Toll样受体信号通路活化与皮瓣缺血再灌注损伤的关系

Relationship between the activation of the oll-like receptor signaling pathway and the ischemia reperfusion injury of the skin flap

目的探讨Toll样受体2(TLR2)、Toll样受体3(TLR3)和Toll样受体4(TLR4)在大鼠皮瓣缺血再灌注损伤模型中的动态表达及介导病程进展中的作用。方法制备大鼠皮瓣缺血再灌注损伤模型,根据再灌注的时间不同分5组(6 h、12 h、24 h、48 h、72 h),同时制备假手术组,每组6只大鼠,采用免疫组化染色和Western blot检测各组大鼠皮瓣组织中TLR2、TLR3和TLR4的表达,同时检测Toll样受体信号通路下游炎性因子TNF-α、IL-6的表达,分析其相关性。结果 HE染色显色在缺血的早期,皮瓣组织呈现不同程度的坏死,在恢复血液灌注后,坏死没有明显好转,大量胶原纤维增生;免疫组化染色显色,与假手术组比较,TLR2、TLR3和TLR4的表达均有不同程度的升高,在再灌注24 h时TLR2和TLR4蛋白表达达高峰,随后逐渐下降,而TLR3在再灌注12 h即达高峰,随后降低,各时间段比较差异有统计学意义(P<0.05);炎性因子TNF-α、IL-6的血清浓度明显高于假手术组,且在再灌注24 h时浓度最高(P<0.05),随后逐渐下降,到72h没有再次升高趋势。结论在皮瓣缺血再灌注早期,皮瓣组织Toll样受体蛋白即迅速升高,同时通过促进下游炎性因子的分泌而介导组织损伤。

Objective To study theTLR2, TLR3 and TI,R4 of in rats in the model of ischemia reperfusion injury. Methods The rats model of flap ischemia-reperfusion injury were made, according to the time of reperfusion, it were divided into 5 groups （ 6 h, 12 h, 24h, 48 h, 72 h）. 6 rats using immunohistochemistry and Western Mot test in the rat of the expression of TLR2, TLR3 and TLR4, while detecting toll-like receptors signaling pathways downstream of inflammatory factor of TNF-α and IL-6. Results HE staining showed different degree of necrosis in the early stage of ischemia. After the hlood peffusion was restored, there was no ohvious improvement in necrosis, and a large number of collagen fibers proliferated. hnmunohistochemical staining show that , comparing to control group , the expression of TLR2 , TLR3 and TLR4 had increased, and in the 24 hours of repeffusion, TLR2 and TLR4 protein expression had to peak and then gradually decline, while TLR3 were in 12 h to peak, then down, each time period is statistically significant difference （P〈0.05） ; The serum concentrations of TNF-α and II-6 were significantly higher than those of the sham operation group, and the concentration was highest at 24 hours, then gradually decreased, and the trend was not increased again in 72 hours. Conclusion In the early stage of ischemia and repeffusion of skin flap, the flap was rapidly elevated, and the tissue damage was mediated hy promoting the secretion of downstream inflammatory factors.