Chlorogenic Acid Inhibits LPS-induced RAW264.7 Cell Inflammation and its Mechanism / 医学研究杂志

ABSTRACT

Objective To clarify the anti-inflammatory effect of chlorogenic acid on lipopolysaccharide(LPS)-induced inflamma-tion in macrophage RAW264.7,and to reveal its possible molecular mechanism.Methods The cellular activity of RAW264.7 after the intervention of different concentrations of chlorogenic acid was detected by CCK-8 method;the macrophage RAW264.7 was stimulated by LPS to preset cellular inflammatory state.The blank control group(K group,n=3),the model control group(L group,n=3),and the experimental group(S group,n=3)were administered separately,and the cell morphology was dynamically observed;cell precipita-tion and supernatants were obtained at 24 h and 48h,respectively.The concentrations of inflammatory cytokine interleukin(IL)-1β,monocyte chemoattractant protein-1(MCP-1),and arginase-1(Arg-1)in the cell supernatants were detected by enzyme-linked immunosorbent assay(ELISA);the mRNA expression of prostaglandin endoperoxide synthase 2(PTGS2),transforming growth factor-β1(TGF-β1),high mobility histone 1(HMGB1),and nuclear transcription factor-κB(NF-κB)were detected by quantitative real-time PCR(RT-qPCR).Results RAW264.7 cellular inflammatory state was successfully preset with 1 μg/ml LPS;12.5～200.0μg/ml chlorogenic acid had no distinct toxicity on RAW264.7.Chlorogenic acid at 50μg/ml and 200μg/ml had obvious proliferation effects on RAW264.7 cells(P＜0.05).Compared with the model control group,the content of IL-1 β protein in the supernatant of the experimental group was significantly decreased,while the content of Arg-1 was significantly increased(P＜0.05).Compared with the model control group,50μg/ml of chlorogenic acid significantly decreased the mRNA expressions of NF-κB,TGF-β1,PTGS2,and HMGB1 in LPS-induced inflammatory cells(P＜0.05).Conclusion Chlorogenic acid has a distinct inhibitory effect on LPS-induced RAW264.7 inflammatory response,which may be achieved by regulating molecular expression of the HMGB1-mediated NF-κB pathway.