SPECIFIC INHIBITION OF CONNEXIN 43 GENE EXPRESSION AFTER ACUPOINTS AND ACUPUNCTURE TREATMENT FOR PRIMARY DYSMENORRHEA  

沉默穴位Cx43的表达后针刺镇痛的实验研究(英文)

作　　者：刘芳[1] 郑翠红[1] 王伟[2] 黄光英[1] 

机构地区：[1]华中科技大学同济医学院附属同济医院中西医结合研究所 [2]华中科技大学同济医学院附属同济医院神经内科

出　　处：《World Journal of Acupuncture-Moxibustion》2008年第2期28-37,共10页世界针灸杂志（英文版）

基　　金：National Basic Research Program of China(973 program, No:2006CB504502);Chinese National Foundation of Natural Sciences .(No.90209009)

摘　　要：Objective To investigate the effect of acupuncture treatment on silencing the expression of Oonnexin 43 （Cx43）, and to study the analgesic mechanism of acupuncture treatment for primary dysmenorrhea （PD） in rats. Methods We used estrostilben to develop the model of primary dysmenorrhea in rat, and RNA interference technology to silence the expression of Cx43 in acupoints. Fifty female rats were randomly divided into five groups （n = 10 in each group） ： normal, model, acupuncture, acupuncture ＋ interference and acupuncture＋ interference control group, pSilencer-Cx43-shRNA and pSilencer-Oon-shRNA were injected locally into the acupoints in interference group and interference-control group, respectively. The incidence rate of writhe reaction over the period of 30 min was evaluated. The expression of the oxytocin receptor （OTR） and vasopressin receptor（VasR） in rat myometrium with Semiquantitative RT-POR and immunohistochemistry. Results （1）The mRNA and protein level of Cx43 in acupoints in interference group were significantly lower those of in the acupuncture group （P〈0.05）. There was no significant difference between acupuncture and interference-control group. （2） Acupuncture could significantly prolong the latency period of writhing body and decrease the number of writhing body as compared with that of model group and interference group. （3）The level of OTR and VasR mRNA and protein in the model group were significantly higher （P〈0.05） as compared to normal group. The results in acupuncture group and interference-control group were similar to the normal group. The results in interference group were similar to the model group. Conclusions Acupuncture may be useful in the treatment of the model of primary dysmenorrhea in the rats. Local injection of Cx43 shRNA expression vetor could silence the expression of Cx43 in acupoint and markedly influence acupuncture effect, demonstrating Cx43 is involved in acupuncture effect.目的:研究沉默痛经大鼠穴位局部Cx43的表达对针刺效应的影响和针刺治疗原发性痛经大鼠的镇痛机制。方法:利用己烯此酚造成原发性痛经的大鼠模型,应用RNAi技术沉默穴位局部Cx43的表达。将50只雌性大鼠随机平均分为5组:正常组(对照),模型组(己烯雌酚),针刺组(己烯雌酚+针刺),针刺+干扰组(己烯雌酚+针刺+pSilencer-Cx43-shRNA)和针刺+干扰对照组(己烯雌酚+针刺+pSilencer-Con-shRNA)。末次注射催产素后,观察痛经大鼠30min的扭体数。半定量RT-PCR和免疫组织化学检测大鼠子宫内膜催产素(OTR)和加压素(VPR)mRNA和蛋白的表达。结果:①干扰组穴位处的Cx43显著低于针刺组(P<0.05),针刺组和干扰对照组之间无明显差别(P>0.05)。②与模型组、干扰组比较,针刺组能显著延长痛经大鼠的扭体潜伏期,减少扭体数。③与正常组比较,模型组OTR和VPR mRNA和蛋白水平显著增高(P<0.05)。与模型组比较,针刺组和干扰对照组的OTR和VPR mRNA和蛋白水平显著降低(P<0.05)。模型组和干扰组之间OTR和VPR mRNA和蛋白水平无显著性差异(P>0.05)。结论:局部注射Cx43特异性shRNA真核表达载体有效地沉默了穴位Cx43的表达,并显著地影响了针刺效应,表明Cx43可能与针刺疗效有密切关系。针刺通过调节子宫内膜OTR和VPR的表达来有效治疗原发性痛经。

关 键 词：Acupuncture Primary dysmenorrhea Oonnexin 43 RNAi shRNA Oxytocin receptor Vasopressin receptor 

分 类 号：R245[医药卫生—针灸推拿学]