过表达或抑制IL-18对大鼠深静脉血栓形成的影响
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国家自然科学基金面上项目(30960389);国家自然基金资助项目(81060151);云南省卫生科技内设研究机构项目(2012ws0007);云南省科技厅重点新产品开发计划项目(2010BC010)


Effects of IL-18 overexpression/inhibition on deep vein thrombosis formation in rats
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    摘要:

    目的:探讨白介素(interleukin,IL)-18对大鼠深静脉血栓(deep vein thrombosis,DVT)模型血栓形成的影响。方法:构建IL18-pCDH-GFP- IL18-LMP-shRNAmir1病毒载体。SD大鼠(n=27)随机均分为3组。过表达组:尾静脉注入IL-18慢病毒过表达载体(IL18-pCDH-GFP,100 μl);抑制组:注入IL-18逆转录病毒抑制载体(IL18-LMP-shRNAmir1,100 μl);对照组:注入无菌生理盐水(100 μl)。注射24 h后,SD大鼠行狭窄法下腔静脉(inferior vena cava,IVC)血栓造模,造模后24 h解剖观察IVC血栓形成情况,并取材称量血栓重量及长度;血栓静脉管壁行实时荧光定量PCR检测IL-18表达情况。结果:IL18-pCDH-GFP- IL18-LMP-shRNAmir1病毒载体的体外细胞实验具备理想的过表达率及抑制率。各组大鼠造模后24 h可见稳定血栓形成。IL-18过表达组的平均成栓长度和重量较IL-18抑制组-对照组明显增加(P < 0.05),real-time PCR结果显示过表达组中IL-18在大鼠静脉壁表达量明显增加(F=3.784,P < 0.05)。结论:IL-18表达量增加,对大鼠DVT造模后IVC血栓形成有促进作用,而表达减少亦对成栓有影响,其表达状况与深静脉血栓的发生-发展有关,IL-18介导的促炎反应在静脉血栓疾病发病机制中可能起重要作用。

    Abstract:

    Objective:To investigate the influence of interleukin-18 (IL-18) on the rat deep vein thrombosis (DVT). Methods:Viral vectors of IL18-pCDH-GFP/IL18-LMP-shRNAmir1 were constructed. SD rats (n=27) were randomly divided into IL-18 overexpression group,IL-18 inhibition group and control group which were injected with 100 μl IL-18 overexpression lentivectors (IL18- pCDH-GFP),100 μl IL-18 inhibition retroviral vectors (IL18-LMP-shRNAmir1),100 μl saline,respectively,into the tail vein. All rats’ inferior vena cavas (IVC) were modeled as “stenosis” to promote IVC thrombosis after 24 h of injection. The weight and length of IVC thrombosis after 24 h of modeled were investigated,and the expression of IL-18 in the venous wall was measured by real-time PCR assay. Results:IL18-pCDH-GFP and IL18-LMP-shRNAmir1 vectors showed the ideal overexpression/inhibition rate in vitro. All groups had stabilized thrombus formation after modeled 24 h. The average length and weight of thrombus in the IL-18 overexpression group were significantly higher than those of other groups (P < 0.05). The level of IL-18 in the overexpression group was remarkably higher than that of other groups in the vessel wall (F=3.784,P < 0.05),which was proved by real-time PCR assay. Conclusion:IL-18 promotes thrombus formation in rats,and inhibition of IL-18 reduces the thrombus formation. IL-18 may be related to the development process of DVT and its proinflammatory effect may play a vital role in the pathogenesis of VTE.

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李光第,白云城,宋 恩,周如丹,王 扬,王宫泽,卜鹏飞,赵学凌.过表达或抑制IL-18对大鼠深静脉血栓形成的影响[J].南京医科大学学报(自然科学版),2015,(3):367-372

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  • 收稿日期:2014-08-11
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  • 在线发布日期: 2015-03-10
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