Effects of emodin on lipopolysaccharide-induced inflammation of biliary epithelial cell and its mechanism
Objective To investigate the effect of emodin on lipopolysaccharide(LPS)-induced biliary epithelial cell and its mechanism.Methods Biliary epithelial cells were randomly divided into control group(normal culture),model group(20 μg·mL-1 LPS),experimental-L,-M,-H groups(20 μg·mL-1 LPS+5,10,20μg·mL-1 emodin),pcDNA-TLR4 group(20 μg·mL-1 LPS+20μg·mL-1 emodin+transfected pcDNA-TLR4 plasmid).Western blot assay was used to detect protein expression of Toll-like receptor 4(TLR4),phosphorylation-nuclear factor kappa B(p-NF-κB),proliferating cell nuclear antigen(PCNA),E-cadherin;cell counting kit 8(CCK-8)assay and 5-Ethynyl-2'-deoxyuridine(Edu)assay were used to detect cell proliferation;enzyme-linked immunosorbent assay(ELISA)assay was used to detect the expression level of inflammatory factors.Results The expression levels of TLR4 protein in control group,model group,experimental-L,-M,-H groups and pcDNA-TLR4 group were 0.21±0.02,0.92±0.11,0.74±0.09,0.61±0.06,0.39±0.04 and 1.37±0.09;the expression levels of p-NF-KB protein were 0.33±0.03,1.03±0.11,0.80±0.08,0.66±0.05,0.41±0.06 and 0.85±0.08;the expression levels of E-cadherin were 1.20±0.10,0.41±0.04,0.58±0.07,0.79±0.06,0.91±0.05 and 0.58±0.05;Edu positive cells rate were(22.11±1.18)%,(51.78±2.35)%,(43.20±3.76)%,(33.19±3.58)%,(26.96±2.31)%and(41.40±2.74)%;IL-6 levels were(9.79±1.35),(38.69±3.99),(29.80±3.41),(25.48±1.50),(15.63±0.91)and(27.75±3.61)pg·mL-1.Compared with control group and model group,compared with experimental-L,-M,-H groups and model group,pcDNA-TLR4 group was compared with experimental-H group,the differences of the above indexes were statistically significant(all P<0.05).Conclusion Emodin may inhibit cell hyperproliferation and EMT by inhibiting TLR4/NF-KB/c-myc pathway,thereby improving the LPS-induced inflammatory response of biliary epithelial cell.