壳寡糖对病理性卵巢衰退小鼠免疫功能和生殖功能的作用

目的：探讨壳寡糖促进病理性卵巢功能衰退小鼠生殖功能和免疫功能恢复的可能性。方法：选用43只生育旺盛期雌性小鼠，除正常对照组（n=8）外，其它通过白消安/环磷酰胺构建病理性卵巢功能衰退模型模拟卵巢功能早衰，随机选取3只，卵巢切片HE染色观察卵泡情况以判断不孕模型。构建成功后将余下32只随机平均分为4组（n=8），经不同剂量壳寡糖（0，100，200，300 mg/（kg·d））灌胃后，比较组间卵巢、脾脏、胸腺脏体比的变化，观察卵泡情况、检测腹腔巨噬细胞吞噬能力、外周血雌二醇（E₂）及孕酮（P）水平，检测卵巢生殖上皮细胞中生殖细胞标志物小鼠血管同源物（MVH）、干细胞标志物OCT-4以及卵巢中免疫因子肿瘤坏死因子α（TNF-α）、白介素-2（IL-2）、白介素-6（IL-6）表达量的变化，并分析生殖干细胞标记物表达水平变化与免疫因子表达水平变化的相关关系。结果：随壳寡糖灌胃剂量的增加，卵巢、脾脏和胸腺脏体比同步增高；卵巢中总卵泡数及各级卵泡数都呈递增趋势；外周血E2水平递增，P水平呈递减趋势；腹腔巨噬细胞吞噬功能随剂量增高而增强；生殖干细胞标记物和免疫因子的表达水平均呈显著递增趋势，表明生殖干细胞标记物的表达水平与免疫因子表达水平的变化呈显著的正相关关系（P < 0.05）。结论：壳寡糖可改善病理性卵巢功能早衰小鼠的免疫功能，促进雌性生殖干细胞增殖、分化，从而促进卵巢病理性早衰机体生殖功能在一定程度上的恢复。

Effects of COS on promoting the pathological ovarian aging mice ovarian function by regulating immune function

Objective: To determine whether the immunopotentiator chitosan oligosaccharide(COS)can recover the reproductive functions of pathological ovarian recession mice and improvetheir immunity. Methods: Forty-three fertile female mice (at around 2 months),in addition to a normal control group (n=8), injected intraperitoneally with busulfan and cyclophosphamide to construct premature ovarian failure mod-els. Three of them were used to test whether the infertility model was constructed successfully by HE staining. Then the models were randomly divided into four groups (n=8) and treated with different dosages of COS by gavage, after which compared different groups' organ ratios (the weight of immune organs and ovary/body weight), ovarian follicles and peritoneal macrophages' phagocytosis as well as estragon(E₂) and pro-gesterone(P) levels in peripheral blood. In addition, we measured the expression dynamics of the ovarian protein reproductive cell marker mouse vasa homolog(MVH), germ stem cell marker OCT-4 in ovarian surface epithelium (OSE) and part of immune factors including tumor necrosis factor (TNF-α),interleukin-2(IL-2)as well as IL-6 to analyze the correlativity between germline stem cells marker dynamics and im-mune factors expression changes. Results: With increasing dosages of COS, organ ratios of ovaries, thymus and spleen both went up syn-chronously; The whole number of follicles and every stages of follicles are all presenting with progressive tendency; E₂ level in peripheral blood ascends, however, progesterone level declined relatively; Neutral red experiment revealed the phagocytosis ability of peritoneal macrophages became stronger with increasing dosages of COS; the results of Western blot had shown that no matter the expression level of germ stem cells marker or immune factors were all presenting increasing tendency, which means that the expression level dynamics of germ stem cell marker has a positive correlation with immune factors expression changes. The results were statistically significant. Conclusion: COS could improve the immunity of mice with pathological ovarian recession and at the same time it would promote the proliferation and differentiation of female germ line stem cells (FGSCs), and then helped saving ovarian functionsto some extent.