电针“足三里”对慢性疲劳综合征大鼠骨骼肌腺苷酸活化蛋白激酶/过氧化物酶体增殖物活化受体γ共激活因子α信号通路基因表达的影响

Electroacupuncture of “Zusanli”(ST 36) Raises Muscular Force by Adjusting AMPK/PGC-1α Signaling in Rats with Chronic Fatigue Syndrome

目的:基于腺苷酸活化蛋白激酶(AMPK)/过氧化物酶体增殖物活化受体γ共激活因子α(PGC-1α)信号通路,观察针刺"足三里"对慢性疲劳综合征(CFS)大鼠骨骼肌细胞线粒体氧化应激的影响,阐释针刺"足三里"防治CFS的部分作用机制。方法:SD大鼠随机分为正常对照组、模型组、足三里组、非经非穴组,每组10只。采用多因素造模法复制CFS模型。足三里组电针双侧"足三里",非经非穴组电针双侧非经非穴,每日1次,每次20min,持续10d。检测造模前后、治疗后大鼠抓力变化;Western blot法检测大鼠骨骼肌三磷酸腺苷(ATP)合酶、AMPK、磷酸化AMPK(p-AMPK)、沉默信息调节因子2相关酶Ⅰ(SIRT 1)以及PGC-1α蛋白的表达;荧光定量PCR技术检测大鼠骨骼肌ATP合酶、SIRT 1以及PGC-1αmRNA的表达。结果:造模后,模型组、足三里组、非经非穴组大鼠抓力较正常对照组明显下降(P<0.05),治疗后足三里组大鼠抓力较模型组明显升高(P<0.05)。与正常对照组比较,模型组大鼠骨骼肌ATP合酶蛋白表达、ATP合酶mRNA表达明显下调(P<0.05,P<0.01),p-AMPK/AMPK差异无统计学意义(P>0.05),SIRT 1蛋白表达明显升高(P<0.01),PGC-1α蛋白及mRNA的表达明显降低(P<0.01)。与模型组相比,足三里组大鼠骨骼肌ATP合酶mRNA表达明显上调(P<0.01),p-AMPK/AMPK上调(P<0.05),SIRT 1蛋白及mRNA、PGC-1α蛋白及mRNA的表达明显上调(P<0.01)。结论:针刺"足三里"可使CFS大鼠骨骼肌AMPK表达明显增加,提高SIRT 1的表达,进一步直接或间接激活PGC-1α,提高线粒体的ATP合成,改善机体线粒体氧化应激反应,维持机体能量代谢,从而起到对CFS的治疗作用。

Objective To observe the effect of electroacupuncture （EA） of ＂Zusanli＂ （ST 36） on mitochondrial oxidative stress of skeletal muscle in rats with chronic fatigue syndrome （CFS） based on adenosine 5＇-monophosphate （AMP）-activated protein kinase （AMPK）/ peroxlsome preliferator-activated receptor-y coactivator-1 α （PGC-1 a） signaling, in order to reveal its mechanism underlying improvement of CFS. Methods Forty SD rats were randomly divided into normal control, CFS model, EA-Zusanli （ST 36） and EA-non-acupoint groups （n = 10 rats in each group）. The CFS model was established by forced exhausted load-bearing swimming （twice daily）, chronic constraint （1 h） and sleep deprivation （20 h/day） for 14 days. Following modeling, EA （2 Hz/100 Hz, 2 V） was applied to bilateral Zusanli （ST 36） or non-acupoint （about 10-- 15 mm superior to the bilateral Iliac creast and about 20 mm lateral to the posterior median line） for 20 min,once a day for 10 days. The expression levels of ATP syn- thase, AMPK, phosphorylated （p）-AMPK, silent mating type information regulation 2 homolog-1 （SIRT 1） and PGC-1 α proteins,and ATP synthase, SIRT 1 and PGO-1 α mRNAs of the quadriceps femoris muscle were detected by Western blot and fluorescence quantitative POR, respectively. The rats＇ grabbing force was detected by using a grabbing-force detector. Results Compared with the normal group,the grabbing force, and the expression levels of ATP synthase and PGO-1 α proteins and mRNAs were sig- nificantly decreased （ P〈0.05, P〈0.01 ）, while the expression of SlRT 1 protein was significantly up-regulated （P〈0.05） in the OFS model group. Following EA intervention, the grabbing force and the expression levels of ATP synthase mRNA, SIRT 1 and PGC-1 α proteins and mRNAs, and p-AMPK/AMPK were significantly up-regulated in the EA-Zusanli （ST 36） group （ P〈0.05, P〈0.01 ）. Conclusion EA of ST 36 can raise the grabbing force of OFS rats, which may be related to its effects in up-regulating the expression of ATP synthase mRNA, SIRT 1 and PGG-1 α proteins and mRNAs, and p-AMPK/AMPK to reduce mitochondrial oxidative stress reaction and in increasing ATP synthesis.