焦孔素D介导的滑膜血管内皮细胞焦亡促进类风湿关节炎关节破坏  

GSDMD-mediated Pyroptosis of Synovial Vascular Endothelial Cells Promotes Joint Destruction in Rheumatoid Arthritis

作　　者：吴滔 张学培 卢烨 邹耀威 欧阳志明[1] 马剑达[1] 戴冽[1] WU Tao;ZHANG Xuepei;LU Ye;ZOU Yaowei;OUYANG Zhiming;MA Jianda;DAI Lie(Department of Rheumatology and Immunology,Sun Yat-Sen Memorial Hospital,Guangzhou 510120,China;Department of Rheumatology and Immunology,The First People's Hospital of Foshan,Foshan 528010,China)

机构地区：[1]中山大学孙逸仙纪念医院风湿免疫科,广东广州510120 [2]佛山市第一人民医院风湿免疫科,广东佛山528010

出　　处：《中山大学学报（医学科学版）》2024年第5期709-718,共10页Journal of Sun Yat-Sen University:Medical Sciences

基　　金：国家自然科学基金(82171780);广东省基础与应用基础研究基金(2022A1515010524,2023A1515030253)。

摘　　要：【目的】了解焦孔素D(GSDMD)在类风湿关节炎(RA)滑膜血管内皮细胞(VEC)中的表达并探讨其介导的细胞焦亡对VEC功能的影响。【方法】收集22例RA及对照组18例非炎性关节病(Orth.A)患者膝关节滑膜,Western blot方法检测活性GSDMD-NT段蛋白水平并分组比较RA患者临床指标的差异;免疫荧光双染检测RA滑膜GSDMD的细胞定位。体外RA滑液干预人脐静脉血管内皮细胞(HUVEC)模拟炎症环境,检测细胞凋亡水平及焦亡通路相关蛋白表达。siRNA转染敲减GSDMD,检测HUVEC细胞焦亡、分泌细胞因子、增殖、迁移和血管生成能力的变化。【结果】RA滑膜组织表达GSDMD-NT显著升高,GSDMD-NT高表达组RA患者关节破坏更严重,滑膜微血管计数更高;RA滑膜VEC表达GSDMD;RA滑液可诱导HUVEC发生GSDMD介导的细胞焦亡,分泌血管内皮生长因子(VEGF)增多,增殖、迁移及血管生成能力增强;下调GSDMD表达可逆转上述效应。【结论】GSDMD介导的少部分滑膜VEC焦亡可能通过分泌VEGF诱导其余VEC增殖、迁移以及血管新生参与RA关节破坏进程,可能是抑制RA关节破坏的新靶点。【Objective】To explore the occurrence of gasdermin D(GSDMD)-mediated pyroptosis and its effect on cell proliferation,migration and tubular formation abilities of synovial vascular endothelial cells(VEC)in rheumatoid arthritis(RA).【Methods】Synovium tissues from knee joints of 22 RA patients and 18 orthopaedic arthropathies(Orth.A)patients were collected.The level of activated GSDMD-NT segment in synovium was detected by Western blot.The clinical characteristics of RA patients were compared between high and low synovial GSDMD-NT groups.The cell localization of GSDMD in RA synovium was detected by immunofluorescence staining.RA synovial fluid was added to the culture of human umbilical vein endothelial cells(HUVEC)in vitro,and the level of apoptosis and expression of pyroptosis pathway proteins were detected.The effects of GSDMD on apoptosis,proliferation,migration and tubule formation of HUVEC cells were analyzed.【Results】GSDMD expression in RA synovium was significantly higher than that in Orth.A,and more severe joint destruction and higher microvascular count score were found in RA patients with high GSDMD-NT expression.Synovial VEC had positive expression of GSDMD.Stimulation with RA synovial fluid could induce GSDMD-mediated pyroptosis in HUVEC,increased the secretion of vascular endothelial growth factor(VEGF)and their abilities of proliferation,migration and tubule formation.Knockdown of GSDMD could reverse the above effects.【Conclusion】GSDMD-mediated pyroptosis of partial synovial VEC aggravates RA joint destruction through VEGF secretion that promotes proliferation,migration and angiogenesis of the remaining VEC,which may be a new target to block neovascularization and inhibit joint destruction in RA.

关 键 词：细胞焦亡 类风湿关节炎 焦孔素D 血管内皮细胞 关节破坏 

分 类 号：R593.22[医药卫生—内科学]