Study On The Mechanism Of Shenluoning Interfering Autophagy Of Diabetic Nephropathy Based On HIF-1?-PI3K/Akt Pathway

Objective:In this study,the effect of shenluoning on the regulation of injury repair of DN podocytes and its mechanism were analyzed by using diabetic nephropathy as the research object and regulating the autophagy process of podocytes as the entry point.Methods:1 To observe the intervention effect of shenluoning on the progression of DN.Rats with DN was induced by single large dose of caudal vein injection of Streptozocin combined with high glucose and high fat diet.The experimental animals were randomly divided into control group,model group and traditional Chinese Medicine Group.At the end of 0,3,6 weeks after intervention treatment,the changes of 24 H Urine Protein excretion,FBG,ALB,BUN and Scr were observed.The pathological changes of the left kidney were observed at the 6 weeks after the intervention.To evaluate the intervention effect of Xintongchangluo compound shenluoning on the progression of DN by observing the results.2 Prediction of the target of shenluoning delaying the progression of DN based on network pharmacology.Using systematic pharmacology database of Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform(TCMSP)to construct the target network of Shenluoning pharmacodynamic substances and screen its active components and action targets.The target protein was transformed by Uniprot database,and then OMIM ?Drugbank?TTD?and DIGSe E were used to search for diabetic nephropathy-related disease targets,the combination of active compounds acts as a target,to identify potential targets for drug interaction and diabetic nephropathy.The results were visualized by Cytoscape and the PPI network analysis was performed to create core subnetworks according to the MCODE score.Finally,the possible molecular mechanism of diabetic nephropathy was explained by using KEGG database and DAVID database for annotation and classification.This study provides an idea for the experimental study of Shenluoning in treating diabetic nephropathy.3 To further analyze the mechanism of shenluoning regulating the process of podocyte autophagy.The drug-containing Serum of Shenluoning was prepared,the podocyte dispersion system and the model of High Glucose injury in vitro were established,and thedrug-containing serum of 10% concentration was used to intervene to observe the effect of HIF-1,PI3 K and Akt on the repair of podocyte filtration barrier injury.The expression of HIF-1 and PI3K/ATK in renal tissues of each group was semi-quantitatively detected by IMMUNOFLUORESCENCE staining.Collected images.And the mechanism of shenluoning regulating autophagy of DN podocyte was elucidated.Results:1 The results showed that shenluoning could improve the general state and body weight of DN model rats,and decrease the urinary protein excretion of DN model rats And it was effective in regulating hyperglycemia,delaying the decrease of ALB lowering the levels of Bun and SCR and protecting renal function in DN model rats.It can improve the glomerular lesion of DN model rats,and has significant difference with model group in delaying glomerulosclerosis.2 The results showed that shenluoning could improve the general state and body weight of DN model rats,and decrease the urinary protein excretion of DN model rats.And it was effective in lowering the Bun and SCR levels,regulating hyperglycemia,delaying the decrease of ALB and protecting renal function in DN model rats.It can improve the glomerular lesion of DN model rats,and has significant difference with model group in delaying glomerulosclerosis.3 The podocyte injury model in vitro and podocyte dispersion system were established.Serum intervention of shenluoning could inhibit the abnormal change of podocyte migration ability and regulate the expression of HIF-1,PI3 K and Akt.Conclusion:1 Shenluoning can delay the progression of renal disease in DN model rats.2 Based on the principle of network pharmacology,it is predicted that the mechanism of diabetic nephropathy intervention is related to autophagy of podocyte regulated by HIF-1,PI3 K and Akt expression.3 Serum containing Shenluoning could inhibit the invasion of podocyte and regulate the expression of HIF-1,PI3 K and p-Akt.The mechanism is related to the regulation of Podocyte Autophagy and the network pharmacology has the authenticity to the target intervention prediction.