ox-LDL/β2GPI/aβ2GPI复合物促进小鼠RAW264.7细胞凋亡和炎症因子表达  

ox-LDL/β2GPI/anti-β2GPI antibody complex promotes apoptosis and inflammatory factors expression in RAW264.7 cells

作　　者：匡铭 周红 张贵婷 何超 吴倩倩 姚雨叶 KUANG Ming;ZHOU Hong;ZHANG Guiting;HE Chao;WU Qianqian;YAO Yuye(Division of Hematology,School of Medicine,Jiangsu University,Zhenjiang 212013,China)

机构地区：[1]江苏大学医学院血液研究室,江苏镇江212013

出　　处：《细胞与分子免疫学杂志》2022年第1期1-8,共8页Chinese Journal of Cellular and Molecular Immunology

基　　金：国家自然科学基金(81370614)。

摘　　要：目的 探讨氧化型低密度脂蛋白/β2糖蛋白I/抗β2糖蛋白I抗体(ox-LDL/β2GPI/aβ2GPI)复合物对小鼠RAW264.7细胞凋亡和炎症因子表达的影响及机制。方法 分别用培养基、ox-LDL、β2GPI/抗β2GPI抗体复合物、ox-LDL/β2GPI复合物、ox-LDL/抗β2GPI抗体复合物和ox-LDL/β2GPI/抗β2GPI抗体复合物处理细胞。用TAK-242阻断Toll样受体4(TLR4)、雷帕霉素(rapamycin)阻断哺乳动物雷帕霉素靶蛋白(mTOR)探究相关通路的作用。最后,用不同浓度的TNF-α刺激细胞。用凋亡试剂盒检测细胞的凋亡,实时定量PCR检测肿瘤坏死因子α(TNF-α)、白细胞介素1β(IL-1β)的mRNA表达,Western blot法检测TNF-α、IL-1β、裂解型胱天蛋白酶3(c-caspase-3)、B细胞淋巴瘤因子2(Bcl2)的蛋白水平。结果 ox-LDL/β2GPI/aβ2GPI复合物能够诱导细胞凋亡、Bcl2表达降低、c-caspase-3表达增加,还可促进TNF-α和IL-1β表达,通路抑制剂可部分减轻凋亡水平和炎症因子表达;一定浓度的TNF-α可促进细胞凋亡。结论 ox-LDL/β2GPI/aβ2GPI复合物通过TLR4、mTOR通路诱导RAW264.7细胞表达炎症因子,促进细胞凋亡。Objective To investigate the effects of ox-LDL/β2GPI/anti-β2GPI antibody complex on apoptosis and inflammatory factors expression in RAW264.7 cells and its mechanism. Methods RAW264.7 cells were treated with medium, ox-LDL, β2GPI/aβ2GPI antibody complex, ox-LDL/β2GPI complex, ox-LDL/aβ2GPI antibody complex, and ox-LDL/β2GPI/aβ2GPI antibody complex separately. The role of signaling pathways was investigated by TLR4 inhibitor TAK-242 and mTOR inhibitor rapamycin. Finally, cells were stimulated with different concentrations of TNF-α. The apoptosis was detected by Annexin Ⅴ-FITC/PI double labeling. The mRNA expressions of TNF-α and IL-1β were detected by real-time quantitative PCR, and the protein expressions of TNF-α, IL-1β, and apoptosis-related proteins cleaved-caspase-3(c-caspase-3) and Bcl2 were detected by Western blot. Results Ox-LDL/β2GPI/aβ2GPI antibody complex promoted RAW264.7 cells’ apoptosis, with a decrease in protein Bcl2 and an increase in protein cleaved-caspase-3, and enhanced the level of inflammatory factors, while apoptosis and the expression of inflammatory factors were partially reduced when macrophages had been pre-treated with inhibitors. TNF-α up-regulated apoptosis in RAW264.7 cells at 100 ng/mL and 200 ng/mL. Conclusion Ox-LDL/β2GPI/aβ2GPI antibody complex induces apoptosis in RAW264.7 cells by increasing the expression of inflammatory factors. TLR4 and mTOR pathways are involved in the process.

关 键 词：氧化型低密度脂蛋白/β2糖蛋白I/抗β2糖蛋白I抗体(ox-LDL/β2GPI/aβ2GPI)复合物 RAW264.7细胞 细胞凋亡 炎症因子 Toll样受体4(TLR4) 哺乳动物雷帕霉素靶蛋白(mTOR) 

分 类 号：R392-33[医药卫生—免疫学] Q255[医药卫生—基础医学]