Vitamin D supplementation alleviates diabetic complications by increasing the amount of irisin in testicular tissues and blood of rats with experimental diabetes.

OBJECTIVE: Diabetes is an important endocrinological disease that has an increasing incidence in the world and affects all biological tissues including testicles. Therefore, this study aimed to reveal the histological and biochemical effects of vitamin D on irisin, apoptosis, total antioxidant status (TAS), and total oxidant status (TOS) in testicular tissues of rats with experimental diabetes. MATERIALS AND METHODS: 41 male Wistar rats, 8-10 weeks old, weighing between 200-220 g, were included in the study as the following groups: control group (n=7; no treatment), sham group [only sodium citrate buffer (SCB)] [n=7; single dose 0.1 Molar (M) SCB given intraperitoneally (i.p)], vitamin D group (n=7; 50 IU/day given orally), diabetes group [n=10; single dose 50 mg/kg Streptozotocin (STZ) dissolved in 0.1 M SCB and given i.p (tail vein blood glucose level above 250 mg/dl after 72 hours)] and diabetes+vitamin D group [n=10, single dose 50 mg/kg STZ, dissolved in 0.1 M SCB and given i.p (tail vein blood glucose level above 250 mg/dl after 72 hours) and when diabetes occurs, oral vitamin D administration of 50 IU/day)]. At the end of the 8 weeks experiment, blood was drawn from the tail vein of all rats, they were sacrificed and testicular tissues were taken. While the amount of irisin in the blood and testicular tissue supernatants was analyzed with the Enzyme-Linked Immunosorbent Assay (ELISA) method, TAS and TOS measurements were analyzed with the REL method, testicular tissues were analyzed histopathologically, immunohistochemically, and with the TUNEL method. RESULTS: When the diabetes group was compared with the control and sham groups, it was reported that the amounts of blood and tissue supernatant irisin and TAS significantly decreased and the TOS was significantly increased; a statistically significant increase in irisin and TAS of blood and tissue supernatants and a significant decrease in TOS were detected when diabetes+vitamin D and diabetes groups were compared among themselves. Similar results were obtained in the immunohistochemical studies. Tissue expressions of irisin decreased in the diabetes group compared to the control and sham groups, while the application of vitamin D increased the tissue expressions of irisin. Additionally, when the numbers of apoptotic cells were compared, it was reported that apoptotic cells in the diabetes group increased significantly compared to the control and sham groups, and vitamin D administration significantly decreased the number of apoptotic cells. CONCLUSIONS: Taken together, vitamin D administration to diabetic rats decreased the number of apoptotic cells and increased the amount of irisin. Vitamin D had an effective role in maintaining the physiological integrity of rat testicular tissues, so vitamin D may be a potent agent to be used in the treatment of diabetes in the future.

Identification of immunohistochemical localization of irisin in the dwarf hamster (Phodopus roborovskii) tissues.

Irisin is mainly secreted by heart and skeletal muscle cells. It is an exercise-induced protein that converts white adipose tissue to brown. Increased irisin expression was lead to weight loss and improved glucose tolerance. We investigated irisin immunoreactivity in various tissues of the dwarf hamsters (Phodopus roborovskii). Tissues were processed, embedded in paraffin, sectioned at 5 µm and stained immunohistochemically for irisin. In the retina, irisin was found almost all layers, except outer nuclear layer. Also, irisin immunoreactivity was observed in the skin, cornea, striated muscle, parotid gland, tongue, oesophagus, stomach and small intestine. The findings from this study support the notion that skeletal muscle is not the primary source of irisin.

Immunohistochemical localization of irisin in mole rats (Spalax leucodon).

Irisin was first identified in skeletal muscle cells. It is an exercise protein that is secreted into the circulation; it causes conversion of white adipose tissue to brown adipose tissue. We investigated irisin immunoreactivity in mole rat (Spalax leucodon) tissues. We examined cerebellum, pituitary, heart, liver, pancreas, spleen, uterus, kidney and striated muscle in female adult mole rats. Tissues were processed, embedded in paraffin, sectioned at 5 µm and stained immunohistochemically for irisin. Irisin immunostaining was detected in the cytoplasm of stained cells; the cytoplasm of Purkinje cells was unstained. We found that irisin may be synthesized in many tissues. The function of locally synthesized irisin currently is unknown.

Immunohistochemical localization of androgen receptors in female mole rat (Spalax leucodon) tissues.

Androgens exert their effects through androgen receptors (AR) in tissues. We investigated the distribution of AR in female mole rat tissues. Tissues were excised, fixed with 10% formalin and embedded in paraffin. Sections were stained after microwave antigen retrieval for immunohistochemistry. Immunostaining of AR immunostaining was detected in the nucleus or cytoplasm of the cells in the cerebral cortex, cerebellum, anterior pituitary, lung, liver, uterus and skin. Granulosa and some thecal cells in the ovary, cardiac muscle cells and adipose cells exhibited a nuclear reaction for AR. In the kidney, labeling of AR was restricted to the cytoplasm of tubule cells. We found that AR could be detected using immunohistochemistry in the nucleus or cytoplasm or both in the presence of androgens.

Immunohistochemical localization of irisin in skin, eye, and thyroid and pineal glands of the crested porcupine (Hystrix cristata).

Irisin was first identified in muscle cells. We detected irisin immunoreactivity in various organs of the crested porcupine (Hystrix cristata). In the epidermis, irisin immunoreactivity was localized mainly in stratum basale, stratum spinosum and stratum granulosum layers; immunoreactivity was not observed in the stratum corneum. In the dermis, irisin was found in the external and internal root sheath, cortex and medulla of hair follicles, and in sebaceous glands. Irisin immunoreactivity was found in the neural retina and skeletal muscle fibers associated with the eye. The pineal and thyroid glands also exhibited irisin immunoreactivity.

Comparative macroanatomical study of the neurocranium in some carnivora.

This study was carried out to investigate the specific anatomical features of the neurocranium of the skull of the dog, cat, badger, marten and otter. Twenty-five animals (five from each species) were used without sexual distinction. The neurocranium consists of os occipitale, os sphenoidale, os pterygoideum, os ethmoidale, vomer, os temporale, os parietale and os frontale. The processus paracondylaris is projected ventrally in the cat, dog, marten and badger, and caudally in the otter. Two foramina were found laterally on each side of the protuberantia occipitalis externa in the otter, and one foramen was found near the protuberantia occipitalis externa in the badger. Foramen was not seen in other species. Paired ossa parietalia joined each other at the midline, forming the sutura sagittalis in the badger, dog, otter and cat while it was separated by the linea temporalis in the marten. The os frontale was small in otters, narrow and long in martens, and quite wide in cats and dogs. The bulla tympanica was rounded in the marten, dog, cat and badger, dorsoventral compressed in otter, and it was very large in all species examined. These observations represented interspecies differences in the neurocranium of marten, otter, badger, cat and dog.