针刺疗法对腹泻型肠易激综合征大鼠Nrf2-Keap1-ARE信号通路的影响
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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。
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.
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.
引文
[1]姚欣,杨云生,赵卡冰.罗马Ⅲ标准研究肠易激综合征临床特点及亚型[J].世界华人消化杂志,2008,16(5):563-566.
[2]Chao GQ,Zhang S.Effectiveness of acupuncture to treat irritable bowel syndrome:a meta-analysis[J].World journal of gastroenterology,2014,20(7):1871-1877.
[3]Zheng L,Lai Y,Lu W,et al.Pinaverium Reduces Symptoms of Irritable Bowel Syndrome in a Multicenter,Randomized,Controlled Trial[J].Clinical gastroenterology and hepatology:the official clinical practice journal of the American Gastroenterological Association,2015,13(7):1285-1292.
[4]Cramer H,Chung VC,Lauche R,et al.Characteristics of acupuncture users among internal medicine patients in Germany[J].Complementary therapies in medicine,2015,23(3):423-429.
[5]Zou N,Lv H,Li J,et al.Changes in brain G proteins and colonic sympathetic neural signaling in chronic-acute combined stress rat model of irritable bowel syndrome(IBS)[J].Translational research:the journal of laboratory and clinical medicine,2008,152(6):283-289.
[6]Huseby NE,Ravuri C,Moens U.The proteasome inhibitor lactacystin enhances GSH synthesis capacity by increased expression of antioxidant components in an Nrf2-independent,but p38 MAPK-dependent manner in rat colorectal carcinoma cells[J].Free radical research,2016,50(1):1-13.
[7]Torres-Rosas R,Yehia G,Pena G,et al.Dopamine mediates vagal modulation of the immune system by electroacupuncture[J].Nature medicine,2014,20(3):291-295.
[8]Trinkley KE,Nahata MC.Medication management of irritable bowel syndrome[J].Digestion,2014,89:253-267.
[9]吴淑娟,潘建春.肠易激综合征发病机制研究进展[J].温州医科大学学报,2015,45(5):387-390.
[10]柯水芳,陆敏.腹泻型肠易激综合征中医药治疗研究进展[J].光明中医,2017,32(4):612-614
[11]徐桂兴,黄碧清,熊俊.针灸治疗肠易激综合征的系统评价[J].中华中医药学刊,2016,34(9):2171-2174
[12]Wang XR,Shi GX,Yang JW,et al.Acupuncture ameliorates cognitive impairment and hippocampus neuronal loss in experimental vascular dementia through Nrf2-mediated antioxidant response[J].Free radical biology&medicine,2015,89:1077-84.
[13]Abed DA,Goldstein M,Albanyan H,et al.Discovery of direct inhibitors of Keap1-Nrf2 protein-protein interaction as potential therapeutic and preventive agents[J].Acta pharmaceutica Sinica B,2015,5(4):285-299.
[14]兰雄华,王旭升.马来酸曲美布汀联合双歧三联活菌治疗肠易激综合征疗效观察[J].中国现代医生,2011,49(1):109-110.
[15]李莉,郭媛,赵晏.神经肽Y参与痛觉调制的研究进展[J].生命科学,2007,19(5):551-556.
[2]Chao GQ,Zhang S.Effectiveness of acupuncture to treat irritable bowel syndrome:a meta-analysis[J].World journal of gastroenterology,2014,20(7):1871-1877.
[3]Zheng L,Lai Y,Lu W,et al.Pinaverium Reduces Symptoms of Irritable Bowel Syndrome in a Multicenter,Randomized,Controlled Trial[J].Clinical gastroenterology and hepatology:the official clinical practice journal of the American Gastroenterological Association,2015,13(7):1285-1292.
[4]Cramer H,Chung VC,Lauche R,et al.Characteristics of acupuncture users among internal medicine patients in Germany[J].Complementary therapies in medicine,2015,23(3):423-429.
[5]Zou N,Lv H,Li J,et al.Changes in brain G proteins and colonic sympathetic neural signaling in chronic-acute combined stress rat model of irritable bowel syndrome(IBS)[J].Translational research:the journal of laboratory and clinical medicine,2008,152(6):283-289.
[6]Huseby NE,Ravuri C,Moens U.The proteasome inhibitor lactacystin enhances GSH synthesis capacity by increased expression of antioxidant components in an Nrf2-independent,but p38 MAPK-dependent manner in rat colorectal carcinoma cells[J].Free radical research,2016,50(1):1-13.
[7]Torres-Rosas R,Yehia G,Pena G,et al.Dopamine mediates vagal modulation of the immune system by electroacupuncture[J].Nature medicine,2014,20(3):291-295.
[8]Trinkley KE,Nahata MC.Medication management of irritable bowel syndrome[J].Digestion,2014,89:253-267.
[9]吴淑娟,潘建春.肠易激综合征发病机制研究进展[J].温州医科大学学报,2015,45(5):387-390.
[10]柯水芳,陆敏.腹泻型肠易激综合征中医药治疗研究进展[J].光明中医,2017,32(4):612-614
[11]徐桂兴,黄碧清,熊俊.针灸治疗肠易激综合征的系统评价[J].中华中医药学刊,2016,34(9):2171-2174
[12]Wang XR,Shi GX,Yang JW,et al.Acupuncture ameliorates cognitive impairment and hippocampus neuronal loss in experimental vascular dementia through Nrf2-mediated antioxidant response[J].Free radical biology&medicine,2015,89:1077-84.
[13]Abed DA,Goldstein M,Albanyan H,et al.Discovery of direct inhibitors of Keap1-Nrf2 protein-protein interaction as potential therapeutic and preventive agents[J].Acta pharmaceutica Sinica B,2015,5(4):285-299.
[14]兰雄华,王旭升.马来酸曲美布汀联合双歧三联活菌治疗肠易激综合征疗效观察[J].中国现代医生,2011,49(1):109-110.
[15]李莉,郭媛,赵晏.神经肽Y参与痛觉调制的研究进展[J].生命科学,2007,19(5):551-556.