电针对肥胖大鼠肠道Toll样受体4和核转录因子κB的影响

Effect of electroacupuncture on intestinal Toll-like receptor 4 and nuclear factor-kappa B in obese rats

目的:观察电针对肥胖大鼠肠道组织Toll样受体4(TLR4)和核转录因子κB(NF-κB)的影响,探讨针灸减肥的作用机制。方法:Wistar雄性大鼠随机分为正常组和模型组,高脂饲料喂养建立肥胖大鼠模型,造模成功的24只大鼠随机分为模型组、抑制剂组和电针组,每组8只。电针治疗选取"关元""中脘""足三里"和"丰隆",每次治疗10 min,抑制剂组采用TAK-242腹腔注射,每周3次,共干预8周。每2周测量1次体质量和血糖,免疫共沉淀法测定肠道组织TLR4和NF-κB p65相互作用,凝胶迁移实验法测定肠道组织NF-κB p65活性,Western blot法测定肠道组织TLR4、NF-κB p65和p-IκBα蛋白表达,实时荧光定量PCR测定肠道组织TLR4、NF-κB p65和IκBαmRNA表达。结果:与对照组比较,模型组体质量、血糖水平、TLR4、NF-κB p65、p-IκBα的蛋白和mRNA水平均显著升高(P<0.01,P<0.05),TLR4与NF-κB p65结合能力及NF-κB p65活性显著增强(P<0.05,P<0.01)。与模型组比较,电针组大鼠体质量显著降低(P<0.05),电针组和抑制剂组干预后餐后血糖水平、TLR4、NF-κB p65、p-IκBα的蛋白和mRNA水平显著降低(P<0.05,P<0.01),NF-κB p65活性显著减弱(P<0.01)。结论:电针能够有效调控肠道TLR4,抑制其与NF-κB p65相互作用,降低NF-κB p65活性,这可能是电针降低肥胖大鼠体质量和血糖水平,从而改善其肥胖状态的潜在机制之一。

Objective To investigate the effect of electroacupuncture(EA)on intestinal Toll-like receptor 4(TLR4)and nuclear factor-kappa B(NF-κB)in obese rats,so as to explore the mechanism of action of acupuncture in losing weight.Me-thods A total of 50 male Wistar rats were randomly divided into control and model groups.High-fat feed was used to establish a rat model of obesity,and after modeling,the 24 rats were randomly divided into model group,TLR4 inhibitor group,and EA group,with 8 rats in each group.The rats in the EA group were given EA at"Guanyuan"(CV4),"Zhongwan"(CV12),"Zusanli"(ST36),and"Fenglong"(ST40),10 minutes each time,3 times a week,and those in the TLR4 inhibitor group were given intraperitoneal injection of TAK-242 three times a week;the course of treatment was 8 weeks for both groups.Body weight and blood glucose were measured every two weeks.Co-immunoprecipitation was used to observe the interaction between TLR4 and NF-κB p65 in the intestinal tissue;electrophoretic mobility shift assay was used to measure the activity of NF-κB p65;Western blot was used to measure the protein expression of TLR4,phosphorylated nuclear factor of kappa light polypeptide gene enhancer in B-cells inhibitor alpha(p-IκBα),and NF-κB p65;quantitative real-time PCR was used to measure the mRNA expression of TLR4,NF-κB p65,and IκBα.Results Compared with the control group,the model group had significant increases in body weight,blood glucose,and protein and mRNA expression of TLR4 and NF-κB p65(P<0.01,P<0.05),as well as significant enhancement in the interaction between TLR4 and NF-κB p65 and activity of NF-κB p65(P<0.05,P<0.01).Compared with the model group,the EA group had a significant reduction in body weight(P<0.05),both of the EA group and the TLR4 inhibitor group had significant reductions in blood glucose,and protein and mRNA expression of TLR4,p-IκBα,and NF-κB p65(P<0.05,P<0.01),as well as significant reductions in the activity of NF-κB p65(P<0.01).Conclusion EA can effectively regulate intestinal TLR4,inhibit the interaction between TLR4 and NF-κB p65,and reduce the activity of NF-κB p65,which may be a potential mechanism of EA in reducing body weight and blood glucose in obese rats.