道地通管汤对卵巢早衰大鼠的影响机制研究

目的:探讨道地通管汤对卵巢早衰模型大鼠卵巢功能的影响机制。方法:将40只动情周期规律的SD雌性大鼠随机分为:空白组、模型组、阳性组(补佳乐0.099 mg/kg)、道地通管汤高剂量组(1.4 g/100g)、道地通管汤中剂量组(0.7 g/100 g)。模型组、阳性组、道地通管汤高剂量组、道地通管汤中剂量组采用雷公藤多苷片混悬液(50 mg/kg)连续灌胃14 d,构建卵巢早衰大鼠模型。造模结束后,各组分别给予相应干预剂,连续28 d。干预结束后,各组均取大鼠血清和卵巢组织。HE染色观察大鼠卵巢组织形态结构;ELISA法检测大鼠血清AMH水平;IHC检测卵巢组织中的Bcl-2、Bax及VEGF蛋白表达;TUNEL染色观察卵巢内的颗粒细胞凋亡情况。结果:与空白组相比,模型组大鼠各级生长卵泡均减少,闭锁卵泡增多;血清AMH水平显著下降(P<0.01);卵巢组织Bax蛋白表达升高(P<0.01);Bcl-2、VEGF蛋白表达降低(P<0.01,P<0.05),卵巢组织内卵泡颗粒细胞凋亡率升高(P<0.01);与模型组相比,各中药剂量组血清AMH水平均有不同程度升高,以高剂量组AMH变化最为突出(P<0.05);中药高剂量组卵巢组织Bax蛋白表达下降(P<0.05),Bcl-2、VEGF蛋白表达升高(P<0.05,P<0.05)。中药高剂量组卵巢组织内卵泡颗粒细胞凋亡率降低(P<0.05)。结论:道地通管汤可改善卵巢组织学形态,缓解卵巢损伤,其机制可能与改善POF模型大鼠性激素水平,减低了Bax蛋白在卵巢组织内的表达水平,升高了Bcl-2及VEGF蛋白在卵巢组织内的表达水平,减低了卵巢组织内卵泡颗粒细胞的凋亡率有关。

Study on the mechanism of effect of Daodi Tongguan Decoction on rats with premature ovarian failure

Objective:To investigate the mechanism of the effect of Daodi Tongguan Decoction on ovarian function in rats with premature ovarian failure.Methods:Forty SD female rats with regular estrous cycle were randomly divided into the blank group,the model group,the positive group(Bujiale 0.099 mg/kg),the Daodigitongguan decoction high-dose group(1.4 g/100 g)and the Daodigitongguan decoction medium-dose group(0.7 g/100 g).The model group,positive group,Daodi Tongguan Decoction high-dose group and Daodi Tongguan decoction medium-dose group were given Tripterygium wilfordii polyglycoside tablets suspension(50 mg/kg)by continuous gavage for 14 days to establish a rat model of premature ovarian failure.After the modeling,each group was given the corresponding intervention agent for 28 consecutive days.After the intervention,serum and ovarian tissue were collected from rats in each group.The morphological structure of rat ovarian tissue was observed after HE staining.Serum AMH level was determined by ELISA.The expressions of Bcl-2,Bax and VEGF in ovarian tissue were detected by IHC.The apoptosis of granulosa cells in ovary was observed by TUNEL.Results:Compared to the normal group,the growth follicles in the model group were reduced and the atretic follicles were increased.Serum AMH level was significantly decreased(P<0.01).The expression of Bax protein in ovarian tissue was increased(P<0.01).The protein expressions of Bcl-2 and VEGF were decreased(P<0.01,P<0.05),and the apoptosis rate of follicular granulosa cells in ovarian tissue was increased(P<0.01).Compared to the molding group,the serum AMH level in the TCM dose groups was increased to varying degrees,and the change of the AMH in high-dose group was the most significant(P<0.05).The expression of Bax protein in ovarian tissue decreased(P<0.05),while the expression of Bcl-2 and VEGF protein increased(P<0.05,P<0.05).The apoptosis rate of follicular granulosa cells in ovarian tissue was decreased in the high dose group(P<0.05).Conclusion:Daodi Tongguan Decoction can improve the histological morphology of ovary and relieve ovarian injury,and its mechanism may be related to improving the sex hormone level of POF model rats,reducing the expression level of Bax protein in ovarian tissue,increasing the expression level of Bcl-2 and VEGF protein in ovarian tissue,and reducing the apoptosis rate of follicular granulosa cells in ovarian tissue.