Abstract:

Objective    To study condyle chondrocyte cell death under circumstances of high TNF-α concentration. Methods    Primary human condyle chondrocytes were cultured in vitro and then 20 μg/L TNF-α was added to induce cell death （T group）. The rate of apoptotic and necrotic cell death were detected by flow cytometry. Before application of TNF-α，chondrocytes were pre-treated with pan-caspase inhibitor Z-VAD-FMK （ZT Group）， or specific programmed necrosis inhibitors Nec-1 （NT group）， or the combination of these two inhibitors （ZNT group）. CM-H2DCFDA staining was used to determine ROS levels. Cell death rates and ROS levels were respectively compared and statistically analyzed. Results    T group showed exacerbated cell death and increased ROS level in remaining living cells ，while cell death and ROS levels of ZT， NT and ZNT groups were significantly alleviated compared to T group （P < 0.05）. Amongst these three groups， ZNT group achieved the lowest cell death rate and ROS levels. Conclusion    Under circumstances of high TNF-α concentration， condylar chondrocytes could go through both apoptosis and necropotosis， and inhibiting both forms of programmed cell death pathways can significantly improve cell survival rate.

Key words: TNF-&alpha, ；necroptosis；condylar chondrocyte；ROS

摘要：

目的    探讨高浓度肿瘤坏死因子-α（TNF-α）环境下髁突软骨细胞的死亡情况。方法    临床收集2012年9月至2014年8月就诊于上海交通大学医学院附属第九人民医院口腔外科的髁突骨折患者无法复位的骨折片段上的软骨组织，体外培养人髁突软骨细胞，加入20 μg/L TNF-α后流式细胞仪分析细胞死亡情况（T组），分别与加入泛caspase抑制剂Z-VAD-FMK（ZT组）、特异性程序性坏死抑制剂Nec-1（NT组）和联合应用抑制剂（ZNT组）的细胞死亡情况进行比较和统计学分析。结果    T组细胞大量死亡，剩余活细胞中活性氧（ROS）水平升高；ZT、NT和ZNT组细胞死亡和ROS水平显著低于T组（P < 0.05），ZNT组细胞死亡和ROS水平最低。结论    高浓度TNF-α刺激下髁突软骨细胞的死亡类型有凋亡和程序性坏死，同时抑制二者可以显著提高软骨细胞的存活率。

关键词: 肿瘤坏死因子-&alpha, 凋亡, 程序性坏死, 髁突软骨细胞, 活性氧