The Mechanism Study Of Self-made Baoshen Mixture On Delaying The Progress Of Diabetic Kidney Disease

Objective:The main target information of Baoshen mixture in the treatment of diabetic kidney disease was screened by the method of network pharmacology,and the main biological processes and signal pathways involved were predicted,and then the efficacy of Baoshen mixture in the treatment of diabetic kidney disease was analyzed by pharmacological experiments.the differentially expressed genes of Baoshen mixture in the treatment of diabetic kidney disease were screened by transcriptional sequencing technique,and its possible mechanism was discussed.Method:The main components of Astragalus membranaceus,rhubarb and saffron were obtained by TCMSP(Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform)database,screened according to toxicopharmacokinetic(ADME),and then obtained the action targets of active components through Swiss Target Prediction database;the related targets of DKD were obtained from Drug Bank,OMIM and Genecards databases,and the network diagram of DKD targets for the treatment of Baoshen mixture was drawn by Cyto Scape3.7.1software,and the PPI network was constructed by using String11.0 database.The biological process,cell components,molecular function and signal pathway involved in the core target of Baoshen mixture in the treatment of DKD were enriched and analyzed by Metascape gene function analysis website,and the map of "Baoshen mixture component-DKD target-pathway" was further constructed.Forty-eight6-week-old male db/db mice were randomly divided into model group and high,middle and low dose groups of Baoshen mixture.The normal control group was db/m male mice of the same litter and the same week.Adaptive feeding for 2 weeks.From the age of 8 weeks,the high,middle and low dose groups of Baoshen mixture were given intragastric administration of 8.1g/kg,5.4g/kg and 2.7g/kg Baoshen mixture every day for 24 weeks.During this period,random blood glucose,body mass and urinary Microalbumin were measured every 4 weeks,and urinary albumin excretion rate(UAER)was calculated.Serum creatinine(Scr),blood urea nitrogen(BUN)and blood lipids were measured before and after intervention(8 weeks old and 32 weeks old).The left kidney was collected and the pathological changes of renal tissue were observed by HE,PAS and Masson staining.The renal tissue of mice was sequenced by Illumina sequencing platform to analyze the difference of gene expression between the treatment group and the model group,and to explore its possible mechanism.Results:1.Through the prediction and analysis of network pharmacology,it was found that there were 44 active components and 640 possible targets in Baoshen mixture.A total of 4163 targets related to DKD were screened,and finally 411 targets of active components of Baoshen mixture and DKD targets were obtained.through further screening of network topology parameters,27 core targets were obtained,including protein kinase B,tumor suppressor gene TP53,vascular endothelial growth factor A,epidermal growth factor receptor,tumor necrosis factor and so on.It is mainly involved in the regulation of cell migration,oxidative stress response,growth and development regulation,epithelial cell proliferation,apoptosis and other biological processes,including insulin resistance,AGE-RAGE signal pathway,MAPK signal pathway,TNF signal pathway,VEGF signal pathway,PI3K-Akt signal pathway and so on.2.After intervention with Baoshen mixture,compared with the normal control group,the body mass,random blood glucose,urinary albumin excretion rate,serum creatinine,blood urea nitrogen,serum cholesterol,triglyceride and low density lipoprotein cholesterol in the model group were significantly higher than those in the normal control group(p < 0.05).The level of high density lipoprotein cholesterol decreased significantly(p < 0.01).Compared with the model group,the urinary albumin excretion rate,serum creatinine,blood urea nitrogen and the levels of serum cholesterol,triglyceride and low density lipoprotein cholesterol decreased in the treatment group,while the level of high density lipoprotein cholesterol increased significantly in the high dose group(p < 0.01).It increased significantly in the middle dose group.Pathological staining showed that after intragastric administration of Baoshen mixture,the pathological injury was alleviated in each dose group,especially in the high dose group.3.The results of transcriptional sequencing showed that compared with the model group,there were 627 differentially expressed genes in kidney tissue of mice in Baoshen mixture treatment group,including Mettl7a1,Igfbp1,Jmjd8,Depp1,Cyp2d12,Mylip,Plcb1,Sesn2,Ugt1a6 b,Atxn1 and so on.Down-regulated genes included Zbtb7 a,Helz,Ppp1r10,Fcgbp,Hba-a2,Hspa1 b,Id2,Ampd3,Hbb-bs,Hba-a1 and so on.The main biological processes of enrichment of differentially expressed genes include cellular oxidative toxicity,regulation of glucose metabolism,rhythmic process,negative regulation of protein localization to nucleolus,cellular process,positive regulation of I-kappa B kinase / NF-kappa B signal transduction,negative regulation of Wnt signal pathway,positive regulation of transforming growth factor β production,insulin metabolism,exercise regulation,Positive regulation of TOR signal,etc.The main signal pathways involved include antigen processing and presentation,steroid synthesis,renin secretion,c AMP signal pathway,TGF-β signal pathway,PI3K-Akt signal pathway,uric acid pathway,MAPK signal pathway,circadian rhythm,glucose metabolism,amino acid biosynthesis,m TOR signal pathway,ECM-receptor interaction,PPAR signal pathway,synthesis and secretion of aldosterone and so on.Conclusion:1.Baoshen mixture in the treatment of DKD may be achieved through multiple targets and signal pathways.2.Baoshen mixture can effectively improve the renal function of db/db mice,reduce the pathological damage of renal tissue,and delay the progress of DKD.3.The mechanism of Baoshen mixture in treating DKD is significantly related to MAPK signal pathway,serine / threonine kinase(PI3K-Akt)signal pathway,central carbon metabolism,Ras-associated protein1(Rap1)signal pathway,insulin resistance,c AMP signal pathway,transforming growth factor-β(TGF-β)signal pathway and so on.