Study on the mechanism of salvianolic acid B in the treatment of renal ischemia/reperfusion-induced acute renal injur
Objective This study aims to explore the potential mechanism of Salvianolic acid B(Sal B)in alleviating ischemia/reperfusion(I/R)-induced acute kidney injury(AKI)through network pharmacology,molecular docking,and in vitro experiments.Methods Network pharmacology was used to find the core target and related pathway of Sal B in the treatment of I/R-AKI,and AutoDock Vina was used for molecular docking of Sal B and core target.The cells were divided into 4 groups[Con group,Con+Sal B group,hypoxia/reoxygenation(H/R)group,Sal B+H/R group].Cell viability was measured by cell counting kit-8(CCK-8)method.Enzyme-linked immunosorbent assay(ELISA)and reverse transcriptase-polymerase chain reaction(RT-PCR)were used to detect the content of inflammation-related factors and the mRNA expression of inflammatory factors in the cell supernatants of each group.The apoptosis rate was de-tected by flow cytometry.The expressions of B cell lymphoma/leukemia-2 associated X protein(bax),B cell lymphoma/leukemia-2(bcl-2)and cleaved cysteinyl aspartate-specific protease-3(cleaved Caspase-3)were detected by Western blotting.T-test was used to compare the two groups.Results The signal path-ways of kyoto encyclopedia of genes and genomes(KEGG)enrichment analysis showed that tumour necrosis factor(TNF),apoptosis and other signal pathways were the key pathways of Sal B treatment of I/R.PPI network analysis showed that TNF and CASP3 were key targets of I/R treatment for Sal B.Molecular doc-king results showed that SalB had good binding force with TNF,CASP3 and other targets.In vitro experi-ments showed that the cell viability of Con group,H/R group and 20,40,80,160 μmol/L Sal B+H/R group(0.42±0.03,0.53±0.02,0.54±0.04,0.48±0.02)was higher than that of H/R group(0.38±0.01).The difference was statistically significant(t=2.8,15.1,8.6,10.2,P＜0.05).Sal B+H/R group of inflammatory factor TNF alpha beta,IL-6,IL-1[(277.06±6.80),(4.69±1.04)ng/L,(0.62±0.53)pg/L]content below H/R group[(388.59±9.01),(22.03±0.56)ng/L,(41.81±1.49)pg/L],higher than in Con group[(217.59±6.26),(15.88±0.62)ng/L,(32.89±1.01)pg/L],statistically significant difference(t=17.4,25.4,45.1,P＜0.05 and t=17.1,16.1,49.4,P＜0.05).The mRNA expression levels of inflammatory factors TNF-α,IL-6 and IL-1β in Sal B+H/R group(2.59±0.39,1.61±0.15,2.09±1.30)were lower than those in H/R group(3.79±0.28,2.36±0.50,1.17±0.59).The difference was statistically significant(t=4.3,2.5,1.2,P＜0.05).The apoptosis rate of HK-2 cells,the expression of cleaved Caspase-3 and bax/bcl-2 protein[(4.43±0.68)％,1.19±0.09,0.99±0.11]in Sal B+H/R group were lower than those in H/R group[(12.06±0.14)％,1.98±0.12,4.16±0.52],the difference was statistically significant(t=19.0,25.4,45.1,P＜0.05).Conclusion To explore the potential mechanism by which Sal B alleviates ische-mia/reperfusion(I/R)-induced AKI through network pharmacology,molecular docking,and in vitro exper-iments.
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