Chen Li, Wang Jing-Zhi, Zhou Guang-Wen, Wu Yong-Gui, Liang Feng-Xia
College of Acupuncture and Orthopedics, Hubei University of Chinese Medicine, Wuhan 430061, China.
School of Basic Medical Sciences, Hubei University of Chinese Medicine, Wuhan 430061, China.
Zhen Ci Yan Jiu. 2020 Jul 25;45(7):541-7. doi: 10.13702/j.1000-0607.190736.
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.
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.
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 (<0.01, <0.05), as well as significant enhancement in the interaction between TLR4 and NF-κB p65 and activity of NF-κB p65 (<0.05,<0.01). Compared with the model group, the EA group had a significant reduction in body weight (<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 (<0.05,<0.01), as well as significant reductions in the activity of NF-κB p65 (<0.01).
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.
探讨电针(EA)对肥胖大鼠肠道Toll样受体4(TLR4)及核因子-κB(NF-κB)的影响,以探究针刺减肥的作用机制。
将50只雄性Wistar大鼠随机分为对照组和模型组。采用高脂饲料建立大鼠肥胖模型,造模成功后,将24只大鼠随机分为模型组、TLR4抑制剂组和电针组,每组8只。电针组大鼠取“关元”(CV4)、“中脘”(CV12)、“足三里”(ST36)和“丰隆”(ST40)进行电针治疗,每次10分钟,每周3次;TLR4抑制剂组大鼠每周腹腔注射TAK-242 3次;两组治疗疗程均为8周。每两周测量大鼠体重和血糖。采用免疫共沉淀法观察肠道组织中TLR4与NF-κB p65的相互作用;采用电泳迁移率变动分析检测NF-κB p65的活性;采用蛋白质免疫印迹法检测TLR4、B细胞κ轻链基因增强子抑制因子α磷酸化(p-IκBα)及NF-κB p65的蛋白表达;采用实时荧光定量PCR检测TLR4、NF-κB p65及IκBα的mRNA表达。
与对照组比较,模型组大鼠体重、血糖及TLR4、NF-κB p65的蛋白和mRNA表达均显著升高(<0.01,<0.05),TLR4与NF-κB p65的相互作用及NF-κB p65的活性显著增强(<0.05,<0.01)。与模型组比较,电针组大鼠体重显著降低(<0.05),电针组和TLR4抑制剂组大鼠血糖及TLR4、p-IκBα、NF-κB p65的蛋白和mRNA表达均显著降低(<0.05,<0.01),NF-κB p65的活性显著降低(<0.01)。
电针可有效调节肠道TLR4,抑制TLR4与NF-κB p65的相互作用,降低NF-κB p65的活性,这可能是电针降低肥胖大鼠体重和血糖的潜在机制。