Selective regulation of IKKβ/NF-κB pathway involved in proliferation inhibition of HFLS-RA cells induced by 1,7-dihydroxyl-3,4-dimethoxylxanthone
Selective regulation of IKKβ/NF-κB pathway involved in proliferation inhibition of HFLS-RA cells induced by 1,7-dihydroxyl-3,4-dimethoxylxanthone
复制标题
DOI：
10.1016/j.kjms.2017.06.015
复制
发表时间：
2017-10-01
影响因子：
3.3
通讯作者：
Zuo, Jian
中科院分区：
文献类型：
--
作者：
Ji, Cong-Lan;Jiang, Hui;Zuo, Jian
类风湿性关节炎是一种常见的自身免疫性疾病，但现有治疗方案对其远期预后影响不大。本研究的目的是研究1，7-二羟基-3，4-二甲双胍-口山酮（XAN）在HFLS-RA细胞中的潜在作用，并描述诱导NF-κ B活性的潜在机制。MTT法检测细胞活力。采用流式细胞术评估促凋亡作用。通过RT-qPCR、Western-blot和免疫荧光方法研究对NF-kB信号传导的调节。结果表明，XAN可诱导HFLSRA细胞增殖抑制和凋亡，且呈浓度依赖性，Pyrrolidinedithiocarbamic acid可增强XAN诱导HFLSRA细胞增殖抑制和凋亡的作用，IKK16可拮抗XAN诱导HFLSRA细胞增殖抑制和凋亡的作用。XAN处理后，NF-κ B B信号通过mRNA表达、磷酸化和核转位等途径被阻断，导致p38蛋白表达上调，X连锁凋亡抑制蛋白表达下调。同时表达p-IKKb、p-IkB和p-p65提示IKK β介导NF-κ B的调节。同时，XAN促进IKK α的表达，这可能与上调的裂解PARP所提示的促凋亡作用有关。这些结果表明IKK β/NF-κ B介导了XAN对HFLS-RA细胞增殖的抑制作用，且IKK β/NF-κ B的不同调节可能对细胞增殖有协同作用。版权所有（C）2017，高雄医学大学.
Rheumatoid arthritis is a common autoimmune disease, however, available regimes exert little influence on it's long-term prognosis. The aim of the current study is to investigate potential effects of 1,7-dihydroxyl-3,4-dimethoxyl-xanthone (XAN) in HFLS-RA cells and describe the underlying mechanisms of induction of NF-kappa B activity. Viability of cells was measured by MTT assay. Flow cytometry was employed to assess the pro-apoptotic effects. Modulation on NF-kB signaling was investigated by RT-qPCR, Western-blot and immunofluorescence methods. It was found that XAN induced proliferation inhibition and apoptosis of HFLSRA cells in the concentration-dependent manner, which were strengthened by pyrrolidinedithiocarbamic acid but antagonized by IKK16. NF-kappa B signaling was abrogated shortly after the treatment of XAN via various means including mRNA expression, phosphorylation and nuclear translocation, which leaded to up-regulation of p38 and down-regulation of X-linked inhibitor of apoptosis protein. Simultaneous suppressions on p-IKKb, p-IkB and p-p65 suggested the regulation on NF-kappa B was IKK beta mediated. Meanwhile, XAN promoted the expression of IKK alpha, which has a possible connection to pro-apoptotic effects suggested by the up-regulated cleaved PARP. These findings indicated IKK beta/NF-kappa B mediates the proliferation of HFLS-RA cells inhibited by XAN, and divergent regulations on IKKs could provide synergic effects on the cells' proliferation. Copyright (C) 2017, Kaohsiung Medical University.