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Dual regulatory roles of HMGB1 in inflammatory reaction of chondrocyte cells and mice

机译：HMGB1对软骨细胞和小鼠炎症反应的双重调节作用

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摘要

Osteoarthritis (OA) is one of the most common bone diseasesas it is reported that the impact of knee osteoarthritis symptomatic form is estimated at 240/100,000 people per year. The inflammation of articular cartilageis thought to be the pathologic drive for development of this disease. HMGB1(high mobility group box-1), a regulatory factor for gene transcription, could stimulate inflammation response. However, theexact regulatory role of HMGB1 in the inflammation of articular cartilage still need to be elucidated. In the current study, we used Quantitative Real-Time PCR(Q-PCR) to detect them RNA levels of Collagen Type II Alpha 1(Col2a1), Aggrecan, MMP3(Matrix Metallopeptidase 3), MMP13, ADAMTs4 and ADAMTs5; Enzyme-Linked Immunosorbent Assay(ELISA) was used to detect the content of IL-1 beta and calpain protein; Cell apoptosis was evaluated by terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end labeling(TUNEL) assay and flow cytometryanalysis; Western blot and immunofluorescence assays were applied to assess the expression of HMGB1; Lastly autophagic activity was mainly verified by monodansylcadaverine (MDC) staining. Our data revealed that in the early stage of chondrocyte inflammation(3 and 6 h of LPS stimulation), cytosolic HMGB1 attenuated inflammation response by facilitating cell autophagy and preventing cell apoptosis. While in the late stage (24 and 48 h of LPS stimulation), the extracellular HMGB1 stimulated inflammation reaction and contributed to the cartilage destruction in OA.
机译：骨关节炎（OA）是最常见的骨质疾病之一，据报道，膝关节骨关节炎的影响症状估计每年240 / 100,000人。关节软骨炎症认为是这种疾病发展的病理驱动。 HMGB1（高迁移率组箱-1），基因转录的调节因子，可以刺激炎症反应。然而，HMGB1在关节软骨炎症中的另一种调节作用仍然需要阐明。在目前的研究中，我们使用定量实时PCR（Q-PCR）来检测胶原IIα1（COL2A1），聚集体，MMP3（基质金属肽酶3），MMP13，ADAMTS4和ADAMTS5的RNA水平。酶联免疫吸附试验（ELISA）用于检测IL-1β和CALPAIN蛋白的含量;通过末端脱氧核苷酸转移酶介导的DUTP-BIOTIN碎片末端标记（TUNEL）测定和流式细胞含量分析来评估细胞凋亡。施用蛋白质印迹和免疫荧光测定以评估HMGB1的表达;最后是通过单丹二烷基酰胺（MDC）染色来验证的自噬。我们的数据显示，在软骨细胞炎症的早期阶段（3和6小时LPS刺激），通过促进细胞自噬和预防细胞凋亡，细胞溶质HMGB1减弱炎症反应。而在晚期（24和48小时的LPS刺激）中，细胞外HMGB1刺激炎症反应，并导致OA中的软骨破坏。

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  • 来源
    《Cell cycle》 |2019年第18期|共13页
  • 作者

    Li Wenzhao; Ni Jiangdong; Song Deye; Ding Muliang; Wang Junjie; Huang Xianzhe; Yan Mingming; Huang Jun;

  • 作者单位

    Cent S Univ Xiangya Hosp 2 Dept Orthoped Changsha Hunan Peoples R China;

    Cent S Univ Xiangya Hosp 2 Dept Orthoped Changsha Hunan Peoples R China;

    Cent S Univ Xiangya Hosp 2 Dept Orthoped Changsha Hunan Peoples R China;

    Cent S Univ Xiangya Hosp 2 Dept Orthoped Changsha Hunan Peoples R China;

    Cent S Univ Xiangya Hosp 2 Dept Orthoped Changsha Hunan Peoples R China;

    Cent S Univ Xiangya Hosp 2 Dept Orthoped Changsha Hunan Peoples R China;

    Cent S Univ Xiangya Hosp 2 Dept Orthoped Changsha Hunan Peoples R China;

    Cent S Univ Xiangya Hosp 2 Dept Orthoped Changsha Hunan Peoples R China;

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  • 原文格式 PDF
  • 正文语种 eng
  • 中图分类 细胞生物学;
  • 关键词

    Osteoarthritis; HMGB1; inflammation; Col2a1; autophagy; apoptosis;

    机译：骨关节炎;HMGB1;炎症;COL2A1;自噬;细胞凋亡;

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