Effects of “Bushen Huoxue Prescription” on hormone levels and granulosa cells apoptosis regulated in premature ovarian Failure rat models

ZHANG Li-na; ZHENG Jin; LIU Te; ZHAO Wei

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Effects of “Bushen Huoxue Prescription” on hormone levels and granulosa cells apoptosis regulated in premature ovarian Failure rat models

更新时间：2022-08-31

- Effects of “Bushen Huoxue Prescription” on hormone levels and granulosa cells apoptosis regulated in premature ovarian Failure rat models
- Shanghai Journal of Traditional Chinese Medicine Vol. 49, Issue 7, Pages: 72-76(2015)
- 作者机构：
  
  1. 上海中医药大学附属龙华医院妇科,上海,200032
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ZHANG Li-na, ZHENG Jin, LIU Te, et al. Effects of “Bushen Huoxue Prescription” on hormone levels and granulosa cells apoptosis regulated in premature ovarian Failure rat models. [J]. Shanghai Journal of Traditional Chinese Medicine 49(7):72-76(2015)
DOI：

ZHANG Li-na, ZHENG Jin, LIU Te, et al. Effects of “Bushen Huoxue Prescription” on hormone levels and granulosa cells apoptosis regulated in premature ovarian Failure rat models. [J]. Shanghai Journal of Traditional Chinese Medicine 49(7):72-76(2015) DOI：

摘要

目的：探讨补肾活血方对卵巢早衰大鼠性激素水平及颗粒细胞凋亡调控因子B淋巴细胞瘤/白血病-2蛋白(Bcl-2)、Bcl-2相关X蛋白(Bax)及半胱氨酸蛋白酶-3(Caspase-3)表达的影响方法：将SD大鼠随机分为6组,包括空白组、模型组、生理盐水对照组(简称对照组)及补肾活血方中药低、中、高剂量组。除空白组外,其余各组均用雷公藤多苷(终浓度50 μg/g)连续灌胃14天建立卵巢早衰大鼠模型。其后各组给予相应干预。观察大鼠动情周期改变及大鼠卵巢组织形态学变化；采用酶联免疫吸附法(ELISA法)检测大鼠血清雌二醇(E,2,)、卵泡刺激素(FSH)、促黄体生成素(LH)水平；采用免疫组化方法检测颗粒细胞凋亡相关因子Bax、Bcl-2、Caspase-3 的表达结果：①与空白组比较,模型组大鼠血清FSH水平明显升高(P,<,0.05),LH水平有一定程度升高,且E,2,水平有所降低,但LH及E,2,的改变差异不显著(P,>,0.05)；中药各剂量组大鼠FSH水平较模型组显著降低(P,<,0.01)；LH水平除中药高剂量组外,低、中剂量组明显低于模型组(P,<,0.01)；补肾活血方组大鼠E,2,水平与模型组比较,除中剂量组外均有明显升高(P,<,0.01)。②免疫组化染色结果显示,模型组大鼠颗粒细胞Bax、Caspase-3蛋白表达较空白组明显上升(P,<,0.01),Bcl-2 蛋白有所下降,但下降幅度不明显(P,>,0.05)；中药各剂量组大鼠卵巢颗粒细胞Bax及caspase-3的表达均显著下降(P,<,0.01),而Bcl-2 蛋白表达仅略有增强(P,>,0.05) 结论：雷公藤多苷可诱发大鼠卵巢颗粒细胞凋亡致卵巢早衰；补肾活血方能降低Bax 及Caspase-3蛋白的表达,减轻颗粒细胞的凋亡,从而对雷公藤多苷所致卵巢早衰起到治疗作用。

Abstract

Objective:To study the effect of “Bushen Huoxue Prescription” on hormone levels and expression levels of apoptosis related factors Bax,Bcl-2 and caspase-3 of granulosa cells in premature ovarian failure (POF) rats model Methods:SD rats were randomly divided into 6 groups:blank control group, model group, control group and low-dose, middle-dose and high-dose “Bushen Huoxue Prescription” groups. Premature ovarian failure rats model were set up by gavaging tripterygium wilfordii (final concentration of 50 mg/kg) for 14 days in all the groups except blank control group. Each group was treated with corresponding measures. Estrous cycle and Ovarian morphologic changes of rats were observed. Levels of E,2,FSH and LH were detected by ELISA, and expression of Bax,Bcl-2 and Caspase-3 were detected by immunohistochemical method Results:① Compared with the black control group,the levels of Follicle-stimulating(FSH) in POF model in the model group were increased obviously (P,<,0.05). Meanwhile,the levels of lactogenic hormone (LH) in POF model increased and Estradiol (E,2,) were decreased too,but neither of them had a statistical difference(P,>,0.05). On “Bushen Huoxue Prescription” treated groups,the levels of FSH(P,<,0.01)and LH (except high-dose group,P,<,0.01) decreased significantly than the model group. E,2, increased obviously (P,<,0.01) in “Bushen Huoxue Prescription” treated groups (but not of middle-dose)compared with POF model. ② Th Results: of immunohistochemical staining indicated that compared to the black control group,the expression levels of Bax and caspase-3 were increased obviously on ovarian granulosa cells of POF model (P,<,0.01),but not in Bcl-2 protein(P,>,0.05). The expression of Bax and caspase-3 were decreased significantly in “Bushen Huoxue Prescription” groups (P,<,0.01), and the expression of Bcl-2 also slightly increased(P,>,0.05) Conclusion:Tripterygium wilfordii glycosides could encourage rats’ ovarian granulosa cell apoptosis and POF. “Bushen Huoxue Prescription” could weaken apoptosis of ovarian granulosa cells on tripterygium wilfordii glycosides induced POF rat models via decreasing the expression level of Bax and caspase-3 proteins.

关键词

卵巢早衰补肾活血方颗粒细胞细胞凋亡B淋巴细胞瘤/白血病-2蛋白Bcl-2相关X蛋白半胱氨酸蛋白酶-3

Keywords

premature ovarian failure“Bushen Huoxue Prescription”granulosa cellsapoptosisBcl-2BaxCaspase-3

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