Dipyridamole Reduces Lung Injury in a Rat Lung Ischemia and Reperfusion Model

Dipyridamole Reduces Lung Injury in a Rat Lung Ischemia and Reperfusion Model | 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 Dipyridamole Reduces Lung Injury in a Rat Lung Ischemia and Reperfusion Model Yücel Özgür, Şenel Altun, Reyhan Işık, Refika Kılıçkaya, Burcu Özcan, and 2 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-3918356/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 : In this study, the effect of Dipyridamole (DIP) on lung tissue in lung ischemia/reperfusion (I/R) injury in rats was investigated. Methods : A total of 24 Wistar rats were divided into four equal groups with six rats in each group: Group Sham, Group I/R (ischemia/reperfusion), and Group I/R+ 10mg/kg DIP, Group I/R+ 100mg/kg DIP. 45 minutes of ischemia and 120 minutes of reperfusion were applied. Results : TNF-α value was decreased in Groups 10mg/kg DIP and I/R+ 100mg/kg DIP compared to Group I/R. TNF-α value value was lower in Groups 10mg/kg DIP and 100mg/kg DIP. The difference in TNF-α value was statistically significant between the groups (p=0.002). Statistically significant results were observed between Group I/R-DIP 100 mg/kg and Group I/R for TNF-α. (p=0.038). MDA value in the blood was higher in Group I/R than in Group Sham. MDA value was decreased in Groups 10mg/kg DIP and 100mg/kg DIP compared to Group I/R. However, the difference in MDA value was not statistically significant between the groups (p=0.071). Catalase level in the blood was higher in Group I/R than Group Sham.). The difference in CAT activity was statistically significant between the groups (p=0.025). Statistically significant results were observed between Group I/R-DIP 100 and Group I/R for CAT. (p=0.047). According to histopathological examination, lung tissue neutrophil infiltration, pulmonary edema, interstitial edema, alveolar hemorrhage and total score levels were significantly higher in Group I/R than Groups I/R-10mg/kg DIP and I/R-100mg/kg DIP(p=0.02, p=0.03, p=0.01, respectively, p=0.03 and p=0.01). Lung tissue pulmonary edema, interstitial edema, neutrophil infiltration, alveolar hemorrhage and total score levels were significantly lower in Group 10mg/kg DIP compared to Group I/R (p=0.03, p=0.01, p=0.02, p=0.03 and p=0.01, respectively). ). Lung tissue pulmonary edema, interstitial edema, neutrophil infiltration, alveolar hemorrhage and total score levels were significantly lower in Group I/R-100mg/kg DIP compared to Group I/R (p=0.009, p=0.002, p=0.001, p=0.002 and p=0.002, respectively). ). Conclusion : Lung tissue may be affected by ischemia/reperfusion injury, and this damage can be reversed with the use of dipyridamole. Dipyridamole Ischemia/Reperfusion Lung Rat Figures Figure 1 Figure 2 Figure 3 Figure 4 Figure 5 INTRODUCTION Cardiopulmonary bypass, pulmonary thromboendarterectomy, lung cancer operations, pulmonary artery resection, cardiopulmonary resuscitation, and lung transplantation often result in lung ischemia-reperfusion (I/R) injury. There is a nearly 25% incidence of lung I/R in lung transplantation cases. [1,2] Various signs of damage occur in the tissue during the reperfusion period, which occurs with blood supply in the tissue after ischemia. This process is called “reperfusion injury”. [3,4] Reperfusion injury formation is directly associated with reactive oxygen species (ROS), endothelial cell damage, increased vascular permeability, and activation of neutrophils and platelets, cytokines, and the complement system. The process exacerbated by reoxygenation during reperfusion is thought to be more harmful than ischemia itself. I/R injury consists of complex pathophysiological processes in which vascular, humoral and cellular factors that require the presence of oxygen are activated. Classically, when arterial nutrition is disrupted by embolism or plug, oxygen demand cannot be met and severe tissue hypoxia occurs. In the subsequent reperfusion period, the inflammatory response exacerbated by the restoration of blood flow initiates tissue damage. [5,6] It is characterized by progressive microvascular obstruction associated with post-ischemic tissue perfusion, thrombus formation, and vasoconstriction. Hypoxia can induce endothelial cells and macrophages to develop procoagulant properties that may contribute to the formation of microvascular thrombosis and compromise blood flow during reperfusion. Clinical and experimental studies have shown that thrombin and pro-inflammatory cytokines such as tumor necrosis factor alpha (TNF-α ), interleukin 1-beta (IL 1-β), IL-6, IL-9 and IL-10, which are potent activators and mediators, release has been shown to be induced. [6,7] Pulmonary edema after reperfusion, changes in vascular balance due to exposure of endothelial cells to ischemia-reperfusion, decreased nitric oxide (NO) during reperfusion, vascular dysfunction resulting from cyclic guanosine monophosphate levels, coagulation, vascular permiability, vasomotor tone. Increased leukocyte adhesion and aggregation function are the most important problems encountered. [7] Dipyridamole (DIP) is an agent from the group of antithrombotic drugs used as a platelet aggregation inhibitor. [8,9] Dipyridamole was introduced to the market as a coronary vasodilator more than half a century ago and is still used as an antithrombotic and vasodilator. It inhibits phosphodiesterases and raises extracellular adenosine levels through inhibition of adenosine reuptake by red blood cells. As a result, intracellular levels of cyclic nucleotides are upregulated. The elevation of cGMP in vascular smooth muscle cells and cAMP in platelets provides the mechanism of vasodilation and antithrombosis, which is further enhanced by the release of PGI2 as a result of the increase in endothelial cell cAMP. DIP also increases the activity of endogenous nitric oxide (NO) via cyclic guanosine 3,5-monophosphate (cGMP), which causes pulmonary vasodilation. Antioxidant and anti-inflammatory effects of DIP have been demonstrated in IR studies performed in the brain, liver and heart. [9–12] It was aimed to investigate its effectiveness in reducing oxidative damage due to I/R damage. METHODOLOGY Materials and Methods 24 Wistar Albino male rats (250 g ± 25 g) were used in the study. For this experimental study, approval was obtained from the Experimental Animals Ethics Committee of the university where the experiment was conducted (2021/240). All mice were cared for in accordance with the Experimental Animal Care Guidelines formulated by the National Society for Medical Research and the Guidelines for the Care and Use of Laboratory Animals prepared by the Institute for Laboratory Animal Resources and published by the National Institute. Health. Our studies were conducted in accordance with the Declaration of Helsinki. Groups The subjects were divided into 4 groups, each of which had 6 rats. In Sham(S) group, only thoracotomy was performed. In I/R group, thoracotomy was performed and I/R period was performed. DIP was applied before ischemia-reperfusion in I/R-DIP 10 mg/kg and I/R-DIP 100 mg/kg groups. The rats underwent 45 minutes of ischemia and 120 minutes of reperfusion. DIP dissolved in physiological saline was administered intraperitoneally (ip) 30 minutes before ischemia. After ischemia, the thorax was closed with sutures. At the end of the experiment, the experimental animals were sacrificed by administering a high dose (150 mg/kg ketamine/30 mg/kg Xylazine) anesthetic substance as ip. A blood sample was taken from the heart for biochemical study. Left lung tissue was fixed in 10% formaldehyde and subjected to pathological examination. Anesthesia and Surgery All procedures were performed under sterile conditions. Anesthesia was provided intraperitoneally with 50 mg/kg ketamine (Ketalar vial, Pfizer Pharma GMBH, Germany) and 10 mg/kg xylazine hydrochloride (Alfazyne 2%, Alfasan International, Holland). When necessary, ketamine (half dose, 25 mg/kg) was repeated in order to keep the depth of anesthesia constant, by looking at reflex responses (painful stimulus to the foot with forceps-pedal reflex). Heparin (50IU) and 0.01 mg atropine were given by ip to the rats before thoracotomy. After tracheostomy was opened, rats were mechanically ventilated with a tidal volume of 10 ml/kg, a respiratory frequency of 70 respirations/min, a peep pressure of 2 cm H 2 O, and 100% O 2 . Arterial monitoring was performed from the right carotid artery. [4,5] Drugs were administered as ip 30 minutes before ischemia. After thoracotomy, the thorax was explored and the inferior ligament was freed by cutting, and the left lung was laterally retracted to expose the left hilar structures. Hilar ischemia performed. Biochemical Examination Blood samples from the rats were taken from the heart just before sacrificing. [4] Blood samples stored at -80 0 C were homogenized in phosphate buffer (PBS, 0.01 M, pH = 7.4). The homogenate was centrifuged at 1600×g for 10 minutes. Malondialdehyde (MDA) and Thiobarbituric Acid (TBA) composition levels (nmol/milliliter) (Micromol), which show lipid peroxidation, were measured by colorimetric method using TBARS Assay kit (Cayman Chemical, Item No: 10009055). TNF-α (Tumor Necrosis Factor-Alpha) levels (pg/ml) were measured by Sandwich–ELISA principle using Rat TNF-α (Tumor Necrosis Factor Alpha) ELISA Kit (Elabscience Biotechnology Inc., Catalog No: E-EL-R0019). Catalase (CAT) activity (nmol/ml), Malondialdehyde (MDA) (nmol/ml) and TNF-α (Tumor Necrosis Factor Alpha) levels (pg/ml) were measured with the measurement methods and kits mentioned above. Histopathological Examination After blood samples were taken, samples were taken from the lung tissue. [4] Samples were evaluated blindly under a light microscope by a pathologist experienced in cytology. Left lung samples taken from all groups were fixed in 10% formol and underwent routine follow-up procedures. Serial sections of 4 µm thickness were taken from the tissues embedded in paraffin blocks. Sections taken were deparaffinized, stained with Hematoxylin-Eosin (H-E) and evaluated under a light microscope. The scoring system used by Tassiopoulos et al. will be modified to determine the activity of damage to the lung tissue. [13] Accordingly, 0 points, no change; 1 point, focal slight changes; 2 points, multifocal moderate changes; 3 points, diffuse marked changes; 4 points; numbered as very heavy changes. Tissue damage score was calculated by summing the scores of 4 indexes (perivascular edema, interstitial edema, neutrophil infiltration, and intraalveolar hemorrhage). Statistical Review The data obtained in the experiment were analyzed using the SPSS 20.0 program for Windows. One Way Anova in the parameters where the data meets the normality assumption for multiple analyses; When the assumption of normality was not met, Kruskal Wallis Test was used and values with p < 0.05 were considered statistically significant. In cases where there is a statistically significant difference between the groups, TUKEY, one of the Post Hock analyzes for normal distribution data, and Tammahane's T2 analysis for non-normally distributed data, was used to examine which group caused the difference between the groups. The Shapiro-Wilk value was analyzed because the sample size was less than 30. Since the data are normally distributed in MDA values, parametric analyzes; Nonparametric analyzes were performed for other data in which the data were not normally distributed. RESULTS TNF-α value decreased in mean values ​​in Group I/R-DIP 10 mg/kg (58.95 pg/ml) and Group I/R-DIP 100 mg/kg (37.44 pg/ml) compared to Group I/R (66.7 pg/ml ). Sham group TNF-α mean value is 8.67 pg/ml. TNF-α value was lower in Group I/R-DIP 10 mg/kg and Group I/R-DIP 100 mg/kg. The difference in TNF-α value was statistically significant between the groups (p = 0.002). Statistically significant results were observed between Group I/R-DIP 100 mg/kg and Group I/R for TNF-α. (p = 0.038). TNF-α (serum) graph with mean values is shown in Fig. 1. MDA value in the blood was higher in Group I/R (5,60 nmol/ml) than in Sham group (3.3 nmol/ml). MDA value in Group I/R-DIP 10 mg/kg (4.78 nmol/ml) and Group I/R-DIP 100 mg/kg (4.15 nmol/ml) decreased in average values ​​compared to Group I/R. However, the difference in MDA value was not statistically significant between the groups (p = 0.071). MDA (serum) graph with mean values is shown in Fig. 2 . CAT level in blood was higher in Group I/R (6.72 nmol/ml) than Sham group (2.42 nmol/ml). CAT activity was decreased in Group I/R-DIP 10 mg/kg (5.70 nmol/ml) and Group I/R-DIP 100 mg/kg (3.26 nmol/ml) compared to Group I/R (6.72 nmol/ml). The difference in CAT activity was statistically significant between the groups (p = 0.025). Statistically significant results were observed between Group I/R-DIP 100 and Group I/R for CAT. (p = 0.047). CAT (serum) graph with mean values is shown in Fig. 3 . The error bar graph is shown in Fig. 4 . Histopathological scores of the lung tissue are shown in table 1. Lung tissue pulmonary edema, interstitial edema, alveolar hemorrhage, neutrophil infiltration, and total score levels were significantly higher in Group I/R than Group I/R-DIP 10 mg/kg and Group I/R-DIP 100 mg/kg (p = 0.03, p = 0.01, p = 0.02, p = 0.03 and p = 0.01). The lung tissue pulmonary edema, interstitial edema, neutrophil infiltration, alveolar hemorrhage and total score levels were observed as follows in Group I/R-DIP 10 mg/kg compared to Group I/R (p = 0.015, p = 0.004, p = 0.009, p = 0.699 and p = 0.002, respectively ). Lung tissue pulmonary edema, interstitial edema, neutrophil infiltration, alveolar hemorrhage and total score levels were statistically significant in Group I/R-DIP 100 mg/kg compared to Group I/R (p = 0.009, p = 0.002, p = 0.004, p = 0.002 and p = 0.002, respectively). Histopathological changes of the lung tissue are shown in Fig. 5 . Lung sections of Sham group stained with H-E showed a normal alveolar histological structure. No infiltration was observed and alveolar structure was normal (Fig. 5 a). However, neutrophil infiltration, alveolar edema and hemorrhage were present in lung sections on I/R (Fig. 5 b). Dense neutrophil infiltrates were shown, alveolar edema and hemorrhage were severely increased. A decrease in histological changes was observed in Group I/R-DIP 10 mg/kg and Group I/R-DIP 100 mg/kg compared to Group I/R (Fig. 5 c and Fig. 5 d). DISCUSSION In this study, we investigated to what extent DIP applied before lung IR prevents histopathological changes in lung tissue. I/R injury to organs such as the pulmonary system, liver, lower extremities, and kidneys may fail after traumatic events and shock. [13–15] Multi organ failure may occur as a result of the inflammatory process activated during I/R injury. I/R injury is characterized by direct accumulation of free oxygen radicals (ROS), endothelial cell damage, increased vascular permeability, activation of cytokine and complement system in parallel with the increase in neutrophil and platelet cell accumulation. [6,7] Acute lung injury may occur in critically ill patients, after cardiopulmonary bypass and lung transplantation. Tissue damage occurs especially during the reperfusion period and it has been observed that this period causes more damage than ischemia. This process is a complex pathophysiological process in which vascular, humoral and cellular factors are involved. In the I/R period, microcirculation is also damaged, blood flow homogeneity is impaired, and local tissue damage may occur. Many studies are carried out to prevent the increase in mortality and morbidity resulting from I/R injury. [16–19] DIP increases the level of adenosine at the interstitial level by inhibiting adenosine reuptake at the cellular level. Adenosine mediates various physiological functions through G protein-bound A1, A2A, A2B and A3 receptors. [8,9] Adenosine plays a role in events such as lipolysis, platelet aggregation, regulation of vascular tone and nucleic acid synthesis in adipose tissue. Inhibition of adenosine uptake increases tissue perfusion by increasing cAMP level and increasing adenosine-induced vasodilation. It causes vasodilation by increasing the protocyclin (PGI2) level. Decreased phosphodiesterase level with inhibition of adenosine transport reduces neutrophil activation and superoxide radical accumulation. [10] DIP shows significant antiplatelet activity when used orally at low doses (300–400 mg/dl) and causes minimal hemodynamic changes. [11] It shows antiplatelet activity by inhibiting platelet aggregation and adhesion. Antioxidant and anti-inflammatory effects of DIP have been demonstrated in I/R studies in the brain, liver and heart. A1 and A2 receptor agonists were found to have protective effects in lung, heart, brain and spinal cord I/R studies. Karagüzel et al, in their study with 10 and 100 mg/kg (ip) DIP in rats, concluded that dipiradomol has a higher anti-inflammatory activity than acetyl salicylic acid. [11] In our study, parallel to the study of Karagüzel et al, we investigated the efficacy of the same doses in lung I/R. By inhibiting the phosphodiesterase enzyme, DIP increases the cyclic nucleotide level and prevents platelet aggregation. In addition to its antiplatelet activity, it has been shown to help prevent ischemic and inflammatory processes related to myocardial and cerebral injury. [10,11] A decrease in IL-6 level was detected in ischemic brain tissue in rats treated with DIP. A decrease in IL-6 level has been shown to reduce ischemic tissue damage. [11] In the early reperfusion period, leukocytes and platelets attacking the postischemic endothelium increase reocclusion. The use of antiplatelet agents has been shown to reduce the formation of microemboli after occlusion. [11,13] It has been shown in various studies that DIP protects the erythrocyte membrane against oxidations. DIP is thought to contribute to the prevention of I/R damage by inhibiting ROS release from PNL. [11] Enzymes such as superoxide dismutase (SOD), CAT and glutathione peroxidase (GPX) play an important role in the prevention of tissue damage due to ROS, which is important in the pathophysiology of I/R injury. The antioxidant enzyme group, oxireductases, plays an important role in the scavenging of free radicals and is important in the protection of the cellular structure. [6,7,20] CAT is one of these enzymes. Although the CAT level was statistically significant in our study, the mean value decreased in the groups given DIP, depending on the dose, compared to Group I/R. During I/R, mediators such as TNF-α, ROS and IL-6 (interleukin-6) are involved in tissue damage, altering cellular protein, lipid and ribonucleic acid structure, causing cellular dysfunction and death. In hypoxic endothelium, vasoconstrictor (endothelin types 1,2 and 3) secretion increases, while vasodilator (nitric oxide) synthesis decreases. It has been shown that TNF-α released from distant organs during ischemia injury causes endothelial damage in the lung. [3] Anti TNF-α antibodies have been shown to reduce 4-hour reperfusion injury. [6] Hong et al, in the liver I/R study, a decrease in ALT and AST activity, and a decrease in endothelin-1 and TNF-α levels in liver tissue were observed with DIP compared to the ischemia group. [20] Cytokines, regulation by signaling capacities on cells, chemotaxis and have a stimulating effect. TNF-α is one of the major mediators of the cytokine cascade and is involved in the production of inflammatory molecules IL-1, IL-6 and IL-9. [3] In our study, although the TNF-α level was statistically significant, it showed a decrease in the mean value in the groups given DIP compared to Group I/R, depending on the dose. In I/R, free oxygen radicals appear after lipid peroxidation as a result of high activity in the cell membrane. These radicals cause damage to the deoxyribonucleic acid (DNA), protein and cellular lipid structure of the cell. [1,2] MDA level is thought to be an indicator of lipid peroxidation of radicals. In previous studies, it has been shown that MDA level increases after I/R and decreases in cases where the agent is applied. Orhan et al found that the increased MDA level in the I/R decreased in the amantadine group. [16] In our study, although the MDA level was not statistically significant, it showed a dose-dependent decrease in the mean value in the DIP groups compared to Group I/R. Due to the direct contact of the lung with the external environment, macrophages are in the largest reservoirs of monocytes and leukocytes (PNL). As a result of reperfusion and reexpansion, lipid mediators, polypeptide mediators and immune complexes increase in the environment. Due to these increased mediators, dysfunction occurs in endothelial cells and monocytes, PNL and macrophages enter the alveolacapillary membrane. These blood cells that come to the environment initiate a series of reactions that cause the formation of superoxide radical. [14–17] Oxidative stress, microproteinuria, regression in serum creatinine and BUN levels were observed in rats who underwent DIP after kidney I/R. [16,17] Also, in the same study, a decrease in DIP-related neutrophil accumulation was demonstrated at the tissue level. In our study, leukocyte accumulation, which was clearly observed in the tissue in Group I/R, showed a dose-dependent decrease in the DIP groups. In the early reperfusion period, leukocytes and platelets attacking the postischemic endothelium increase reocclusion. The use of antiplatelet agents has been shown to reduce the formation of microemboli after occlusion. [16] It has been shown in various studies that DIP protects the erythrocyte membrane against oxidations. [16] DIP is thought to contribute to the prevention of I/R damage by inhibiting the release of ROS from PNL. [14,17] Microvascular obstruction caused by thrombus and vasoconstriction occurs in the lung during the post-ischemic perfusion period. Increased platelet and endothelium interaction causes pulmonary arterial constriction and decreased alveolar blood flow. Vascular abnormality causes pulmonary hypertension, vascular reality variability, vascular obstruction, intrapulmonary shunt, increased vascular permeability, and ventilation/perfusion change. [19,21] Histopathological indicators of lung injury are thickening of the alveolar wall, interstitial edema, neutrophil and lymphocyte infiltration. An increase in extravascular albumin accumulation was observed after 30 and 45 minutes of ischemia. [18] In another study, they found an increase in perivascular edema during the 30-minute reperfusion period, and an increase in alveolar edema, leukocyte accumulation, intraalveolar bleeding, and interstitial edema at the end of the 4-hour reperfusion period. [15] In our study, an increase in perivascular edema, interstitial edema and alveolar hemorrhage was observed in Group I/R compared to the control group. In these parameters, dose-related improvement was found in the DIP groups. The limitations of our study are the limited number of animals used and the fact that we clamp not only the pulmonary artery but also the hilum in the lung. In addition, the normal daily dose of DIP in the clinic is 3–4 mg/kg. The doses we applied were determined as 10 and 100 mg/kg doses based on other studies. Conclusion In this study, we showed, for the first time in the literature, that the antioxidant activity of DIP is evident in lung I/R. Histopathologically poor results due to increased inflammation and oxidative stress after I/R were significantly reversed with DIP. Although there was a decrease in the mean values of TNF-α, MDA and CAT from the biochemical results in both groups given DIP compared to the I/R group, statistically significant difference was observed. Future studies will contribute to the possibility of clinical use of the drug. Declarations Author Contribution Y.U., Ş.A., B.O., R.I. and R.K. conceived and planned the study. Y.U. , N.S. and S.A. performed the analysis. Y.U., B.O, and R.I. wrote the original draft with input from all authors. Y.U., B.K. and R.K. visualized the results. S.A., R.I., B.K. and Y.Ö. validated the study. B.O, N.Ş. and R.K. contributed to the interpretation of the results. Y.U. supervised the entire process. All of the authors read and approved the final manuscript.All authors reviewed the manuscript. References Weicun L, Xuan L, Fang L, Yunqiu L. Salvianolic acid B attenuates lung ischemia-reperfusion injury in rat possibly by inhibiting the P2X7/NLRP3 signaling pathway. Int J Clin Exp Med 2018;11(4):3273-3280. Ferrari RS, Andrade CF. Oxidative Stress and Lung Ischemia-Reperfusion Injury. Oxid Med Cell Longev. 2015;2015:590987. De Perrot M, Liu M, Waddell TK, Keshavjee S. Ischemia-reperfusion-induced lung injury. Am J Respir Crit Care Med. 2003 Feb 15;167(4):490-511. Eppinger MJ, Deeb GM, Bolling SF, Ward PA. Mediators of ischemia-reperfusion injury of rat lung. 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Tables Table 1 Histopathological scores of the lung tissue I/R (n = 6) I/R-DIP 10mg/kg (n = 6) I/R-DIP 100 mg/kg (n = 6) p Mean Std. Deviation Std. Error Min Max Mean Std. Deviation Std. Error Min Max Mean Std. Deviation Std. Error Min Max Pulmonary Edema 3,83 0,41 0,17 3,00 4,00 3,00 0,00 0,00 3,00 3,00 2,67 0,52 0,21 2,00 3,00 0,003* İnterstitial Edema 3,50 0,55 0,22 3,00 4,00 2,17 0,41 0,17 2,00 3,00 1,83 0,41 0,17 1,00 2,00 0,001* Neutrophil Accumulation 3,83 0,41 0,17 3,00 4,00 2,67 0,52 0,21 2,00 3,00 2,50 0,55 0,22 2,00 3,00 0,005* Alveolar Hemorrhage 3,83 0,41 0,17 3,00 4,00 3,67 0,52 0,21 3,00 4,00 2,17 0,41 0,17 2,00 3,00 0,002* Total İschemia Score 15,00 0,63 0,26 14,00 16,00 11,50 0,55 0,22 11,00 12,00 10,00 0,89 0,37 9,00 11,00 0,001* *p < 0,05 Kruskall Wallis H Test Additional Declarations No competing interests reported. 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Özgür","email":"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAAA80lEQVRIie3PIY/CMBTA8TZNOjOmt0zAR9iCIsexr9KmCWp4FBk0KYZD36n7Cij0oc40aJKazaCLWwiCbWApkyT0L94TfT9RAGy2Vy1vJpznulrYaUNIMxGPv2uC2hNHhG5Dn1xHB1bkZDobbdROhMPztushAPUpNZFxPyISs82e8o/JWsUCART8bE0kxT4VLosknKvJSsGKYNQxE6ekwq9JFg5WKmlDMKAiGlVkEYJS0ackkMe+TyQhgYQ8/soUEwhy41+8f1ZoPZ0lnnSKvLyoz98l3+mTgfT+bptm9YSimdnj+6ru/Tm5rYvx2Gaz2d60K4nsU/6W0FfwAAAAAElFTkSuQmCC","orcid":"","institution":"Bahcelievler State Hospital, Anesthesiology and Reanimation Clinic","correspondingAuthor":true,"prefix":"","firstName":"Yücel","middleName":"","lastName":"Özgür","suffix":""},{"id":274007497,"identity":"9f71bb4b-adce-4d90-920f-f2b7207cf30f","order_by":1,"name":"Şenel Altun","email":"","orcid":"","institution":"Bahcelievler State Hospital, Cardiovascular Surgeon 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Clinic","correspondingAuthor":false,"prefix":"","firstName":"Burcu","middleName":"","lastName":"Özcan","suffix":""},{"id":274007501,"identity":"b8950cbb-01c0-4453-beb0-59e5ac7ab6c8","order_by":5,"name":"Nurettin Şahin","email":"","orcid":"","institution":"Sadi Konuk Training and Research Hospital, General Surgery Clinic","correspondingAuthor":false,"prefix":"","firstName":"Nurettin","middleName":"","lastName":"Şahin","suffix":""},{"id":274007502,"identity":"4c2f255f-2537-47c1-b4eb-f1efa42c6823","order_by":6,"name":"Bülent Köksal","email":"","orcid":"","institution":"Samsun Bafra State Hospital, Orthopedics and Traumatology Clinic","correspondingAuthor":false,"prefix":"","firstName":"Bülent","middleName":"","lastName":"Köksal","suffix":""}],"badges":[],"createdAt":"2024-02-01 18:30:38","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-3918356/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-3918356/v1","draftVersion":[],"editorialEvents":[],"editorialNote":"","failedWorkflow":false,"files":[{"id":51495909,"identity":"039642c5-9db3-4038-9675-f948563bb09f","added_by":"auto","created_at":"2024-02-22 15:37:56","extension":"png","order_by":1,"title":"Figure 1","display":"","copyAsset":false,"role":"figure","size":5713,"visible":true,"origin":"","legend":"\u003cp\u003eBox chart of TNF-α ( serum) with mean deviation.\u003c/p\u003e","description":"","filename":"1.png","url":"https://assets-eu.researchsquare.com/files/rs-3918356/v1/74dce3c084aa11748922e937.png"},{"id":51495908,"identity":"baad3217-0e05-43ef-ad1e-050d7a080b7c","added_by":"auto","created_at":"2024-02-22 15:37:56","extension":"png","order_by":2,"title":"Figure 2","display":"","copyAsset":false,"role":"figure","size":67548,"visible":true,"origin":"","legend":"\u003cp\u003eBox chart of MDA ( serum) with mean deviation.\u003c/p\u003e\n\u003cp\u003eIntergroup comparisons of the serum malondialdehyde (MDA) levels\u003c/p\u003e","description":"","filename":"2.png","url":"https://assets-eu.researchsquare.com/files/rs-3918356/v1/650c392b29619ad625bb0e78.png"},{"id":51495907,"identity":"a5c0a99f-84e7-4ee0-80da-d5596777140d","added_by":"auto","created_at":"2024-02-22 15:37:56","extension":"png","order_by":3,"title":"Figure 3","display":"","copyAsset":false,"role":"figure","size":63934,"visible":true,"origin":"","legend":"\u003cp\u003eBox chart of CAT ( serum) with mean deviation.\u003c/p\u003e\n\u003cp\u003eIntergroup comparisons of the serum catalase (CAT) levels.\u003c/p\u003e","description":"","filename":"3.png","url":"https://assets-eu.researchsquare.com/files/rs-3918356/v1/fb0aba9fc1323d208d1c9c81.png"},{"id":51496231,"identity":"fe5827fd-8b73-4a22-ba17-0eafb04d71b3","added_by":"auto","created_at":"2024-02-22 15:45:57","extension":"png","order_by":4,"title":"Figure 4","display":"","copyAsset":false,"role":"figure","size":21733,"visible":true,"origin":"","legend":"\u003cp\u003eCAT and MDA error bar charts\u003c/p\u003e","description":"","filename":"4.png","url":"https://assets-eu.researchsquare.com/files/rs-3918356/v1/94a74eb5622ca5eb87276d68.png"},{"id":51495911,"identity":"a57ca327-3f98-4fdc-94e8-d4d47bbfe586","added_by":"auto","created_at":"2024-02-22 15:37:57","extension":"png","order_by":5,"title":"Figure 5","display":"","copyAsset":false,"role":"figure","size":2126077,"visible":true,"origin":"","legend":"\u003cp\u003eLung tissue preparations, hematoxylin-eosin : (a) Normal lung tissue parenchyma; grup Sham (b) Intense neutrophil infiltration and alveolar hemorrhage alveolar edema in lung tissue, Group I/R \u0026nbsp;(c) Neutrophil infiltration, alveolar hemorrhage alveolar edema and hemorrhage in grup I/R-DIP 10 mg/kg; (d) Neutrophil infiltration, alveolar hemorrhage, alveolar edema and decreased hemorrhage in grup I/R-DIP 100 mg/kg.\u003c/p\u003e","description":"","filename":"5.png","url":"https://assets-eu.researchsquare.com/files/rs-3918356/v1/23c12b8ed29fbf37842a9d81.png"},{"id":55265543,"identity":"67828614-36d8-447f-9dbc-8dcfbc1267f5","added_by":"auto","created_at":"2024-04-25 02:06:24","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":2422518,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-3918356/v1/6e0195ac-020c-4172-84fa-2142755a94d7.pdf"},{"id":51495910,"identity":"9ac57797-9746-473f-a66f-f3585449c3d8","added_by":"auto","created_at":"2024-02-22 15:37:56","extension":"sav","order_by":1,"title":"","display":"","copyAsset":false,"role":"supplement","size":2874,"visible":true,"origin":"","legend":"","description":"","filename":"DIP1.sav","url":"https://assets-eu.researchsquare.com/files/rs-3918356/v1/7aa45d4712351e10166df447.sav"}],"financialInterests":"No competing interests reported.","formattedTitle":"Dipyridamole Reduces Lung Injury in a Rat Lung Ischemia and Reperfusion Model","fulltext":[{"header":"INTRODUCTION","content":"\u003cp\u003eCardiopulmonary bypass, pulmonary thromboendarterectomy, lung cancer operations, pulmonary artery resection, cardiopulmonary resuscitation, and lung transplantation often result in lung ischemia-reperfusion (I/R) injury. There is a nearly 25% incidence of lung I/R in lung transplantation cases. \u003csup\u003e[1,2]\u003c/sup\u003e\u003c/p\u003e \u003cp\u003eVarious signs of damage occur in the tissue during the reperfusion period, which occurs with blood supply in the tissue after ischemia. This process is called \u0026ldquo;reperfusion injury\u0026rdquo;. \u003csup\u003e[3,4]\u003c/sup\u003e Reperfusion injury formation is directly associated with reactive oxygen species (ROS), endothelial cell damage, increased vascular permeability, and activation of neutrophils and platelets, cytokines, and the complement system. The process exacerbated by reoxygenation during reperfusion is thought to be more harmful than ischemia itself. I/R injury consists of complex pathophysiological processes in which vascular, humoral and cellular factors that require the presence of oxygen are activated. Classically, when arterial nutrition is disrupted by embolism or plug, oxygen demand cannot be met and severe tissue hypoxia occurs. In the subsequent reperfusion period, the inflammatory response exacerbated by the restoration of blood flow initiates tissue damage. \u003csup\u003e[5,6]\u003c/sup\u003e It is characterized by progressive microvascular obstruction associated with post-ischemic tissue perfusion, thrombus formation, and vasoconstriction. Hypoxia can induce endothelial cells and macrophages to develop procoagulant properties that may contribute to the formation of microvascular thrombosis and compromise blood flow during reperfusion. Clinical and experimental studies have shown that thrombin and pro-inflammatory cytokines such as tumor necrosis factor alpha (TNF-α ), interleukin 1-beta (IL 1-β), IL-6, IL-9 and IL-10, which are potent activators and mediators, release has been shown to be induced. \u003csup\u003e[6,7]\u003c/sup\u003e Pulmonary edema after reperfusion, changes in vascular balance due to exposure of endothelial cells to ischemia-reperfusion, decreased nitric oxide (NO) during reperfusion, vascular dysfunction resulting from cyclic guanosine monophosphate levels, coagulation, vascular permiability, vasomotor tone. Increased leukocyte adhesion and aggregation function are the most important problems encountered. \u003csup\u003e[7]\u003c/sup\u003e\u003c/p\u003e \u003cp\u003eDipyridamole (DIP) is an agent from the group of antithrombotic drugs used as a platelet aggregation inhibitor.\u003csup\u003e[8,9]\u003c/sup\u003e Dipyridamole was introduced to the market as a coronary vasodilator more than half a century ago and is still used as an antithrombotic and vasodilator. It inhibits phosphodiesterases and raises extracellular adenosine levels through inhibition of adenosine reuptake by red blood cells. As a result, intracellular levels of cyclic nucleotides are upregulated. The elevation of cGMP in vascular smooth muscle cells and cAMP in platelets provides the mechanism of vasodilation and antithrombosis, which is further enhanced by the release of PGI2 as a result of the increase in endothelial cell cAMP. DIP also increases the activity of endogenous nitric oxide (NO) via cyclic guanosine 3,5-monophosphate (cGMP), which causes pulmonary vasodilation. Antioxidant and anti-inflammatory effects of DIP have been demonstrated in IR studies performed in the brain, liver and heart. \u003csup\u003e[9\u0026ndash;12]\u003c/sup\u003e It was aimed to investigate its effectiveness in reducing oxidative damage due to I/R damage.\u003c/p\u003e"},{"header":"METHODOLOGY","content":"\u003ch3\u003eMaterials and Methods\u003c/h3\u003e\n\u003cp\u003e24 Wistar Albino male rats (250 g\u0026thinsp;\u0026plusmn;\u0026thinsp;25 g) were used in the study. For this experimental study, approval was obtained from the Experimental Animals Ethics Committee of the university where the experiment was conducted (2021/240). All mice were cared for in accordance with the Experimental Animal Care Guidelines formulated by the National Society for Medical Research and the Guidelines for the Care and Use of Laboratory Animals prepared by the Institute for Laboratory Animal Resources and published by the National Institute. Health. Our studies were conducted in accordance with the Declaration of Helsinki.\u003c/p\u003e \u003cp\u003eGroups\u003c/p\u003e \u003cp\u003eThe subjects were divided into 4 groups, each of which had 6 rats. In Sham(S) group, only thoracotomy was performed. In I/R group, thoracotomy was performed and I/R period was performed. DIP was applied before ischemia-reperfusion in I/R-DIP 10 mg/kg and I/R-DIP 100 mg/kg groups. The rats underwent 45 minutes of ischemia and 120 minutes of reperfusion. DIP dissolved in physiological saline was administered intraperitoneally (ip) 30 minutes before ischemia. After ischemia, the thorax was closed with sutures. At the end of the experiment, the experimental animals were sacrificed by administering a high dose (150 mg/kg ketamine/30 mg/kg Xylazine) anesthetic substance as ip. A blood sample was taken from the heart for biochemical study. Left lung tissue was fixed in 10% formaldehyde and subjected to pathological examination.\u003c/p\u003e \u003cp\u003eAnesthesia and Surgery\u003c/p\u003e \u003cp\u003eAll procedures were performed under sterile conditions. Anesthesia was provided intraperitoneally with 50 mg/kg ketamine (Ketalar vial, Pfizer Pharma GMBH, Germany) and 10 mg/kg xylazine hydrochloride (Alfazyne 2%, Alfasan International, Holland). When necessary, ketamine (half dose, 25 mg/kg) was repeated in order to keep the depth of anesthesia constant, by looking at reflex responses (painful stimulus to the foot with forceps-pedal reflex). Heparin (50IU) and 0.01 mg atropine were given by ip to the rats before thoracotomy. After tracheostomy was opened, rats were mechanically ventilated with a tidal volume of 10 ml/kg, a respiratory frequency of 70 respirations/min, a peep pressure of 2 cm H\u003csub\u003e2\u003c/sub\u003eO, and 100% O\u003csub\u003e2\u003c/sub\u003e. Arterial monitoring was performed from the right carotid artery. \u003csup\u003e[4,5]\u003c/sup\u003e Drugs were administered as ip 30 minutes before ischemia. After thoracotomy, the thorax was explored and the inferior ligament was freed by cutting, and the left lung was laterally retracted to expose the left hilar structures. Hilar ischemia performed.\u003c/p\u003e \u003cp\u003eBiochemical Examination\u003c/p\u003e \u003cp\u003eBlood samples from the rats were taken from the heart just before sacrificing. \u003csup\u003e[4]\u003c/sup\u003e Blood samples stored at -80 \u003csup\u003e0\u003c/sup\u003e C were homogenized in phosphate buffer (PBS, 0.01 M, pH\u0026thinsp;=\u0026thinsp;7.4). The homogenate was centrifuged at 1600\u0026times;g for 10 minutes. Malondialdehyde (MDA) and Thiobarbituric Acid (TBA) composition levels (nmol/milliliter) (Micromol), which show lipid peroxidation, were measured by colorimetric method using TBARS Assay kit (Cayman Chemical, Item No: 10009055). TNF-α (Tumor Necrosis Factor-Alpha) levels (pg/ml) were measured by Sandwich\u0026ndash;ELISA principle using Rat TNF-α (Tumor Necrosis Factor Alpha) ELISA Kit (Elabscience Biotechnology Inc., Catalog No: E-EL-R0019). Catalase (CAT) activity (nmol/ml), Malondialdehyde (MDA) (nmol/ml) and TNF-α (Tumor Necrosis Factor Alpha) levels (pg/ml) were measured with the measurement methods and kits mentioned above.\u003c/p\u003e \u003cp\u003eHistopathological Examination\u003c/p\u003e \u003cp\u003eAfter blood samples were taken, samples were taken from the lung tissue. \u003csup\u003e[4]\u003c/sup\u003e Samples were evaluated blindly under a light microscope by a pathologist experienced in cytology. Left lung samples taken from all groups were fixed in 10% formol and underwent routine follow-up procedures. Serial sections of 4 \u0026micro;m thickness were taken from the tissues embedded in paraffin blocks. Sections taken were deparaffinized, stained with Hematoxylin-Eosin (H-E) and evaluated under a light microscope. The scoring system used by Tassiopoulos et al. will be modified to determine the activity of damage to the lung tissue. \u003csup\u003e[13]\u003c/sup\u003e Accordingly, 0 points, no change; 1 point, focal slight changes; 2 points, multifocal moderate changes; 3 points, diffuse marked changes; 4 points; numbered as very heavy changes. Tissue damage score was calculated by summing the scores of 4 indexes (perivascular edema, interstitial edema, neutrophil infiltration, and intraalveolar hemorrhage).\u003c/p\u003e \u003cp\u003eStatistical Review\u003c/p\u003e \u003cp\u003eThe data obtained in the experiment were analyzed using the SPSS 20.0 program for Windows. One Way Anova in the parameters where the data meets the normality assumption for multiple analyses; When the assumption of normality was not met, Kruskal Wallis Test was used and values with p\u0026thinsp;\u0026lt;\u0026thinsp;0.05 were considered statistically significant. In cases where there is a statistically significant difference between the groups, TUKEY, one of the Post Hock analyzes for normal distribution data, and Tammahane's T2 analysis for non-normally distributed data, was used to examine which group caused the difference between the groups. The Shapiro-Wilk value was analyzed because the sample size was less than 30. Since the data are normally distributed in MDA values, parametric analyzes; Nonparametric analyzes were performed for other data in which the data were not normally distributed.\u003c/p\u003e"},{"header":"RESULTS","content":"\u003cp\u003eTNF-α value decreased in mean values ​​in Group I/R-DIP 10 mg/kg (58.95 pg/ml) and Group I/R-DIP 100 mg/kg (37.44 pg/ml) compared to Group I/R (66.7 pg/ml ). Sham group TNF-α mean value is 8.67 pg/ml. TNF-α value was lower in Group I/R-DIP 10 mg/kg and Group I/R-DIP 100 mg/kg. The difference in TNF-α value was statistically significant between the groups (p\u0026thinsp;=\u0026thinsp;0.002). Statistically significant results were observed between Group I/R-DIP 100 mg/kg and Group I/R for TNF-α. (p\u0026thinsp;=\u0026thinsp;0.038). TNF-α (serum) graph with mean values is shown in Fig.\u0026nbsp;1.\u003c/p\u003e \u003cp\u003eMDA value in the blood was higher in Group I/R (5,60 nmol/ml) than in Sham group (3.3 nmol/ml). MDA value in Group I/R-DIP 10 mg/kg (4.78 nmol/ml) and Group I/R-DIP 100 mg/kg (4.15 nmol/ml) decreased in average values ​​compared to Group I/R. However, the difference in MDA value was not statistically significant between the groups (p\u0026thinsp;=\u0026thinsp;0.071). MDA (serum) graph with mean values is shown in Fig.\u0026nbsp;\u003cspan refid=\"Fig4\" class=\"InternalRef\"\u003e2\u003c/span\u003e.\u003c/p\u003e \u003cp\u003eCAT level in blood was higher in Group I/R (6.72 nmol/ml) than Sham group (2.42 nmol/ml). CAT activity was decreased in Group I/R-DIP 10 mg/kg (5.70 nmol/ml) and Group I/R-DIP 100 mg/kg (3.26 nmol/ml) compared to Group I/R (6.72 nmol/ml). The difference in CAT activity was statistically significant between the groups (p\u0026thinsp;=\u0026thinsp;0.025). Statistically significant results were observed between Group I/R-DIP 100 and Group I/R for CAT. (p\u0026thinsp;=\u0026thinsp;0.047). CAT (serum) graph with mean values is shown in Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e3\u003c/span\u003e. The error bar graph is shown in Fig.\u0026nbsp;\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e4\u003c/span\u003e.\u003c/p\u003e \u003cp\u003eHistopathological scores of the lung tissue are shown in table 1. Lung tissue pulmonary edema, interstitial edema, alveolar hemorrhage, neutrophil infiltration, and total score levels were significantly higher in Group I/R than Group I/R-DIP 10 mg/kg and Group I/R-DIP 100 mg/kg (p\u0026thinsp;=\u0026thinsp;0.03, p\u0026thinsp;=\u0026thinsp;0.01, p\u0026thinsp;=\u0026thinsp;0.02, p\u0026thinsp;=\u0026thinsp;0.03 and p\u0026thinsp;=\u0026thinsp;0.01). The lung tissue pulmonary edema, interstitial edema, neutrophil infiltration, alveolar hemorrhage and total score levels were observed as follows in Group I/R-DIP 10 mg/kg compared to Group I/R (p\u0026thinsp;=\u0026thinsp;0.015, p\u0026thinsp;=\u0026thinsp;0.004, p\u0026thinsp;=\u0026thinsp;0.009, p\u0026thinsp;=\u0026thinsp;0.699 and p\u0026thinsp;=\u0026thinsp;0.002, respectively ).\u003c/p\u003e \u003cp\u003eLung tissue pulmonary edema, interstitial edema, neutrophil infiltration, alveolar hemorrhage and total score levels were statistically significant in Group I/R-DIP 100 mg/kg compared to Group I/R (p\u0026thinsp;=\u0026thinsp;0.009, p\u0026thinsp;=\u0026thinsp;0.002, p\u0026thinsp;=\u0026thinsp;0.004, p\u0026thinsp;=\u0026thinsp;0.002 and p\u0026thinsp;=\u0026thinsp;0.002, respectively).\u003c/p\u003e \u003cp\u003eHistopathological changes of the lung tissue are shown in Fig.\u0026nbsp;\u003cspan refid=\"Fig3\" class=\"InternalRef\"\u003e5\u003c/span\u003e. Lung sections of Sham group stained with H-E showed a normal alveolar histological structure. No infiltration was observed and alveolar structure was normal (Fig.\u0026nbsp;\u003cspan refid=\"Fig3\" class=\"InternalRef\"\u003e5\u003c/span\u003ea). However, neutrophil infiltration, alveolar edema and hemorrhage were present in lung sections on I/R (Fig.\u0026nbsp;\u003cspan refid=\"Fig3\" class=\"InternalRef\"\u003e5\u003c/span\u003eb). Dense neutrophil infiltrates were shown, alveolar edema and hemorrhage were severely increased. A decrease in histological changes was observed in Group I/R-DIP 10 mg/kg and Group I/R-DIP 100 mg/kg compared to Group I/R (Fig.\u0026nbsp;\u003cspan refid=\"Fig3\" class=\"InternalRef\"\u003e5\u003c/span\u003ec and Fig.\u0026nbsp;\u003cspan refid=\"Fig3\" class=\"InternalRef\"\u003e5\u003c/span\u003ed).\u003c/p\u003e"},{"header":"DISCUSSION","content":"\u003cp\u003eIn this study, we investigated to what extent DIP applied before lung IR prevents histopathological changes in lung tissue.\u003c/p\u003e \u003cp\u003eI/R injury to organs such as the pulmonary system, liver, lower extremities, and kidneys may fail after traumatic events and shock. \u003csup\u003e[13\u0026ndash;15]\u003c/sup\u003e Multi organ failure may occur as a result of the inflammatory process activated during I/R injury. I/R injury is characterized by direct accumulation of free oxygen radicals (ROS), endothelial cell damage, increased vascular permeability, activation of cytokine and complement system in parallel with the increase in neutrophil and platelet cell accumulation. \u003csup\u003e[6,7]\u003c/sup\u003e Acute lung injury may occur in critically ill patients, after cardiopulmonary bypass and lung transplantation. Tissue damage occurs especially during the reperfusion period and it has been observed that this period causes more damage than ischemia. This process is a complex pathophysiological process in which vascular, humoral and cellular factors are involved. In the I/R period, microcirculation is also damaged, blood flow homogeneity is impaired, and local tissue damage may occur. Many studies are carried out to prevent the increase in mortality and morbidity resulting from I/R injury. \u003csup\u003e[16\u0026ndash;19]\u003c/sup\u003e\u003c/p\u003e \u003cp\u003eDIP increases the level of adenosine at the interstitial level by inhibiting adenosine reuptake at the cellular level. Adenosine mediates various physiological functions through G protein-bound A1, A2A, A2B and A3 receptors. \u003csup\u003e[8,9]\u003c/sup\u003e Adenosine plays a role in events such as lipolysis, platelet aggregation, regulation of vascular tone and nucleic acid synthesis in adipose tissue. Inhibition of adenosine uptake increases tissue perfusion by increasing cAMP level and increasing adenosine-induced vasodilation. It causes vasodilation by increasing the protocyclin (PGI2) level. Decreased phosphodiesterase level with inhibition of adenosine transport reduces neutrophil activation and superoxide radical accumulation. \u003csup\u003e[10]\u003c/sup\u003e DIP shows significant antiplatelet activity when used orally at low doses (300\u0026ndash;400 mg/dl) and causes minimal hemodynamic changes. \u003csup\u003e[11]\u003c/sup\u003e It shows antiplatelet activity by inhibiting platelet aggregation and adhesion. Antioxidant and anti-inflammatory effects of DIP have been demonstrated in I/R studies in the brain, liver and heart. A1 and A2 receptor agonists were found to have protective effects in lung, heart, brain and spinal cord I/R studies. Karag\u0026uuml;zel et al, in their study with 10 and 100 mg/kg (ip) DIP in rats, concluded that dipiradomol has a higher anti-inflammatory activity than acetyl salicylic acid. \u003csup\u003e[11]\u003c/sup\u003e In our study, parallel to the study of Karag\u0026uuml;zel et al, we investigated the efficacy of the same doses in lung I/R.\u003c/p\u003e \u003cp\u003eBy inhibiting the phosphodiesterase enzyme, DIP increases the cyclic nucleotide level and prevents platelet aggregation. In addition to its antiplatelet activity, it has been shown to help prevent ischemic and inflammatory processes related to myocardial and cerebral injury. \u003csup\u003e[10,11]\u003c/sup\u003e A decrease in IL-6 level was detected in ischemic brain tissue in rats treated with DIP. A decrease in IL-6 level has been shown to reduce ischemic tissue damage. \u003csup\u003e[11]\u003c/sup\u003e In the early reperfusion period, leukocytes and platelets attacking the postischemic endothelium increase reocclusion. The use of antiplatelet agents has been shown to reduce the formation of microemboli after occlusion. \u003csup\u003e[11,13]\u003c/sup\u003e It has been shown in various studies that DIP protects the erythrocyte membrane against oxidations. DIP is thought to contribute to the prevention of I/R damage by inhibiting ROS release from PNL. \u003csup\u003e[11]\u003c/sup\u003e\u003c/p\u003e \u003cp\u003eEnzymes such as superoxide dismutase (SOD), CAT and glutathione peroxidase (GPX) play an important role in the prevention of tissue damage due to ROS, which is important in the pathophysiology of I/R injury. The antioxidant enzyme group, oxireductases, plays an important role in the scavenging of free radicals and is important in the protection of the cellular structure. \u003csup\u003e[6,7,20]\u003c/sup\u003e CAT is one of these enzymes. Although the CAT level was statistically significant in our study, the mean value decreased in the groups given DIP, depending on the dose, compared to Group I/R.\u003c/p\u003e \u003cp\u003eDuring I/R, mediators such as TNF-α, ROS and IL-6 (interleukin-6) are involved in tissue damage, altering cellular protein, lipid and ribonucleic acid structure, causing cellular dysfunction and death. In hypoxic endothelium, vasoconstrictor (endothelin types 1,2 and 3) secretion increases, while vasodilator (nitric oxide) synthesis decreases. It has been shown that TNF-α released from distant organs during ischemia injury causes endothelial damage in the lung. \u003csup\u003e[3]\u003c/sup\u003e Anti TNF-α antibodies have been shown to reduce 4-hour reperfusion injury. \u003csup\u003e[6]\u003c/sup\u003e Hong et al, in the liver I/R study, a decrease in ALT and AST activity, and a decrease in endothelin-1 and TNF-α levels in liver tissue were observed with DIP compared to the ischemia group. \u003csup\u003e[20]\u003c/sup\u003e Cytokines, regulation by signaling capacities on cells, chemotaxis and have a stimulating effect. TNF-α is one of the major mediators of the cytokine cascade and is involved in the production of inflammatory molecules IL-1, IL-6 and IL-9. \u003csup\u003e[3]\u003c/sup\u003e In our study, although the TNF-α level was statistically significant, it showed a decrease in the mean value in the groups given DIP compared to Group I/R, depending on the dose.\u003c/p\u003e \u003cp\u003eIn I/R, free oxygen radicals appear after lipid peroxidation as a result of high activity in the cell membrane. These radicals cause damage to the deoxyribonucleic acid (DNA), protein and cellular lipid structure of the cell. \u003csup\u003e[1,2]\u003c/sup\u003e MDA level is thought to be an indicator of lipid peroxidation of radicals. In previous studies, it has been shown that MDA level increases after I/R and decreases in cases where the agent is applied. Orhan et al found that the increased MDA level in the I/R decreased in the amantadine group. \u003csup\u003e[16]\u003c/sup\u003e In our study, although the MDA level was not statistically significant, it showed a dose-dependent decrease in the mean value in the DIP groups compared to Group I/R.\u003c/p\u003e \u003cp\u003eDue to the direct contact of the lung with the external environment, macrophages are in the largest reservoirs of monocytes and leukocytes (PNL). As a result of reperfusion and reexpansion, lipid mediators, polypeptide mediators and immune complexes increase in the environment. Due to these increased mediators, dysfunction occurs in endothelial cells and monocytes, PNL and macrophages enter the alveolacapillary membrane. These blood cells that come to the environment initiate a series of reactions that cause the formation of superoxide radical. \u003csup\u003e[14\u0026ndash;17]\u003c/sup\u003e Oxidative stress, microproteinuria, regression in serum creatinine and BUN levels were observed in rats who underwent DIP after kidney I/R. \u003csup\u003e[16,17]\u003c/sup\u003e Also, in the same study, a decrease in DIP-related neutrophil accumulation was demonstrated at the tissue level. In our study, leukocyte accumulation, which was clearly observed in the tissue in Group I/R, showed a dose-dependent decrease in the DIP groups.\u003c/p\u003e \u003cp\u003eIn the early reperfusion period, leukocytes and platelets attacking the postischemic endothelium increase reocclusion. The use of antiplatelet agents has been shown to reduce the formation of microemboli after occlusion. \u003csup\u003e[16]\u003c/sup\u003e It has been shown in various studies that DIP protects the erythrocyte membrane against oxidations. \u003csup\u003e[16]\u003c/sup\u003e DIP is thought to contribute to the prevention of I/R damage by inhibiting the release of ROS from PNL. \u003csup\u003e[14,17]\u003c/sup\u003e Microvascular obstruction caused by thrombus and vasoconstriction occurs in the lung during the post-ischemic perfusion period. Increased platelet and endothelium interaction causes pulmonary arterial constriction and decreased alveolar blood flow. Vascular abnormality causes pulmonary hypertension, vascular reality variability, vascular obstruction, intrapulmonary shunt, increased vascular permeability, and ventilation/perfusion change. \u003csup\u003e[19,21]\u003c/sup\u003e Histopathological indicators of lung injury are thickening of the alveolar wall, interstitial edema, neutrophil and lymphocyte infiltration. An increase in extravascular albumin accumulation was observed after 30 and 45 minutes of ischemia. \u003csup\u003e[18]\u003c/sup\u003e In another study, they found an increase in perivascular edema during the 30-minute reperfusion period, and an increase in alveolar edema, leukocyte accumulation, intraalveolar bleeding, and interstitial edema at the end of the 4-hour reperfusion period. \u003csup\u003e[15]\u003c/sup\u003e In our study, an increase in perivascular edema, interstitial edema and alveolar hemorrhage was observed in Group I/R compared to the control group. In these parameters, dose-related improvement was found in the DIP groups.\u003c/p\u003e \u003cp\u003eThe limitations of our study are the limited number of animals used and the fact that we clamp not only the pulmonary artery but also the hilum in the lung. In addition, the normal daily dose of DIP in the clinic is 3\u0026ndash;4 mg/kg. The doses we applied were determined as 10 and 100 mg/kg doses based on other studies.\u003c/p\u003e"},{"header":"Conclusion","content":"\u003cp\u003eIn this study, we showed, for the first time in the literature, that the antioxidant activity of DIP is evident in lung I/R. Histopathologically poor results due to increased inflammation and oxidative stress after I/R were significantly reversed with DIP. Although there was a decrease in the mean values of TNF-α, MDA and CAT from the biochemical results in both groups given DIP compared to the I/R group, statistically significant difference was observed. Future studies will contribute to the possibility of clinical use of the drug.\u003c/p\u003e "},{"header":"Declarations","content":"\u003ch2\u003eAuthor Contribution\u003c/h2\u003e\u003cp\u003eY.U., Ş.A., B.O., R.I. and R.K. conceived and planned the study. Y.U. , N.S. and S.A. performed the analysis. Y.U., B.O, and R.I. wrote the original draft with input from all authors. Y.U., B.K. and R.K. visualized the results. S.A., R.I., B.K. and Y.\u0026Ouml;. validated the study. B.O, N.Ş. and R.K. contributed to the interpretation of the results. Y.U. supervised the entire process. All of the authors read and approved the final manuscript.All authors reviewed the manuscript.\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\n\u003cli\u003eWeicun L, Xuan L, Fang L, Yunqiu L. Salvianolic acid B attenuates lung ischemia-reperfusion injury in rat possibly by inhibiting the P2X7/NLRP3 signaling pathway. Int J Clin Exp Med 2018;11(4):3273-3280.\u003c/li\u003e\n\u003cli\u003eFerrari RS, Andrade CF. Oxidative Stress and Lung Ischemia-Reperfusion Injury. Oxid Med Cell Longev. 2015;2015:590987. \u003c/li\u003e\n\u003cli\u003eDe Perrot M, Liu M, Waddell TK, Keshavjee S. Ischemia-reperfusion-induced lung injury. Am J Respir Crit Care Med. 2003 Feb 15;167(4):490-511. \u003c/li\u003e\n\u003cli\u003eEppinger MJ, Deeb GM, Bolling SF, Ward PA. Mediators of ischemia-reperfusion injury of rat lung. Am J Pathol. 1997 May;150(5):1773-84. \u003c/li\u003e\n\u003cli\u003eOvechkin AV, Lominadze D, Sedoris KC, Robinson TW, Tyagi SC, Roberts AM. Lung ischemia-reperfusion injury: implications of oxidative stress and platelet-arteriolar wall interactions. Arch Physiol Biochem. 2007 Feb;113(1):1-12. \u003c/li\u003e\n\u003cli\u003eScully M, Gang C, Condron C, Bouchier-Hayes D, Cunningham AJ. Protective role of cyclooxygenase (COX)-2 in experimental lung injury: evidence of a lipoxin A4-mediated effect. J Surg Res. 2012 Jun 1;175(1):176-84. \u003c/li\u003e\n\u003cli\u003eLu YT, Hellewell PG, Evans TW. Ischemia-reperfusion lung injury: contribution of ischemia, neutrophils, and hydrostatic pressure. Am J Physiol. 1997 Jul;273(1 Pt 1):L46-54. \u003c/li\u003e\n\u003cli\u003eCiacciarelli M, Zerbinati C, Violi F, Iuliano L. Dipyridamole: a drug with unrecognized antioxidant activity. Curr Top Med Chem. 2015;15(9):822-9. \u003c/li\u003e\n\u003cli\u003eYe Y, Long B, Qian J, Perez-Polo JR, Birnbaum Y. Dipyridamole with low-dose aspirin augments the infarct size-limiting effects of simvastatin. Cardiovasc Drugs Ther. 2010 Dec;24(5-6):391-9. \u003c/li\u003e\n\u003cli\u003ePuri N, Mohey V, Singh M, Kaur T, Pathak D, Buttar HS, Singh AP. Dipyridamole attenuates ischemia reperfusion induced acute kidney injury through adenosinergic A1 and A2A receptor agonism in rats. Naunyn Schmiedebergs Arch Pharmacol. 2016 Apr;389(4):361-8. \u003c/li\u003e\n\u003cli\u003eGarc\u0026iacute;a-Bonilla L, Sosti V, Campos M, Penalba A, Boada C, Sumalla M, Hern\u0026aacute;ndez-Guillamon M, Rosell A, Montaner J. Effects of acute post-treatment with dipyridamole in a rat model of focal cerebral ischemia. Brain Res. 2011 Feb 10;1373:211-20. \u003c/li\u003e\n\u003cli\u003eKarag\u0026uuml;zel E, Kutlu \u0026Ouml;, Yuluğ E, Mungan S, Kazaz İO, Tok DS, \u0026Ouml;zg\u0026uuml;r GK. Comparison of the protective effect of dipyridamole and acetylsalicylic acid on long- term histologic damage in a rat model of testicular ischemia-reperfusion injury. J Pediatr Surg. 2012 Sep;47(9):1716-23. \u003c/li\u003e\n\u003cli\u003eTassiopoulos AK. 1997. Role of nitric oxide and tumor necrosis factor on lung injury caused by ischemia/reperfusion of the lower extremities, J Vasc Surg;26(4):647-5.\u003c/li\u003e\n\u003cli\u003eSaito M, Chen-Yoshikawa TF, Suetsugu K, Okabe R, Takahagi A, Masuda S, Date H. Pirfenidone alleviates lung ischemia-reperfusion injury in a rat model. J Thorac Cardiovasc Surg. 2019 Jul;158(1):289-296. \u003c/li\u003e\n\u003cli\u003eJiang T, Yang W, Zhang H, Song Z, Liu T, Lv X. Hydrogen Sulfide Ameliorates Lung Ischemia-Reperfusion Injury Through SIRT1 Signaling Pathway in Type 2 Diabetic Rats. \u003cem\u003eFront Physiol\u003c/em\u003e. 2020;11:596. \u003c/li\u003e\n\u003cli\u003eLiao WI, Wu SY, Tsai SH, Pao HP, Huang KL, Chu SJ. 2-Methoxyestradiol Protects Against Lung Ischemia/Reperfusion Injury by Upregulating Annexin A1 Protein Expression. \u003cem\u003eFront Immunol\u003c/em\u003e. 2021;12.\u003c/li\u003e\n\u003cli\u003eOrhan M, Tuna Taş A, \u0026Uuml;nal Y, Arslan M, Yazar H, Sezen Cem Ş, et al. The effects of amantadine on lung tissue in lower limb ischemia/reperfusion injury model in rats. Turk Gogus Kalp Dama 2021;29:077-083\u003c/li\u003e\n\u003cli\u003eXiao K, Song L, Hu Y, et al. Novel Role of miR-18a-5p and Galanin in Rat Lung Ischemia Reperfusion-Mediated Response. \u003cem\u003eOxid Med Cell Longev\u003c/em\u003e. 2021;2021:6621921. Published 2021 Aug 14. \u003c/li\u003e\n\u003cli\u003eZhou Y, Zhou X, Zhou W, Pang Q, Wang Z. The protective effect of dexmedetomidine in a rat ex vivo lung model of ischemia-reperfusion injury. Acta Cir Bras. 2018 Jan;33(1):1-13. \u003c/li\u003e\n\u003cli\u003eYe Y, Lin Y, Perez-Polo R, Huang MH, Hughes MG, McAdoo DJ, Manickavasagam S, Uretsky BF, Birnbaum Y. Enhanced cardioprotection against ischemia-reperfusion injury with a dipyridamole and low-dose atorvastatin combination. Am J Physiol Heart Circ Physiol. 2007 Jul;293(1):H813-8. \u003c/li\u003e\n\u003cli\u003eHong Y, Liao WS, Mo RX. Dipyridamole preconditioning protects against ischemia/reperfusion injury of rat liver. Zhongguo Wei Zhong Bing Ji Jiu Yi Xue. 2006 Jul;18(7):425-7. \u003c/li\u003e\n\u003cli\u003eYe Y, Lin Y, Perez-Polo R, Huang MH, Hughes MG, McAdoo DJ, Manickavasagam S, Uretsky BF, Birnbaum Y. Enhanced cardioprotection against ischemia-reperfusion injury with a dipyridamole and low-dose atorvastatin combination. Am J Physiol Heart Circ Physiol. 2007 Jul;293(1):813-8. \u003c/li\u003e\n\u003c/ol\u003e"},{"header":"Tables","content":"\u003cdiv class=\"gridtable\"\u003e\n \u003ctable id=\"Tab1\" border=\"1\"\u003e\n \u003ccaption language=\"En\"\u003e\n \u003cdiv class=\"CaptionNumber\"\u003eTable 1\u003c/div\u003e\n \u003cdiv class=\"CaptionContent\"\u003e\n \u003cp\u003eHistopathological scores of the lung tissue\u003c/p\u003e\n \u003c/div\u003e\n \u003c/caption\u003e\n \u003cthead\u003e\n \u003ctr\u003e\n \u003cth align=\"left\"\u003e\u0026nbsp;\u003c/th\u003e\n \u003cth align=\"left\" colspan=\"5\"\u003e\n \u003cp\u003eI/R (n\u0026thinsp;=\u0026thinsp;6)\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\" colspan=\"5\"\u003e\n \u003cp\u003eI/R-DIP 10mg/kg (n\u0026thinsp;=\u0026thinsp;6)\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\" colspan=\"5\"\u003e\n \u003cp\u003eI/R-DIP 100 mg/kg (n\u0026thinsp;=\u0026thinsp;6)\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\" rowspan=\"2\"\u003e\n \u003cp\u003ep\u003c/p\u003e\n \u003c/th\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003cth align=\"left\"\u003e\u0026nbsp;\u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eMean\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eStd. Deviation\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eStd. Error\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eMin\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eMax\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eMean\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eStd. Deviation\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eStd. Error\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eMin\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eMax\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eMean\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eStd. Deviation\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eStd. Error\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eMin\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eMax\u003c/p\u003e\n \u003c/th\u003e\n \u003c/tr\u003e\n \u003c/thead\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003ePulmonary Edema\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e3,83\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e0,41\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e0,17\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e3,00\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e4,00\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e3,00\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e0,00\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e0,00\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e3,00\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e3,00\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e2,67\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e0,52\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e0,21\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e2,00\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e3,00\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003e0,003*\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003eİnterstitial Edema\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e3,50\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e0,55\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e0,22\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e3,00\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e4,00\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e2,17\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e0,41\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e0,17\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e2,00\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e3,00\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e1,83\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e0,41\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e0,17\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e1,00\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e2,00\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003e0,001*\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003eNeutrophil Accumulation\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e3,83\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e0,41\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e0,17\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e3,00\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e4,00\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e2,67\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e0,52\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e0,21\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e2,00\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e3,00\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e2,50\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e0,55\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e0,22\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e2,00\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e3,00\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003e0,005*\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003eAlveolar Hemorrhage\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e3,83\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e0,41\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e0,17\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e3,00\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e4,00\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e3,67\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e0,52\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e0,21\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e3,00\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e4,00\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n 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align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003e0,001*\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n \u003ctfoot\u003e\n \u003ctr\u003e\n \u003ctd colspan=\"17\"\u003e*p\u0026thinsp;\u0026lt;\u0026thinsp;0,05 Kruskall Wallis H Test\u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tfoot\u003e\n \u003c/table\u003e\n\u003c/div\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":"Dipyridamole, Ischemia/Reperfusion, Lung, Rat","lastPublishedDoi":"10.21203/rs.3.rs-3918356/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-3918356/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003e\u003cstrong\u003eBackground\u003c/strong\u003e: In this study, the effect of Dipyridamole (DIP) on lung tissue in lung ischemia/reperfusion (I/R) injury in rats was investigated.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eMethods\u003c/strong\u003e: A total of 24 Wistar rats were divided into four equal groups with six rats in each group: Group Sham, Group I/R (ischemia/reperfusion), and Group I/R+ 10mg/kg DIP, Group I/R+ 100mg/kg DIP. 45 minutes of ischemia and 120 minutes of reperfusion were applied.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eResults\u003c/strong\u003e: TNF-α value was decreased in Groups 10mg/kg DIP and I/R+ 100mg/kg DIP compared to Group I/R. TNF-α value value was lower in Groups 10mg/kg DIP and 100mg/kg DIP. The difference in TNF-α value was statistically significant between the groups (p=0.002). Statistically significant results were observed between Group I/R-DIP 100 mg/kg and Group I/R for TNF-α. (p=0.038).\u003c/p\u003e\n\u003cp\u003eMDA value in the blood was higher in Group I/R than in Group Sham. MDA value was decreased in Groups 10mg/kg DIP and 100mg/kg DIP compared to Group I/R. However, the difference in MDA value was not statistically significant between the groups (p=0.071). Catalase level in the blood was higher in Group I/R than Group Sham.). The difference in CAT activity was statistically significant between the groups (p=0.025). Statistically significant results were observed between Group I/R-DIP 100 and Group I/R for CAT. (p=0.047).\u003c/p\u003e\n\u003cp\u003eAccording to histopathological examination, lung tissue neutrophil infiltration, pulmonary edema, interstitial edema, alveolar hemorrhage and total score levels were significantly higher in Group I/R than Groups I/R-10mg/kg DIP and I/R-100mg/kg DIP(p=0.02, p=0.03, p=0.01, respectively, p=0.03 and p=0.01). Lung tissue pulmonary edema, interstitial edema, neutrophil infiltration, alveolar hemorrhage and total score levels were significantly lower in Group 10mg/kg DIP compared to Group I/R (p=0.03, p=0.01, p=0.02, p=0.03 and p=0.01, respectively). ). Lung tissue pulmonary edema, interstitial edema, neutrophil infiltration, alveolar hemorrhage and total score levels were significantly lower in Group I/R-100mg/kg DIP compared to Group I/R (p=0.009, p=0.002, p=0.001, p=0.002 and p=0.002, respectively). ).\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eConclusion\u003c/strong\u003e: Lung tissue may be affected by ischemia/reperfusion injury, and this damage can be reversed with the use of dipyridamole.\u003c/p\u003e","manuscriptTitle":"Dipyridamole Reduces Lung Injury in a Rat Lung Ischemia and Reperfusion Model","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2024-02-22 15:37:52","doi":"10.21203/rs.3.rs-3918356/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":"108f1e0a-50a7-4915-a630-77ae0ca98955","owner":[],"postedDate":"February 22nd, 2024","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"posted","subjectAreas":[],"tags":[],"updatedAt":"2024-04-24T11:01:25+00:00","versionOfRecord":[],"versionCreatedAt":"2024-02-22 15:37:52","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-3918356","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-3918356","identity":"rs-3918356","version":["v1"]},"buildId":"qtupq5eGEP_6zYnWcrvyt","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}