【摘要】 目的:观察针刺通过Nrf2-Keap1-ARE信号通路对IBS-D大鼠的作用机制。方法:50只Wistar大鼠随机分为5组,每组10只。大鼠实施强行应激法（CAS）刺激获得IBS-D模型;大鼠CAS刺激前1 h灌胃番木液（6 g/kg）,其后实施CAS刺激获得药物致泻+CAS刺激模型;大鼠CAS刺激后,取ST25（天枢）、ST36（足三里）和LR3（太冲）针刺,获得CAS+针刺组模型;体外构建siRNA,进行鼠尾静脉注射后进行CAS刺激和针刺治疗得到CAS刺激+针刺+Nrf2 siRNA(Nrf2^sh)模型;正常组正常饲养。检测各组5-HT、NPY、MDA、GSH/GSSG、SOD和ROS水平以及Nrf2信号通路中基因表达水平和蛋白表达水平。结果:IBS模型组、药物致泻模型+CAS刺激组和CAS+针刺+Nrf2^sh组的ROS和MDA水平高于CAS+针刺组（P<0.01）和正常组（P<0.01）,GSH水平均低于CAS+针刺组（P<0.01）和正常对照组（P<0.01）,而GSSG水平呈相反趋势（P<0.01）;正常组和CAS+针刺组的SOD水平均与其他组有差异（P<0.01）,5-HT和NPY的水平以及JNK的基因和蛋白质表达均低于其他组（P<0.01）,Nrf2、Keap-1和HO-1的基因和蛋白质表达均与其他组有差异（P<0.01）。结论:Nrf2在IBS-D治疗中具有重要作用,针刺可上调Nrf2的表达,从而抑制肠道氧化性损伤和脂质过氧化,并能在针刺后上调HO-1的表达,为针刺疗法治疗IBS-D的分子机制做出了一定的理论基础。更多还原

【Abstract】 Objective: To observe the mechanism of Nrf2-Keap1-ARE signaling pathway in rats with diarrhea-predominant IBS（ IBS-D） treated by acupuncture.Methods: The IBS-D model was established by chronic and acute stress（ CAS）.50 Wistar rats were randomly divided into five groups,i.e.group A（ normal rats）,group B（ CAS rats）,group C（ drug-induced diarrhea + CAS rats）,group D（ CAS + acupuncture rats）,and group E( CAS + acupuncture + Nrf2^sh rats),with 10 rats in each group.The levels of 5-HT,NPY,MDA,GSH/GSSG,SOD and ROS,as well as gene and protein expression of Nrf2 signaling pathway were detected in the groups.Results: The levels of ROS and MDA were significantly higher in group B,group C,and group E,compared to those in group A and group D（ P < 0.01）.The level of GSH was significantly lower in group B,group C,and group E,compared to those in group A and group D（ P < 0.01）,with the opposite trend of GSSG（ P < 0.01）.The levels of 5-HT and NPY,as well as the gene and protein expressions of JNK were significantly lower,however the level of SOD was significantly higher in group A and group D compared to those in other groups（ P < 0.01）.The gene and protein expressions of Nrf2,Keap-1 and HO-1 were greatly higher in group A and group D（ P < 0.01）.Conclusion: Nrf2 plays a key role in the treatment of IBS-D.Acupuncture can up-regulate Nrf2 expression and restrain intensional oxidative damage and lipid peroxidation.It can also enhance the expression of HO-1 and inhibit defecation frequency.This study provided certain theoretical basis for acupuncture treating IBS-D in terms of molecular mechanism.更多还原