TNF-α在构建大鼠骨性关节炎模型中对内质网应激的影响探讨

发布时间：2018-06-12 16:25

本文选题：肿瘤坏死因子-α + 骨性关节炎　；参考：《中国骨质疏松杂志》2017年10期

【摘要】：目的观察不同剂量肿瘤坏死因子-α(tumor necrosis factor,TNF-α)在构建大鼠骨性关节炎(osteoarthritis,OA)模型中对内质网应激的影响,初步探讨大鼠骨性关节炎病理过程中内质网应激的作用。方法将20只SPF级SD雄性大鼠随机分为健康对照组、TNF-α1ng/m L干预组、TNF-α5ng/m L干预组和TNF-α10ng/m L干预组,分别给予不同剂量TNF-α处理,4周后取材,培养提取的各组原代软骨细胞,采用MTT法检测不同剂量TNF-α对软骨细胞活力的影响;ELISA法检测内皮细胞粘附分子-1(endothelial cell adhesion molecule-1,s ICAM-1)含量变化,TUNEL法检测各组软骨细胞凋亡率,蛋白印迹法(Western blot)检测炎症因子MMP9、ICAM1的表达以及内质网应激指标IRE1、p-IRE1、GRP78的水平变化。结果 MTT法检测提示加入不同剂量的TNF-α后软骨细胞活力未见明显改变;TUNEL法检测结果提示,与对照组相比,TNF-α干预组软骨细胞凋亡率较高(P0.01),并呈浓度依赖性促进细胞凋亡;Western blot结果表明,与对照组相比,软骨细胞的p-IRE1、IRE1、GRP78、MMP9及ICAM1的表达水平呈TNF-α浓度依赖性升高(P0.01)。结论在TNF-α构建大鼠骨性关节炎模型中,炎症因子与内质网应激指标均发生显著改变,内质网应激可能在大鼠骨性关节炎的发生发展过程中发挥重要影响,且当TNF-α浓度为10 ng/m L时,其对炎性因子及内质网指标的作用效果最为显著。
[Abstract]:Objective to observe the effects of different doses of tumor necrosis factor- 伪 on endoplasmic reticulum stress in rat osteoarthritis model, and to explore the role of endoplasmic reticulum stress in the pathological process of rat osteoarthritis. Methods Twenty Sprague-SPF male rats were randomly divided into two groups: TNF- 伪 1ng/m L intervention group and TNF- 伪 10ng/m L intervention group. The primary chondrocytes of each group were cultured after 4 weeks of treatment with different doses of TNF- 伪 10ng/m. The effect of different doses of TNF- 伪 on chondrocyte viability was detected by MTT assay. The changes of the content of endothelial cell adhesion molecule cell adhesion molecule-1and ICAM-1 were detected by Elisa. The apoptosis rate of chondrocytes in each group was detected by Tunel method. Western blot was used to detect the expression of ICAM1 and the level of endoplasmic reticulum stress (IRE1) -IRE1 / GRP78. Results MTT assay showed that the activity of chondrocytes did not change after adding different doses of TNF- 伪. Compared with the control group, the apoptotic rate of chondrocytes in the TNF- 伪 intervention group was higher than that in the control group. The results showed that the expression of p-IRE1, IRE1, GRP78, MMP9 and ICAM1 in chondrocytes was increased in a concentration-dependent manner in a concentration dependent manner (P0.01a), and the results showed that the expression levels of p-IRE1, IRE1, GRP78, MMP9 and ICAM1 in chondrocytes were increased in a concentration-dependent manner. Conclusion the inflammatory factors and endoplasmic reticulum stress were significantly changed in rat osteoarthritis induced by TNF- 伪. Endoplasmic reticulum stress may play an important role in the development of osteoarthritis in rats. When the concentration of TNF- 伪 was 10 ng/m / L, the effect of TNF- 伪 on inflammatory factors and endoplasmic reticulum was the most significant.
【作者单位】：兰州军区总医院全军骨科中心关节外科;宁夏医科大学;第四军医大学西京医院老年病科;
【基金】：国家自然科学基金(81371983) 甘肃省青年科技基金(1606RJYA300) 甘肃省自然科学基金(1606RJZA208) 甘肃省科技支撑计划(S04671)
【分类号】：R-332;R684.3

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