双氢青蒿素通过抑制炎症反应在血管重塑中的作用机制研究

目的 探究双氢青蒿素通过抑制炎症反应对血管重塑的抑制作用。方法 采用贴块法培养小鼠血管平滑肌细胞(vascular smooth muscle cells，VSMCs)，以TNF-α诱导建立体外平滑肌细胞增殖模型，通过手术剥夺动脉方式建立小鼠股动脉损伤模型，并采用双氢青蒿素干预细胞及股动脉损伤小鼠。采用CCK-8法检测细胞增殖；H&E染色评价股动脉组织病理变化情况并测量中膜厚度；ELISA法检测细胞中IL-1β、TNF-α、IL-6、CCR8及小鼠股动脉组织中IL-1β、TNF-α、IL-6、TGF-β1的含量；免疫组织化学法检测股动脉组织NF-κB p65、CCR8表达；qRT-PCR分析细胞及股动脉组织中IL-1β、TNF-α、IL-6、IL-8、CCR8、NF-κB p65、TGF-β1、MCP-1 mRNA表达；Western blotting分析细胞及股动脉组织中IL-1β、IL-6、TNF-α、IL-8、p-NF-κB p65、NF-κB p65、CCR8、TGF-β1、MCP-1蛋白表达。结果 在TNF-α诱导的VSMCs增殖及股动脉损伤小鼠模型中，VSMCs增殖及TNF-α、IL-6含量明显升高，IL-1β、IL-6、IL-8 mRNA表达明显升高，IL-6、CCR8、NF-κB p65蛋白表达明显升高(P<0.05或P<0.01)；双氢青蒿素可明显降低TNF-α诱导的VSMCs中IL-1β、IL-8 mRNA及股动脉损伤小鼠IL-1β mRNA表达和IL-1β、p-NF-κB p65蛋白表达(P<0.05或P<0.01)。结论 双氢青蒿素能降低血管炎症，延缓损伤血管的重塑，表明其为经皮冠状动脉介入治疗(如支架植入)的潜在治疗药物。

Study on the Mechanism of Dihydroartemisinin in Vascular Remodeling by Inhibiting Inflammation

OBJECTIVE To explore the inhibitory effect of dihydroartemisinin on vascular remodeling by inhibiting inflammation. METHODS The patch method was used to culture mouse vascular smooth muscle cells(VSMCs), an in vitro smooth muscle cell proliferation model was established by TNF-α induction, a mouse femoral artery injury model was established by surgical deprivation of the artery, and dihydroartemisinin was used to interfere with cell and femoral artery injury in mice. The cell proliferation was detected by CCK-8 method, the pathological changes of femoral artery was evaluated by H&E staining and the thickness of media was measured, and the contents of IL-1β, TNF-α, IL-6, CCR8 in cells and IL-1β, TNF-α, IL-6, TGF-β1 in mouse femoral artery tissue were detected by ELISA method. The expression of NF-κB p65 and CCR8 in femoral artery tissue was detected by immunohistochemical method, and the expression of IL-1β, TNF-α, IL-6, IL-8, CCR8, NF-κB p65, TGF-β1 and MCP-1 mRNA in cells and femoral artery tissue was analyzed by qRT-PCR. Western blotting was used to analyze the expression of IL-1β, IL-6, TNF-α, IL-8, p-NF-κB p65, NF-κB p65, CCR8, TGF-β1, MCP-1 protein in cells and femoral artery tissue. RESULTS In the models of TNT-α-induced VSMCs proliferation and femoral artery injury of mice, the proliferation of VSMCs, the content of TNF-α and IL-6, the expression of IL-1β, IL-6, IL-8, NF-κB p65 and TGF-β1 mRNA were significantly increased, and the expression of IL-6, CCR8 and NF-κB p65 protein was significantly increased(P<0.05 or P<0.01). Dihydroartemisinin could significantly reduce the expression of IL-1β and IL-8 mRNA of VSMCs induced by TNF-α, and reduce the expression of IL-1β mRNA and IL-1β, p-NF-κB p65 protein of femoral artery injury in mice(P<0.05 or P<0.01). CONCLUSION Dihydroartemisinin can reduce vascular inflammation and delay remodeling of injured vessels, indicating that dihydroartemisinin can be used as a potential therapeutic drug for percutaneous coronary intervention(such as stent implantation).