电针“中髎”(BL 33)和“天枢”(ST 25)对卵巢早衰大鼠卵巢功能的影响
[Effects of electroacupuncture at "Zhongliao" (BL 33) and "Tianshu" (ST 25) on ovarian function in rats with premature ovarian insufficiency].
作者信息
Wang Weiming, Wang Yang, Wu Jiani, Yang Likun, Liu Zhishun
机构信息
Department of Acupuncture and Moxibustion, Guang'anmen Hospital, China Academy of Chinese Medical Sciences, Beijing 100053, China.
出版信息
Zhongguo Zhen Jiu. 2018 May 12;38(5):5193-26. doi: 10.13703/j.0255-2930.2018.05.018.
OBJECTIVE
To explore the effects of electroacupuncture (EA) at "Zhongliao" (BL 33) and "Tianshu" (ST 25) on ovarian function in rats with premature ovarian insufficiency (POI).
METHODS
A total of 48 SD female rats with regular estrus were divided into a blank group (=8), a model group (=10), an EA group (=10), a binding group (=10) and a tamoxifen (TAM) group (=10). The rats in the model group, EA group, binding group and TAM group were all treated with intraperitoneal injection of 4-vinylcyclohexene diepoxide (VCD, 160 mg/kg) for 15 consecutive days to establish the model of POI; the rats in the blank group were treated with normal diet. After the model was established successfully, the rats in the EA group were treated with EA at "Zhongliao" (BL 33) and "Tianshu" (ST 25) with continuous wave (1 to 3 Hz, 0.1 to 1 mA) for 20 minutes, once a day (five times a week) for the first two weeks and once every other day (three times a week) for the following two weeks. The rats in the TAM group were treated with subcutaneous injection of tamoxifen (1mg/kg), once a day (five times a week) for the first two weeks and once every other day (three times a week) for the following two weeks. The rats in the binding group were bound by a small sack as the EA group. The rats in the blank group and the model group were treated with normal diet. After four weeks, the sexual gland weight and index were tested in each group; the ELISA method was applied to test the level of anti-mllerian hormone (AMH) and inhibin B; the morphology of ovary was observed; the number of primordial follicles, primary follicle, antral follicle and atretic follicle was counted; the expression of insulin-like growth factor-1 (IGF-1) and insulin-like growth factor-1 receptor (IGF-1R) were measured.
RESULTS
(1) Compared with the blank group, the ovary weight, ovary index, uterus weight and uterus index were significantly decreased after treatment in the model group, EA group, binding group and TAM group (all <0.01); but the differences between the model group and the EA group, binding group, TAM group were not significant (all >0.05). (2) Compared with the blank group, the levels of serum AMH, inhibin B and E were significantly reduced; the levels of FSH and LH were significantly increased in the model group; EA group, binding group and TAM group (all <0.01). Compared with the model group, the levels of serum AMH, inhibin B and E were significantly increased, the level of FSH and LH were significantly reduced in the EA group and TAM group (all <0.01). (3) Compared with the blank group, in the model group, EA group, binding group and TAM group the ovary was dark red and pale, surrounded by particle or not; the morphology was small and atrophic; the primordial follicles was reduced even vanished; the structure of primary follicle was damaged and loosely arranged; the mature follicle was few; the atretic follicle and interstitial gland were increased. (4) Compared with the blank group, the expressions of IGF-1 mRNA and IGF-1R mRNA were increased in the model group (all <0.01); compared with the blank group, the expression of IGF-1 mRNA was increased in the binding group (<0.05), but that of IGF-1R mRNA was not significantly different (>0.05); compared with the model group, the expression of IGF-1 mRNA was not significantly different in the EA group, binding group and TAM group (all >0.05), but that of IGF-1R mRNA was reduced (<0.05, <0.01).
CONCLUSION
EA at "Zhongliao" (BL 33) and "Tianshu" (ST 25) has improvement effect on ovarian function in rats with VCD-induced POI, which is likely to be related to regulating the IGF-1R mRNA expression to improve the IGF-1/ IGF-1R axis.
目的
探讨电针“中髎”(BL 33)和“天枢”(ST 25)对卵巢早衰(POI)大鼠卵巢功能的影响。
方法
将48只处于正常发情期的SD雌性大鼠分为空白组(n = 8)、模型组(n = 10)、电针组(n = 10)、捆绑组(n = 10)和他莫昔芬(TAM)组(n = 10)。模型组、电针组、捆绑组和TAM组大鼠均连续15天腹腔注射4 - 乙烯基环己烯二环氧化物(VCD,160 mg/kg)以建立POI模型;空白组大鼠给予正常饮食。成功建立模型后,电针组大鼠采用连续波(1至3 Hz,0.1至1 mA)电针“中髎”(BL 33)和“天枢”(ST 25)20分钟,每天1次(每周5次),前两周如此,后两周隔天1次(每周3次)。TAM组大鼠皮下注射他莫昔芬(1mg/kg),每天1次(每周5次),前两周如此,后两周隔天1次(每周3次)。捆绑组大鼠像电针组一样用小布袋捆绑。空白组和模型组大鼠给予正常饮食。四周后,检测各组大鼠性腺重量和指数;采用ELISA法检测抗苗勒管激素(AMH)和抑制素B水平;观察卵巢形态;计数原始卵泡、初级卵泡、窦状卵泡和闭锁卵泡数量;检测胰岛素样生长因子-1(IGF - 1)和胰岛素样生长因子-1受体(IGF - 1R)的表达。
结果
(1)与空白组相比,模型组、电针组、捆绑组和TAM组治疗后卵巢重量、卵巢指数、子宫重量和子宫指数均显著降低(均P < 0.01);但模型组与电针组、捆绑组、TAM组之间差异无统计学意义(均P > 0.05)。(2)与空白组相比,模型组血清AMH、抑制素B和E水平显著降低;卵泡刺激素(FSH)和黄体生成素(LH)水平显著升高;电针组、捆绑组和TAM组情况相同(均P < 0.01)。与模型组相比,电针组和TAM组血清AMH、抑制素B和E水平显著升高,FSH和LH水平显著降低(均P < 0.01)。(3)与空白组相比,模型组、电针组、捆绑组和TAM组卵巢暗红色、苍白,周围有无颗粒;形态小且萎缩;原始卵泡减少甚至消失;初级卵泡结构受损且排列松散;成熟卵泡少;闭锁卵泡和间质腺增多。(4)与空白组相比,模型组IGF - 1 mRNA和IGF - 1R mRNA表达升高(均P < 0.01);与空白组相比,捆绑组IGF - 1 mRNA表达升高(P < 0.05),但IGF - 1R mRNA表达差异无统计学意义(P > 0.05);与模型组相比,电针组、捆绑组和TAM组IGF - 1 mRNA表达差异无统计学意义(均P > 0.05),但IGF - 1R mRNA表达降低(P < 0.05,P < 0.01)。
结论
电针“中髎”(BL 33)和“天枢”(ST 25)对VCD诱导的POI大鼠卵巢功能有改善作用,可能与调节IGF - 1R mRNA表达以改善IGF - 1/IGF - 1R轴有关。