TNFα1型受体敲除对小鼠急性后肢缺血的作用(英文)  

The protective effects of tumor necrosis factor α receptor 1 deletion on hindlimb ischemia

作　　者：蒋峻[1] 李长岭[1] 魏玲 王建安[3] 

机构地区：[1]浙江大学医学院附属邵逸夫医院心内科,杭州310016 [2]美国拿卡罗来纳医科大学病理系 [3]浙江大学医学院附属第二医院心内科

出　　处：《中华急诊医学杂志》2006年第3期197-202,共6页Chinese Journal of Emergency Medicine

摘　　要：目的研究肿瘤坏死因子α1型受体(tumornecrosisfactorαreceptor1,TNFαR1)信号通路在小鼠急性后肢缺血中的作用,并探讨其作用机制。方法通过结扎TNFαR1/和野生型小鼠股动静脉及其主要分支建立后肢急性缺血模型,激光多普勒测手术前后后肢血液灌流,TUNEL染色观察细胞凋亡,凝胶电泳观察DNA凋亡梯带,免疫蛋白印迹法测定Caspase3、Bax蛋白的表达。结果术后1天TNFαR1/组缺血侧后肢血液灌流显著高于野生型组,腓肠肌TUNEL阳性细胞显著低于野生型组。TNFαR1/组缺血评分则显著低于野生型组,两组的自发截肢率分别为50%、0%。TNFαR1/组DNA凋亡梯带、Caspase3和Bax蛋白表达减弱。结论TNFαR1敲除可抑制TNFα信号通路下游死亡相关蛋白的激活,减少细胞死亡和调亡,对急性后肢缺血有保护作用。Objective Signals in the tumor necrosis factor α （TNFα） pathway, including TNFα and tumor necrosis factor a receptor 1 （TNFR1）, are upregulatecl after acute hindlimb ischemia. It is unclear, however, whether this upregulation is a pathophysiological consequence or a compensatory response. In the present study, the effect of TNFR1 deletion in a mouse model of hindlimb ischemia was investigated. Methods Hindlimb isehemia was produced through ligation of the proximal and distal left femoral artery and its branches in TNFR1 knockout （TNFR1^-/-） and wild type （WT） mice. Laser Doppler blood flow measurement showed that limb perfusion was significantly higher in TNFR1^-/- mice than that in the WT mice at 1 day after operation. Results The calculated ischemie scores were （3.67±0.52） in WT group and （1.83±0.41） in TNFR1^-/- group （P 〈 0.05）. The rate of amputation was 50% in WT mice versus 0% in TNFR1^-/- mice. There were less TUNEL positive cells in the calf muscle of TNFR1^-/- mice than that of WT mice. Furthermore, DNA fragmentation was more obvious in WT mice. Western blot showed less expression of Bax and cleaved easpase 3 in TNFR1^-/- mice 1 day after isehemia, suggesting a reduced apoptotic process in the absence of TNFR1. Conclusion In mice with hindlimb isehemia, knockout of TNFR1 prevents the activation of death-related proteins down streaming to TNF α and attenuated cell death including apoptosis. Systemic block of the TNFR1 might be a new interventional methods to preserve the limbs from acute iscbemic attack in patients with peripheral arterial obstructive disease.

关 键 词：TNFΑ 后肢缺血 1型受体 急性 小鼠 Caspase-3 TUNEL阳性细胞 敲除 Bax蛋白表达 receptor 

分 类 号：R363[医药卫生—病理学]