针刺研究

2021, v.46(11) 907-913

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电针调控骨骼肌AdipoR1/AMPK/PPARα通路改善糖尿病肥胖大鼠糖脂代谢紊乱的机制探讨
Electroacupuncture improves glucose and lipid metabolism by regulating APN/AMPK/PPARα signaling of skeletal muscle in Zucker diabetic obese rats

段浩茹;李瑞;宋姗姗;胡素芹;庄舒婷;李秋艳;
DUAN Hao-ru;LI Rui;SONG Shan-shan;HU Su-qin;ZHUANG Shu-ting;LI Qiu-yan;School of Acupuncture-moxibustion and Tuina, Beijing University of Chinese Medicine;Department of Respirology and Allergy, the Third Affiliated Hospital of Shenzhen University;

摘要(Abstract):

目的:观察电针疗法对Zucker糖尿病肥胖(ZDF)大鼠血清脂联素(APN)及骨骼肌脂联素受体(AdipoR1)/腺苷酸活化蛋白激酶(AMPK)/过氧化物酶体增殖物激活受体α(PPARα)信号通路及骨骼肌形态的影响,探讨电针调节2型糖尿病(T2DM)糖脂代谢紊乱、改善骨骼肌胰岛素抵抗(IR)的机制。方法:将12只2月龄SPF级雄性ZDF大鼠随机分为模型组、电针组,每组6只;6只2月龄SPF级雄性Zucker瘦型大鼠为空白组。采用高脂饲料喂养4周诱导T2DM大鼠模型。电针组予双侧"脾俞""胰俞""足三里""三阴交"电针治疗,每日1次,每周6次,连续4周。检测治疗前后各组大鼠空腹血糖(FBG),治疗后放射免疫法检测各组大鼠血清胰岛素(INS)、C肽(C-P)、APN水平,酶比色法检测各组大鼠血清游离脂肪酸(FFA)、低密度脂蛋白(LDL)、胆固醇(TC)、甘油三酯(TG)含量,计算胰岛素抵抗指数(HOMA-IR),Western blot法检测各组大鼠股四头肌AMPK、PPARα、AdipoR1蛋白表达量,HE染色法观察各组大鼠股四头肌形态学变化。结果:治疗前,模型组、电针组FBG含量明显高于空白组(P<0.01)。治疗后,模型组FBG含量高于空白组(P<0.01);电针组FBG含量低于模型组(P<0.05)。与空白组比较,模型组血清INS、C-P、 HOMA-IR、FFA、TC、LDL、TG含量均明显升高(P<0.01,P<0.05),血清APN含量及骨骼肌AMPK、PPARα、AdipoR1蛋白表达量显著降低(P<0.01,P<0.05);与模型组比较,电针组血清INS、C-P、HOMA-IR、FFA、LDL、TC含量明显降低(P<0.01),血清APN含量及骨骼肌AMPK、PPARα、AdipoR1蛋白表达量显著升高(P<0.01)。HE染色示,空白组股四头肌肌细胞未见明显异常;模型组股四头肌肌细胞排列紊乱,肌纤维断裂、不清晰,肌纤维间隙变大,局部可见少量的脂肪细胞浸润;电针组肌纤维虽也有断裂现象和局部少量脂肪细胞浸润,但肌纤维排列较模型组有一定改善。结论:电针可以降低ZDF大鼠血糖、血脂,改善IR,其机制可能与电针通过APN上调骨骼肌AdipoR1/AMPK/PPARα信号通路,改善骨骼肌纤维排列有关。
Objective To observe the effect of electroacupuncture(EA) on skeletal muscle adiponectin receptor(Adipor1)/adenylate activated protein kinase(AMPK)/peroxisome proliferator-activated receptor α(PPARα) signaling pathway and skeletal muscle morphology by the secretion of serum adiponectin in Zucker diabetic obese(ZDF) rats, so as to explore its mechanism underlying regulating glucose and lipid metabolism of type 2 diabetes mellitus(T2 DM) and improving skeletal muscle insulin resistance(IR). Methods Twelve male ZDF rats and six Zucker thin(ZL) rats were selected. The rats were fed with Purina#5008 high-fat diet for four weeks to induce T2 DM model after adaptive feeding with normal diet for one week. The ZDF rats were randomly divided into model group and EA group according to blood glucose level after modeling and 6 ZL rats were used as the blank control group. Rats in the EA group were treated with “Pishu”(BL20), EA stimulation of “Yishu”(EX-B3), “Zusanli”(ST36) and “Sanyinjiao”(SP6), once a day and 6 times a week for 4 weeks. Rats of the model and blank control groups were grabbed and fixed in the same way as EA group. Fasting blood glucose(FBG) was measured before and after EA intervention. Serum levels of insulin(INS), C-peptide(C-P), adiponectin(APN) were measured by radioimmunoassay, and those of free fatty acid(FFA), low-density lipoprotein(LDL), cholesterol(TC), triglyceride(TG) content determined by enzyme colorimetry and insulin resistance index(HOMA-IR) was calculated. The protein expression levels of AdipoR1, AMPK and PPARα proteins in the quadriceps femoris tissues were detected by Western blot and histopathological changes of quadriceps femoris muscle were observed by H.E. staining. Results Compared with the blank control group, the levels of FBG, serum INS, C-P, FFA, LDL, TC, TG and HOMA-IR in the model group were significantly increased(P<0.01, P<0.05), while the levels of serum APN and the expressions of AdipoR1, AMPK and PPARα proteins in the skeletal muscle were significantly decreased(P<0.01, P<0.05). Compared with the model group, the levels of FBG, serum INS, C-P, FFA, LDL, TC and HOMA-IR in the EA group were significantly decreased(P<0.01), and those of serum APN and expression levels of AdipoR1, AMPK and PPARα proteins in the skeletal muscle significantly increased(P<0.01), but the serum TG level had no remarkable change in the EA group(P>0.05). In addition, H.E. staining showed disordered arrangement of skeletal muscle cells, rupture and fuzziness of muscle fibers, enlargement of the space between muscular fibers and infiltration of small number of adipose cells which were relatively milder in the EA group. Conclusion EA can reduce blood glucose and lipid levels, and improve IR in ZDF rats, which may be related to its functions in up-regulating AdipoR1/AMPK/PPARα signaling and in promoting APN secretion.

关键词(KeyWords): 电针;2型糖尿病;糖脂代谢紊乱;脂联素信号通路;胰岛素抵抗
Electroacupuncture;Type 2 diabetes;Disorder of glucose and lipid metabolism;Adiponectin signaling pathway;Insulin resistance

Abstract:

Keywords:

基金项目(Foundation): 国家自然科学基金面上项目(No.81973935)

作者(Author): 段浩茹;李瑞;宋姗姗;胡素芹;庄舒婷;李秋艳;
DUAN Hao-ru;LI Rui;SONG Shan-shan;HU Su-qin;ZHUANG Shu-ting;LI Qiu-yan;School of Acupuncture-moxibustion and Tuina, Beijing University of Chinese Medicine;Department of Respirology and Allergy, the Third Affiliated Hospital of Shenzhen University;

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DOI: 10.13702/j.1000-0607.20210201

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