Harpagoside suppresses IL-6 expression in primary human osteoarthritis chondrocytes.
Harpagoside suppresses IL-6 expression in primary human osteoarthritis chondrocytes.
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DOI：
10.1002/jor.23262
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发表时间：
2017-02
影响因子：
2.8
通讯作者：
Haqqi, Tariq M.
中科院分区：
文献类型：
--
作者：
Haseeb, Abdul;Ansari, Mohammad Yunus;Haqqi, Tariq M.
越来越多的证据表明炎症反应参与了骨关节炎的发病机制。哈帕皂苷是哈帕根草的生物活性成分之一，具有抗炎活性。在这里，我们使用体外炎症模型来研究哈帕皂苷抑制炎性细胞因子/趋化因子如IL-6和基质降解酶的产生的可能性。我们进一步研究了哈帕皂苷在原代人骨性关节炎软骨细胞中的可能靶点。在用IL-1β刺激前，先用哈巴豆苷处理骨性关节炎软骨细胞。应用TaqMan人趋化因子聚合酶链式反应芯片检测92种细胞因子/趋化因子的基因表达谱。用TaqMan实验证实了所选mRNAs的表达水平。用ELISA法和免疫印迹法检测IL-6和MMP13的蛋白水平。免疫印迹法检测细胞总蛋白水平和信号蛋白的磷酸化水平。免疫荧光和共聚焦显微镜观察IL-6和c-Fos的细胞定位。用酶联免疫吸附试验检测c-fos/AP-1的DNA结合活性。哈帕糖苷显著改变了IL-1β刺激的骨性关节炎软骨细胞的全局趋化因子表达谱。IL-1β可显著诱导骨性关节炎软骨细胞IL-6的表达，而哈帕苷可显著抑制IL-6的表达。哈巴豆苷不能抑制IL-1诱导的核转录因子κB和C/EBPDNA转录因子的激活，但可抑制AP-1转录因子的主要成分c-fos在IL-1β诱导下的磷酸化和β结合活性。此外，在病理条件下，哈帕皂苷显著抑制了骨关节炎软骨细胞中基质金属蛋白酶-13的表达。SiRNA介导的IL-6基因敲除导致基质金属蛋白酶-13的表达和分泌受到抑制，直接将IL-6的作用与基质金属蛋白酶-13的表达联系起来。综上所述，本研究提示哈帕皂苷通过抑制病理条件下骨关节炎软骨细胞c-fos/AP-1的活性而发挥显著的抗炎作用。
There is growing evidence in support of the involvement of inflammatory response in the pathogenesis of osteoarthritis (OA). Harpagoside, one of the bioactive components of Harpagophytum procumbens (Hp), has been shown to possess anti-inflammatory properties. Here we used an in vitro model of inflammation in OA to investigate the potential of harpagoside to suppress the production of inflammatory cytokines/chemokines such as IL-6 and matrix degrading proteases. We further investigated the likely targets of harpagoside in primary human OA chondrocytes. OA chondrocytes were pre-treated with harpagoside before stimulation with IL-1β. mRNA expression profile of 92 cytokines/chemokines was determined using TaqMan Human Chemokine PCR Array. Expression levels of selected mRNAs were confirmed using TaqMan assays. Protein levels of IL-6 and MMP-13 were assayed by ELISA and immunoblotting. Total protein levels and phosphorylation of signaling proteins were determined by immunoblotting. Cellular localization of IL-6 and c-Fos was performed by immunofluorescence and confocal microscopy. DNA binding activity of c-FOS/AP-1 was determined by ELISA. Harpagoside significantly altered the global chemokine expression profile in IL-1β-stimulated OA chondrocytes. Expression of IL-6 was highly induced by IL-1β, which was significantly inhibited by pre-treatment of OA chondrocytes with harpagoside. Harpagoside did not inhibit the IL-1-induced activation of NF-κB and C/EBPβ transcription factors but suppressed the IL-1β-triggered induction, phosphorylation and DNA binding activity of c-FOS, one of the main components of AP-1 transcription factors. Further, harpagoside significantly inhibited the expression of MMP-13 in OA chondrocytes under pathological conditions. siRNA-mediated knockdown of IL-6 resulted in suppressed expression and secretion of MMP-13 directly linking the role of IL-6 with MMP-13 expression. Taken together, the present study suggests that harpagoside exert a significant anti-inflammatory effect by inhibiting the inflammatory stimuli mediated by suppressing c-FOS/AP-1 activity in OA chondrocytes under pathological conditions.
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DOI：
10.1016/j.clim.2012.12.011
发表时间：
2013-03
期刊：
Clinical immunology (Orlando, Fla.)
影响因子：
--
作者：
Haseeb A;Haqqi TM
通讯作者：
Haqqi TM
DOI：
10.1073/pnas.251194298
发表时间：
2001-11-20
影响因子：
11.1
作者：
Bennett, BL;Sasaki, DT;Anderson, DW
通讯作者：
Anderson, DW
影响因子：
5.5
作者：
Haseeb, Abdul;Chen, Dongxing;Haqqi, Tariq M.
通讯作者：
Haqqi, Tariq M.
影响因子：
3.3
作者：
Inaba, Kazunori;Murata, Kazuya;Matsuda, Hideaki
通讯作者：
Matsuda, Hideaki
影响因子：
158.5
作者：
Felson, DT
通讯作者：
Felson, DT