青蒿琥酯对结合型胆汁酸诱导小鼠原代肝细胞IL-6产生的机制研究

Artesunate reduces IL-6 production in primary mouse hepatocytes induced by conjugated bile acids through inhibiting p-p65 signaling pathway

目的探讨青蒿琥酯(Artesunate, AS)对结合型胆汁酸诱导的小鼠原代肝细胞IL-6表达水平的影响及机制。方法体外培养小鼠原代肝细胞,并用25μmol/L的结合型胆汁酸处理,CCK-8法检测青蒿琥酯对小鼠原代肝细胞的毒性影响,RT-PCR法检测细胞因子IL-6的mRNA表达水平,蛋白印迹法检测肝细胞IL-6,p65, p-p65的水平。结果 25μmol/L结合型胆汁酸处理小鼠原代肝细胞能够诱导小鼠原代肝细胞IL-6的mRNA和蛋白质表达;青蒿琥酯减轻了TCA诱导的p65磷酸化和IL-6的蛋白水平;进一步研究发现青蒿琥酯是通过抑制p-p65(phosphorylated-p65)信号通路减轻小鼠原代肝细胞IL-6的表达。结论青蒿琥酯通过抑制p-p65信号通路减轻结合型胆汁酸诱导的原代小鼠肝细胞IL-6的产生。

Objective To investigate the effect and underlying mechanism of artesunate(AS) on the expression of interleukin-6(IL-6) in mouse primary hepatocytes induced by conjugate bile acids. Methods Mouse hepatocytes were isolated and primarily cultured, and then treated with 25 μmol/L conjugated bile acids. CCK-8 assay was used to determine the toxic effects of AS on the liver cells. The mRNA level of IL-6 in the cells was measured by RT-qPCR, and the protein levels of IL-6, p-65 and phosphorylated-p65(p-p65) were determined by Western blotting. Results Conjugated bile acids of 25 μmol/L significantly promoted the expression of IL-6 at mRNA and protein levels in mouse primary hepatocytes, while, AS reduced the taurocholic acid(TCA)-induced phosphorylation of p65 and IL-6 level. Further studies revealed that AS suppressed the IL-6 expression through inhibiting p-p65 signaling pathway in mouse primary hepatocytes. Conclusion AS reduces IL-6 expression in mouse primary hepatocytes induced by conjugate bile acids via inhibiting the p-p65 signaling pathway.