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基于磷脂酰肌醇3激酶信号通路探究针刺对2型糖尿病大鼠胰岛素抵抗的影响

陈宏1 张智龙1 王栩1 杨元庆1

(1.天津市中医药研究院附属医院, 天津 300120)

【摘要】目的:通过观察磷脂酰肌醇3激酶 (PI 3K) 信号转导通路上、下游关键蛋白的表达变化以及针刺干预效应, 探讨“调理脾胃针法”改善2型糖尿病胰岛素抵抗的分子机制。方法:Wistar大鼠采用随机数字表法分为空白组10只, 造模组25只, 造模组大鼠以高脂高糖饲料喂养2个月后, 采用小剂量链脲佐菌素腹腔注射法建立2型糖尿病大鼠模型。造模成功的20只大鼠按照血糖值随机分为模型组及针刺组各10只。针刺组大鼠采用“调理脾胃针法”针刺双侧“曲池”“合谷”“血海”“足三里”等穴, 每次30min, 每日1次, 每周5次, 共针刺4周。测量各组大鼠空腹血糖及空腹血清胰岛素值, 计算胰岛素敏感指数 (ISI) 。采用Western blot及qPCR法检验股四头肌胰岛素受体底物-1 (IRS-1) 、IRS-2、葡萄糖转运蛋白-4 (GLUT-4) 的蛋白及基因表达。结果:干预前, 与空白组比较, 模型组和针刺组ISI明显降低 (P<0.01) ;治疗后, 与空白组比较, 模型组ISI仍然明显降低 (P<0.01) , 针刺组较模型组ISI明显升高 (P<0.01) ;针刺组治疗后ISI较干预前明显升高 (P<0.01) 。模型组与空白组比较, IRS-1、IRS-2、GLUT-4蛋白相对表达量明显下降 (P<0.01) , 针刺组IRS-1、IRS-2、GLUT-4蛋白相对表达量明显高于模型组 (P<0.01) 。模型组与空白组比较, IRS-1、IRS-2、GLUT-4基因相对表达量明显下降 (P<0.01) , 针刺组IRS-1、IRS-2、GLUT-4基因相对表达量明显高于模型组 (P<0.01) 。结论:“调理脾胃针法”可改善2型糖尿病胰岛素抵抗, 提高肌肉组织PI 3K信号通路上IRS-1、IRS-2、GLUT-4基因及蛋白表达的水平, 其改善胰岛素抵抗与此相关。

【关键词】 2型糖尿病;手针;胰岛素抵抗;胰岛素受体底物;葡萄糖转运蛋白-4;股四头肌;

【DOI】

【基金资助】 国家自然科学基金项目 (No.81373784) ; 天津市中医药管理局中医、中西医结合科研专项课题 (No.13026) ;

Effect of “Spleen-Stomach Harmonizing” Needling on Insulin Resistance and Expression of Insulin Receptor Substrate-1, -2, and Glucose Transporter-4 in Insulin Resistance Type 2 Diabetes Rats

CHEN Hong1 ZHANG Zhi-long1 WANG Xu1 YANG Yuan-qing1

(1.The Affiliated Hospital of Tianjin Academy of Traditional Chinese Medicine, Tianjin, China 300120)

【Abstract】Objective To observe the effect of manual acupuncture stimulation of “Quchi” (LI 11), “Hegu” (LI 4), “Zusanli” (ST 36), etc. with “spleen-stomach harmonizing” technique on insulin resistance index (ISI) and the expression of insulin receptor substrate-1 (IRS-1), IRS-2 (two upstream key proteins) and glucose transporter-4 (GluT-4, downstream key protein) of the phosphatidylinositol 3-kinase (PI 3K)/Akt signaling pathway in insulin resistance type 2 diabetes (IRT 2D) rats, so as to explore its mechanism underling improvement of type 2 diabetes. Methods A total of 35 male Wistar rats were used in the present study. The IRT 2Dmodel was established by feeding the animal with high-fat and high-sugar diet for 8 weeks, and subsequent intraperitoneal injection (i.p.i.) of a low dose of streptozotocin (25 mg/kg). Using the stratified random sampling method, 20 successful IRT 2Drats were equally divided into model and acupuncture groups. Ten normal rats were used as the control group. The bilateral acupoints of LI 11, LI 4, ST 36, “Xuehai” (SP 10), “Fenglong” (ST 40), “Yinlingquan” (SP 9), “Diji” (SP 8), “Sanyinjiao” (SP 6) and “Taichong” (LR 3) were punctured with filiform needles, followed by manipulating the needles with “spleen-stomach harmonizing” technique. The treatment was given once daily for 4 weeks except the weekends. Fasting blood glucose and serum insulin levels were measured using ELISA. The quadriceps muscle of thigh of each rat was sampled to test the expression levels of IRS-1, IRS-2, and GluT-4 proteins and genes with Western blot and quantitative Real-time PCR, respectively. Results Following four-week acupuncture intervention, the modeling-induced remarkable increase of ISI, and considerable down-regulation of expression levels of IRS-1, IRS-2 and GluT-4 proteins and genes in the quadriceps femoris were significantly reversed relevant to the model group (P < 0.01), suggesting a possible activation of PI 3 K/Akt signaling after acupuncture administration. Conclusion Spleen-stomach regulation needling can improve insulin resistance and up-regulate the expression of IRS-1, IRS-2 and GluT-4 in the quadriceps femoris in type 2 diabetes rats.

【Keywords】 Type 2 diabetes; Hand acupuncture; Insulin resistance; Insulin receptor substrate; Glucose transporter type 4; Quadriceps femoris;

【DOI】

【Funds】 National Natural Science Foundation of China (No. 81373784); Special Research Projects of Traditional Chinese Medicine and Integrated Chinese and Western medicine by Tianjin Administration of Traditional Chinese Medicine (No. 13026);

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This Article

ISSN:1000-0607

CN: 11-2274/R

Vol 42, No. 03, Pages 197-201

June 2017

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Article Outline

Abstract

  • 1 Materials and methods
  • 2 Results
  • 3 Discussions
  • References