TRPA1沉默抑制IL-1β诱导的软骨细胞炎性损伤和软骨基质降解-现代医学

TRPA1沉默抑制IL-1β诱导的软骨细胞炎性损伤和软骨基质降解

单位：1. 西安交通大学附属红会医院小儿骨科, 陕西西安 710054;
2. 西安交通大学附属红会医院手外科, 陕西西安 710054;
3. 甘肃省定西市人民医院骨二科, 甘肃定西 743000

关键词：骨关节炎软骨细胞瞬时感受器电位锚蛋白-1 炎症反应软骨退化 p38 MAPK信号通路

分类号：R684

出版年·卷·期（页码）：2019·47·第八期（1001-1006）

摘要：

目的：探讨瞬时感受器电位锚蛋白-1（TRPA1）沉默对IL-1β诱导的软骨细胞炎性损伤和软骨退化的影响。方法：小鼠软骨细胞ATDC5分为空白对照组、白细胞介素-1β（IL-1β）组、scramble组、siTRPA1组、IL-1β+scramble组和IL-1β+siTRPA1组。MTT法检测细胞活力，qRT-PCR检测肿瘤坏死因子-α（TNF-α）、白细胞介素6（IL-6）、基质金属蛋白酶-13（MMP-13）、解整链蛋白金属蛋白酶5（ADAMTS5）、Ⅱ胶原（Col Ⅱ）及蛋白聚糖（ACAN）mRNA的表达水平，Western blot检测细胞Col Ⅱ、ACAN及p38丝裂原活化蛋白激酶（MAPK）磷酸化（p-p38 MAPK）水平。结果：IL-1β刺激后，ATDC5细胞TRPA1 mRNA和蛋白的表达显著升高（P<0.05）。与空白对照组相比，IL-1β组中细胞活性明显下降，IL-6、TNF-α、MMP-13和ADAMTS5 mRNA表达显著升高，Col Ⅱ、ACAN mRNA和蛋白表达降低（P<0.05）；而与IL-1β组相比，IL-1β+siTRPA1组中细胞活性显著升高，IL-6、TNF-α、MMP-13和ADAMTS5 mRNA表达降低，Col Ⅱ、ACAN mRNA和蛋白表达增加（P<0.05）。此外，TRPA1沉默抑制IL-1β诱导的p-p38 MAPK蛋白表达的升高（P<0.05）。结论：TRPA1沉默能够抑制IL-1β诱导的软骨细胞炎症反应和软骨基质降解，其机制可能是通过阻断p38 MAPK信号通路。

Objective:To investigate the effect of TRPA1 silencing on IL-1β-induced chondrocyte inflammatory injury and cartilage matrix degeneration.Methods:Mice chondrocytes ATDC5 were divided into blank control group, IL-1β group, scramble group, siTRPA1 group, IL-1β+scramble group, and IL-1β+siTRPA1 group. Cell viability was evaluated by MTT assay. The mRNA level of TNF-α, IL-6, MMP-13, ADAMTS5, Col Ⅱ and ACAN was detected by qRT-PCR and the protein level of Col Ⅱ, ACAN and p-p38 MAPK was detected by Western blot.Results:After stimulation of IL-1β, the mRNA and protein expression of TRPA1 in ATDC5 cells were significantly increased (P<0.05). Compared with control blank group, the decreased cell viability, increased IL-6 and TNF-α level,upregulated mRNA level of MMP-13 and ADAMTS5,and downregulated mRNA and protein level of Col Ⅱ and ACAN were observed in IL-1β group (P<0.05). Cell viability and expression of Col Ⅱ and ACAN were obviously increased in IL-1β+siTRPA1 group, whereas the level of IL-6, TNF-α, MMP-13 and ADAMTS5 was markedly decreased in IL-1β+siTRPA1 group compared with IL-1β group (P<0.05). Furthermore, TRPA1 silencing inhibited the increased protein expression of p-p38 MAPK induced by IL-1β (P<0.05).Conclusion:TRPA1 silencing inhibits chondrocyte inflammation and cartilage degeneration induced by IL-1β, which may be through blocking p38 MAPK signaling pathway.

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