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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 To investigate the role of tumor necrosis factor α receptor 1 (TNFαR1) signaling pathway in acute hindlimb ischemia in mice and its mechanism of action. Methods Acute ischemia model of hindlimb was established by ligation of femoral artery and its main branches of TNFαR1 / wild type mice. Hepatic perfusion of hind limbs was measured by laser Doppler sonography. Cell apoptosis was observed by TUNEL staining. DNA apoptosis ladder was observed by gel electrophoresis. The expression of Caspase3 and Bax proteins were detected by Western blot. Results At 1 day after operation, the blood flow of ischemic hindlimb in TNFαR1 / group was significantly higher than that in wild type group, and the TUNEL positive cells in gastrocnemius muscle were significantly lower than those in wild type group. TNFαR1 / group ischemia score was significantly lower than the wild-type group, spontaneous amputation rates were 50%, 0%. TNFαR1 / DNA apoptotic ladder, Caspase3 and Bax protein expression decreased. Conclusion TNFαR1 knockdown can inhibit the activation of death-related proteins downstream of TNFα signaling pathway, decrease cell death and apoptosis, and protect against acute hindlimb ischemia.