六味地黄丸抑制巨噬细胞激活抗肝纤维化作用机制研究
Liuwei Dihuang Pills Attenuates Liver Fibrosis by Inhibiting Macrophage Activation in CCl4 Induced Liver Injury
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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。Abstract: 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 of α-SMA significantly increased， and LWDHP potently 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 factors and chemotactic factors. CONCLUSION LWDHP shows a potent inhibition of macrophage activation in CCl4-induced liver fibrosis.
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Keywords:
- liver fibrosis /
- Liuwei Dihuang Pills /
- macrophage
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期刊类型引用(6)
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