Investigation of effect of hippophae rhamnoides L. (Seabuckthorn) on distribution of irisin and asprosin in kidney and testicular injury in rats with experimentally induced diabetes

Diabetes is a disease which increased amount of glucose in blood and urine affects the whole organism, especially heart, liver, brain, kidney and testicular tissues due to insufficiency of insulin hormone secreted from pancrease or as a result of insufficient use of insulin in the body. Studies have shown that oxidative stress, which occurs due to inbalance of free radicals, plays in etiology of diabetes. Studies which sea buckthorn was used to improve free radical/antioxidant balance drew attention. In this study, examination of biochemical, spermatogenical and histopathological effects of sea buckthorn against oxidative stress in kidney and testicular tissues after STZ-induced diabetes was aimed. In addition, examination of immunoreactivity of Irisin and Asprosin, whose relationship with energy and glucose metabolism has been recovered in recent years, in kidney and testicular tissues, and investigation of possible protective effects of antioxidant-rich sea buckthorn was aimed. For this purpose; 39 Sprague-Dawley male rats at 7 weeks of weeks of age were used. These animals were divided into 5 groups as Control group, Citrate group, SBT group, Diabetes group and Diabetes + SBT group. At the beginning of experiment, blood-glucose levels and body weights of all rats were measured. No procedure was applied to control group rats (n:7); Citrate groups rats (n:7) received 0,01 M citrate buffer intraperitonally. SBT group animals (n:7) received seabuckthorn oil via oral gavage with dose of 70 mg/kg every other day for 8 weeks. Diabetes group rats (n:9) injected intraperitonally with STZ with dose of 50 mg/kg. After 72 hours, rats with blood-glucose level higher than 250 mg/dL were considered diabetic. Diabetes+SBT group rats (n:9) injected with STZ with dose of 50 mg/kg. After 72 hours, animals with blood-glucose level higher than 250 mg/dL were considered diabetic. The rats considered as diabetic were received seabuckthorn oil with dose of 70 mg/kg every other day via oral gavage method. And the end of 8 weeks, blood-glucose levels, body weight were measured and all rats were sacrificed and testicular and kidney weight of animals were measured. Serum samples for biochemical examinations and sperm samples for spermatological examination were taken. Testicular and kidney tissues examined with histopathological (Hematoxylin-Eosin, Periodic Acid-Schiff and Crossman stains) and immunohistochemical methods. As a result of experiment, body weight values decreased in diabetes group and increased in Diabetes+SBT group. Also blood glucose levels where higher in Diabetes group compared to Control group. In diabetes+SBT group, blood-glucose levels were decreased when compared to Diabetes group. As a result of Hematoxylin-Eosin, Periodic Acid Schiff and Crossman stains in kidney tissues, SBT has been shown to reducing effect against changes such as hyperemia, narrowing of Bowman's space, thickening of basement membrane and increase of connective tissue. It was observed that the kidney weight decreased in the Diabetes + SBT group against increased in the Diabetes group. Irisin immunoreactivity in kidney was observed to be decreased in diabetes group, whereas increased in Diabetes+SBT group when compared to Control group. It was observed that kidney Asprosin immunoreactivity was increased in Diabetes group and was similar in Diabetes + SBT group when compared to control group. When oxidative stress associated with diabetes was examined, lowest TOS values were detected in Diabetes group and highest TAS values were detected in SBT group. Also, highest and lowest oxidative stress index were observed in Diabetes+SBT and Diabetes groups, respectively. According to results of Hematoxylin-Eosin, Periodic Acid Schiff and Crossman stains in testicular tissues, SBT has shown to reducing effect against histopathological changes such as degeneration, interstitial hyperemia, increase of connective tissue and thickening of the basement membrane which occured in the seminiferous tubules and Leydig cells. However, no effect was observed in testicular weight as a result of SBT treatment. In immunohistochemical stains of testicular tissue, it was observed that Irisin level decreased in the diabetes group compared to the control group, whereas it increased in the Diabetes + SBT group. Immunohistochemical staining of Asprosin in testis showed decreased levels of immunoreactivity in Diabetes group when compared to control group, and increased immunoreactivity in Diabetes+SBT group. Sperm motility and density were increased in the Diabetes + SBT group compared to the Diabetes group, but there was no effect on the abnormality. However, the lowest abnormality value was found in animals treated with only SBT. As a result, SBT oil treatment to animals with induced diabetes, has been found to reduce weight loss as a result of diabetes, decrease blood-glucose levels and it was observed that SBT reduced the harmful effects of oxidative stress after diabetes. In addition, it was determined that SBT oil had a healing effect on result of spermatogenic examination of diabetic animals. Finally, it was observed that SBT had a decreasing effect on histopathological damage in testicular and kidney tissues caused by diabetes, and had a positive effect on Irisin and Asprosin.