The Study Of Electroacupuncture On AMPK/mTORC1 Expression And Energy Metabolism Disorders In ZDF Rats

ObjectiveThis research adopts the Zucker diabetic fatty(ZDF)hereditary diabetes in obese rats,with electroacupuncture as the intervention methods,selecting "Zusanli(ST36)" and"Sanyinjiao(SP6)",and observing the therapeutic effect of electroacupuncture for ZDF rats,hepatic AMP activated protein kinase(AMPK)and mammalian target of rapamycin(mTOR)protein’s multiprotein complex expression of mTORC1,liver cell ultrastructure of mitochondria and the effect of skeletal muscle oxidative stress related products,and trying to interpret the curative may be affected by hepatic AMPK/mTORC1 expression and ultrastructure of mitochondria,thus regulating energy metabolism disorder and improving the mechanism of insulin resistance.MethodsTwenty-two 7-week-old male ZDF(Leprfa/fa)and six of the same litter Zucker lean(Leprfa/+,ZL)rats were chosen.Twenty-four ZDF rats were randomly divided into electroacupuncture group(n=8),western medicine group(n=8)and model group(n=6).Six ZL rats were control group.Following the principle of "treating both the symptoms and the root causes" of traditional Chinese medicine,rats in the electroacupuncture group took therapy on bilateral "Zusanli" and "Sanyinjiao" acupoints andwere inserted by the needle vertically about 3mm.Huatuo SDZ-V electroacupuncture instrument was used,with density wave,2Hz/100Hz,intensity 2mA,20min each time,3 times a week,once every other day,6 weeks,a total of 18 times;Rats in control group and model group were restrained for the same time every day without electroacupuncture stimulation.The drug group was given 10mg/kg pioglitazone gavage,once a day,for 6 weeks,a total of 42 days.During and after treatment,sampling and testing were performed:① Basic and biochemical indexes were detected:Body weight and fasting plasma glucose(FPG)were tested once every two weeks during and after treatment.Fasting insulin(FINS)were calculated.② Morphological detection:the structure,morphology and number of liver mitochondria were observed under transmission electron microscope.③Protein detection:Western Blot was used to detect the expression of AMPK and mTORCl in liver.④Oxidative stress-related indicators:The activity of glutathione peroxidase(GSH-Px),the expression level of malondialdehyde(MDA)and the activity of superoxide dismutase(SOD),which inhibate oxygen free radical,were detected by spectrophotometer colorimetry.Results1.Compared with the control group,the body weight of the model group increased and the fasting blood glucose increased.After 6 weeks of electroacupuncture treatment,body weight and fasting blood glucose in the electroacupuncture group decreased compared with the model group(P<0.05).2.Compared with the control group,the ultrastructure of liver mitochondria in the model group was different,and the structure was abnormal,and the number of mitochondria decreased.After 6 weeks of electroacupuncture treatment,compared with the model group,the ultrastructure of liver mitochondria in the electroacupuncture group was more complete,without swelling and deformation,and the number of liver mitochondria increased.3.Compared with the control group,the expression of AMPK in the liver of model group was significantly decreased(P<0.05),and the expression of mTORC1 was significantly increased(P<0.05).After 6 weeks of electroacupuncture treatment.the expression of AMPK increased and the expression of mTORC1 decreased in the liver of the electroacupuncture group compared with the model group,and the difference was significant(P<0.05).4.The activities of SOD,GSH-Px and MDA in skeletal muscle of model group were decreased compared with those of the control group.There was significant difference(P<0.05).After 6 weeks of electroacupuncture treatment,the activities of SOD and GSH-Px in skeletal muscle of electroacupuncture group were increased,while the content of MDA was decreased,which was significantly different from that of model group rats(P<0.05).Conclusion1.6-week electroacupuncture treatment effectively reduces weight gain and FPG in ZDF rats.2.Electroacupuncture promotes liver damage recovery of ZDF rats of mitochondrial structure and fusion,raised and lowered AMPK expression mTORC1 expression,enhance the antioxidant enzymes SOD and GSH-Px activity,removing oxygen free radicals,reducing lipid peroxide synthesis of MDA,and making the body from oxygen free radical damage,which may be the mechanism of electroacupuncture lowering blood sugar and improving IR.