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目的分析免疫性血管炎家兔血清可溶性CD40配体(sCD40L)水平变化,探讨其在血管炎发病机制中的意义及药物丹参酮ⅡA和阿司匹林治疗对其水平的影响。方法日本大耳幼年家兔48只分为正常对照组、模型对照组、丹参酮ⅡA治疗组和阿司匹林治疗组。牛血清清蛋白静脉推注诱导法建立免疫性血管炎模型。检测其外周血小板数量、血小板聚集功能,采用流式细胞仪检测其血小板活性标志Annexin V,ELISA双抗体夹心法检测治疗第7天及第28天各组血清sCD40L水平。HE染色、弹力纤维染色和电镜观察其血管病变。结果免疫性血管炎家兔急性期血清sCD40L水平较正常对照组显著升高(P<0.05),血小板数量、血小板聚集功能及Annexin V检测的血小板活性也显著增高。血清sCD40L水平与血小板数量、血小板聚集功能和血小板活性呈正相关。丹参酮ⅡA和阿司匹林治疗均可显著降低sCD40L水平,降低血小板数量和活性,并能显著改善免疫性血管炎家兔的血管病理损害。恢复期模型对照组家兔血小板数量、血小板聚集功能和活性已恢复正常,但血清sCD40L水平仍显著高于正常对照组。结论CD40-CD40L信号系统可能参与了免疫性血管炎的发病机制,丹参酮ⅡA和阿司匹林可能通过下调sCD40L水平及降低血小板活性从而达到抗感染、抗凝作用。
Objective To analyze the changes of serum soluble CD40 ligand (sCD40L) level in rabbits with immune vasculitis, and to explore its significance in pathogenesis of vasculitis and the effects of tanshinone ⅡA and aspirin on its level. Methods 48 young rabbits were divided into normal control group, model control group, tanshinone Ⅱ A treatment group and aspirin treatment group. Bovine serum albumin intravenous injection induced immune vasculitis model. The number of peripheral platelets and platelet aggregation were measured. The activity of Annexin V was detected by flow cytometry. The level of sCD40L in serum of each group was detected by double antibody sandwich enzyme-linked immunosorbent assay (ELISA) on day 7 and day 28. HE staining, elastic fiber staining and electron microscopy of vascular lesions. Results The level of sCD40L in acute phase of immune vasculitis in rabbits was significantly higher than that in normal control group (P <0.05). The number of platelets, platelet aggregation and the activity of Annexin V were also significantly increased. Serum sCD40L levels and the number of platelets, platelet aggregation and platelet activity was positively correlated. Tanshinone IIA and aspirin treatment can significantly reduce sCD40L levels, reduce platelet number and activity, and can significantly improve the immune vasculitis in rabbits vascular pathological damage. In the convalescent model rabbits, the platelet count, platelet aggregation function and activity returned to normal in the control group, but the level of sCD40L in serum was still significantly higher than that in the normal control group. Conclusion CD40-CD40L signaling system may be involved in the pathogenesis of immune vasculitis. Tanshinone IIA and aspirin may achieve anti-infective and anticoagulant effects by down-regulating sCD40L levels and decreasing platelet activity.