The Network Pharmacology And Experimental Study Of Rhubarb-salvia Miltiorrhiza In The Treatment Of Renal Interstitial Fibrosis

Objective: To study the therapeutic effect and possible mechanism of rhubarb-salvia miltiorrhiza on renal interstitial fibrosis by network pharmacology and UUO model.Methods: The effective components of rhubarb-salvia miltiorrhiza in the treatment of renal interstitial fibrosis were screened in TCMSP and Sym Map databases by network pharmacology,and the potential targets of rhubarb-salvia miltiorrhiza were screened in SEA,ETCM,STITCH and Sym Map databases.The disease targets of renal interstitial fibrosis were screened in OMIM,Gene Cards,Dis Ge NET and Pub-Gene databases,and then the drug targets and disease targets were intersected.The common target was analyzed in the STRING database,and a protein-protein interaction network was constructed.Analyze GO function and KEGG pathway enrichment to predict the main components,key targets and main signal pathways of rhubarb-salvia miltiorrhiza in the treatment of renal interstitial fibrosis.Then the UUO model was used for experimental verification.Thirty-two 8-week-old male C57BL/6mice were used to verify the therapeutic effect of luteolin on renal interstitial fibrosis.Randomly divide mice into sham operation group,model group,luteolin low-dose group and luteolin high-dose group,with 8 mice in each group.The model of unilateral ureteral obstruction was established by feeding for 3 days.After 7 days of intragastric administration,the kidneys were harvested for Masson and H&E staining.The expression levels of collagenΙ,α-SMA,TGF-β1,Smad3 and E-cadherin were detected by real-time quantitative PCR and immunohistochemistry to evaluate the therapeutic effect of luteolin on renal interstitial fibrosis.Results: A total of 50 compounds from rhubarb-salvia miltiorrhiza were selected and included in the follow-up study,among which quercetin,luteolin,kaempferol and β-sitosterol were the main effective components.1226 targets of salvia miltiorrhiza,406 targets of rhubarb and 34 targets of renal interstitial fibrosis were obtained.Among them,TGF-β1,VEGFA,MMP9,TNF and IL6 are the key targets of rhubarb-salvia miltiorrhiza in the treatment of renal interstitial fibrosis,mainly involving proteoglycan,AGE-RAGE,IL-17 and hypoxia inducible factor-1 signaling pathways.These results indicate that the effective components of rhubarbsalvia miltiorrhiza may act on the above pathways to achieve the purpose of treating renal interstitial fibrosis.The results of Masson and H&E staining showed that luteolin can reduce the area of renal interstitial fibrosis and inflammatory cell infiltration,which has a certain renal protective effect.The results of immunohistochemistry and real-time quantitative PCR showed that luteolin could reduce the expression levels of collagen Ι,α-SMA,TGF-β 1 and p-Smad2/3,and increase the expression level of E-cadherin.Therefore,it is speculated that rhubarb-salvia miltiorrhiza has the effect of improving renal interstitial fibrosis,and it may play a role mainly by inhibiting the TGF-β/Smads signaling pathway.Conclusion: Rhubarb-salvia miltiorrhiza has a certain effect on improving renal interstitial fibrosis,and it may be mainly related to the inhibition of TGF-β/Smads signaling pathway.Its clinical application is wide,but the pharmacological research reports are relatively few.In this study,the network pharmacology method was used to explore the mechanism of rhubarb-salvia miltiorrhiza in the treatment of renal interstitial fibrosis,reflecting the complex network relationship of multiple components,multiple targets and multiple channels,provided the basis for its clinical application,and provided a new idea for further research on the mechanism of rhubarb-salvia miltiorrhiza in the treatment of renal interstitial fibrosis.