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[针刺干预通过调节肥胖糖尿病大鼠的PKCβ/P66shc信号通路诱导氧化应激改善]

[Acupuncture intervention induced improvement of oxidative stress by regulating PKCβ/P66shc signaling in obese diabetic rats].

作者信息

Yin Gang, Shen Guo-Ming, Jiang Ai-Juan, Li Jing-Ya

机构信息

College of Integrated Traditional Chinese and Western Medicine, Anhui University of Chinese Medicine, Hefei 230000, China.

Graduate School of Anhui University of Chinese Medicine, Hefei 230000.

出版信息

Zhen Ci Yan Jiu. 2021 Aug 25;46(8):642-8. doi: 10.13702/j.1000-0607.201318.

Abstract

OBJECTIVE

To observe the effect of electroacupuncture (EA) at "Zusanli"(ST36)-"Sanyinjiao"(SP6) on glucose and lipid metabolism and insulin resistance (IR) in obese diabetic rats, so as to explore its mechanism underlying improvement of obesity diabetes.

METHODS

SPF male rats were randomly divided into normal control, model, meridian-acupoint EA (acupoint), non-meridian non-acupoint EA (non-acupoint), and medication (metformin) groups, with 10 rats in each group. The diabetes model was established by feeding the rats with high-fat diet for 8 weeks. EA (1.5 mA, 10 Hz/100 Hz) was applied to unilateral ST36 and SP6 for 20 min, once daily (except Sundays) for 4 weeks. Rats of the medication group were treated by gavage of metformin (300 mg/kg) once daily for 4 weeks (except Sundays). The body weight and length were measured and the Lee's index was calculated. The contents of total cholesterol (TC), triglyceride (TG), low density lipoprotein-cholesterol (LDL-C), high density lipoprotein-cholesterol (HDL-C) in the plasma were detected by using a full-automatic biochemical analyzer. The content of fasting serum insulin (FINS) was assayed by using radioimmunoassay, the fasting blood glucose (FBG) was measured, and serum superoxide dismutase (SOD) activity by using xanthine oxidase method, serum malondialdehyde (MDA) by color method, serum glutathione peroxidase (GSH-Px) activity by indirect method, reactive oxygen species (ROS) by Dithio-bis-nitrobenzoic acid (DTNB) direct method, and the homeostasis model assessment of IR (HOMA-IR) and insulin sensitive index (ISI) were calculated. The expression levels of pancreatic tissue P66shc mRNA and PKCβ mRNA were detected by using RT-PCR, and the histopathological changes of the liver and adipose tissues were observed after H.E. staining.

RESULTS

Compared with the normal control group, the Lee's index, levels of FBG, FINS, HOMA-IR, TC, TG, LDL-C, MDA, ROS, and P66shc mRNA and PKCβ mRNA expressions were significantly increased (<0.05,<0.01), and ISI, HDL-C, SOD, GSH-Px significantly decreased (<0.05, <0.01) in the model group. After the interventions, the levels of Lee's index,levels of FBG, FINS, HOMA-IR, TC, TG, LDL-C, MDA, ROS, and expressions of P66shc mRNA and PKCβ mRNA were remarkably down-regulated (<0.05, <0.01), and those of ISI, HDL-C, SOD, and GSH-Px up-regulated (<0.05, <0.01) in both EA and medication groups. H.E. staining showed many white adipocytes in the adipose tissue, radial and cord-like arrangement of liver cells, and many of them with vacuoles in the cytoplasm of small vesicular lipid droplets in the model group; and relative reduction of white adipocytes in number, smaller in cell body, and no obvious abnormal changes of structure and arrangement of liver cells in the EA and medication groups.

CONCLUSION

EA of ST36 and SP6 can improve glucose and lipid metabolism and IR in obese diabetic rats, which may be related to its function in suppressing PKCβ/P66shc signaling and oxidative stress.

摘要

目的

观察电针“足三里”(ST36)-“三阴交”(SP6)对肥胖型糖尿病大鼠糖脂代谢及胰岛素抵抗(IR)的影响,以探讨其改善肥胖型糖尿病的作用机制。

方法

将SPF级雄性大鼠随机分为正常对照组、模型组、经穴电针组(穴位组)、非经非穴电针组(非穴位组)和药物组(二甲双胍组),每组10只。采用高脂饮食喂养大鼠8周建立糖尿病模型。电针(1.5 mA,10 Hz/100 Hz)刺激单侧ST36和SP6,每次20分钟,每日1次(周日除外),共4周。药物组大鼠每日灌胃二甲双胍(300 mg/kg),共4周(周日除外)。测量大鼠体重、体长并计算李氏指数。采用全自动生化分析仪检测血浆中总胆固醇(TC)、甘油三酯(TG)、低密度脂蛋白胆固醇(LDL-C)、高密度脂蛋白胆固醇(HDL-C)含量。采用放射免疫法测定空腹血清胰岛素(FINS)含量,测量空腹血糖(FBG),采用黄嘌呤氧化酶法检测血清超氧化物歧化酶(SOD)活性,采用比色法检测血清丙二醛(MDA)含量,采用间接法检测血清谷胱甘肽过氧化物酶(GSH-Px)活性,采用二硫代双硝基苯甲酸(DTNB)直接法检测活性氧(ROS),并计算IR的稳态模型评估(HOMA-IR)和胰岛素敏感指数(ISI)。采用RT-PCR检测胰腺组织P66shc mRNA和PKCβ mRNA表达水平,苏木精-伊红(H.E.)染色后观察肝脏和脂肪组织的组织病理学变化。

结果

与正常对照组比较,模型组大鼠李氏指数、FBG、FINS、HOMA-IR、TC、TG、LDL-C、MDA、ROS水平及P66shc mRNA和PKCβ mRNA表达显著升高(<0.05,<0.01),ISI、HDL-C、SOD、GSH-Px显著降低(<0.05,<0.01)。干预后,电针组和药物组大鼠李氏指数、FBG、FINS、HOMA-IR、TC、TG、LDL-C、MDA、ROS水平及P66shc mRNA和PKCβ mRNA表达均显著下调(<0.05,<0.01),ISI、HDL-C、SOD、GSH-Px水平上调(<0.05,<0.01)。H.E.染色显示,模型组脂肪组织中有大量白色脂肪细胞,肝细胞呈放射状和条索状排列,许多肝细胞胞质内有小泡状脂滴空泡;电针组和药物组白色脂肪细胞数量相对减少,细胞体积变小,肝细胞结构和排列无明显异常改变。

结论

电针ST36和SP6可改善肥胖型糖尿病大鼠的糖脂代谢及IR,其机制可能与抑制PKCβ/P66shc信号通路及氧化应激有关。

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