亲环素A在二氧化硅诱导巨噬细胞泡沫化中的表达
|
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( SiO_2). 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 SiO_2 group were treated with 50 mg/L of SiO_2 and ox-LDL. The cells of 100 mg/L SiO_2 group were treated with 100 mg/L of SiO_2 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 SiO_2 was 100 mg/L compared with the control and other four SiO_2groups( P < 0. 01). The cell foaming change in the 100 mg/L SiO_2 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 SiO_2 group and 100 mg/L SiO_2group( 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 SiO_2.
Objective To investigate the role of cyclophilin A in the foaming process of macrophages induced by free silica( SiO_2). 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 SiO_2 group were treated with 50 mg/L of SiO_2 and ox-LDL. The cells of 100 mg/L SiO_2 group were treated with 100 mg/L of SiO_2 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 SiO_2 was 100 mg/L compared with the control and other four SiO_2groups( P < 0. 01). The cell foaming change in the 100 mg/L SiO_2 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 SiO_2 group and 100 mg/L SiO_2group( 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 SiO_2.
引文
[1]ROMERO F,SHAH D,DUONG M,et al.A pneumocytemacrophage paracrine lipid axis drives the lung toward fibrosis[J].Am J Respir Cell Mol Biol,2015,53(1):74-86.
[2]FAN A,WU X,WU H,et al.Atheroprotective effect of oleoylethanolamide(OEA)targeting oxidized LDL[J].PLo S One,2014,9(1):e85337-e85346.
[3]MAISA A,HEARPS A C,ANGELOVICH T A,et al.Monocytes from HIV-infected individuals show impaired cholesterol efflux and increased foam cell formation after transendothelial migration[J].AIDS,2015,29(12):45-57.
[4]DEROSA G,SAHEBKAR A,MAFFIOLI P.The role of various peroxisome proliferator-activated receptors and their ligands in clinical practice[J].J Cell Physiol,2017,233(1):153-161.
[5]GU H F,LI H Z,TANG Y L,et al.Nicotinate-curcumin impedes foam cell formation from THP-1 cells through restoring autophagy flux[J].PLo S ONE,2016,11(4):e0154820.
[6]NI J,LI Y,LI W,et al.Salidroside protects against foam cell formation and apoptosis,possibly via the MAPK and AKT signaling pathways[J].Lipids Health Dis,2017,16(1):198.
[7]NIE Y,SUN L,WU Y,et al.AKT2 regulates pulmonary inflammation and fibrosis via modulating macrophage activation[J].JImmunol,2017,198(11):4470-4480.
[8]章述军,雷青松,李麟,等.两种PMA诱导方案对THP-1巨噬细胞M1和M2亚型相关基因表达的影响[J].中国细胞生物学学报,2017,39(6):765-771.
[9]TANDON A,HARIOUDH M K,ISHRAT N,et al.An MD2-derived peptide promotes LPS aggregation,facilitates its internalization in THP-1 cells,and inhibits LPS-induced pro-inflammatory responses[J].Cell Mol Life Sci,2018,75(13):2431-2446.
[10]林韬琦,卢德赵,沃兴德.巨噬细胞内胆固醇平衡机制研究进展[J].中国动脉硬化杂志,2010,18(11):919-921.
[11]候晓敏,陈紫莺,田盈,等.游离Si O2对肺泡巨噬细胞泡沫化的作用研究[J].医学研究生学报,2018,31(1):19-24.
[12]候晓敏,陈紫莺,田盈,等.抑制PPARγ对游离SiO2诱导肺泡巨噬细胞CD36表达及脂质蓄积的调节作用[J].环境与职业医学,2018,35(1):55-59.
[13]张舒宁,乔燕,胡志乔,等.熊果酸对矽肺大鼠肺泡巨噬细胞数量和波形蛋白表达的影响[J].中国职业医学,2018,45(5):572-576,580.
[14]LIU H,CHENG Y,YANG J,et al.BBC3 in macrophages promoted pulmonary fibrosis development through inducing autophagy during silicosis[J].Cell Death Dis,2017,8(3):e2657.
[15]杨萌,姚三巧.胶原样结构巨噬细胞受体和上皮间质转化在矽肺发病机制中作用研究进展[J].中国职业医学,2017,44(3):371-375.
[16]张秀峰,朱丽,刘舒鹏,等.矽肺患者肺泡巨噬细胞中胆固醇及ox-LDL水平分析[J].环境与职业医学,2016,33(12):57-61.
[17]FU Y,ZHAO Y,HUANG B.Tribbles homolog 1 enhances cholesterol efflux from oxidized low-density lipoprotein-loaded THP-1macrophages[J].Exp Ther Med,2017,14(1):862-866.
[18]LI E,WANG T,WANG F,et al.FGF21 protects against ox-LDLinduced apoptosis through suppressing CHOP expression in THP1macrophage derived foam cells[J].Bmc Cardiovascular Disorders,2015,15(1):1-7.
[19]REISS A B,PATEL C A,RAHMAN M M,et al.Interferon-gamma impedes reverse cholesterol transport and promotes foam cell transformation in THP-1 human monocytes/macrophages[J].Med Sci Monit,2004,10(11):420-425.
[20]PARKS B W,BLACK L L,ZIMMERMAN K A.et al.CD36,but not G2A,modulates efferocytosis,inflammation and fibrosis following bleomycin-induced lung injury[J].J Lipid Res,2013,54(4):1114.
[21]KOVANEN P T,Pentikinen M O.Circulating lipoproteins as proinflammatory and anti-inflammatory particles in atherogenesis[J].Curr Opin Lipidol,2003,14(5):411-419.
[22]RAMACHANDRAN S,VINITHA A.Cyclophilin A enhances macrophage differentiation and lipid uptake in high glucose conditions:a cellular mechanism for accelerated macro vascular disease in diabetes mellitus[J].Cardiovasc Diabetol,2016,15(1):152.
[23]LIU S,EIJKELENKAMP R,DUVIGNEAU J,et al.Silica assisted nucleation of polymer foam cells with nanoscopic dimensions:impact of particle size,line tension and surface functionality[J].Acs Appl Materi Interfaces,2017,9(43):37929-37940.
[24]XUE Z,YUAN W,LI J,et al.Cyclophilin A mediates the ox-LDL-induced activation and apoptosis of macrophages via autophagy[J].Int J Cardiol,2017,230:142-148.
[25]CHEN Y H,LIN W W,LIU C S,et al.Caveolin-1 expression ameliorates nephrotic damage in a rabbit model of cholesterol-induced hypercholesterolemia[J].PLo S One,2016,11(4):e0154210.
[26]NIGRO P,SATOH K,O'Dell M R,et al.Cyclophilin A is an inflammatory mediator that promotes atherosclerosis in apolipoprotein E-deficient mice[J].J Exp Med,2011,208(1):53-66.
[27]TIANTIAN Z,JUNFENG Z,HENG G.Functions of cyclophilin A in atherosclerosis[J].Experimental&Clinical Cardiology,2013,18(2):e118.
[2]FAN A,WU X,WU H,et al.Atheroprotective effect of oleoylethanolamide(OEA)targeting oxidized LDL[J].PLo S One,2014,9(1):e85337-e85346.
[3]MAISA A,HEARPS A C,ANGELOVICH T A,et al.Monocytes from HIV-infected individuals show impaired cholesterol efflux and increased foam cell formation after transendothelial migration[J].AIDS,2015,29(12):45-57.
[4]DEROSA G,SAHEBKAR A,MAFFIOLI P.The role of various peroxisome proliferator-activated receptors and their ligands in clinical practice[J].J Cell Physiol,2017,233(1):153-161.
[5]GU H F,LI H Z,TANG Y L,et al.Nicotinate-curcumin impedes foam cell formation from THP-1 cells through restoring autophagy flux[J].PLo S ONE,2016,11(4):e0154820.
[6]NI J,LI Y,LI W,et al.Salidroside protects against foam cell formation and apoptosis,possibly via the MAPK and AKT signaling pathways[J].Lipids Health Dis,2017,16(1):198.
[7]NIE Y,SUN L,WU Y,et al.AKT2 regulates pulmonary inflammation and fibrosis via modulating macrophage activation[J].JImmunol,2017,198(11):4470-4480.
[8]章述军,雷青松,李麟,等.两种PMA诱导方案对THP-1巨噬细胞M1和M2亚型相关基因表达的影响[J].中国细胞生物学学报,2017,39(6):765-771.
[9]TANDON A,HARIOUDH M K,ISHRAT N,et al.An MD2-derived peptide promotes LPS aggregation,facilitates its internalization in THP-1 cells,and inhibits LPS-induced pro-inflammatory responses[J].Cell Mol Life Sci,2018,75(13):2431-2446.
[10]林韬琦,卢德赵,沃兴德.巨噬细胞内胆固醇平衡机制研究进展[J].中国动脉硬化杂志,2010,18(11):919-921.
[11]候晓敏,陈紫莺,田盈,等.游离Si O2对肺泡巨噬细胞泡沫化的作用研究[J].医学研究生学报,2018,31(1):19-24.
[12]候晓敏,陈紫莺,田盈,等.抑制PPARγ对游离SiO2诱导肺泡巨噬细胞CD36表达及脂质蓄积的调节作用[J].环境与职业医学,2018,35(1):55-59.
[13]张舒宁,乔燕,胡志乔,等.熊果酸对矽肺大鼠肺泡巨噬细胞数量和波形蛋白表达的影响[J].中国职业医学,2018,45(5):572-576,580.
[14]LIU H,CHENG Y,YANG J,et al.BBC3 in macrophages promoted pulmonary fibrosis development through inducing autophagy during silicosis[J].Cell Death Dis,2017,8(3):e2657.
[15]杨萌,姚三巧.胶原样结构巨噬细胞受体和上皮间质转化在矽肺发病机制中作用研究进展[J].中国职业医学,2017,44(3):371-375.
[16]张秀峰,朱丽,刘舒鹏,等.矽肺患者肺泡巨噬细胞中胆固醇及ox-LDL水平分析[J].环境与职业医学,2016,33(12):57-61.
[17]FU Y,ZHAO Y,HUANG B.Tribbles homolog 1 enhances cholesterol efflux from oxidized low-density lipoprotein-loaded THP-1macrophages[J].Exp Ther Med,2017,14(1):862-866.
[18]LI E,WANG T,WANG F,et al.FGF21 protects against ox-LDLinduced apoptosis through suppressing CHOP expression in THP1macrophage derived foam cells[J].Bmc Cardiovascular Disorders,2015,15(1):1-7.
[19]REISS A B,PATEL C A,RAHMAN M M,et al.Interferon-gamma impedes reverse cholesterol transport and promotes foam cell transformation in THP-1 human monocytes/macrophages[J].Med Sci Monit,2004,10(11):420-425.
[20]PARKS B W,BLACK L L,ZIMMERMAN K A.et al.CD36,but not G2A,modulates efferocytosis,inflammation and fibrosis following bleomycin-induced lung injury[J].J Lipid Res,2013,54(4):1114.
[21]KOVANEN P T,Pentikinen M O.Circulating lipoproteins as proinflammatory and anti-inflammatory particles in atherogenesis[J].Curr Opin Lipidol,2003,14(5):411-419.
[22]RAMACHANDRAN S,VINITHA A.Cyclophilin A enhances macrophage differentiation and lipid uptake in high glucose conditions:a cellular mechanism for accelerated macro vascular disease in diabetes mellitus[J].Cardiovasc Diabetol,2016,15(1):152.
[23]LIU S,EIJKELENKAMP R,DUVIGNEAU J,et al.Silica assisted nucleation of polymer foam cells with nanoscopic dimensions:impact of particle size,line tension and surface functionality[J].Acs Appl Materi Interfaces,2017,9(43):37929-37940.
[24]XUE Z,YUAN W,LI J,et al.Cyclophilin A mediates the ox-LDL-induced activation and apoptosis of macrophages via autophagy[J].Int J Cardiol,2017,230:142-148.
[25]CHEN Y H,LIN W W,LIU C S,et al.Caveolin-1 expression ameliorates nephrotic damage in a rabbit model of cholesterol-induced hypercholesterolemia[J].PLo S One,2016,11(4):e0154210.
[26]NIGRO P,SATOH K,O'Dell M R,et al.Cyclophilin A is an inflammatory mediator that promotes atherosclerosis in apolipoprotein E-deficient mice[J].J Exp Med,2011,208(1):53-66.
[27]TIANTIAN Z,JUNFENG Z,HENG G.Functions of cyclophilin A in atherosclerosis[J].Experimental&Clinical Cardiology,2013,18(2):e118.