Experimental Study On The Effects Of Electroacupuncture At "Zusanli" And "Shenshu" On GLUT2 And GCK In T2DM Rats

Objective:This study was designed to observe electroacupuncture at Zusanli and Shenshu for T2DM rats,the changes of body weight,blood glucose,insulin,islet B cell function index and islet B cell morphology,to summarize the effects of different intervention methods on the efficacy of rats.The effects of glucose transporter-2(GLUT2),glucokinase(GCK)protein and mRNA expression in rat pancreas by electroacupuncture at Zusanli and Shenshu for T2DM rats,to explore the possible mechanism of electroacupuncture at Zusanli and Shenshu on T2DM rats.Methods:①Forty male SD rats were randomly selected as blank group,the remaining rats were fed with high glucose and high fat diet for 50 days,by intraperitoneal injection of STZ 35mg/kg(2%concentration)for T2DM modeling.Then,32 T2DM rat models were randomly divided into different groups,electro-acupuncture ST-36 group,electro-acupuncture BL-23 group and glimepiride group,8 rats in each group.②electro-acupuncture ST-36 group and electro-acupuncture BL-23 group took both side acupunctures,the electro-acupunctureparameters were 2Hz,2mA continuous wave 20 min/time,1 time/d,6d/week.In the same time,glimepiride group were given glimepiride 10ml/kg,all interventions were conducted for 4 weeks.③Before the intervention,the first,second,third,and fourth week of intervention,the body mass of each group was weighed.Before and after the intervention,we detected rats fasting blood glucose(FBG)and oral glucose tolerance(OGTT).④After the last intervention,collect specimens,took the serum of each group,serum insulin(FINS)was measured by radioimmunoassay,calculated the insulin secretion index(HOMA-B).Taking part of the rat pancreas tissue in the Bonn solution,the morphological changes of the pancreas were observed by HE staining,in addition to taking part of the rat pancreas tissue in liquid nitrogen frozen.The expression of GLUT2 and GCK in pancreas was detected by Western blot,and Real-time PCR was used to detect the expression of GLUT2 and GCK mRNA in pancreas.Result:Efficacy of indicators on results show:①Body mass:During the intervention process,the body mass of the normal group was significantly increased(P<0.01),the body mass of the model group was gradually reduced(P<0.01 or P<0.05).Body mass of electro-acupuncture ST-36 group,electro-acupuncture BL-23 group and glimepiride gavage group was still reduced of intervention in the first and second weeks,the third week began to gradually increase,after 4 weeks of intervention,there was a significant increase over the previous week(P<0.01 or P<0.05).②FBG:After 4 weeks of intervention,the FBG of the model group was significantly higher than before(P<0.01).The FBG of electro-acupuncture ST-36 group,electro-acupuncture BL-23 group and glimepiride gavage group was significantly lower than the model group(P<0.01 or P<0.05).③FINS:Compared with the model group,electro-acupuncture ST-36 group,electro-acupuncture BL-23 group and glimepiride gavage group was significantly higher.④ OGTT 2h blood glucose under the area:After 4 weeks of intervention,the model group was significantly higher than before(P<0.05).electro-acupuncture ST-36 group,electro-acupuncture BL-23 group and glimepiride gavage group were significantly lower than the model group(P<0.01 or P<0.05).⑤HOMA-B:The model group was significantly lower than the normal group.electro-acupuncture ST-36 group,electro-acupuncture BL-23 group and glimepiride gavage group were significantly higher than the model group(P<0.01 or P<0.05).⑥Islet B cell morphology:Islet structure of control group was full and large,oval or cylindrical-shaped,uniformly distributed within the islet cell nucleus,tightly packed,and abundant of cytoplasm.That of the model group,however,was irregular,smaller size,sparse arrangement of nuclei within islets,and there were decreased cell,cell swelling,necrosis and degeneration in the cytoplasm.Compared with the model group,in three intervention groups the volume increased,the number of islet eel 1 nucleus increased,and cytoplasmic vacuoles degeneration reduced.The mechanism of indicators on results show:①GLUT2 protein:Compared with the model group,electro-acupuncture ST-36 group,electro-acupuncture BL-23 group were significantly increased in the pancreas GLUT2 protein expression(P<0.01 or P<0.05).②GCK protein:Compared with the model group,electro-acupuncture ST-36 group,electro-acupuncture BL-23 group and glimepiride gavage group were significantly increased in the pancreas GLUT2 protein expression(P<0.01 or P<0.05),and electro-acupuncture ST-36 group was significantly higher than glimepiride gavage group(P<0.05).③GLUT2 mRNA:Compared with the model group,electro-acupuncture ST-36 group and electro-acupuncture BL-23 group were significantly increased in the level of GLUT2 mRNA(P<0.01).④GCK mRNA:Compared with the model group,electro-acupuncture ST-36 group,electro-acupuncture BL-23 group were significantly increased in the level of GCK mRNA(P<0.01 or P<0.05).Conclusion:electro-acupuncture ST-36 group,electro-acupuncture BL-23 group①Electro-acupuncture ST-36 and electro-acupuncture BL-23 can increase the body weight of T2DM rats,and enough acupuncture effect is needed.They can reduce the FBG,OGTT 2h blood glucose area,improve the FINS,HOMA-B level,islet B cell morphology,and up-regulate the expression of GLUT2,GCK protein and mRNA in islet B cells.②Glimepiride gavage can increase the body weight of T2DM rats,and enough effect is needed.It can reduce the FBG,OGTT 2h blood glucose area,improve the FINS,HOMA-B level,islet B cell morphology,and up-regulate the expression of GCK protein③Electro-acupuncture ST-36,electro-acupuncture BL-23 and glimepiride gavage can improve the FINS,HOMA-B level,and reduce the area under OGTT 2h blood glucose,improve islet B cell morphology,and the effect of electro-acupuncture ST-36 is better than glimepiride gavage and electro-acupuncture BL-23.Electro-acupuncture ST-36 can adjust the body weight of T2DM rats,FBG,OGTT 2h blood glucose area,FINS,HOMA-B and islet B cell morphology that is better than glimepiride gavage and electro-acupuncture BL-23,and glimepiride gavage is better than electro-acupuncture BL-23.Electro-acupuncture ST-36 can up-regulate the expression of GLUT2,GCK protein and mRNA in islet B cells that is better than electro-acupuncture BL-23 and glimepiride gavage,and electro-acupuncture BL-23 is better than glimepiride gavage.④Electro-acupuncture ST-36 and electro-acupuncture BL-23 can improve the symptoms and signs of T2DM rats,may be achieved by upregulating GLUT2,GCK protein and mRNA expression in islet B cells,and electro-acupuncture ST-36 is better than electro-acupuncture BL-23.It is suggested that Zusanli points regulate the glucose metabolism of T2DM rats more prominent and have more comprehensive intervention effect,and specific mechanisms still need further study.