, Moodi H, Hosseini M (2019) The impact of long-term consumption of diets enriched with olive, cottonseed or sesame oils on kidney morphology: A stereological study. An Acad Bras Cienc 9(1):1\u0026ndash;11\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eIjeoma AD, Tochukwu DM (2020) Profiling and comparison of fatty acids in the oils from the fruits of Dacryodes edulis and Canarium schweinfurthii. J Med Plants Stud 8(5):213\u0026ndash;217\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eKiin-Kabari DB, Umunna PS, Giami SY (2020) Physicochemical properties and fatty acid profile of African elemi fruit pulp oil compared with kernel oil. Eur J Agric Food Sci 2(6):1\u0026ndash;5\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eMishnev M, Korolev A, Zadorin A, Astashkin V (2024) Cyclic Thermomechanical Loading of Epoxy Polymer: Modeling with Consideration of Stress Accumulation and Experimental Verification. \u003cem\u003ePolymers\u003c/em\u003e, 16: 910\u0026ndash;939\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eNyam MA, Makut MD, Itelima UJ, Daniel AM (2014) Nutritional Potential of the Fruits of Black Olive (Canarium schweinfurthii Linn) from Plateau State. Nigeria Pakistan J Nutrition 13(6):335\u0026ndash;339\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eOghenesuvwe EE, Ekene NE, Ajaghaku DL (2014) Guidelines on dosage calculation and stock solution preparation in experimental animals\u0026rsquo; studies. J Nat Sci Res 4(18):100\u0026ndash;106\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eOlivier TT, Armel JS, Francis NT (2016) A Review on Traditional Uses, Phytochemical Composition and Pharmacological Profile of Canarium scweinfurthii Eng. Nat Sci 14(11):17\u0026ndash;22\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eShenghua Z, Ruoyang S, Junhui W, Jun-Feng W, Xin-Min L (2014) Unpredictable chronic mild stress not chronic restraint stress induces depressive behaviours in mice. NeuroReport 25:1151\u0026ndash;1155\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eTang N, Liu XT, Lin LX, Yang WX, Li LQ, Guo-En W, Yan-Hua W (2024) Erzhiwan Ameliorates Restraint Stress- and Monobenzone-Induced Depigmentation in Mice by Inhibiting Macrophage Migration Inhibitory Factor and 8-Hydroxy-2-Deoxyguanosine. Clin Cosmet Invest Dermatology 17:147\u0026ndash;158\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eTeixeira-Santos L, Albino- Teixeira A, Pinho D (2021) An alternative method for oral drug administration by voluntary intake in male and female mice. Lab Anim 55(1):76\u0026ndash;80\u003c/span\u003e\u003c/li\u003e\u003c/ol\u003e"}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":true,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":true,"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":"Stress, renal dysfunction, canarium schweinfurhii","lastPublishedDoi":"10.21203/rs.3.rs-6874801/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-6874801/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003ch2\u003eBackground\u003c/h2\u003e \u003cp\u003eChronic stress regardless of its origin has server repercussion on health, notably impacting the renal system. the kidney are vulnerable to prolonged stress, during stress the renal tissue are exposed to oxidative damage. This study aim to evaluate the protective effect of canarium schweinfurhii oil on renal dysfunction of Wistar rats subjected to restraint stress on the following biomarkers Creatinine, Urea, Blood urea nitrogen and Total protein respectively.\u003c/p\u003e\u003ch2\u003eResult\u003c/h2\u003e \u003cp\u003eA total of twenty-eight (28) apparently healthy adult male Wistar rat weighing between 150\u0026minus;170g were used for this study. They were allocated into four groups of seven rat (n\u0026thinsp;=\u0026thinsp;7 rat/group). Group I: control group and administered with 10ml/kg/bw of water, Group II: stress group\u0026thinsp;+\u0026thinsp;10ml/kg/bw of water, Group III: this group under goes stress and treated with 10ml/kg (1ml/100g) of canarium schweinfurhii oil, Group IV: this group under goes stress and treated with 20ml/kg (2ml/100g) of canarium schweinfurhii oil. All group were treated for 21 days. All data were expressed as mean\u0026thinsp;\u0026plusmn;\u0026thinsp;SEM. The data obtained were statistically analyzed using analysis of variance (ANOVA) with Tukey\u0026rsquo;s multiple comparison post hoc tests to compare the level of significance between control and experimental groups. All statistical analysis were evaluated using SPSS version 17.0 software. The values of P\u0026thinsp;\u0026lt;\u0026thinsp;0.05 were considered as significant.\u003c/p\u003e\u003ch2\u003eConclusion\u003c/h2\u003e \u003cp\u003eThe findings concluded canarium schweinfurhii oil at a dose of 10ml/kg (1ml/100g) provided some protection against stress-induced renal dysfunction\u003c/p\u003e","manuscriptTitle":"Effect of Canarium Schweinfurthii (Africa Black Olive) Oil on Restraint Stressed Induced Renal Dysfunction in Wistar Rat","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2025-06-13 15:32:12","doi":"10.21203/rs.3.rs-6874801/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":"5a5a579c-6135-4a8c-bba1-6c4c405fb837","owner":[],"postedDate":"June 13th, 2025","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"posted","subjectAreas":[{"id":50021853,"name":"General Cell Biology \u0026 Physiology"}],"tags":[],"updatedAt":"2025-06-13T15:32:12+00:00","versionOfRecord":[],"versionCreatedAt":"2025-06-13 15:32:12","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-6874801","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-6874801","identity":"rs-6874801","version":["v1"]},"buildId":"XKTyCvWXoU3ODBz1xrDgd","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}