Wang Weiming, Wang Yang, Wu Jiani, Yang Likun, Liu Zhishun
Departmentof Acupuncture and Moxibustion, Guang'anmen Hospital of China Academy of Chinese Medical Sciences, Beijing 100053, China.
Zhongguo Zhen Jiu. 2018 Apr 12;38(4):405-11. doi: 10.13703/j.0255-2930.2018.04.016.
To explore the protective effect and possible mechanism of electroacupuncture (EA) pretreatment at "Zhongliao" (BL 33) and "Tianshu" (ST 25) on ovarian in rats with premature ovarian insufficiency (POI).
A total of 32 SD female rats with regular estrus were randomly divided into a blank group, a model group, an EA group and an immobilization group, 8 rats in each one. The rats in the model group, EA group and immobilization group were treated with intraperitoneal injection of 4-vinylcyclohexene diepoxide (160 mg/kg) for 15 days to establish the model. From the beginning of model establishment, the rats in the EA group were treated with EA pretreatment at "Zhongliao" (BL 33) and "Tianshu" (ST 25), once a day for the first two weeks (5 treatments per week) and once every two days for the last two weeks (3 treatments per week). EA was given 20 min per treatment for four weeks. The rats in the immobilization group were treated with immobilization with identical duration, frequency and total amount as the EA group. No treatment was given in the blank group and model group. When model was successfully established in the model group, rats were sacrificed to test the weight of sexual gland and the sexual gland index; the ELISA method was applied to test the level of anti-müllerian hormone (AMH) and inhibin B; the radioimmunoassay method was applied to test the level of follicle-stimulating hormone (FSH), luteinizing hormone (LH), estradiol (E), progesterone (P) and testosterone (T). In addition, the morphology and the number of ovarian follicles were observed; and western blot method was applied to test the expression of B-cell lymphoma-2 (Bcl-2) and Bcl-2-associated X protein (Bax) in ovarian tissue.
Compared with the blank group, the bilateral ovary weight, ovary index, uterus weight and uterus index decreased (<0.05, <0.01); serum AMH, inhibin B, E and P reduced, while serum FSH, LH and T increased in the model group (all <0.01); primordial follicles and primary follicles decreased, while atretic follicles increased in the model group (all <0.01); the expression of Bax protein increased, while the expression of Bcl-2 protein decreased in the model group (both <0.01). Compared with the model group, the bilateral ovary weight, ovary index and uterus weight increased (<0.05, <0.01); serum AMH, inhibin B, E and P increased, while serum FSH and LH reduced (<0.05, <0.01); primordial follicles and primary follicles increased, while atretic follicles reduced (<0.05, <0.01); the expression of Bax protein reduced, while the expression of Bcl-2 protein increased in the EA group (<0.05, <0.01). The differences of each outcome indexes were not significant between the model group and immobilization group (all >0.05).
EA pretreatment at "Zhongliao" (BL 33) and Tianshu (ST 25) for 4 weeks has certain protective effect on ovarian in POI rats, which is likely to be related with up-regulating ovarian granular cells, inhabiting the expression of Bcl-2 protein and down-regulating the expression of Bax protein.
探讨电针“中髎”（BL 33）和“天枢”（ST 25）预处理对卵巢早衰（POI）大鼠卵巢的保护作用及可能机制。
将32只处于正常发情期的SD雌性大鼠随机分为空白组、模型组、电针组和束缚组，每组8只。模型组、电针组和束缚组大鼠腹腔注射4-乙烯基环己烯二环氧化物（160 mg/kg），连续15天以建立模型。从造模开始，电针组大鼠接受“中髎”（BL 33）和“天枢”（ST 25）电针预处理，前两周每天1次（每周5次），后两周每两天1次（每周3次）。每次电针20分钟，共4周。束缚组大鼠接受与电针组相同持续时间、频率和总量的束缚处理。空白组和模型组不做处理。模型组大鼠造模成功后处死，检测性腺重量和性腺指数；采用酶联免疫吸附测定法（ELISA）检测抗苗勒管激素（AMH）和抑制素B水平；采用放射免疫分析法检测促卵泡生成素（FSH）、促黄体生成素（LH）、雌二醇（E）、孕酮（P）和睾酮（T）水平。此外，观察卵巢卵泡形态及数量；采用蛋白质免疫印迹法检测卵巢组织中B细胞淋巴瘤-2（Bcl-2）和Bcl-2相关X蛋白（Bax）的表达。
与空白组相比，模型组大鼠双侧卵巢重量、卵巢指数、子宫重量和子宫指数降低（<0.05，<0.01）；血清AMH、抑制素B、E和P降低，而血清FSH、LH和T升高（均<0.01）；原始卵泡和初级卵泡减少，闭锁卵泡增加（均<0.01）；模型组Bax蛋白表达增加，Bcl-2蛋白表达降低（均<0.01）。与模型组相比, 电针组大鼠双侧卵巢重量、卵巢指数和子宫重量增加（<0.05，<0.01）；血清AMH、抑制素B、E和P增加，而血清FSH和LH降低（<0.05，<0.01）；原始卵泡和初级卵泡增加，闭锁卵泡减少（<0.05，<0.01）；电针组Bax蛋白表达降低，Bcl-2蛋白表达增加（<0.05，<0.01）。模型组和束缚组各观察指标差异均无统计学意义（均>0.05）。
电针“中髎”（BL 33）和“天枢”（ST 25）预处理4周对POI大鼠卵巢具有一定保护作用，其机制可能与上调卵巢颗粒细胞、抑制Bax蛋白表达及下调Bcl-2蛋白表达有关。
