Inhibitory effects of baicalin on LPS-induced inflammation and pyroptosis of cardiomyocytes and their mechanism
Objective To observe the inhibitory effects of baicalin on lipopolysaccharide(LPS)-induced pyroptosis and inflammation of cardiomyocytes and to analyze their mechanism.Methods The rat cardiomyocytes H9C2 were culti-vated in vitro and we divided cells in the logarithmic growth phase into the control group,LPS group,and baicalin group.The control group did not receive intervention.In the LPS group,H9C2 cells were stimulated by LPS to construct the cell injury models in vitro.In the baicalin group,cells were induced by LPS and then were added with 10,20,40,and 80 μmol/L baicalin,respectively.The cell viability in each group was detected by CCK-8.The concentration of baicalin with the highest cell viability was selected as the optimal concentration for subsequent experiments.Then,H9C2 cells in the logarithmic phase were divided into the control group,LPS group,baicalin group,inhibitor group,baicalin+inhibitor group,and baicalin+activator group.Except the control group without intervention,LPS stimulation was conducted to con-struct the cell injury models in the other groups;on this basis,the baicalin group was treated with baicalin,while the in-hibitor group was treated with phosphatidylinositol 3-kinase(PI3K)/serine protein kinase(AKT)pathway inhibitor LY294002,the baicalin+inhibitor group was treated with baicalin and LY294002,and the baicalin+activator group was treated with baicalin and PI3K/AKT pathway activator insulin-like growth factor 1(IGF-1).The inflammatory cytokines interleukin(IL)-1β and IL-6 in the cell supernatant were detected by ELISA.Western blotting was used to detect the rela-tive expression levels of pyroptosis-related proteins pyroporin D(GSDMD),nucleotide oligomeric domain-like receptor protein 3(NLRP3),Caspase-1,and PI3K/AKT pathway-related proteins PI3K,AKT,phosphorylation(p-)PI3K,and p-AKT.Results Compared with the control group,the expression levels of inflammatory cytokines IL-6,IL-1β,pyropto-sis-related proteins GSDMD,NLRP3,Caspase-1,p-PI3K and p-AKT increased in the LPS group.Compared with the LPS group,the expression levels of IL-6,IL-1β,GSDMD,NLRP3,Caspase-1,p-PI3K and p-AKT decreased in the baicalin group and inhibitor group.Compared with the baicalin group,the expression levels of IL-6,IL-1β,GSDMD,NLRP3,Caspase-1,p-PI3K and p-AKT decreased in the baicalin + inhibitor group,while the trend was opposite in the baicalin + activator group(all P<0.05).Conclusion Baicalin could inhibit the inflammatory response and pyroptosis of rat cardio-myocytes in vitro induced by LPS,and the mechanism may be related to inhibiting the activation of PI3K/AKT signaling pathway.
baicalinmyocardial damageinflammatory responsepyroptosisphosphatidylinositol 3-kinaseseronine protein kinase
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