不同腧穴配伍电针对肥胖大鼠脂代谢和肝脏Toll样受体4/核转录因子κB信号通路的影响

Effect of electroacupuncture stimulation of different acupoint groups on lipid metabolism and liver TLR4/NF-κB signaling in obese rats

目的:观察不同腧穴配伍电针对肥胖大鼠脂代谢和肝脏Toll样受体4(TLR4)/核转录因子κB (NF-κB)信号通路的影响,探讨不同腧穴配伍电针对肥胖的调节效应是否具有差异性。方法:Wistar大鼠随机分为正常组、模型组、下肢电针组、腹部电针组、标本配穴组,每组10只。采用高脂饮食喂养诱导肥胖大鼠模型。下肢电针组选取"足三里""丰隆",腹部电针组选取"中脘""天枢""关元",标本配穴组选取上述所有腧穴,分别给予电针治疗,10 min/次,3次/周,共治疗8周。治疗后,观察各组大鼠体质量、进食量,全自动生化分析仪检测各组大鼠血清总胆固醇(TC)、甘油三酯(TG)、游离脂肪酸(NEFA)含量,Western blot法检测大鼠肝脏组织TLR4、NF-κB p65蛋白表达,实时荧光定量PCR法检测大鼠肝脏组织TLR4、NF-κB p65 mRNA表达。结果:与正常组比较,模型组大鼠体质量、进食量、血清脂代谢指标(TC、TG、NEFA)及肝脏TLR4、NF-κB p65蛋白和mRNA表达均升高(P<0.05,P<0.01)。治疗后,与模型组比较,3个干预组体质量、进食量、血清脂代谢指标(TC、TG、NEFA)及肝脏TLR4、NF-κB p65蛋白和mRNA表达降低(P<0.05,P<0.01)。与下肢电针组比较,腹部电针组血清TC、TG、NEFA升高(P<0.01,P<0.05);肝脏TLR4 mRNA表达降低(P<0.05);标本配穴组肝脏TLR4、NF-κB p65蛋白和mRNA表达降低(P<0.01,P<0.05)。与腹部电针组比较,标本配穴组血清TC、TG、NEFA及肝脏TLR4、NF-κB p65蛋白表达和NF-κB p65 mRNA表达降低(P<0.01,P<0.05)。结论:标本配穴电针和下肢电针对血脂的调节作用优于腹部电针,这可能与"足三里""丰隆"穴对脂代谢相关基因的调控有关;不同腧穴配伍电针对肝脏TLR4、NF-κB p65调节作用的差异可能与对炎性相关信号因子的调控有关。

Objective To observe the effect of electroacupuncture(EA) of different acupoint combinations on lipid metabolism and liver Toll-like receptor 4(TLR4)/nuclear factor kappa B(NF-κB) signaling in obese rats, so as to explore the specificity of regulatory effects of different acupoints for obesity.Methods Male SD rats were randomly divided into normal control, model, hindlimb acupoint EA ["Fenglong"(ST40)-"Zusanli"(ST36), hindlimb EA], abdominal acupoint EA ["Zhongwan"(CV12)-"Tianshu"(ST25), "Guanyuan"(CV4)-the opposite ST25, abdominal EA], BiaoBen-acupoint EA [ST40-ST36, CV12-CV4, ST25, BiaoBen EA] groups, with 10 rats in each group. The obesity model was induced by feeding the rats with high fat diet. EA(2 Hz, 1 mA) was applied to the acupoint groups mentioned above for 10 min(per acupoint group), 3 times a week for 8 weeks. After the treatment, the body weight and food intake in each group were recorded. The contents of serum total cholesterol(TC), triglyceride(TG), non-esterified fatty acid(NEFA) were detected by using an automatic biochemical analyzer. The expression levels of TLR4 and NF-κB p65 mRNAs and proteins in the liver tissue were detected by quantitative real-time PCR and Western blot, separately. Results Compared with the normal group, the body weight and food intake, serum TC, TG and NEFA contents, hepatic TLR4 and NF-κB p65 protein and gene expression were significantly increased in the model group(P<0.05, P<0.01). After the treatment, compared with the model group, the body weight, food intake, TC, TG, NEFA, TLR4 and NF-κB p65 protein and mRNA expression were significantly decreased in the hindlimb EA, abdominal EA and BiaoBen EA groups(P<0.05, P<0.01). Comparison among the 3 treatment groups showed that the serum TC, TG and NEFA contents were significantly lower in the hindlimb EA and BiaoBen EA groups than in the abdominal EA group(P<0.01,P<0.05). The expression levels of TLR4 and NF-κB p65 mRNAs and proteins were significantly lower in the BiaoBen EA group than in the hindlimb EA and abdominal EA groups(P<0.01,P<0.05).Conclusion BiaoBen EA is superior to abdominal EA in improving lipid metabolism in obesity rats, which may be related to its stronger effectiveness in down-regulating hepatic TLR4/NF-κB p65 signaling.