| Objectives:Premature ovarian failure (POF) meant non-physiological menopause after puberty and before40years old. The incidence of POF was one to three percent, while the incidence in secondary amenorrhea was two to ten percent. The pathogene-sis of this disease was not very clear so far, but it’s quite certain that the disease could be caused by iatrogenic factors, and it’s caused widely concerned recent years. Espe-cially for the young malignant tumor patients through chemotherapy, the quality of life could be influenced seriously by the irreversible damage of reproductive system. And it’s a tough rock for the scientists to help these patients to regain their endocrine and reproductive ability. It’s indicated by lots of researches that there was an oxida-tion and anti-oxidation balance in the microenvironment of ovaries and follicles. The specific performance was that enzymatic antioxidant and non-enzymatic antioxidant expressed significantly high in follicular fluid. Hydrogen did have some reducibility, and have protective effect on ischemia reperfusion injury, sepsis, low-estrogen in-duced osteopenia. This experiment use cyclophosphamide (CTX) to induce ovarian damage in rats. Investigating the influence towards the damage by hydrogen rich sa-line, and seek the theoretical basis for clinical application.Methods:Animals were randomly divided into five groups:group control; group control+H2:hydrogen rich saline was injected10ml/kg twice at8am and8pm each day; group ovarian injure (OI):a loading dose of CTX (50mg/kg) was received at the first day followed by daily intraperitoneal CTX injection at8mg/kg for the next con-secutive14days; group OI+0.6H2:the same method was used to inject CTX, while hydrogen rich saline was injected10ml/kg twice at8am and8pm each day during the same time CTX of8mg/kg was injected; group OI+0.3H2:the same method was used to inject CTX, while hydrogen saline at concentration of0.3mmol/L was injected10ml/kg twice at8am and8pm each day during the same time CTX of8mg/kg was injected.After the last injection, the animals were executed in batches on the day15, day30and day45.The pathology of ovaries was observed by hematoxylin-eosin staining. The levels of follicle-stimulating hormone (FSH), estradiol (E2),superoxide dismutase(SOD), catalase (CAT) and malondialdehyde (MDA) in serum were de-tected. And the levels of f SOD, CAT and MDA besides Nrf2expression was detected in collected ovarian protein. Results:Comparing to group control, sexual hormone had a significant change at15days,30days,45days after intraperitoneal injection of CTX as high serous level of FSH and low serous level of E2. But in the groups given hydrogen rich saline, the ab-normal expression of sexual hormone was much better in the ovarian damaged rats, and showed a dose-dependent effect. After injection of CTX, the changes of patho-logical manifestations and oxidation-deoxidization systems emerged in the ovaries: MDA as oxidative products expresses high, SOD and CAT as anti-oxidases showed a high activity. While after hydrogen rich saline was given, the expression of MDA was much lower, but the activity of SOD and CAT was higher. The expression of ovarian Nrf2was increased in the damaged ovaries compared to group control, but the ex-pression was even more increased after hydrogen rich saline was given to induce an increased expression of the anti-oxidases induced by Nrf2.Conclusion:Hydrogen rich saline did have protective effect on chemotherapy in-duced ovarian oxidative damage, and showed a dose-dependent effect. Hydrogen rich saline could change the expression of sexual hormone and pathological manifestations in ovarian tissues. Hydrogen rich saline could reduce the expression of MDA, while increase the activity of SOD and CAT. It could also increase the expression of ovarian Nrf2, this may be related to the stimulation of endogenous antioxidant system (keap1-Nrf2-ARE). |
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