Based On The Method Of Cell Culture Technology Research The Intervention Mechanism Of Baicalin On Lipopolysaccharide-induced Inflammation In HBE16 Cells

Objective: To explore the molecular mechanisms of anti-inflammatory of baicalin by lipopolysaccharide-induced inflammation in HBE16 cells.Method: According to lipopolysaccharide-induced inflammation in HBE16 cells,the groups were sectionalized blank group,model group,intervention group of various doses of baicalin,after 6,12,24 hours collecting the cell supernatant and cell lysis buffer.The expression of leukin-6(IL-6),interleukin-8(IL-8),and tumor necrosis factor-α(TNF-α)were quantitative detection by enzyme linked immuno-sorbent assay(ELISA).The messenger RNA expressions of IL-6,IL-8,and TNF-αwere detected by real-time reverse transcription-polymerase chain reaction(RT-PCR).The expression of NF-?B p65 、 IKKα/βand I?B-αof HE16 cells treated with lipopolysaccharide various doses of baicalin for 24 hours were detected by Western Blotting to determine whether the signaling pathway NF-?B was involved in the anti-inflammatory action of baicalin.Result:(1)The expression of IL-6,IL-8 and TNF-α were quantitative tested by ELISA assay.Compared with blank group,we found that treating cells with lipopolysaccharide significantly increased the productions of IL-6,IL-8and TNF-α(P<0.05);Baicalin group compared with lipopolysaccharide group,baicalin at all doses significantly decreased the productions of IL-6,IL-8 and TNF-α(P< 0.05);(2)The expressions of IL-6’s m RNA,IL-8’s m RNA and TNF-α’s m RNA were detected by RT-PCR.We found that the messenger RNA expressions results were similar to ELISA findings.Compared with blank group,treatment with lipopolysaccharide significantly increased the messenger RNA expressions of IL-6,IL-8 and TNF-α(P<0.01);Baicalin group compared with lipopolysaccharide group,baicalin at all doses significantly decreased the expressions of IL-6,IL-8 and TNF-α(P< 0.01);(3)The expression of NF-?B p65、IKKα/βand I?B-αof HE16 cells treated with lipopolysaccharide various doses of baicalin were detected by Western Blotting to determine whether the signaling pathway NF-?B was involved in the anti-inflammatory action of baicalin.Compared with blank group,we found that treating cells with lipopolysaccharide significantly increased the expressions of NF-?B p65,IKKα/β and that the expressions of I?B-α was decreased(P<0.01);Baicalin at all doses significantly decreased the expressions of NF-?B p65、IKKα/β and that the expressions of I?B-α was increased(P<0.05).However The dosses of 50 and 100 ug/ml group is more obvious.Conclusion: Baicalin can decrease the expressions of NF-?B p65、IKKα/βand increase the expressions of I?B-α to inhibit the lipopolysaccharide activated NF-?B signaling pathway and the messenger RNA expressions of IL-6,IL-8 and TNF-α of lipopolysaccharide-induced inflammation in HBE16 cells,leading to decrease expressions of IL-6,IL-8 and TNF-α.