目的:探討冠心康通過活化ERK5抗動脈粥樣硬化的潛在分子機制.方法:用ERK5抑制劑(ERK5-IN-1和XMD8-92)干預RAW264.7細胞,探討ERK5失活對巨噬細胞胞葬作用的影響.用ox-LDL干預RAW264.7細胞建立巨噬細胞胞葬作用功能受損的細胞模型,然后在存在或不存在XMD8-92干預的情況下,用冠心康含藥血清處理該細胞模型.流式細胞儀檢測巨噬細胞的胞葬率,RT-qPCR和Western blot法檢測巨噬細胞ERK5和C1qA的mRNA和蛋白的表達.結(jié)果:ERK5抑制劑XMD8-92和ERK5-IN-1均可抑制RAW264.7細胞的胞葬作用,抑制ERK5的活化和C1q A mRNA和蛋白的表達.冠心康含藥血清可增強ox-LDL導致的受損的巨噬細胞胞葬作用,這一作用與冠心康活化ERK5和上調(diào)C1q A mRNA和蛋白的表達有關.結(jié)論:ERK5激酶的失活可通過下調(diào)C1qA的表達損傷巨噬細胞胞葬作用;冠心康可通過活化ERK5上調(diào)C1qA的表達,促進ox-LDL負載的巨噬細胞胞葬作用.

【文章頁數(shù)】：14 頁

【文章目錄】：
1 Materials and methods
    1.1 Materials
        1.1.1 Animals and cells
        1.1.2 Reagents and drugs instruments
    1.2 Experimental design
        1.2.1 Preparation of GXK decoctions and rat medicine serum
        1.2.2 Treatment of RAW264.7 cells
        1.2.3 Effect of ERK5 inhibitors on RAW264.7 cells
        1.2.4 Complement component C1q effect of ox-LDL on RAW264.7 macrophage efferocytosis
        1.2.5 Effect of medicated serum on ox-LDL-loaded macrophages
    1.3 Experimental methods
        1.3.1 Efferocytosis assay
        1.3.2 RT-qPCR
        1.3.3 Western Blot analysis
    1.4 Statistical analysis
2 Results
    2.1 ERK5 inhibitors suppressed macrophage efferocytosis in vitro
    2.2 ERK5 inhibitors suppressed ERK5 activation and expression of C1qA in macrophages
    2.3 Ox-LDL weakened macrophage efferocytosis
    2.4 GXK improved efferocytosis of ox-LDL-loaded macrophages by activating ERK5
    2.5 GXK upregulated C1qA expression via ERK5 activation in ox-LDL-loaded macrophages
3 Discussion
4 Conclusion



本文編號：3895587