Probing Into The Effect Of Erchen Decoction On Treating Obesity From The Spleen From The Mechanism Of AQP7 Regulating Fat Decomposition

Objective:The aim of this study is to explore effects of differents doses Erchen decoction(ECD)in treating obesity from the spleen;research the possible targets and pathways by network pharmacology,and explore the regulatory effect of ECD on fat decomposition from the perspective of AQP7 and its upstream pathway.Methods:Thirty SPF male ZDF rats were randomly separate into four groups of 10 rats each.Respectively,Zucker Diabetes Fatty(Z)group,low dose ECD(LE)group,middle dose ECD(ME)group and high dose ECD(HE)group.The rats were fed with Purina5008 diet.There were 10 LZ rats in the normal control(L)group,which were fed with growth maintenance diet.The rats in the Erchen decoction group were treated with Erchen decoction every day,and the same volume of normal saline was given to the L group and Z group.1.The indexes related to obesity such as body weight,abdominal circumference,blood glucose,blood lipids and FSI were observed.After 11 weeks of drug intervention,the rats were killed to obtain visceral white adipose tissue,and the changes of adipocyte area were observed by HE staining.2.The active compounds and their structures of Erchen decoction were obtained by TCMSP and PubChem database.The structure information was input into STITCH and SwissTargetPrediction database,and the potential targets were predicted by chemical similarity principle.Use "Obesity" as the keyword to query the treatment targets of obesity in OMIM and DisGeNet databases,and select the key nodes in Erchentang-obesity coincidence target PPI network by Sring database and PageRank algorithm.Finally,the information of er Chen Tang-obesity coincidence target was imported into DAVID database for KEGG and GO enrichment analysis to predict the possible pathway and biological process of er Chen Tang in the treatment of obesity.3.The expression of PI3K/Akt and JNK-MAPK pathway in white adipose tissue was detected by Western blotting and the expression and localization of AQP7 in white adipose tissue was detected by immunohistochemical method.Results:1.Intervention effect of Erchen decoction on related Indexes of Obesity.Erchen decoction could significantly inhibit the increasing trend of body weight and abdominal circumference,improve the levels of serum TG and TC,and significantly reduce the level of FSI and HOMA-IR index.Erchen decoction had no significant effect on the function of liver and kidney in rats.HE staining results showed that the middle dose of Erchen decoction could significantly reduce the volume of white adipocytes in rats.2.Network Pharmacological study on the Therapeutic Mechanism of Erchen decoction on Obesity.The main results are as follows:(1)135 active components of Erchen decoction,106 potential targets for treating obesity and 94 significantly enriched pathways were obtained through database search.(2)Erchen decoction may treat obesity by acting on the targets of TP53,MAPK8,AKT1,IL6 and MAPK1,as well as PI3K-Akt signal pathway,conversion between pentose and glucuronic acid,linoleic acid metabolism,MAPK signal pathway,arachidonic acid metabolism,fat digestion and absorption.3.Effects of Erchen decoction on JNK-MAPK,PI3K/Akt signal pathway and AQP7 expression in white adipose tissue of ZDF rats.The results of Western blotting showed that the level of p-JNK/JNK in ZDF group was significantly higher than that in LZ group,while the levels of PI3K and p-Akt/Akt were significantly lower than those in LZ group.The level of p-JNK/JNK in ME group was significantly lower than that in ZDF group,while the levels of PI3K and p-Akt/Akt were significantly higher than those in ZDF group.The results of Western blotting and immunohistochemistry showed that the expression of AQP7 in ZDF rats was significantly lower than that in LZ,and the expression of AQP7 in ME group was significantly higher than that in ZDF group.Conclusion:1.Erchen decoction can improve the obesity state of ZDF rats,reduce the levels of TG and TC,and improve the insulin resistance caused by obesity,among which the middle dose of Erchen decoction has the best effect.2.The results of network pharmacology show that Erchen decoction may treat obesity by regulating lipid metabolism,participating in lipid transport and relieving adipose tissue inflammation,and its effect has the characteristics of "multi-pathway","multi-target" and "multi-pathway".3.JNK-MAPK pathway is activated and PI3K/Akt pathway is inhibited in obese rats.Erchen decoction can up-regulate the expression of AQP7 by inhibiting JNK-MAPK pathway and activating PI3K/Akt pathway,restore the ability of white adipocytes to transfer glycerol,improve lipid metabolism,and thus treat obesity.