电针不同穴位对多囊卵巢综合征大鼠高雄激素血症及卵巢雄激素受体表达的影响
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摘要
目的:观察电针不同穴位对多囊卵巢综合征(PCOS)大鼠高雄激素血症和卵巢雄激素受体(AR)表达影响的作用差异,探讨电针治疗PCOS的可能机制。方法:SD大鼠随机分为对照组、模型组、足三里组、关元组、三阴交组、综合组,每组10只。采用来曲唑灌胃诱导PCOS模型。足三里组、关元组、三阴交组电针干预相应穴位,综合组同时电针所有穴位,每次20min,每日1次,连续14d。治疗后测量大鼠体质量、卵巢质量,HE染色观察卵巢形态结构,酶联免疫吸附法检测血清性激素水平和性激素结合球蛋白(SHBG),计算游离雄激素指数(FAI),免疫组化法检测卵巢卵泡AR的表达。结果:与对照组相比,模型组大鼠体质量和卵巢质量明显增加(P<0.01),血清睾酮(T)和FAI水平明显升高(P<0.01),雌二醇(E2)和SHBG水平明显降低(P<0.01),卵巢晚期卵泡AR蛋白表达增加(P<0.05)。各电针组与模型组比较,体质量均明显减轻(P<0.05),卵巢形态改善,T和FAI显著降低(P<0.01),足三里组、关元组和综合组E2、SHBG明显升高(P<0.01,P<0.05),关元组卵巢质量明显降低(P<0.01)。关元组和三阴交组晚期卵泡AR蛋白表达较模型组降低(P<0.05,P<0.01)。结论:电针不同穴位对PCOS大鼠高雄激素血症和卵巢多囊形态均有改善作用,不同穴位对PCOS大鼠的干预特点不同,临床可根据PCOS患者不同的临床表现选穴。
Objective To observe the effect of electroacupuncture(EA)at different acupoints on hyperandrogenism and ovarian androgen receptor(AR)expression in rats with polycystic ovary syndrome(PCOS),so as to explore its mechanisms underlying improvement of PCOS.Methods Sixty female SD rats were randomly divided into control,model,Zusanli(ST 36),Sanyinjiao(SP 6),Guanyuan(CV 4)and comprehensive groups(10 rats/group).The PCOS model was established by giving(gavage)the animals with Letrozole solution(1.0 mg/kg),once daily for 21 consecutive days.EA stimulation was applied to ST 36,SP 6,or CV 4 respectively,once daily for 14 days.The comprehensive group was treated by EA treatment of the abovementioned three acupoints in the meantime.Before and after the treatment,the body mass and the ovary weight were measured.The morphological structure of the ovary was observed under microscope after H.E.staining.The levels of serum hormone and sex hormone binding globulin(SHBG)were assayed by ELISA.Free androgen index(FAI)was calculated and the expression of AR in the ovary was detected by immunohistochemistry.Results After modeling,the body mass,ovary weight,serum testosterone(T)and FAI levels and the expression of AR protein in late ovarian follicles were significantly increased(P<0.01,P<0.05),and serum estradiol(E2)and SHBG contents were significantly decreased relevant to the control group(P<0.01).Following EA intervention and compared with the model group,the body weight of each EA group was significantly reduced(P<0.05),the ovarian pathological changes were improved,and serum T and FAI levels were significantly decreased(P<0.01),and serum E2 and SHBG contents significantly increased in the ST 36,CV 4 and comprehensive groups(P<0.01,P<0.05).The ovary weight of only CV 4 group was significantly lower than that of the model group(P<0.01).The expression of AR protein in late follicles of both CV 4 and SP 6 groups was markedly decreased(P<0.05,P<0.01).Conclusion EA at ST 36,SP 6,and CV 4 all can improve the hyperandrogenism and ovarian polycystic morphological changes of PCOS rats,while different acupoints have different functional characteristics.
Objective To observe the effect of electroacupuncture(EA)at different acupoints on hyperandrogenism and ovarian androgen receptor(AR)expression in rats with polycystic ovary syndrome(PCOS),so as to explore its mechanisms underlying improvement of PCOS.Methods Sixty female SD rats were randomly divided into control,model,Zusanli(ST 36),Sanyinjiao(SP 6),Guanyuan(CV 4)and comprehensive groups(10 rats/group).The PCOS model was established by giving(gavage)the animals with Letrozole solution(1.0 mg/kg),once daily for 21 consecutive days.EA stimulation was applied to ST 36,SP 6,or CV 4 respectively,once daily for 14 days.The comprehensive group was treated by EA treatment of the abovementioned three acupoints in the meantime.Before and after the treatment,the body mass and the ovary weight were measured.The morphological structure of the ovary was observed under microscope after H.E.staining.The levels of serum hormone and sex hormone binding globulin(SHBG)were assayed by ELISA.Free androgen index(FAI)was calculated and the expression of AR in the ovary was detected by immunohistochemistry.Results After modeling,the body mass,ovary weight,serum testosterone(T)and FAI levels and the expression of AR protein in late ovarian follicles were significantly increased(P<0.01,P<0.05),and serum estradiol(E2)and SHBG contents were significantly decreased relevant to the control group(P<0.01).Following EA intervention and compared with the model group,the body weight of each EA group was significantly reduced(P<0.05),the ovarian pathological changes were improved,and serum T and FAI levels were significantly decreased(P<0.01),and serum E2 and SHBG contents significantly increased in the ST 36,CV 4 and comprehensive groups(P<0.01,P<0.05).The ovary weight of only CV 4 group was significantly lower than that of the model group(P<0.01).The expression of AR protein in late follicles of both CV 4 and SP 6 groups was markedly decreased(P<0.05,P<0.01).Conclusion EA at ST 36,SP 6,and CV 4 all can improve the hyperandrogenism and ovarian polycystic morphological changes of PCOS rats,while different acupoints have different functional characteristics.
引文
[1]任丽娜,郭丽红,马文珠,等.针刺治疗多囊卵巢综合征:随机对照试验的系统综述和Meta分析[J].针刺研究,2014,39(3):238-246.
[2]杨宝芝,崔薇,李静.电针对多囊卵巢综合征患者卵子质量及妊娠结局的影响[J].针刺研究,2015,40(2):151-156.
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[4]兰蕾,刘迈兰,曾芳,等.针灸治疗不孕症的古代文献分析[J].辽宁中医杂志,2009,36(11):1968-1969.
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[7]黄冠青.游离睾酮与游离雄激素指数在PCOS临床诊断中的应用[D].衡阳:南华大学,2015.
[8]赖毛华,马红霞,姚红,等.腹针对肥胖型多囊卵巢综合征患者内分泌及糖脂代谢的影响[J].针刺研究,2010,35(4):298-302.
[9]王昕,赵明亮,吉长福,等.针刺“足三里”穴对脾虚证大鼠血清中睾酮和雌二醇水平的影响[J].针刺研究,2011,36(4):268-271.
[10]彭艳,李琴华,吴效科,等.电针对多囊卵巢综合征模型系统的调控[J].科技导报,2008,26(12):34-42.
[11]郑艳华,丁涛,叶丹凤,等.低频电针对多囊卵巢综合征大鼠糖代谢及氧化应激的影响[J].针刺研究,2015,40(2):125-130.
[12]WEIL S J,VENDOLA K,ZHOU J,et al.Androgen receptor gene expression in the primate ovary:cellular localization,regulation and functional correlations[J].J Clin Endocrinol Metab,1998,83(7):2479-2485.
[13]HAMPTON J H,MANIKKAM M,LUBAHN D B,et al.Androgen receptor mRNA expression in the bovine ovary[J].Domest Anim Endocrinol,2004,27(1):81-88.
[14]潘光鑫,石良艳,罗爱月.雄激素在卵巢中的研究进展[J].中国妇幼保健,2015,30(33):5931-5933.
[15]朱桂金,徐蓓,聂睿.重新认识雄激素在卵泡发育中的作用[C].中华医学会第六次全国生殖医学学术会议专刊,2012.
[16]PRIZANT H,GLEICHER N,SEN A.Androgen actions in the ovary:balance is key[J].J Endocrinol,2014,222(3):R 141-R 151.
[17]WALTERS K A,ALLAN C M,HANDELSMAN D J.Androgen actions and the ovary[J].Biol Reprod,2012,78(3):380-389.
[18]WALTERS K A.Role of androgens in normal and pathological ovarian function[J].Reproduction,2015,149(4):R 193-R 218.
[19]赵玲.雄激素受体在多囊卵巢综合征卵巢表达的研究[D].合肥:安徽医科大学,2011.
[20]王晶晶.益肾化痰活血方联合达英35对PCOS大鼠T、LH及其受体表达的影响[D].郑州:河南中医学院,2015.
[2]杨宝芝,崔薇,李静.电针对多囊卵巢综合征患者卵子质量及妊娠结局的影响[J].针刺研究,2015,40(2):151-156.
[3]孙洁,赵继梦,季蓉,等.电针对多囊卵巢综合征大鼠卵巢高雄激素症的影响[J].针刺研究,2013,38(6):465-472.
[4]兰蕾,刘迈兰,曾芳,等.针灸治疗不孕症的古代文献分析[J].辽宁中医杂志,2009,36(11):1968-1969.
[5]KAFALI H,IRIADAM M,OZARDALI I,et al.Letrozoleinduced polycystic ovaries in the rat:a new model for cystic ovarian disease[J].Arch Med Res,2004,35(2):103-108.
[6]余曙光,徐斌.实验针灸学[M].北京:人民卫生出版社,2012:269-272.
[7]黄冠青.游离睾酮与游离雄激素指数在PCOS临床诊断中的应用[D].衡阳:南华大学,2015.
[8]赖毛华,马红霞,姚红,等.腹针对肥胖型多囊卵巢综合征患者内分泌及糖脂代谢的影响[J].针刺研究,2010,35(4):298-302.
[9]王昕,赵明亮,吉长福,等.针刺“足三里”穴对脾虚证大鼠血清中睾酮和雌二醇水平的影响[J].针刺研究,2011,36(4):268-271.
[10]彭艳,李琴华,吴效科,等.电针对多囊卵巢综合征模型系统的调控[J].科技导报,2008,26(12):34-42.
[11]郑艳华,丁涛,叶丹凤,等.低频电针对多囊卵巢综合征大鼠糖代谢及氧化应激的影响[J].针刺研究,2015,40(2):125-130.
[12]WEIL S J,VENDOLA K,ZHOU J,et al.Androgen receptor gene expression in the primate ovary:cellular localization,regulation and functional correlations[J].J Clin Endocrinol Metab,1998,83(7):2479-2485.
[13]HAMPTON J H,MANIKKAM M,LUBAHN D B,et al.Androgen receptor mRNA expression in the bovine ovary[J].Domest Anim Endocrinol,2004,27(1):81-88.
[14]潘光鑫,石良艳,罗爱月.雄激素在卵巢中的研究进展[J].中国妇幼保健,2015,30(33):5931-5933.
[15]朱桂金,徐蓓,聂睿.重新认识雄激素在卵泡发育中的作用[C].中华医学会第六次全国生殖医学学术会议专刊,2012.
[16]PRIZANT H,GLEICHER N,SEN A.Androgen actions in the ovary:balance is key[J].J Endocrinol,2014,222(3):R 141-R 151.
[17]WALTERS K A,ALLAN C M,HANDELSMAN D J.Androgen actions and the ovary[J].Biol Reprod,2012,78(3):380-389.
[18]WALTERS K A.Role of androgens in normal and pathological ovarian function[J].Reproduction,2015,149(4):R 193-R 218.
[19]赵玲.雄激素受体在多囊卵巢综合征卵巢表达的研究[D].合肥:安徽医科大学,2011.
[20]王晶晶.益肾化痰活血方联合达英35对PCOS大鼠T、LH及其受体表达的影响[D].郑州:河南中医学院,2015.