六味地黄丸抑制巨噬细胞激活抗肝纤维化作用机制研究

摘要

目的：观察六味地黄丸对四氯化碳(CCl4)小鼠肝纤维化过程中巨噬细胞激活的影响。方法每周3次腹腔注射 CCl4共6周制备肝纤维化模型,六味地黄丸在 CCl4造模同时灌胃给药。免疫荧光检测α-SMA 表达,免疫组化检测巨噬细胞标志物 CD68表达,qPCR 检测α-SMA、TNF-α、IL-1β、MCP1、CXCR3表达,Western blot 检测α-SMA、MCP1和 CXCR3。结果造模6周,α-SMA 表达显著升高(P <0.01),六味地黄丸显著抑制α-SMA 表达(P <0.01)；CCl4造模后 CD68主要分布在纤维间隔呈强阳性表达；与正常组相比,模型组 TNF-α、IL-1β、MCP1、CXCR3表达显著升高(P <0.01)；与模型组相比,六味地黄丸显著降低 CD68及促炎症因子、趋化因子的表达(P <0.01)。结论六味地黄丸对 CCl4肝纤维化过程中的巨噬细胞激活有显著抑制作用。%OBJECTIVE To observe the inhibitory effects of Liuwei Dihuang Pills(LWDHP) on macrophages activation in CCl4-induced liver fibrosis in mice.METHODS C57BL/6 mice were induced liver fibrosis by CCl4 exposure and administered with LWDHP for 6 weeks simultaneously.Liver tissue was investigated by HE and Sirius red staining.α-SMA was analyzed by immunofluorescence,qPCR and Western blot.Liver macrophages were observed by immunohistochemistry of CD68.The pro-inflammatory cytokines and chemokine such as TNF-α,IL-1β,CXCR3 and MCP1 were detected by qPCR or Western blot analysis.RESULTS After 6 weeks of CCl4 administration,the expression ofα-SMA significantly increased,and LWDHP po-tently inhibited the α-SMA expression.Immunohistochemistry showed that the expression of CD68 was very weak in normal group,CD68 was distributed mainly between fibrotic septa with strong positive expression in CCl4 model group;Real-time quantitative PCR showed that TNF-α,IL-1β,MCP1 and CXCR3 expression significantly increased in model group compared with normal group.Compared with the model group,LWDHP significantly reduced the expression of CD68,inflammatory fac-tors and chemotactic factors.CONCLUSION LWDHP shows a potent inhibition of macrophage activation in CCl4-induced liv-er fibrosis.