补肾活血方对卵巢早衰小鼠颗粒细胞凋亡的影响

摘要

目的：探讨补肾活血方抑制卵巢早衰(POF)小鼠卵泡颗粒细胞凋亡的作用机制。方法以小鼠透明带3为抗原,皮下多点注射免疫BALB/c雌性小鼠建立免疫性POF模型。取40只清洁级雌性小鼠,随机分为空白组、模型组、阳性药组、补肾活血方组。补肾活血方组给予补肾活血方药液灌胃,阳性药组给予戊酸雌二醇药液灌胃,模型组和空白组给予等量生理盐水灌胃,2次/d,连续15 d。取卵巢、子宫、胸腺进行称重,计算其脏器指数；HE染色观察颗粒细胞凋亡情况；采用放射免疫法检测雌二醇(E2)、促黄体生成激素(LH)和促卵泡生成激素(FSH)的水平；采用RT-PCR分析卵巢颗粒细胞中Fas、Fas-L mRNA表达情况。结果与空白组比较,模型组卵巢、胸腺、子宫质量和E2水平显著降低(P＜0.05,P＜0.01),FSH、LH水平和Fas、Fas-L mRNA表达显著升高(P＜0.01)；与模型组比较,给药组颗粒细胞凋亡明显减少,卵巢指数、子宫指数、胸腺指数增加(P＜0.05),E2水平升高(P＜0.01),FSH、LH水平及Fas、Fas-L mRNA表达明显降低(P＜0.05,P＜0.01)。结论补肾活血方可调节下丘脑-垂体-卵巢轴的激素水平,通过降低Fas、Fas-L的mRNA表达抑制细胞凋亡。%Objective To discuss Bushen Huoxue Prescription mechanism of action on follicular granulosa cell apoptosis of premature ovarian failure (POF) mice. Methods The mouse zona pellucida 3 as the antigen, multi-point injection of subcutaneous immunization BALB/c to female mice was used to establish autoimmune POF model. Forty clean female mice were randomly divided into blank group, model group, Estradiol Valerate group and Bushen Huoxue Prescription group. All medicine groups were given relevant medicine for gavage, while model and blank groups were given normal saline with the same amount for gavage for 15 days. After the treatment, ovary, uterus, and thymus were weighed, and the viscera index was calculated. An ovarian slice was taken and HE staining was used to observe granulosa cell apoptosis. The levels of E2, LH, and FSH were detected by using the radioimmunoassay method. RT-PCR was used to analyze Fas and Fas-L mRNA expressions in ovarian granulosa cells. Results Compared with blank group, the weight of ovary, thymus, and uterus decreased significantly (P<0.05, P<0.01);Serum E2 level dropped (P<0.01);levels of FSH and LH, and expressions of Fas and Fas-L mRNA increased significantly (P<0.01). Compared with model group, granulosa cell apoptosis significantly decreased, and ovary index, uterus index, and thymus index increased (P<0.05);serum E2 level increased (P<0.01);levels of FSH and LH, and expressions of Fas and Fas-L mRNA significantly decreased (P<0.05). Conclusion Bushen Huoxue Prescription can decrease expressions of Fas and Fas-L mRNA to inhibit cell apoptosis by adjusting the hormone level of hypothalamus-pituitary-ovarianaxis.