Effect Of Hydrogen On The Reproductive Function Of Male Rats In Hypobaric Hypoxia Environment

OBJECTThe aim of this study is using the advantage of hydrogen selective resistance to oxidation to research the effect of Reactive Oxygen Species during the injury of reproductive health of male induced by hypobaric hypoxia environment, and the effect and mechanism of hydrogen on protection of njury of reproductive system of male.METHODS1. 48 male wistar rats were randomly divided into 4 groups: blank control group(C)、Hypobaric hypoxia group(H)、Hypobaric hypoxia with Physiological saline group(HS)、Hypobaric hypoxia with saturated hydrogen saline group(HH). All the rats were raised in low pressure oxygen cabin which simulating 6000 m environment except group C. Group HS injected with physiological saline(10ml/Kg/d), Group HH injected with saturatedhydrogen saline(10ml/Kg/d). All the samples were collected at 15 th days and 30 th days.2. The testicular tissue of rats were collected and the specimens were embedded in paraffin and each block was cut into sections and stained with hematoxylin and eosin. the morphological changes of rat testis was observed by optical microscope. The ultrastructural change of rat testis was observed by Transmission Electron Microscope.3. The sperm suspension was made by bilateral epididymis tail, and the concentration and vitality of sperm was analysed by phase contrast microscope. The survival rate of sperm was detected by Hypo-osmotic Swelling experiment. The proportion of abnormal sperm was detected by rapid staining of Diff-Quik.4. The expression level of Testosterone, Luteinizing Hormones and Follicle Stimulating Hormone in Rat serum was detected by solid-phase radioimmunoassay5. The generate capacity of Hydroxyl radical, Total Antioxidant Capacity and the level of Malondialdehyde was detected according to the instruction of kit.RESULTS1. Paraffin-section hematoxylin-eosin staining showed that, compared with group C15 d,group H15 d the tube wall of seminiferous tubule folds, seminaferous epithelium thins, spermatogenic cell disorganization in group H15 d. Seminaferous epithelium of group H30 d is thicker than group H15 d, and the number of s Spermatogenic cell also increases, vacuolar degeneration appears intraepithelial. HH15 d group compared with H15 d group and HS15 d group, the number of spermatogenic cells increased, cell polarity is in order, seminaferous epithelium of HH15 d is becoming thicker, the number of sperm increases.2. The result of Transmission Electron Microscope indicates that, H15 d compared with group C15 d, sertoli cell degeneration and necrosis, has unclear boundary with spermatogonial cells, the nucleus of spermatogonia is irregular, mitochondria were vacuolated changes, many of ntercellular tight junctions is destroyed; H15 d compared with group C15 d, the nucleus of sertoli cell arranges neatly and clearly. But, primary spermatocytes mitochondria was still vacuolated, with endoplasmic reticulum expansion.There is no signifance difference between HS group and H group. Morphology and structure of sertoli cell are integrity. Density of spermatogonial cells mitochondrial is normal. Intercellular boundaries were clear. Testicular sertoli cells and spermatogenic cells connected tightly in group HH30 d. but mitochondria vacuoles was still visible in the primary spermatocyte.3. Rat epididymal sperm suspension test results shows that group H sperm concentration, vitality, survival rates were lower than those in group C, more abnormal sperm(P<0.01), there was no significant difference between HS group and H group. Compared with HH15 d group and H15 d group, sperm concentration, activity rate and the survival rate was significantly higher(P<0.01), the percentage of abnormal sperm decreased significantly(P<0.01). HH30 d indexes were no significant difference compared with H30 d, but compared with the HS30 d group, sperm concentration and sperm activity rate was significantly higher(P<0.05).4. Rat serum sex hormone detection results show, Group H15 d FSH levels were significantly reduced compared with the C15d(P<0.01). In group H30 d, T and FSH levels were significantly lower(P<0.05). In group HS, no significant difference compared with H, HH group compared with H group and HS group, there were also no significant differences5. Serum·OH, MDA, T-AOC detection results show H group compared with C group, ·OH, MDA, T-AOC were increased obviously. There was no significant difference compared with HS group and H group. Compared with HH group and H group, ·OH, MDA, T-AOC significantly reduced.CONCLUSIONS1. Hypobaric hypoxia environment can lead to injury and dysfunction of male reproductive system.2. The increase of ROS in hypobaric hypoxia environment is one of the manchansims that induce injury and dysfunction of male reproductive system.3. The male reproductive system has certain adaptability to the injury caused byhypobaric hypoxia environment, however, this adaptability can not completely repaired spermatogenesis dysfunction caused by hypobaric hypoxia environment, so effective measures of protection is necessary.4. Hydrogen can effectively remove the ROS such as ·OH in rats, reduce the injury caused by oxidative stress, effectively protect the injury of reproductive function induced by hypobaric hypoxia environment, especial for the early stage of acute injury...