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[电针干预对胰岛素抵抗模型大鼠下丘脑PI 3 K和p - PI 3 K蛋白表达的影响]

[Effects of electroacupuncture intervention on expression of hypothalamic PI 3 K and p-PI 3 K Proteins in insulin resistance model rats].

作者信息

Wang Jing-Zhi, Wang Hua, Zhou Huan-Jiao, Liu Jian-Min, Tang Hong-Tu, Yang Feng, Li Nian, Liang Feng-Xia

出版信息

Zhen Ci Yan Jiu. 2014 Feb;39(1):46-51, 57.

Abstract

OBJECTIVE

To observe the effect of "Shuanggu Yitong" (Double-reinforcing and one-unblocking) needling [electroacupuncture (EA) of "Guanyuan" (CV 4), "Housanli" (ST 36), "Fenglong" (ST 40) and "Zhongwan" (CV 12)] on expression of hypothalamic phosphatidylinositol 3-kinase (PI 3 K) and p-PI 3 K proteins in insulin resistance rats.

METHODS

Sixty Wistar rats were randomized into normal control, model, EA prevention, EA, cerebrospinal fluid (CSF), and Wortmannin (PI 3 K antagonist) groups (n = 10 rats/group). Insulin resistance model was established by feeding the animal with high fat forage continuously for 8 weeks. For rats of the EA group, EA (2 Hz, 1 mA) was applied to ipsilateral "Housanli"(ST 36)-"Fenglong" (ST 40), and "Guanyuan" (CV 4)-"Zhongwan" (CV 12) for 15 min, once daily, 5 times a week and for 8 weeks, beginning after modeling. For rats of the EA prevention group, EA was conducted simultaneously with the modeling. For rats of the Wortmannin group and CSF group, cerebroventricular microinjection of Wortmannin (50 nmol/L, 3 mg/kg) or artificial CSF was given through an implanted steel cannel. Fasting plasma glucose (FPG) and fasting serum insulin (FINS) contents were assayed using glucose-tested instrument and ELISA respectively, insulin activity index (IAI) was calculated [IAI = Ln (1/FPG x FINS)], and the expres- sion of hypothalamic PI 3 K and p-PI 3 K proteins was detected by Western blot.

RESULTS

In comparison with the normal control group, the body weight (BW), FPG and FINS contents at time-points of 8 and 16 weeks were considerably increased (P < 0.01), and IAI, hypothalamic PI 3 Kp 110 and p-PI 3 Kp 110 protein expression levels were obviously decreased in the model group (P < 0.01). Compared with the model group, the FPG and FINS contents at the 8th week after modeling in the EA prevention group, the BW and FPG levels at the 16th week in the EA prevention, EA and CSF groups, FINS levels at the 16th week in the EA prevention and EA groups were remarkably decreased (P<0.05, P<0.01), IAI, and hypothalamic PI 3 Kp 110 and p-PI 3 Kp 110 protein expression levels were significantly up-regulated in the EA prevention, EA and CSF groups (P < 0.05, P < 0.01). No significant differences were found among the EA prevention, EA and CSF groups, and between the model and Wortmannin groups in the BW, FPG, FINS, IAI and hypothalamic PI 3 Kp 110 and p-PI 3 Kp 110 expression levels (P > 0.05).

CONCLUSION

EA intervention can reduce insulin resistance by suppressing the increase of body weight, blood glucose and insulin contents, and increasing hypothalamic PI 3 K expression levels in insulin resistance rats.

摘要

目的

观察“双固一通”针法[针刺“关元”(CV 4)、“后三里”(ST 36)、“丰隆”(ST 40)和“中脘”(CV 12)并接电针]对胰岛素抵抗大鼠下丘脑磷脂酰肌醇3激酶(PI 3 K)及p-PI 3 K蛋白表达的影响。

方法

将60只Wistar大鼠随机分为正常对照组、模型组、电针预防组、电针组、脑脊液组和渥曼青霉素(PI 3 K拮抗剂)组(每组10只)。通过连续8周喂饲高脂饲料建立胰岛素抵抗模型。电针组大鼠于造模后开始,采用2 Hz、1 mA电流针刺同侧“后三里”(ST 36)-“丰隆”(ST 40)及“关元”(CV 4)-“中脘”(CV 12)15分钟,每日1次,每周5次,共8周。电针预防组大鼠在造模的同时进行电针干预。渥曼青霉素组和脑脊液组大鼠通过植入的钢套管进行脑室微量注射渥曼青霉素(50 nmol/L,3 mg/kg)或人工脑脊液。分别采用血糖仪和酶联免疫吸附测定法检测空腹血糖(FPG)和空腹血清胰岛素(FINS)含量,计算胰岛素活性指数(IAI)[IAI = Ln(1/FPG×FINS)],并采用蛋白质免疫印迹法检测下丘脑PI 3 K和p-PI 3 K蛋白的表达。

结果

与正常对照组比较,模型组大鼠在8周和16周时体重、FPG及FINS含量显著增加(P < 0.01),IAI、下丘脑PI 3 Kp 110及p-PI 3 Kp 110蛋白表达水平明显降低(P < 0.01)。与模型组比较,电针预防组造模后8周时FPG和FINS含量,电针预防组、电针组和脑脊液组在16周时体重和FPG水平,电针预防组和电针组在16周时FINS水平均显著降低(P < 0.05,P < 0.01);电针预防组、电针组和脑脊液组IAI、下丘脑PI 3 Kp 110及p-PI 3 Kp 110蛋白表达水平均显著上调(P < 0.05,P < 0.01)。电针预防组、电针组和脑脊液组之间以及模型组与渥曼青霉素组之间在体重、FPG、FINS、IAI及下丘脑PI 3 Kp 110和p-PI 3 Kp 110表达水平方面差异均无统计学意义(P > 0.05)。

结论

电针干预可通过抑制胰岛素抵抗大鼠体重、血糖及胰岛素含量的升高,增加下丘脑PI 3 K表达水平,从而降低胰岛素抵抗。

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