LPS刺激RAW264.7、THP-1细胞HMGB1释放及共刺激分子表达  

LPS promotes HMGB1 release and costimulatory molecule expression in RAW264.7 and THP-1 cells

作　　者：苏静 孔庆玲 李淼 祁继金 仲青 周竟心 高玲 胡娜[1] 陈伟[1,3,4,5] SU Jing;KONG Qingling;LI Miao;QI Jijin;ZHONG Qing;ZHOU Jingxin;GAO Ling;HU Na;CHEN Wei(Department of Hematology,the First People′s Hospital of Suqian,Suqian,Jiangsu 223800,China;Xuzhou Children′s Hospital,Xuzhou,Jiangsu 221002;Department of Hematology,the Affiliated Hospital of Xuzhou Medical University,Xuzhou,Jiangsu 221002;Blood Diseases Institute,Xuzhou Medical University,Xuzhou,Jiangsu 221002;Key Laboratory of Bone Marrow Stem Cells,Xuzhou,Jiangsu 221002)

机构地区：[1]南京医科大学附属宿迁第一人民医院血液科,江苏宿迁223800 [2]徐州市儿童医院,江苏徐州221002 [3]徐州医科大学附属医院血液科,江苏徐州221002 [4]徐州医科大学血液病研究所,江苏徐州221002 [5]江苏省骨髓干细胞重点实验室,江苏徐州221002

出　　处：《徐州医科大学学报》2022年第11期799-804,共6页Journal of Xuzhou Medical University

基　　金：江苏省科技厅自然科学基金面上项目(BK20221218);宿迁市血液病学重点实验室建设项目资助(M202111);徐州医科大学优秀人才基金项目(XYFY2021001)。

摘　　要：目的探讨细菌脂多糖(LPS)对巨噬细胞高迁移率族蛋白B1(HMGB1)及共刺激分子表达的影响。方法培养THP-1、RAW264.7细胞,予100、200、1000μg/L LPS刺激24 h,CCK8法检测细胞增殖的变化;流式细胞术检测细胞凋亡情况;免疫印迹法检测培养上清以及细胞内HMGB1的水平;定量聚合酶链反应(Q-PCR)检测细胞内HMGB1转录水平;流式细胞术检测细胞表面CD80、CD83及CD86的表达变化。结果LPS促进RAW264.7增殖,低浓度LPS抑制THP-1细胞增殖,高浓度LPS促进THP-1细胞增殖;LPS促进THP-1、RAW264.7细胞的凋亡;THP-1及RAW264.7细胞培养上清中的HMGB1均增加(P<0.05),细胞内HMGB1水平下降(P<0.05)。THP-1、RAW264.7细胞内HMGB1的mRNA转录水平升高(P<0.05);流式细胞术检测THP-1、RAW264.7细胞表面CD80、CD83、CD86的表达示LPS促进细胞表面CD80、CD83和CD86的表达(P<0.05)。结论LPS影响巨噬细胞增殖和凋亡,引起细胞内HMGB1分泌至胞外,促进细胞表面共刺激分子表达增加。Objective To investigate the effect of bacterial lipopolysaccharide(LPS)on the expression of macrophage high mobility group protein B1(HMGB1)and costimulatory molecules.Methods THP-1 and RAW264.7 cells were cultured and stimulated with 100,200 and 1000μg/L LPS for 24 h.The cell proliferation was detected by CCK8 assay.The cell apoptosis was detect by flow cytometry.The level of HMGB1 in the culture supernatant and within the cells was detected by Western blot.The transcription of HMGB1 was examined by Q-PCR.The expression of CD80,CD83,and CD86 were detected by flow cytometry.Results LPS promoted the proliferation of RAW264.7 cells.LPS inhibited the proliferation of THP-1 cells at a low concentration,but promoted THP-1 cell proliferation at a high concentration(P<0.05).LPS promoted the apoptosis of both THP-1 and RAW264.7 cells(P<0.05).The expression of HMGB1 increased in THP-1 and RAW264.7 cell culture supernatant(P<0.05),but intracellular HMGB1 decreased(P<0.05).The transcriptional level of HMGB1 went up in THP-1 and RAW264.7 cells(P<0.05).The expression of CD80,CD83,and CD86 increased on these two cells(P<0.05).Conclusions LPS affects the proliferation and apoptosis of THP-1 and RAW264.7 cells,promotes HGMB1 release,and increases the expression of CD80,C83,and CD86 on cell surface.

关 键 词：高迁移率族蛋白B1 移植物抗宿主病 细菌脂多糖 

分 类 号：R-332[医药卫生]