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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
作者:梁小菊1  张丽君2  成德亮2  梁小弟3 
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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