Effects of ethyl acetate extract of Gentianopsis paludosa on the apoptosis of NCM460 cells via NF-κB signaling pathway
AIM To investigate the effects of ethyl acetate extract of Gentianopsis paludosa (Hook.f.) Ma on the apoptosis of human normal colon epithelial cell NCM460.METHODS The NCM460 inflammatory cell model induced by 10 g/mL lipopolysaccharide (LPS) was administered with the ethyl acetate extract of G.paludosa (50,100,200 g/mL) for 24 hours.The cells had their survival rate detected by CCK-8 kit;their levels of inflammatory factors IL-1β and IL-6 detected by ELISA;their apoptosis detected by TUNEL staining;their expressions of apoptosis-related proteins Bcl-2,Bax,caspase-3,caspase-9 and NF-κB signaling pathway-related proteins NF-κB,p-NF-κB,IKKα/β,p-IKKα/β,IκBαand p-IκBαdetected by Western blot;and their nucleus NF-κB expression observed using the immunofluorescence technique.RESULTS Compared with the control group,the NCM460 cells exposed to LPS displayed decreased cell viability ( P<0.01);increased cell apoptosis and levels of IL-1βand IL-6 (P<0.01);decreased Bax,caspase-3,caspase-9,p-IKKα/β/IKKα/β,p-IκBα/IκBα,p-NF-κB/NF-κB protein expressions ( P<0.01 );and increased Bcl-2 protein expression ( P<0.01 ).Compared with the model group,after 24 hours treatment of ethyl acetate extract of G.paludosa,the trial group displayed increased cell viability (P<0.05,P<0.01);decreased apoptosis (P<0.01);decreased levels of IL-1β and IL-6 (P<0.01);decreased protein expressions of Bax,caspase-3,caspase-9,p-IKKα/β/IKKα/β,p-IκBα/IκBα,p-NF-κB/NF-κB ( P<0.05,P<0.01);increased Bcl-2 protein expression ( P<0.01);and inhibited NF-κB translocation into the nucleus.CONCLUSION Ethyl acetate extract of G.paludosa may improves the apoptosis of LPS-induced NCM460 cells via NF-κB signaling pathway.
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