To Explore the Molecular Mechanism of Jiawei Shengjiang San(加味升降散)in Treating Diabetic Kidney Disease Based on Network Pharmacology and Animal Experiments
Objective:To predict the potential mechanism of action of Jiawei Shengjiang san in the treatment of diabetic kidney disease(DKD)through network pharmacological research,and to verify the efficacy and mechanism of the drug through db/db mouse animal model,so as to expand the clinical application of classical prescription.Methods:The main chemical components and potential targets of Jiawei Shengjiang san were collected by TCMSP.Then the disease targets of DKD were collected through Drug Bank database,GenCards database and OMIM database.The intersection target of drugs and diseases was obtained using the Diagram platform of Draw Venn,and the PPI network system was established using the String platform to obtain the core action target of drugs and diseases.Metascape was used to analyze the"TCM ingredient-Target-pathway"and the main biological processes involved and related pathways.Then,a network system of"Ingredients-Target-pathway of Diabetic Kidney Disease"was established by Cytoscape 3.7.2 software system.Totally 30 8-week-old db/db mice were randomly divided into model group,Perindopril group and Jiawei Shengjiang san low-dose,medium-dose and high-dose groups by body weight stratification method,with 6 mice in each group.And 6 db/m mice at the same age of 8 weeks formed their own group as the blank control group.The blank control group and model group were intragastric with 7 g/(kg·d)pure water every day.Jiawei Shengjiang san low-dose,medium-dose and high-dose groups were respectively intragastric with low[3.50 g(kg·d)],medium[7.00 g/(kg·d)],high[14.00 g/(kg·d)]dose Jiawei Shengjiang san for 12 weeks.Perindopril control group was intragastric with perindopril[0.48 mg/(kg·d)]for 12 weeks.After the experiment,blood creatinine(SCr)and blood urea nitrogen(BUN)were detected by automatic biochemical analyzer.And 24-hour urinary albumin,NGAL,TNF-α,IL-1β,VCAM-1,MCP-1 and HbA1c were detected by enzyme-linked immunosorbent assay(ELISA).The protein expression of p-PI3K,PI3K,p-Akt,Akt,p-NF-κB p65 and NF-κB p65 was measured by Western blotting.Results:A total of 85 main drug active ingredients,976 targets,and 872 main targets of DKD were obtained.Totally 226 intersection targets of drugs and diseases were obtained,among which the core active ingredients of drugs were quercetin,luteolin,[3-glusterol,kaempferol,buttercup flavin,palmitoleic acid,etc.The core targets were PIK3CA,PIK3R1,AKT1,AKT2,MAPK14,IL1β,TNF,IL6,etc.The main pathway involved was the role of AGE-RAGE signaling pathway in diabetic complications.The results of animal experiments showed that compared with the blank control group,the SCr and BUN of the model group were significantly increased,and that the 24-hour urinary albumin was increased;The serum levels of NGAL,TNF-α,IL-1β,VCAM-1,MCP-1 and HbA1c were increased in model group;The p-PI3K,p-PI3K/PI3K,p-Akt,p-Akt/Akt decreased in model group,and p-NF-κB p65,p-NF-κB p65/NF-κB p65 increased in model group.Compared with model group,Jianwei Shengjiang san could decrease SCr,BUN,24-hour urinary microalbumin,NGAL,HbA1c,TNF-α,IL-1β,VCAM-1,MCP-1 and other inflammatory factors in db/db mice.It can also up-regulate the expression levels of p-PI3K and p-Akt(P＜0.05),and inhibited the activation of p-NF-κB p65 protein(P＜0.05).Conclusion:Jiawei Shengjiang san can regulate diabetic kidney disease through multi-target and multi-pathway,and its mechanism may be mainly through the regulation of PI3K/Akt/NF-κB signaling pathway.
万方数据
维普
