电针调节TLR4、NLRP3改善功能性消化不良大鼠胃黏膜炎症反应机制研究

Mechanism of Electroacupuncture Regulation of TLR4 and NLRP3 on Improving Gastric Mucosal Inflammation in Rats with Functional Dyspepsia

目的观察电针“足三里”“太冲”穴是否通过调节TLR4、NLRP3而改善大鼠胃黏膜炎症反应,从而起到干预功能性消化不良(FD)的作用。方法将48只10 d龄幼年SD大鼠随机分为正常组、模型组、针刺组、非穴位组,每组各12只,雌雄各半。采用碘乙酰胺联合夹鼠尾法造制备FD大鼠肝郁脾虚证模型。造模成功后,针刺组和非穴位组分别电针大鼠双侧“足三里”穴、“太冲”穴、“非经非穴”,1次/d,每次20 min,连续14 d。Elisa法检测胃组织中白介素1β(IL-1β)的含量表达;RT-PCR法检测各组大鼠胃组织中Toll样受体4(TLR4)、NOD样受体热蛋白3(NLRP3)的mRNA表达;免疫组化染色法检测各组大鼠胃组织中TLR4、NLRP3的蛋白表达。结果针刺组干预后,FD大鼠胃黏膜炎症反应得到了明显缓解;与正常组比较,其中模型组中FD大鼠胃组织中TLR4蛋白及mRNA表达、NLRP3蛋白表达及IL-1β水平均明显升高(P<0.01),NLRP3 mRNA表达水平增高(P<0.05)。与模型组比较,针刺组FD大鼠胃组织中TLR4蛋白表达、NLRP3蛋白及mRNA表达、IL-1β水平均降低(P<0.05),其中TLR4 mRNA表达显著降低(P<0.01);与针刺组比较,非穴位组大鼠胃组织中TLR4、NLRP3蛋白及mRNA表达升高,IL-1β水平升高(P<0.05)。结论电针“足三里”“太冲”穴能够下调FD大鼠胃组织中TLR4、NLRP3的表达,降低炎症因子IL-1β的表达水平,最终缓解了炎症反应,起到了电针对FD的干预作用。

Objective To observe whether electroacupuncture at Zusanli(ST36)and Taichong(LR3)points can improve gastric mucosa inflammation in rats by regulating TLR4 and NLRP3,thus playing a role in the intervention of functional dyspepsia(FD).Methods Forty-eight 10-day-old young SD rats were randomly divided into normal group,model group,acupuncture group and non-acupoint group,with 12 rats in each group and half male and half female.The model of LIVER depression and spleen deficiency in FD rats was established by iodoacetamide combined with rat tail clamping method.After modeling,electroacupuncture at Zusanli(ST36),Taichong(LR3)and non-acupoint points was performed on both sides of rats in acupuncture group and non-acupoint group,once a day,20 min each time,for consecutive 14 d.The expression of interleukin 1β(IL-1β)in gastric tissue was determined by Elisa.The mRNA expressions of Toll-like receptor 4(TLR4)and Nod-like receptor thermoprotein 3(NLRP3)in gastric tissues of rats were detected by RT-PCR.TLR4 and NLRP3 protein expression in gastric tissues of rats were detected by immunohistochemical staining.Results After acupuncture intervention,the inflammatory response in gastric mucosa of FD rats was significantly relieved.Compared with normal group,TLR4 protein and mRNA expression,NLRP3 protein expression and IL-1βlevel in gastric tissue of FD rats in model group were significantly increased(P<0.01),and NLRP3 mRNA expression level was increased(P<0.05).Compared with model group,TLR4 protein expression,NLRP3 protein and mRNA expression and IL-1βlevel in gastric tissue of FD rats in acupuncture group were decreased(P<0.05),and TLR4 mRNA expression was significantly decreased(P<0.01).Compared with acupuncture group,the expression of TLR4 and NLRP3 protein and mRNA in gastric tissue of non-acupoint group was increased,and the level of IL-1βwas increased(P<0.05).Conclusion Electroacupuncture at Zusanli(ST36)and Taichong(LR3)points could down-regulate the expression of TLR4 and NLRP3 in the gastric tissue of FD rats,reduce the level of inflammatory factors IL-1β,and finally alleviate the inflammatory response,thus playing an intervention role in FD.