Effect of Honokiol on Autophagy and Apoptosis of Cardiomyocytes Induced by Lipopolysaccharide
Objective:To investigate the effect of honokiol on lipopolysaccharide(LPS)-induced myocardial cell injury,autophagy,and apoptosis,and to elucidate the potential underlying mechanisms.Methods:A myocardial cell injury model was established by treating H9C2 cells with 10 μg/mL of LPS.Grouping was performed in combination with autophagy inhibitor(3-MA):blank control,model control,honokiol(12.5,25.0,50.0 µmol/L)groups,and honokiol(50.0 μmol/L)+3-MA group.Cell viability was determined using the CCK-8 assay,and levels of inflammatory factors were measured by ELISA.ROS activity was assessed using the DCFH method,and autophagy was observed with LC3 im-munofluorescence staining.Apoptosis was evaluated by flow cytometry,and Western blot was used to detect the protein levels related to auto-phagy,apoptosis,and the PI3K/Akt/mTOR pathway.Results:Compared with the blank control group,the model group demonstrated reduced cell viability and LAMP2 protein levels(P＜0.01),and increased levels of inflammatory factors,ROS activity,number of autophagic vesicles,apoptosis,and expressions of LC3Ⅱ/Ⅰ,Beclin 1,p62,Cleaved caspase-3/Caspase-3,phosphorylated PI3K/PI3K,and phosphorylated Akt/Akt(P＜0.01).Honokiol treatment resulted in a significant decrease in inflammatory factors,ROS activity,apoptosis,and expressions of p62 and Cleaved caspase-3/Caspase-3,as well as a decrease in PI3K/Akt/mTOR pathway-related proteins(P＜0.01).It also increased cell viability,number of autophagic vesicles,and levels of LC3Ⅱ/Ⅰ,Beclin 1,and LAMP2(P＜0.01),showing a concentration-dependent effect.Compared with the honokiol(50.0 μmol/L)group,the honokiol(50.0 µmol/L)+3-MA group exhibited higher levels of inflammatory factors,ROS ac-tivity,apoptosis,and expressions of p62 and Cleaved caspase-3/Caspase-3(P＜0.01),and down-regulated cell viability,number of autophagic vesicles,and expressions of LC3Ⅱ/Ⅰ,Beclin 1,LAMP2,phosphorylated PI3K/PI3K,and phosphorylated Akt/Akt(P＜0.05 or P＜0.01).Conclusion:Honokiol could induce autophagy and attenuate LPS-induced myocardial cell injury,ROS activity,and apoptosis in a concentra-tion-dependent manner.These effects were associated with the modulation of the PI3K/Akt/mTOR signaling pathway,enhancing autophagy and promoting autophagic flux.
HonokiolLipopolysaccharideAutophagyApoptosisPhosphatidylinositol 3-kinase/protein kinase B/mammalian target of rapa-mycin(PI3K/Akt/mTOR)signaling pathway
万方数据
维普
