背俞指针疗法对GERD大鼠食管下端组织的ICC内质网AMPK/mTOR自噬通路的影响

目的研究背俞指针疗法对胃食管反流病(gastroesophageal reflux disease,GERD)大鼠食管下端组织的ICC内质网中腺苷酸活化蛋白激酶(AMPK)和哺乳动物雷帕霉素靶蛋白(mTOR)mRNA及蛋白表达的影响,探讨背俞指针疗法的效应机制。方法将76只大鼠随机分为空白组、空白指针组、模型组、指针组。采用贲门钢圈固定法制作GERD动物模型,指针组予背俞指针疗法干预,疗程均为2周。干预结束后,切取大鼠食道下段组织制作标本。采用Western Blot检测食管下端组织ICC内质网中的AMPK和mTOR蛋白表达情况;采用逆转录聚合酶链反应(RT-PCR)检测食管下端组织ICC内质网的AMPK mRNA、mTOR mRNA水平。结果Western Blot检测结果表明:模型组AMPK蛋白表达低于空白组,指针组AMPK蛋白表达显著高于模型组,差异具有统计学意义(P<0.05);空白指针组AMPK蛋白表达与模型组比较,差异无统计学意义(P>0.05);mTOR蛋白表达情况组间比较差异均无统计学意义(P>0.05)。RT-PCR检测结果表明,mTOR mRNA及AMPK mRNA水平组间比较均无统计学差异(P>0.05)。结论GERD的发病机制可能与食管下段组织ICC AMPK蛋白水平下调有关,与AMPK/mTOR自噬信号通路可能不相关。背俞指针疗法治疗GERD的起效机制可能通过提高ICC细胞AMPK蛋白表达而实现。

Effect of Finger Needle on Back-Shu Points on AMPK/mTOR Signaling Pathway of Ulrtasture of Interstitial Cell of Cajal Gastroesophageal Reflux Disease Rats

Objective To investigate the effects of finger needle on back-Shu points on protein expression of adenylate activated protein kinase(AMPK)and mammalian target protein of rapamycin(mTOR)in the ICC endoplasmic reticulum in rats with gastroesophageal reflux disease(GERD),exploring the effect mechanism of back-Shu points finger needle therapy.Method A total of 76 SD rats were randomly divided into blank group,blank-with finger needle on back-Shu points group,model group and finer needle treatment group.The rat model of GERD was established with cardia fixed with steel ring method.The blank-with finger needle on back-Shu points group were given finger needle on back-Shu points.The treatment course was 2 weeks.After the intervention,the tissue of the lower esophagus of the rats was cut for specimen.Expressions of AMPK and mTOR proteins in the ICC endoplasmic reticulum in the lower esophagus were detected by Western Blot.AMPKmRNA and mTOR mRNA levels in the ICC endoplasmic reticulum of the lower esophagus were detected by RT-PCR.Results Western Blot results showed that the protein expression of AMPK was higher in the finger needle treatment group and the blank group than that in the model group(P<0.05).There was no difference in AMPK protein expression between the blank pointer group and the model group(P>0.05).There was no significant difference in mTOR protein expression among groups(P>0.05).RT-PCR results showed that there was no significant difference in levels of mTORmRNA or AMPKmRNA among groups(P>0.05).Conclusion The pathogenesis of GERD may be related to the down-regulation of ICC AMPK protein level in lower esophageal tissues,and may not be related to AMPK/mTOR autophagy signaling pathway.The mechanism of efficacy of finger needle on back-Shu points therapy in GERD treatment may be achieved by increasing AMPK protein expression in ICC cells.