To Explore The Protective Effect And Mechanism Of Jiedu Tongluo Yishen Formula On Kidney Of Experimental Diabetic Rats Based On PI3K/Akt/mTOR Pathway Pathway

Objective: By studying the effects of Jiedu Tongluo Yishen Recipe on the general state,glucose and lipid metabolism,renal function,pathological changes of kidney tissue,autophagy related protein expression and PI3K/Akt/m TOR protein pathway expression of experimental diabetic rats fed with high fat and high sugar combined with streptozotocin,the mechanism of its kidney protection was preliminarily discussed,and the theory of "toxic damage to kidney collaterals" was verified and enriched,which provided a theoretical basis for treating diabetic nephropathy with traditional Chinese medicine.Methods:1.One hundred and twenty male SPF grade Sprague dawley(SD)normal rats(100g–120 g)were adaptively fed for one week,and then 110 rats were randomly selected to be fed with high-fat and high-glucose diet for six weeks.A single intraperitoneal injection of low-dose streptozotocin(STZ)35mg/kg was performed to induce the experimental diabetic rat model,and then the model determination period of two weeks was passed.Seventy-five rats with fasting blood glucose ≥ 16.7mmol/L and in good condition were randomly divided into a model group(15 rats),a positive drug irbesartan group(15 rats),a high dose of Jiedu Tongluo Yishen decoction group(15 rats),a dose of Jiedu Tongluo Yishen decoction group(15 rats),and a low dose of Jiedu Tongluo Yishen decoction group(15 rats).In addition to a normal group(10 rats),a total of six groups were included.The rats in the normal control group and the model group were intragastrically administered with an equal volume of purified water every day.The rats in the Chinese medicine group were intragastrically administered with Jiedu Tongluo Yishen Decoction,and those in the positive control group were intragastrically administered with irbesartan suspension for successive 8weeks.During the treatment,the general state,body weight and 24-hour urine volume of rats were recorded.After eight weeks of intervention,the rats were anesthetized and the blood samples were taken from the kidneys and abdominal aorta.Renal function,blood lipid and 24 h urine protein quantification were detected,and renal weight/body weight was recorded at the end of the experiment.2.The pathomorphological changes of kidney tissues were observed under light microscope and transmission electron microscope to study the effects of Jiedu Tongluo Yishen Decoction on the pathomorphology of kidney tissues of diabetic rats.3.The effects of Jiedu Tongluo Yishen Decoction on Nephrin and α-SMA in renal tissue were detected by immunohistochemistry and Westernblot.The effect of Jiedu Tongluo Yishen Decoction on renal tissue Beclin-1,LC3-II/I,P62,p-PI3 K,PI3K,p-Akt,Akt,p-m TOR,and m TOR was detected by Westernblot.Results:1.Jiedu Tongluo Yishen Decoction can improve the general conditions of experimental diabetic rats and correct the disorders of glucose and lipid metabolism.Jiedu Tongluo Yishen Decoction can reduce the pothological proteinuria of experimental diabetic rats,reduce the elevated serum Scr and BUN,and improve renal function.2.Jiedu Tongluo Yishen Decoction can improve the glomerular enlargement of the kidney of diabetic rats,the pathological morphology of glomerular basement membrane,mesangium,and renal tubular epithelial cells,the ultrastructural changes of renal podocyte marker proteins,renal tissue fibrosis marker proteins,and the changes of autophagosomes and autophagosomes to exert the effect of protecting the kidney.3.Jiedu Tongluo Yishen Decoction can up-regulate the expression of nephrin protein in the kidney tissue of diabetic rats and down-regulate the expression of α-SMA protein.JTDLS was able to up-regulate the protein expressions of LC3II/I and Beclin-1,and down-regulate the protein expression of P62.JTDLS was able to down-regulate the protein expressions of PI3 K,Akt,and m TOR.Conclusion: JTDLS has the effects of improving the general condition of experimental diabetic rats,improving glucolipid metabolism,inhibiting pathological damage of renal tissue structure,increasing the level of autophagy of renal tissue,and down-regulating the expression of PI3K/Akt/m TOR pathway,thereby protecting the function of the kidney.Finally,the theory of "toxin damaging kidney and collaterals" was enriched,and the biological connotation of Jiedu Tongluo Yishen Formula was explored.