摘要
目的探讨亲环素A在矽尘诱导巨噬细胞泡沫化中的作用。方法取对数生长期人周围血单核细胞株TPH-1,用质量浓度为100μg/L的豆蔻酸佛波醇乙酸酯诱导分化为巨噬细胞,培养48 h后,分为4组。空白对照组巨噬细胞不予任何处理,氧化型低密度脂蛋白(ox-LDL)对照组巨噬细胞予含终质量浓度为50 mg/L ox-LDL的培养基,50 mg/L二氧化硅(SiO_2)组巨噬细胞予含终质量浓度为50 mg/L的SiO_2和50 mg/L ox-LDL的培养基,100 mg/L SiO_2组巨噬细胞予含终质量浓度为100 mg/L SiO_2和50 mg/L ox-LDL的培养基,培养48 h后,采用MTS法检测细胞活力,以油红O染色法和比色法检测巨噬细胞脂质蓄积,酶联免疫吸附实验法检测巨噬细胞上清液中亲环素A表达,以蛋白质印迹法检测巨噬细胞中亲环素A表达。结果与对照组及其余4个SiO_2组比较,SiO_2质量浓度为100 mg/L时巨噬细胞活力最强(P <0. 01)。油红O染色结果显示,与空白对照组比较,100 mg/L SiO_2组细胞泡沫样改变显著。与空白对照组比较,ox-LDL对照组、50 mg/L SiO_2组、100 mg/L SiO_2组巨噬细胞内总胆固醇和游离胆固醇表达水平均升高(P <0. 05),胆固醇酯比重增加(P <0. 05),亲环素A在巨噬细胞上清液中表达水平均升高(P <0. 05),在巨噬细胞内相对表达水平均降低(P <0. 05)。结论亲环素A可能参与了矽尘所致的巨噬细胞泡沫化过程。
Objective To investigate the role of cyclophilin A in the foaming process of macrophages induced by free silica( SiO2). Methods The human peripheral blood mononuclear cell THP-1 in the logarithmic growth phase was induced and differentiated into human macrophages with phorbol 12-myristate 13-acetate at 100 μg/L for 48 hours. The cells were divided into 4 groups. The cells in the blank control group were not treated. The cells in the oxidized low-density lipoprotein( ox-LDL) control group were treated with a final concentration of 50 mg/L ox-LDL. The cells of 50 mg/L SiO2 group were treated with 50 mg/L of SiO2 and ox-LDL. The cells of 100 mg/L SiO2 group were treated with 100 mg/L of SiO2 and 50 mg/L of ox-LDL. After treatment of cells for 48 h,cell viability was measured by MTS method. The lipid accumulation of cells was observed by oil red O staining and colorimetric method;the expression of cyclophilin A in cell supernatant was detected by enzyme-linked immunosorbent assay,and the expression of cyclophilin A in the cells was detected by Western blotting. Results The cell viability was the highest when the concentration of SiO2 was 100 mg/L compared with the control and other four SiO2groups( P < 0. 01). The cell foaming change in the 100 mg/L SiO2 group observed by oil red O staining significantly increased compared with the blank control group. The expression of total cholesterol,free cholesterol increased in the ox-LDL control group,50 mg/L SiO2 group and 100 mg/L SiO2group( P <0. 05),and the cholesterol specific gravity increased( P < 0. 05) compared with the blank control group,meanwhile the expression of cyclophilin A in the cell supernatant increased( P < 0. 05),and the expression in the macrophages cells decreased( P < 0. 05). Conclusion Cyclophilin A is involved in the foaming process of macrophages induced by SiO2.
作者
田盈
徐秋敏
王爽
郝小慧
郭灵丽
刘和亮
TIAN Yin;XU Qiumin;WANG Shuang;HAO Xiaohui;GUO Lingli;LIU Heliang(School of Public Health,North China University of Science and Technology,Tangshan,Hebei 063210,China)
出处
《中国职业医学》
CAS
北大核心
2018年第6期681-685,共5页
China Occupational Medicine
基金
国家自然科学基金(81602814)
河北省卫计委医学科学研究重点课题计划(20160720)
唐山市科技计划项目(15130212C)
