淫羊藿苷通过上调 TRIB1 抑制核转录因子 -κB 缓解类风湿关节炎成纤维样滑膜细胞炎症

吴志明　李慧　周易芬　曹正柳　张鹏　万欣　郭健

【摘 要】目的：探討淫羊藿苷（ICA）是否通过上调TRIB1表达抑制核转录因子-κB（NF-κB）缓解类风湿关节炎成纤维样滑膜细胞（RA-FLS）炎症。方法：收集南昌大学第一附属医院接受膝关节置换手术的活动期类风湿关节炎（RA）患者的滑膜组织，分离培养RA-FLS后，用肿瘤坏死因子-α（TNF-α，10 ng·mL^-1）处理RA-FLS建立炎症细胞模型。首先，为明确ICA抑制RA-FLS增殖和炎症，促进细胞凋亡，分为空白对照组、TNF-α组、TNF-α + ICA组；然后，为明确TRIB1过表达可抑制RA-FLS的增殖和炎症，促进细胞凋亡，空白vector或oe-TRIB1转染TNF-α处理的RA-FLS，分为空白对照组、TNF-α组、TNF-α + vector组、TNF-α + TRIB1组；接着，为明确ICA以依赖于TRIB1的方式抑制NF-κB，siTRIB1转染TNF-α处理的RA-FLS细胞，分为空白对照组、TNF-α组、TNF-α + ICA组、TNF-α + ICA + siTRIB1组。分别采用qRT-PCR、Western blot检测基因和蛋白水平；ELISA检测炎性细胞因子；CCK-8、流式细胞术检测细胞增殖和凋亡。结果：ICA处理和TRIB1过表达均抑制TNF-α诱导的RA-FLS的增殖和炎症反应，且促进其凋亡。TNF-α处理后RA-FLS中p-p65和p-IκB水平显著上调，但ICA显著降低p-p65和p-IκB水平；且与siTRIB1联合治疗时，ICA的抑制作用消失。结论：ICA通过上调TRIB1抑制NF-κB缓解RA，提示ICA可能是一种有前景的治疗RA药物。

【关键词】 类风湿关节炎；淫羊藿苷；TRIB1；核转录因子-κB；成纤维样滑膜细胞

Icariin Alleviating Fibroblastic Synovial Cell Inflammation in Rheumatoid Arthritis by Upregulating TRIB1 to Inhibit NF-κB

WU Zhi-ming，LI Hui，ZHOU Yi-fen，CAO Zheng-liu，ZHANG Peng，WAN Xin，GUO Jian

【ABSTRACT】Objective：To investigate whether icariin（ICA）relieves inflammation of rheumatoid arthritis fibroblastic synovial（RA-FLS）cells by upregulating TRIB1 to inhibit nuclear transcription factors-κB（NF-κB）.Methods：The synovial tissue of patients with RA who underwent knee Joint replacement surgery in the First Affiliated Hospital of Nanchang University was collected，and RA-FLS cells were isolated and cultured.TNF-α（10 ng·mL^-1）was used to deal with RA-FLS cells to establish inflammatory cell models.Firstly，to clarify that ICA inhibits the proliferation and inflammation of RA-FLS cells and promote cell apoptosis，the blank control group，the TNF-α processing group，and the TNF-α + ICA group were established.Then，to clarify that overexpression of TRIB1 can inhibit the proliferation and inflammation of RA-FLS，and promote cell apoptosis，and blank vector or oe-TRIB1 was transfected into TNF-α processed RA-FLS cells，the Control group，the TNF-α processing group，the TNF-α + Vector group，TNF-α + TRIB1 group were established.Next，to clarify that ICA inhibits NF-κB dependent on TRIB1，and siTRIB1 transfecting TNF-α processed RA-FLS cells，the blank control group the TNF-α processing group，the TNF-α + ICA group，the TNF-α + ICA + siTRIB1 group were established.Gene and protein levels were detected using qRT-PCR and Western blot，respectively;ELISA was used to detect inflammatory cytokines;and CCK-8 and flow cytometry were used to detect cell proliferation and apoptosis.Results：Both ICA treatment and TRIB1 overexpression inhibited TNF-α induced proliferation and inflammatory response of RA-FLS cells，and promoted their apoptosis.The levels of p-p65 and p-IκB in the TNF-α processed RA-FLS cells significantly upregulated，but ICA significantly reduced the levels of p-p65 and p-IκB.When combined with sitRIB1 in the treatment，the inhibitory effect of ICA disappears.Conclusion：ICA inhibits NF by upregulating TRIB1-κB to alleviate RA，suggesting that ICA may be a promising therapeutic drug for RA.

【Keywords】 rheumatoid arthritis;icariin;TRIB1;nuclear transcription factor-κB;fibroblastic synovial cells

类风湿关节炎（rheumatoid arthritis，RA）是一种免疫性疾病，表现为对称性关节炎，常伴有软骨破坏、骨侵蚀［1］，是患者残疾和生活质量下降的常见原因［2-3］。RA发病涉及各类免疫细胞和成纤维样滑膜细胞（FLS）功能异常［4］。目前，免疫抑制剂（如甲氨蝶呤、来氟米特）和生物制剂等治疗有一定的疗效［5-6］，但患者面临着巨大的经济和身体负担。

淫羊藿苷（ICA）是中草药淫羊藿的重要单体，具有骨愈合及骨保护作用［7］。……