基于Bax/Bcl-2/Caspase-3通路探讨健脾益肾化浊方对多囊卵巢综合征大鼠卵巢功能的改善作用

Improving Effect of Jianpi Yishen Huazhuo Recipe on Ovarian Function in Rats with Polycystic Ovary Syndrome Based on Bax/Bcl-2/Caspase-3 Pathway

关键词:  多囊卵巢综合征  健脾益肾化浊方  Bcl-2  Bax  Caspase-3

Key words:  polycystic ovarian syndrome  Jianpi Yishen Huazhuo recipe  Bcl-2  Bax  Caspase-3

摘要:

目的 观察健脾益肾化浊方通过Bax/Bcl-2/Caspase-3通路调节多囊卵巢综合征（polycystic ovary syndrome，PCOS）大鼠卵巢功能的作用机制。方法 SD大鼠采用高脂饮食联合来曲唑造模，将造模成功的大鼠随机分为空白对照组，模型组，健脾益肾化浊方高、中、低剂量组及阳性药二甲双胍组。应用ELISA法检测各组大鼠血清LH、T和LH/FSH水平；HE染色观察大鼠卵巢组织的形态；免疫组化法和Western blotting法检测各组大鼠Bcl-2相关蛋白（Bax）、B淋巴细胞瘤-2（Bcl-2）、含半胱氨酸的天冬氨酸蛋白水解酶（Caspase-3）的蛋白表达。结果 ELISA结果显示，健脾益肾化浊方各剂量治疗组LH、T和LH/FSH水平明显下降，FSH水平明显升高；HE染色结果显示，健脾益肾化浊方各剂量组的卵巢组织可得到明显改善；免疫组化结果显示，Bax在卵巢颗粒细胞、间质及黄体细胞中的表达升高，Bcl-2在卵母细胞、黄体和卵泡液中表达升高，Caspase-3在颗粒细胞、卵母细胞膜和间质中表达升高；Western blotting结果显示，健脾益肾化浊方各治疗组Bax和Caspase-3蛋白相对表达量下调，Bcl-2蛋白相对表达量上调，Bax/Bcl-2比值降低。结论 健脾益肾化浊方可通过下调PCOS大鼠卵巢组织Bax、Caspase-3蛋白的表达，降低Bax/Bcl-2比值，上调Bcl-2蛋白表达，调整卵巢细胞增殖凋亡过程，改善卵巢微环境，调节内分泌水平，促进卵泡的发育与成熟，从而达到改善卵巢排卵功能的作用。

Abstract:

OBJECTIVE To investigate the mechanism of Jianpi Yishen Huazhuo Recipe in regulating ovarian function in rats with polycystic ovary syndrome(PCOS) through the Bax/Bcl-2/Caspase-3 pathway. METHODS SD rats were fed with a high-fat diet combined with letrozole to establish a model of PCOS. The successfully modeled rats were randomly divided into blank control group, model group, high-, medium-, and low-dose Jianpi Yishen Huazhuo Recipe groups, as well as positive drug metformin group. ELISA was used to detect the serum levels of LH, T, and LH/FSH in each group of rats. HE staining was used to observe the morphology of ovarian tissue. Immunohistochemistry and Western blotting was used to detect the protein expression of Bax, Bcl-2, and Caspase-3 in ovarian tissues of rats. RESULTS ELISA results showed that the levels of LH, T, and LH/FSH in each dose group of Jianpi Yishen Huazhuo Recipe significantly decreased, while the level of FSH significantly increased. HE staining results showed that ovarian tissue in each dose group of Jianpi Yishen Huazhuo Recipe was significantly improved. Immunohistochemistry results showed that Bax expression increased in granulosa cells, interstitial cells, and luteal cells, while Bcl-2 expression increased in oocytes, luteal cells, and follicular fluid. Caspase-3 expression increased in granulosa cells, oocyte membranes, and interstitial cells. Western blotting results showed that the relative expression of Bax and Caspase-3 proteins decreased in each dose group of Jianpi Yishen Huazhuo Recipe, while the relative expression of Bcl-2 protein increased, and the ratio of Bax/Bcl-2 decreased. CONCLUSION Jianpi Yishen Huazhuo Recipe can improve ovarian ovulation function by downregulating the expression of Bax and Caspase-3 proteins in ovarian tissue of PCOS rats, reducing the ratio of Bax/Bcl-2, upregulating the expression of Bcl-2 protein, adjusting the process of ovarian cell proliferation and apoptosis, improving the ovarian microenvironment, regulating endocrine levels, and promoting follicular development and maturation.