世界中医药

目的:探讨电针不同穴位对2型糖尿病大鼠的疗效差异及其机制。方法:40只雄性SD大鼠随机选取8只作为空白组,其余大鼠予高糖高脂饲料喂养50 d后,按35 mg/kg质量比腹腔注射2%链脲佐菌素(Streptozotocin,STZ)溶液进行T2DM造模,并将造模成功的32只大鼠随机分为模型组、电针足三里组、电针肾俞组、格列美脲灌胃组,每组８只。各电针组分别取双侧“足三里”“肾俞”穴进行电针治疗,电针参数为2 Hz,2 mA连续波20 min/次,1次/d,6 d/周,连续4周。于干预前及干预后,检测各组大鼠灌胃前空腹血糖(Fasting Blood Glucose,FBG)、体质量及OGTT实验。末次干预结束,处死大鼠取血,取胰腺组织,采用放免法测血清胰岛素(FINS),计算胰岛素分泌指数(HOMA-B),Real-time PCR检测胰腺中葡萄糖转运蛋白-2(Glucose Transporter-2,GLUT2)、葡萄糖激酶(Glucokinase,GCK)的mRNA表达水平,HE染色观察胰腺的形态变化。结果:干预4周后,除空白组体质量显著增加(P<0.01),其余各组体质量均显著减轻(P<0.01);模型组FBG显著高于干预前、空白组(P<0.01),格列美脲灌胃组显著低于模型组(P<0.01),两电针组比较模型组均有下降趋势(P>0.05);葡萄糖耐量实验(Oralglucose Tolerance,OGTT)实验结果与FBG同步;电针足三里组、格列美脲灌胃组FINS显著低于空白组、模型组及电针肾俞组(P<0.01);模型组HOMA-B显著低于空白组(P<0.01),各干预组比较模型组均有上升趋势(P>0.05);模型组GLUT2的mRNA表达水平显著低于空白组(P<0.01),电针足三里组、电针肾俞组显著高于模型组(P<0.05,P<0.01);模型组GCK的mRNA表达水平显著低于空白组(P<0.01),各干预组均显著高于模型组(P<0.01,P<0.05),且电针足三里组显著高于格列美脲灌胃组(P<0.01)。HE染色:模型组胰岛边界不清,体积明显变小,胰岛细胞数量减少且排列杂乱,细胞肿胀、坏死,可见细胞空泡变性。3个干预组与模型组比较,有不同程度的改善,胰岛边界较模型组清晰,体积增大,胰岛内细胞数量增加,排列也较均匀,细胞空泡变性减少。结论:电针足三里、肾俞穴可能是通过上调GLUT2、GCK mRNA的表达水平而改善T2DM大鼠的胰岛B细胞的分泌功能、保护胰岛B细胞的形态、降低FINS水平,其中足三里穴的效果更优。

English Summary:

To observe the effects and mechanism of different acupuncture points on rats who were infected with type-2 diabetes mellitus(T2DM).Methods:Among forty male SD rats, 8 rats were randomly selected into a control group, and the rest rats were fed on high fat and sugar diet in 50 days.T2DM model was established by intraperitoneal perfusion with 2% STZ solution according to the mass ratio of 35 mg/kg after the diet. A total of 32 rats who were made as model successfully were randomly divided into the model group, the group of electroacupuncture at Zusanli(ST36), electroacupuncture at Shenshu (BL23) group and Glimepiride group, 8 rats in each. The rats in all the electroacupuncture groups were treated with electroacupuncture on Zusanli and Shenshu respectively. Electro-acupuncture (2 Hz, 2 mA) was applied as the intervention, 6 times per week, lasting for 4 weeks. Fasting blood glucose (FBG) and body mass were measured and OGTT experiment was conducted before intervention and after intervention respectively. After the last intervention, the rats were killed, and blood samples and pancreatic tissue were collected. Radioimmunoassay was used to test FINS, calculating the homeostasis model assessment-B(HOMA-B). The mRNA level of GLUT2 and GCK in the pancreas was tested with Real-time PCR. And the morphological changes of the pancreas were stained with HE. Results:After the 4 weeks' intervention, except that the rat's mass body in the control group continued increasing (P<0.01), body mass in other groups faced significant reduction (P<0.01). FBG of the model group was significantly higher than that before intervention and that of the blank group (P<0.01). FBG of the Glimepiride group was prominently lower than that of the model group(P<0.01).Compared with the model group, the two electroacupuncture groups showed a decrease tendency(P>0.05).The results of the OGTT experiment was in accord with FBG. FINS of electroacupuncture at Zusanli(ST36) group and Glimepiride group was markedly lower than the control group,model group and electroacupuncture at Shen-shu group(P<0.01).HOMA-B of the model group was significantly lower than the blank group(P<0.01) and all groups after intervention had an increase tendency in comparison with the model group(P>0.05). The mRNA level of GLUT2 in the blank group(P<0.01) was markedly higher than that of the model group(P<0.01),whose mRNA level of GLUT2 was significantly lower than that of electroacupuncture at Zusanni group and electroacupuncture at Shen-shu group(P<0.05, P<0.01).The mRNA level of GCK in the blank group(P<0.01) was significantly higher than that of the model group, whose GCK level was markedly lower than that of all other groups after intervention. And the GCK level of electroacupuncture at Zusanni group was significantly higher than that of Glimepiride group(P<0.01). HE staining：the islet boundary of the model group was not clear and the volume of the islet markedly became smaller.The islet cells whose number reduced arrayed disordered and appeared swelling and necrosis, which proved vacuolar degeneration.In comparison with the model group, the three groups after intervention all got improved to some degree. The islet boundary of theirs was clear and the volume of the islet appeared bigger.The islet cells whose number increased arrayed evenly and degeneration of which got reduced.Conclusion:Electroacupuncture at Zusanli(ST36) and Shenshu(BL23) could enhance the mRNA level of GLUT2 and GCK to improve islet B cell secretory function in rats with T2DM, protect islet B cell morphology and reduce the level of FINS, especially Zusanli point.

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