Study On The Therapeutic Effect And Mechanism In Type 2 Diabetic Nephropathy Rats By Tiaolipiwei Acupuncture

ObjectiveDynamically detecting the therapeutic effect of"Tiaolipiwei Acupuncture"on Type 2diabetic nephropathy rats’from the changes of blood glucose,body weight,urine protein,blood function and other indicators.Study the mechanism of action of"Tiaolipiwei Acupuncture"on diabetic nephropathy,from the Changes of the protein expression levels of PI3K,AKT,mTOR,LKB1,AMPK,SIRT1.Methods1.Pre-rearing:During the experiment the rats in the Blank（NC）group were basic feeding and the Model group with high-sugar and high-fat feeding,for 8 weeks.2.Modeling:Establish the type 2 diabetic nephropathy rats model.3.Grouping:The successfully modeled Type 2 diabetic nephropathy rats were divided into the Acupuncture（AT）group,and the Type 2 diabetic nephropathy（T2DN）group.Then divide each group and the NC group into 4-weekth group,8-weekth group,12-weekth group.4.Experiment:The rats in the AT group acupunctured for 30 minutes with its acupuncture overclothes on.Model groups adopted the same method to grab and dress,but not take any treatment.Adopted the same method to grab the NC groups,but without any intervention.5.Detection:From the 0^thweek of experiment,the body weight and fasting blood glucose（FBG）were measured at 8:00 on the 6th day of each week and each group.After the4^th,8^th,and 12^thweeks’experiment,test the 24-hour urine protein（24hUPro）,collecting the blood of each group to determin serum biochemical indexes:Blood urea nitrogen（BUN）,Serum creatinine（Scr）,Total protein（TP）,Albumin（ALB）,Triglyceride（TG）,High-density lipoprotein（HDL）,and Cholesterol（CHO）.Western blot detected the protein expression levels of nephrin,P62,Lc3-Ⅰ,Lc3-Ⅱ,PI3K,AKT,mTOR,LKB1,AMPK and SIRT1.Result1.Rat modeling:After 8 weeks of high-fat and high-sugar feeding,Compared with one-time fasting injection of Streptozotocin（STZ）（35 mg/kg）,the group of two consecutive days（STZ）（25 mg/kg）intraperitoneal injection method can increased the survival rate to>68%.2.Changes in body weight:After the 4^th,8^th,and 12^thweeks of acupuncture,between the Experimental group and the NC group the difference was significant（P<0.05）.While the AT group and the T2DN group,there was no significant difference（P>0.05）.3.Changes of FBG:After the 4^th,8^th,and 12^thweeks of acupuncture,between the Experimental group and the NC group,the difference was extremely significant（P<0.01）.Compared with the T2DN group,the AT group showed a decreasing trend,after the 12^thweek,the difference was significant（P<0.05）.4.24-hour urine protein and serum functional indicators:After the 4^th,8^th,and 12^thweeks of acupuncture treatment,There were extremely significant differences between the Experimental group and the NC group in each index（P<0.01）.The comparison between the T2DN group and the AT group:the blood indexes of the rats in each group showed the trend overall.The levels of 24hUPRO,BUN,SCR,CHO showed,NC group<AT group<T2DN group;The levels of TP,ALB,HDL showed:NC group>AT group>T2DN group.In addition to the TG indicator,others showed significant difference（P<0.05）.5.Western-Blot test results:After the 4^th,8^th,and 12^thweeks of Acupuncture treatment,the protein content of each Experimental group was compared with that of the NC group,the difference was extremely significant（P<0.01）.The AT group Compared with T2DN group of each protein,the levels of P62,Lc3-Ⅰ,PI3K,AKT and mTOR were decreased,and the levels of Lc3-Ⅱ,nephrin,P-LKB1,P-AMPKA2 and SIRT1 were all increased,the difference was significant（p<0.05）or extremely significant（p<0.01）.Conclusion1."Tiaolipiwei Acupuncture"can effectively control and lower the blood sugar in T2DN rats and slow down the weight loss;2."Tiaolipiwei Acupuncture"can reduce urine protein,blood fat and blood viscosity,improve the function of kidneys and the pathological changes,and play a benign maintenance role in the diabetic nephropathy disease.3.It is preliminarily judged that after"Tiaolipiwei Acupuncture"treatment,can inhibit the PI3K/AKT/mTOR signaling pathway,activate the LKB1/AMPK/SIRT1 signaling pathway,and protect the kidney and maintain their physiological functions.