沉默狼疮相关肺动脉高压高表达的HMGB1通过TLR4/NF-κB途径抑制低氧诱导的PASMC增殖、迁移及胶原合成  

Silencing high expression of HMGB1 in lupus associated pulmonary hypertension via TLR4/NF-κB pathway inhibits proliferation,migration and collagen synthesis of PASMC induced by hypoxia

作　　者：李鸣远[1] 孟岩[2] 武云[1] LI Mingyuan;MENG Yan;WU Yun(Department of General Medicine,the First Affiliated Hospital of Xinjiang Medical University,Urumqi 830054,China)

机构地区：[1]新疆医科大学第一附属医院全科医学科,乌鲁木齐830054 [2]新疆医科大学第一附属医院风湿免疫科,乌鲁木齐830054

出　　处：《中国免疫学杂志》2022年第17期2123-2128,共6页Chinese Journal of Immunology

基　　金：国家自然科学基金项目(81860096)。

摘　　要：目的:探讨沉默狼疮相关肺动脉高压(SLE-PAH)高表达的HMGB1通过TLR4/NF-κB途径抑制低氧诱导的PASMC增殖、迁移及胶原合成的作用机制。方法:分别测定SLE-PAH患者血浆中HMGB1含量与低氧诱导PASMC内HMGB1表达的变化;siRNA沉默PASMC内HMGB1表达,MTT法与EdU实验测定细胞增殖,Boyden小室试验测定细胞迁移,胶原酶消化法与Western blot测定胶原合成能力,同时检测TLR4/NF-κB信号通路的表达情况;采用siRNA-HMGB1与TLR4激活剂处理PASMC,低氧环境下培养,再次测定上述指标的变化。结果:与正常受试者及SLE患者相比,SLE-PAH患者血浆中HMGB1含量显著升高(P<0.05),且SLE-PAH患者中处于SLE活动期与合并重度PAH血浆HMGB1含量高于非活动期与合并轻中度PAH(P<0.05);低氧诱导PASMC同样促进了HMGB1表达(P<0.05);而沉默HMGB1则能抑制低氧诱导的PASMC增殖、迁移及胶原合成(P<0.05),同时抑制TLR4/NF-κB信号通路的表达(P<0.05),但该变化能被TLR4激活剂LPS所逆转(P<0.05)。结论:HMGB1通过TLR4/NF-κB途径抑制低氧诱导的PASMC增殖、迁移及胶原合成,是SLE-PAH可能的治疗靶标。Objective:To investigate the mechanism of silencing high expression of HMGB1 in lupus associated pulmonary hypertension(SLE-PAH)by TLR4/NF-κB pathway inhibiting PASMC proliferation,migration and collagen synthesis induced by hypoxia.Methods:The content of HMGB1 in plasma of SLE-PAH patients and the expression of HMGB1 in PASMC induced by hypoxia were determined respectively. The expression of HMGB1 in PASMC was silenced by siRNA. Cell proliferation was determined by MTT and EdU assay. Cell migration was determined by Boyden chamber assay. Collagen synthesis was determined by collagenase digestion and Western blot. The expression of TLR4/NF-κB signal pathway was detected. PASMC was treated with siRNA-HMGB1 and TLR4activator,cultured under hypoxia environment,and the changes of above indexes were measured again.Results:Compared with normal and SLE patients,the content of HMGB1 in SLE-PAH patients was significantly higher(P<0.05),and the plasma HMGB1levels in patients with active and severe PAH were higher than those in patients with inactive and mild to moderate PAH(P<0.05);hypoxia induced PASMC also promoted HMGB1 expression(P<0.05);HMGB1 silencing inhibited PASMC proliferation,migration and collagen synthesis induced by hypoxia(P<0.05),and inhibited the expression of TLR4/NF-κB pathway. However,this change could be reversed by TLR4 activator LPS(P<0.05).Conclusion:HMGB1 through TLR4/NF-κB pathway inhibits the proliferation, migration and collagen synthesis of PASMC induced by hypoxia,which is a possible therapeutic target for SLE-PAH.

关 键 词：系统性红斑狼疮 肺动脉高压 肺动脉平滑肌肉细胞 高迁移率族蛋白 Toll样受体4 

分 类 号：R593.241[医药卫生—内科学]