补肾活血方对免疫性卵巢早衰小鼠卵泡凋亡相关蛋白的影响

摘要

Objective To study the efffects of Bushen Huoxue decoction on premature ovarian failure (POF) of follicle apoptosis-related protein expression in mice and provide ideas for clinical prevention and treatment of POF with Chinese medicine.Methods The zona pellucida 3 as the antigen,the BALB/c female mice,were given multi-point injectionin subcutaneous immunization to induce the models of autoimmune POF.Then the mice were treated with Bushen Huoxue decoction in low-dose,medium-dose,high-dose and Premarin(as control),respectively.Ovarian index,thymus index,spleen index and the morphology of ovary tissue were studied.The follicle apoptosis-related regulatory protein (Bcl-2/Bax) expression was detected by immunohistochemistry SABC and serum AzpAb content was measured by ELISA.Results Compared with model group,the ovarian index and thymus index were increased (P<0.01,P<0.05),and spleen index was decreased (P<0.05) in Bushen Huoxue decoction high-dose group and Premarin group; the ovarian index and thymus index were increased and spleen index was decreased in Bushen Huoxue prescription median-dose group and low-dose group,but there were no significant differences.Bcl-2 protein expression in the control group,Bushen Huoxue prescription all dose groups,Premarin group were significantly higher than that of modeling group (all P<0.05).While Bax protein and AzpAb expressions in the control group,Bushen huoxue decoction in all dose groups,Premarin group were significantly lower than that in model group(P<0.05).Conclusion Bushen Huoxue decoction may influence the apoptosis-related regulatory protein (such as Bel-2 and Bax) expressions,decrease AzpAb expression to improve ovarian function.%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.