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、si TRPA1组、IL-1β+scramble组和IL-1β+si TRPA1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ADAMTS5mRNA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+si TRPA1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-1βgroup,scramble group,si TRPA1 group,IL-1β+scramble group,and IL-1β+si TRPA1 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-αlevel,upregulated mRNA level of MMP-13 and ADAMTS5,and downregulated mRNA and protein level of Col II and ACAN were observed in IL-1βgroup(P<0.05).Cell viability and expression of Col II and ACAN were obviously increased in IL-1β+si TRPA1 group,whereas the level of IL-6,TNF-α,MMP-13 and ADAMTS5 was markedly decreased in IL-1β+si TRPA1 group compared with IL-1βgroup(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.
作者
梁小菊
张丽君
成德亮
梁小弟
LIANG Xiaoju;ZHANG Lijun;CHENG Deliang;LIANG Xiaodi(Department of Pediatric Orthopedics,Red Cross Hospital,Xi’an Jiaotong University,Xi’an 710054,China;Department of Hand Surgery,Red Cross Hospital,Xi’an Jiaotong University,Xi’an 710054,China;Department of Orthopaedics,Dingxi City People’s Hospital,Dingxi 743000,China)
出处
《现代医学》
2019年第8期1001-1006,共6页
Modern Medical Journal
