养阴舒肝颗粒对VCD诱导卵巢功能下降大鼠Bcl-2,Bax及Caspase-3蛋白表达的影响  

Effect of Yangyin Shugan Granule on Expression of Bcl-2,Bax and Caspase-3 Protein in Rats with VCD-induced Diminished Ovarian Function

作　　者：王吉菊 刘建[2] 黄旭春[2] 曾玉燕[2] 王小云[2] WANG Ji-ju;LIU Jian;HUANG Xu-chun;ZENG Yu-yan;WANG Xiao-yun(The Second Clinical College,Guangzhou University of Chinese Medicine,Guangzhou 510405,China;Guangdong Hospital of Traditional Chinese Medicine,Guangzhou 510120,China)

机构地区：[1]广州中医药大学第二临床医学院,广州510405 [2]广东省中医院,广州510120

出　　处：《中国实验方剂学杂志》2018年第20期159-163,共5页Chinese Journal of Experimental Traditional Medical Formulae

基　　金：广东省自然科学基金项目(2017A030313728)

摘　　要：目的：探讨养阴舒肝颗粒对卵巢功能下降模型大鼠卵巢功能及;B淋巴细胞瘤-2（Bcl-2）,Bcl-2相关X蛋白（Bax）,半胱氨酸天冬氨酸蛋白酶-3（Caspase-3）蛋白的影响。方法：选取动情周期规则的雌性SD大鼠,取10只作为正常组,其余大鼠均采用160 mg·kg^（-1）去氧乙烯基环己烯（VCD）腹腔注射诱导卵巢功能下降模型,取造模成功的56只大鼠按体质量随机分为模型组,养阴舒肝颗粒低（2.1 g·kg^（-1））,中（4.2 g·kg^（-1））,高剂量组（8.4 g·kg^（-1））和坤泰胶囊组,每组11只（模型组12只）。采用养阴舒肝颗粒和坤泰胶囊进行灌胃给药,干预治疗4周后进行卵巢称质量,计算各级卵泡的数目,酶联免疫吸附测定法（ELISA）检测血清性激素水平卵泡刺激素（FSH）,雌二醇（E_2）,抗缪勒氏管激素（AMH）,蛋白免疫印迹法（Western blot）检测卵巢组织Bcl-2,Bax,Caspase-3的表达量。结果：与正常组比较,模型组大鼠的卵巢质量和卵巢指数下降（P〈0.01）,闭锁卵泡数目增多（P〈0.01）,FSH水平升高（P〈0.01）,E_2和AMH水平下降（P〈0.05）,卵巢组织Bcl-2蛋白的表达减少（P〈0.01）,Bax,Caspase-3蛋白的表达（P〈0.01）;与模型组比较,养阴舒肝颗粒可以增加模型大鼠的双侧卵巢质量（P〈0.05,P〈0.01）;养阴舒肝颗粒和坤泰胶囊可以增加卵巢指数（P〈0.05,P〈0.01）;养阴舒肝颗粒可以减少闭锁卵泡的数目（P〈0.05）;养阴舒肝颗粒和坤泰胶囊均可以降低FSH水平（P〈0.05）,升高E_2和AMH水平（P〈0.05）,增加卵巢组织Bcl-2的表达（P〈0.01）,同时抑制Bax,Caspase-3的表达（P〈0.01）。结论：养阴舒肝颗粒可以提高卵巢功能下降模型大鼠的卵巢功能,其机制可能与抗细胞凋亡有关。Objective： To investigate the effects of Yangyin Shugan granule（ YYSG） on ovarian function in rats with diminished ovarian function,and explore its effect on B-cell lymphoma-2（ Bcl-2）,Bcl-2 associated X protein（ Bax）,and cysteine-aspartic acid protease-3（ Caspase-3） protein expression. Method： SD female rats with estrous cycle rules were selected,10 used as blank group,and the others were used to establish diminished ovarian function model by injecting 160 mg·kg^（-1）4-vinylcyclohexenediepoxide（ VCD） into enterocoelia. 56 model rats with diminished ovarian function were randomly divided into 5 groups： model group,low-,middle-,highdosage YYSG groups,and Kuntai capsule group,n = 11 on each group（ n = 12 in model group）. The YYSG groups（ 2. 1,4. 2,8. 4 g·kg^（-1）） and Kuntai capsule group had intragastric administration for 4 weeks after the model establishment. Then the ovarian weight was measured and follicle count was calculated. The levels of serum sex hormones [follicule-stimulating hormone（ FSH）,estradiol（ E_2）,and anti-mullerian hormone（ AMH） ]were detected by enzyme-linked immunosorbent assay（ ELISA）. The expression levels of Bcl-2,Bax,Caspase-3 in ovarian tissue were detected by Western blot. Result： As compared with the normal group,the ovary weight and ovary index were decreased（ P〈0. 01）,Atresia follicle count was decreased（ P〈0. 01）,FSH level was increased（ P〈0. 01）,E_2 and AMH levels were decreased（ P〈0. 05）,Bcl-2 protein expressions were decreased（ P〈0. 01）,and Bax and Caspase-3 protein expressions were increased（ P〈0. 01） in model group. As compared with the model group,the ovary weights of YYSG groups were increased（ P〈0. 05,P〈0. 01）; YYSG granule and Kuntai capsule could increase the ovarian index（ P〈0. 05,P〈0. 01）; YYSG granule could reduce the numbers of atresia follicles（ P〈0. 05）; YYSG granule and Kuntai capsule could decrease FSH level but increase E_2 and AMH levels（ P〈0.

关 键 词：养阴舒肝颗粒 卵巢功能下降 去氧乙烯基环己烯(VCD) 细胞凋亡 

分 类 号：R2-0[医药卫生—中医学] R22