逍遥散加减方对高催乳激素血症大鼠卵巢颗粒细胞增殖和细胞周期的影响

Effect of Modified Xiaoyao San Powder on Ovarian Granulose Cell Proliferation and Cell Cycle of Hyperprolactinemia Rats

目的:探讨以肝脾为核心,运用逍遥散加减方治疗高催乳素血症(HPRL)模型大鼠,观察逍遥散加减方对体外培养HPRL大鼠卵巢颗粒细胞增殖及细胞周期的影响。方法:选用SD雌性大鼠120只,随机分为正常组、模型组、阳性药溴隐亭组(1 mg·kg-1)、逍遥散加减方高、中、低剂量组(55,22.5,13.75 g·kg-1)。采用ip甲氧氯普胺,制作HPRL大鼠模型。给药30 d后,采用MTT检测各组细胞培养24,48,72,96 h细胞增殖情况;采用流式细胞仪分析细胞周期各时相的变化及细胞凋亡指数。结果:与正常组比较,模型组24,48,72,96 h大鼠卵巢颗粒细胞增殖明显降低,细胞凋亡指数显著增高,S期的细胞增殖明显降低,G0/G1期细胞相对比例明显增加(P<0.05);与模型组比较,溴隐亭组、逍遥散加减方高、中、低剂量组明显增加大鼠卵巢颗粒细胞增殖(P<0.05),逍遥散加减方高剂量组降低卵巢细胞凋亡指数,使S期的细胞增殖显著增多,G0/G1期细胞相对比例减少(P<0.05),其中加味逍遥散高剂量组的作用与溴隐亭组相近。结论:逍遥散加减方促进卵巢颗粒细胞增殖,降低细胞凋亡指数,其作用机制可能是通过促进颗粒细胞从G0期向S期转化,増加S期细胞比例完成。

Objective： To using establish the hyperprolactinemia （HPRL） rat model by using modified Xiaoyao San, to observe the effect of modified Xiaoyao San on ovarian granulosa cell proliferation and cell cycle of HPRL rat in vitro, with the liver and spleen as the core, Intraperitoneal injection of metoclopramide was adopted to make the rat model of HPRL. Method： Totally 120 female SD rats were randomly divided into six groups： the blank group, the negative control group （model group） , the positive control group （Bromocriptine group, 1 mg. kg-1 ） , and the modified Xiaoyao San powder high, middle and low dose groups （55, 22.5, 13.75 g .kg-1 ）. The intervention period was 30 days. MTT assay was used to detect the cell proliferation of cultured cells in each group at 24, 48, 72, 96 h. The change in each phase of the cell cycle was analyzed by flow cytometry and apoptotic index. Result： Compared with the model group, the model group showed significant decreases in granulose cell proliferation at 24, 48, 72, 96 h and S phase and increase in cell ratio in G0/G1 phase （P 〈 0.05）. Compared with the model group, modified Xiaoyao San powder group and Bromocriptine group showed significant reduce apoptotic index and increase in cell proliferation in S phase and decreases in cell ratio in G0/G1 phase （P 〈 0. 05 ）. The effect of modifid Xiaoyao San high dose group was similar with the Bromocnptine group. Conclusion： Modified Xiaoyao San has a significant effect in promoting proliferation of ovarian granulose ceils and reduce apoptotic index, and its mechanism may be correlated with the transformation of granulose ceils to S phase from GO period and the increase in ratio of cells in S phase.