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目的了解肿瘤坏死因子(TNF)相关的凋亡诱导配体(TRAIL)及其受体在严重烧伤大鼠胸腺组织细胞异常凋亡中的作用。方法将50只Wistar大鼠随机分为假伤组(模拟烧伤)10只和烧伤组40只(设伤后4、12、24、48 h 4个时相点)。应用膜联蛋白A5-异硫氰酸荧光素/碘化丙啶双染法,观察大鼠胸腺组织中细胞凋亡的情况;反转录-聚合酶链反应和蛋白质印迹法检测TRAIL的死亡受体5(DR5)、DR4、诱骗受体1(DcR-1)、DcR2在大鼠胸腺组织中的表达。结果与假伤组大鼠细胞凋亡率[(6.7±0.8)%]比较,烧伤组于伤后4 h[(17.1±0.4)%]起增高,12 h时[(25.2±1.1)%]达高峰,48 h时仍明显高于假伤组(P<0.05)。烧伤组大鼠胸腺组织中DR5的表达显著高于假伤组,DcR2的表达则显著低于假伤组;其余受体的表达组问相似。结论严重烧伤后早期大鼠胸腺组织的细胞凋亡明显增加,且胸腺组织中DR5和DcR2的表达异常,提示TRAIL凋亡途径可能参与了病理性细胞凋亡过程。
Objective To investigate the role of tumor necrosis factor (TNF) related apoptosis inducing ligand (TRAIL) and its receptor in abnormal apoptosis of thymocytes in severely burned rats. Methods Fifty Wistar rats were randomly divided into sham group (simulated burns) 10 and burned group 40 (4, 12, 24, 48 h after injury). Using Annexin A5-fluorescein isothiocyanate / propidium iodide double staining method to observe the apoptosis of rat thymus tissue; reverse transcription-polymerase chain reaction and Western blot analysis of TRAIL death by Body 5 (DR5), DR4, decoy receptor 1 (DcR-1), DcR2 expression in rat thymus tissue. Results Compared with the sham injury group, the apoptotic rate in the burn group was significantly higher than that in the sham injury group [(6.7 ± 0.8)%], and the burn group increased from 4 h [(17.1 ± 0.4)%] [(25.2 ± 1.1)%] Reaching the peak at 48 h still significantly higher than the sham group (P <0.05). The expression of DR5 in the thymus tissue of burn group was significantly higher than that of the sham group, while the expression of DcR2 was significantly lower than that of the sham injury group. The expressions of other receptors were similar. Conclusion The apoptotic rate of thymus in thymus rats increased significantly after severe burns and the expression of DR5 and DcR2 in thymus was abnormal, suggesting that TRAIL apoptotic pathway may be involved in the pathological apoptosis.