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目的研究补肾活血方对卵巢早衰(POF)小鼠卵泡凋亡调控相关基因的表达,为临床中医药预防和治疗卵巢早衰提供思路。方法采用透明带3作抗原,对BALB/c雌性小鼠皮下多点注射,建立免疫性POF模型。用补肾活血方低、中、高剂量进行治疗,设倍美力为阳性对照药,采用ELISA法测定外周血清抗透明带抗体(AzpAb)的含量,进行脏器指数及卵巢病理学观察,免疫组织化学SABC法检测卵泡凋亡调控基因(bcl-2/Bax)表达变化。结果与空白组比较,模型组AzpAb表达明显增高,差异有统计学意义(P<0.01);与模型组比较,各剂量组、倍美力组表达均明显下降,差异有统计学意义(P<0.01);与空白组比较,模型组卵巢指数、胸腺指数均明显下降,脾脏指数明显增加,差异有统计学意义(P<0.01),与模型组比较,高剂组和倍美力组卵巢指数(P<0.01)、胸腺指数(P<0.05)增加,脾脏指数降低(P<0.05);与空白组比较,模型组bcl-2蛋白表达明显下降、Bax蛋白表达显著增加,差异有统计学意义(P<0.01),与模型组比较,各剂量组、倍美力组bcl-2蛋白表达明显增高(P<0.01),Bax蛋白表达显著降低(P<0.01)。结论补肾活血方可能通过调节凋亡调控基因(bcl-2/Bax)的表达,使AzpAb表达明显下降,从而调节免疫反应,抑制抗体集聚,阻止卵泡的过度凋亡,改善卵巢的免疫功能,使有生机卵泡增加,淋巴细胞、浆细胞浸润减少,达到抑制自身免疫损伤、保护卵巢、改善卵巢功能的目的。
Objective To study the expression of the related genes regulating the apoptosis of follicles in Bushen Huoxue Recipe for premature ovarian failure (POF) mice, and to provide ideas for the prevention and treatment of premature ovarian failure in clinical Chinese medicine. Methods BALB / c female mice were injected subcutaneously with zona pellucida 3 as an antigen to establish an immune POF model. The patients were treated with Bushen Huoxue Recipe in low, medium and high dose. The concentration of AzpAb was determined by ELISA, and the visceral index and ovarian pathology were observed. The immune tissues Changes of bcl-2 / Bax expression in ovarian follicles detected by chemical SABC method. Results Compared with the blank group, the expression of AzpAb in the model group was significantly increased (P <0.01). Compared with the model group, the expression of AzpAb in each dose group and the BeiMeiLi group was significantly decreased (P < 0.01). Compared with the blank group, the ovarian index and thymus index of the model group were significantly decreased, the spleen index was significantly increased (P <0.01), compared with the model group, ovarian index (P <0.01), the thymus index (P <0.05) and the spleen index decreased (P <0.05). Compared with the blank group, the expression of bcl-2 protein and the protein expression of Bax in model group were significantly increased, the difference was statistically significant (P <0.01). Compared with the model group, the expressions of bcl-2 protein and Bax protein were significantly decreased (P <0.01). Conclusion Bushen Huoxue prescription may regulate the expression of bcl-2 / Bax and decrease the expression of AzpAb in order to regulate the immune response, inhibit the antibody agglutination, prevent excessive apoptosis of ovarian follicles, and improve the immune function of the ovary. Increased viable follicles, lymphocytes, decreased plasma cell infiltration, to inhibit the autoimmune damage, protect the ovary, the purpose of improving ovarian function.