肺炎衣原体通过上调酰基辅酶A:胆固醇酰基转移酶1诱导人单核细胞株THP-1源性泡沫细胞形成的实验研究

目的 观察酰基辅酶A:胆固醇酰基转移酶1(ACAT1)在肺炎衣原体(C.pn)诱导泡沫细胞形成中的作用,初步探讨C.pn诱导泡沫细胞形成的机制.方法 人单核细胞株(THP-1)给予160 mmol/L佛波酯(PMA)孵育48 h,诱导分化为巨噬细胞后随机分为4组:阴性对照组、阳性对照组、C.pn感染组和ACAT抑制剂58-035+C.pn感染组.分别运用RT-PCR和Western blot检测各组ACAT1 mRNA和蛋白表达.运用油红O染色观察细胞浆内脂滴的变化,用酶荧光学法检测细胞内胆固醇酯含量的变化.结果 C.pn感染呈浓度和时间依赖性地上调ACAT1 mRNA和蛋白表达.高浓度的C.pn(5×105和1×106IFU)感染THP-1源性巨噬细胞48 h后,细胞浆内脂滴明显增多,胆固醇酯与总胆固醇的比值明显增加(＞50%).ACAT抑制剂58-035可以抑制C.pn诱导的细胞浆内脂滴的增多.随着58-035浓度的增加,逐渐抑制C.pn诱导的细胞内胆固醇酯含量的增加.结论 ACAT1表达上调是C.pn诱导泡沫细胞形成的机制之一,这可能为进一步阐明C.pn感染促进动脉粥样硬化发生发展提供一个新的理论依据.     

肺炎衣原体通过上调酰基辅酶A:胆固醇酰基转移酶1诱导人单核细胞株THP-1源性泡沫细胞形成的实验研究

Objective To investigate the expression changes of acyl-coenzyme A: cholesterol acyhransferase 1 (ACAT1) on Chlamydia pneumoniae (C. pn) induced foam cell formation. Methods Human monocytic cell line (THP-1) was induced into macrophages by 160 nmol/L phorbol myristate acetate (PMA)for 48 h, and were randomly allocated into four groups: negative control group (50 μg/ml LDL for 48 h) ; positive control group (50 μg/ml ox-LDL for48 h) ; C. pn infection group (50 μg/ml LDL plus 1× 105,4×105,5×105and 1×106 IFU C.pn for 48 h or 1×106 IFU C.pn for 0,24,48 and 72h); ACAT inhibitor 58-035 plus C. pn infection group (1, 5, 10 μg/ml ACAT inhibitor 58-035 pretreatment for 1 h,50 μg/ml LDL and 1 × 106 IFU C. pn for 48 h). The mRNA and protein expressions of ACATI were determined by RT-PCR and Western blot, respectively. Lipid droplets in cytoplasm were observed by oil red 0 staining. The contents of intracellular cholesteryl esters were detected by enzyme-fluorescence. Results The mRNA and protein expressions of ACATI were significantly up-regulated in positive control cells compared those in negative control cells and further upregulated by C. pn infection in a time-dependent and concentration-dependent manner (all P ＜ 0.05). There were significantly increases in the accumulation of lipid droplets and the ratio of cholesteryl ester to total cholesterol in positive control cells as compared with negative control cells and these were further aggravated by C. pn (at the concentrations of 5× 105 and 1×106IFU for 48 h) and C. pn infection induced increases in the accumulation of lipid droplets and the ratio of cholesteryl ester to total cholesterol could he significantly attenuated by ACAT inhibitor 58-035 (all P ＜ 0.05). Conclusion Chlamydia pneumoniae induces THP-1-derived foam cell formation by up-regulating the expression of ACATI.     

肺炎衣原体通过上调酰基辅酶A:胆固醇酰基转移酶1诱导人单核细胞株THP-1源性泡沫细胞形成的实验研究

Objective To investigate the expression changes of acyl-coenzyme A: cholesterol acyhransferase 1 (ACAT1) on Chlamydia pneumoniae (C. pn) induced foam cell formation. Methods Human monocytic cell line (THP-1) was induced into macrophages by 160 nmol/L phorbol myristate acetate (PMA)for 48 h, and were randomly allocated into four groups: negative control group (50 μg/ml LDL for 48 h) ; positive control group (50 μg/ml ox-LDL for48 h) ; C. pn infection group (50 μg/ml LDL plus 1× 105,4×105,5×105and 1×106 IFU C.pn for 48 h or 1×106 IFU C.pn for 0,24,48 and 72h); ACAT inhibitor 58-035 plus C. pn infection group (1, 5, 10 μg/ml ACAT inhibitor 58-035 pretreatment for 1 h,50 μg/ml LDL and 1 × 106 IFU C. pn for 48 h). The mRNA and protein expressions of ACATI were determined by RT-PCR and Western blot, respectively. Lipid droplets in cytoplasm were observed by oil red 0 staining. The contents of intracellular cholesteryl esters were detected by enzyme-fluorescence. Results The mRNA and protein expressions of ACATI were significantly up-regulated in positive control cells compared those in negative control cells and further upregulated by C. pn infection in a time-dependent and concentration-dependent manner (all P ＜ 0.05). There were significantly increases in the accumulation of lipid droplets and the ratio of cholesteryl ester to total cholesterol in positive control cells as compared with negative control cells and these were further aggravated by C. pn (at the concentrations of 5× 105 and 1×106IFU for 48 h) and C. pn infection induced increases in the accumulation of lipid droplets and the ratio of cholesteryl ester to total cholesterol could he significantly attenuated by ACAT inhibitor 58-035 (all P ＜ 0.05). Conclusion Chlamydia pneumoniae induces THP-1-derived foam cell formation by up-regulating the expression of ACATI.     

肺炎衣原体通过上调酰基辅酶A:胆固醇酰基转移酶1诱导人单核细胞株THP-1源性泡沫细胞形成的实验研究

Objective To investigate the expression changes of acyl-coenzyme A: cholesterol acyhransferase 1 (ACAT1) on Chlamydia pneumoniae (C. pn) induced foam cell formation. Methods Human monocytic cell line (THP-1) was induced into macrophages by 160 nmol/L phorbol myristate acetate (PMA)for 48 h, and were randomly allocated into four groups: negative control group (50 μg/ml LDL for 48 h) ; positive control group (50 μg/ml ox-LDL for48 h) ; C. pn infection group (50 μg/ml LDL plus 1× 105,4×105,5×105and 1×106 IFU C.pn for 48 h or 1×106 IFU C.pn for 0,24,48 and 72h); ACAT inhibitor 58-035 plus C. pn infection group (1, 5, 10 μg/ml ACAT inhibitor 58-035 pretreatment for 1 h,50 μg/ml LDL and 1 × 106 IFU C. pn for 48 h). The mRNA and protein expressions of ACATI were determined by RT-PCR and Western blot, respectively. Lipid droplets in cytoplasm were observed by oil red 0 staining. The contents of intracellular cholesteryl esters were detected by enzyme-fluorescence. Results The mRNA and protein expressions of ACATI were significantly up-regulated in positive control cells compared those in negative control cells and further upregulated by C. pn infection in a time-dependent and concentration-dependent manner (all P ＜ 0.05). There were significantly increases in the accumulation of lipid droplets and the ratio of cholesteryl ester to total cholesterol in positive control cells as compared with negative control cells and these were further aggravated by C. pn (at the concentrations of 5× 105 and 1×106IFU for 48 h) and C. pn infection induced increases in the accumulation of lipid droplets and the ratio of cholesteryl ester to total cholesterol could he significantly attenuated by ACAT inhibitor 58-035 (all P ＜ 0.05). Conclusion Chlamydia pneumoniae induces THP-1-derived foam cell formation by up-regulating the expression of ACATI.     

肺炎衣原体通过上调酰基辅酶A:胆固醇酰基转移酶1诱导人单核细胞株THP-1源性泡沫细胞形成的实验研究

Objective To investigate the expression changes of acyl-coenzyme A: cholesterol acyhransferase 1 (ACAT1) on Chlamydia pneumoniae (C. pn) induced foam cell formation. Methods Human monocytic cell line (THP-1) was induced into macrophages by 160 nmol/L phorbol myristate acetate (PMA)for 48 h, and were randomly allocated into four groups: negative control group (50 μg/ml LDL for 48 h) ; positive control group (50 μg/ml ox-LDL for48 h) ; C. pn infection group (50 μg/ml LDL plus 1× 105,4×105,5×105and 1×106 IFU C.pn for 48 h or 1×106 IFU C.pn for 0,24,48 and 72h); ACAT inhibitor 58-035 plus C. pn infection group (1, 5, 10 μg/ml ACAT inhibitor 58-035 pretreatment for 1 h,50 μg/ml LDL and 1 × 106 IFU C. pn for 48 h). The mRNA and protein expressions of ACATI were determined by RT-PCR and Western blot, respectively. Lipid droplets in cytoplasm were observed by oil red 0 staining. The contents of intracellular cholesteryl esters were detected by enzyme-fluorescence. Results The mRNA and protein expressions of ACATI were significantly up-regulated in positive control cells compared those in negative control cells and further upregulated by C. pn infection in a time-dependent and concentration-dependent manner (all P ＜ 0.05). There were significantly increases in the accumulation of lipid droplets and the ratio of cholesteryl ester to total cholesterol in positive control cells as compared with negative control cells and these were further aggravated by C. pn (at the concentrations of 5× 105 and 1×106IFU for 48 h) and C. pn infection induced increases in the accumulation of lipid droplets and the ratio of cholesteryl ester to total cholesterol could he significantly attenuated by ACAT inhibitor 58-035 (all P ＜ 0.05). Conclusion Chlamydia pneumoniae induces THP-1-derived foam cell formation by up-regulating the expression of ACATI.     

肺炎衣原体通过上调酰基辅酶A:胆固醇酰基转移酶1诱导人单核细胞株THP-1源性泡沫细胞形成的实验研究

Objective To investigate the expression changes of acyl-coenzyme A: cholesterol acyhransferase 1 (ACAT1) on Chlamydia pneumoniae (C. pn) induced foam cell formation. Methods Human monocytic cell line (THP-1) was induced into macrophages by 160 nmol/L phorbol myristate acetate (PMA)for 48 h, and were randomly allocated into four groups: negative control group (50 μg/ml LDL for 48 h) ; positive control group (50 μg/ml ox-LDL for48 h) ; C. pn infection group (50 μg/ml LDL plus 1× 105,4×105,5×105and 1×106 IFU C.pn for 48 h or 1×106 IFU C.pn for 0,24,48 and 72h); ACAT inhibitor 58-035 plus C. pn infection group (1, 5, 10 μg/ml ACAT inhibitor 58-035 pretreatment for 1 h,50 μg/ml LDL and 1 × 106 IFU C. pn for 48 h). The mRNA and protein expressions of ACATI were determined by RT-PCR and Western blot, respectively. Lipid droplets in cytoplasm were observed by oil red 0 staining. The contents of intracellular cholesteryl esters were detected by enzyme-fluorescence. Results The mRNA and protein expressions of ACATI were significantly up-regulated in positive control cells compared those in negative control cells and further upregulated by C. pn infection in a time-dependent and concentration-dependent manner (all P ＜ 0.05). There were significantly increases in the accumulation of lipid droplets and the ratio of cholesteryl ester to total cholesterol in positive control cells as compared with negative control cells and these were further aggravated by C. pn (at the concentrations of 5× 105 and 1×106IFU for 48 h) and C. pn infection induced increases in the accumulation of lipid droplets and the ratio of cholesteryl ester to total cholesterol could he significantly attenuated by ACAT inhibitor 58-035 (all P ＜ 0.05). Conclusion Chlamydia pneumoniae induces THP-1-derived foam cell formation by up-regulating the expression of ACATI.     

肺炎衣原体通过上调酰基辅酶A:胆固醇酰基转移酶1诱导人单核细胞株THP-1源性泡沫细胞形成的实验研究

Objective To investigate the expression changes of acyl-coenzyme A: cholesterol acyhransferase 1 (ACAT1) on Chlamydia pneumoniae (C. pn) induced foam cell formation. Methods Human monocytic cell line (THP-1) was induced into macrophages by 160 nmol/L phorbol myristate acetate (PMA)for 48 h, and were randomly allocated into four groups: negative control group (50 μg/ml LDL for 48 h) ; positive control group (50 μg/ml ox-LDL for48 h) ; C. pn infection group (50 μg/ml LDL plus 1× 105,4×105,5×105and 1×106 IFU C.pn for 48 h or 1×106 IFU C.pn for 0,24,48 and 72h); ACAT inhibitor 58-035 plus C. pn infection group (1, 5, 10 μg/ml ACAT inhibitor 58-035 pretreatment for 1 h,50 μg/ml LDL and 1 × 106 IFU C. pn for 48 h). The mRNA and protein expressions of ACATI were determined by RT-PCR and Western blot, respectively. Lipid droplets in cytoplasm were observed by oil red 0 staining. The contents of intracellular cholesteryl esters were detected by enzyme-fluorescence. Results The mRNA and protein expressions of ACATI were significantly up-regulated in positive control cells compared those in negative control cells and further upregulated by C. pn infection in a time-dependent and concentration-dependent manner (all P ＜ 0.05). There were significantly increases in the accumulation of lipid droplets and the ratio of cholesteryl ester to total cholesterol in positive control cells as compared with negative control cells and these were further aggravated by C. pn (at the concentrations of 5× 105 and 1×106IFU for 48 h) and C. pn infection induced increases in the accumulation of lipid droplets and the ratio of cholesteryl ester to total cholesterol could he significantly attenuated by ACAT inhibitor 58-035 (all P ＜ 0.05). Conclusion Chlamydia pneumoniae induces THP-1-derived foam cell formation by up-regulating the expression of ACATI.     

肺炎衣原体通过上调酰基辅酶A:胆固醇酰基转移酶1诱导人单核细胞株THP-1源性泡沫细胞形成的实验研究

Objective To investigate the expression changes of acyl-coenzyme A: cholesterol acyhransferase 1 (ACAT1) on Chlamydia pneumoniae (C. pn) induced foam cell formation. Methods Human monocytic cell line (THP-1) was induced into macrophages by 160 nmol/L phorbol myristate acetate (PMA)for 48 h, and were randomly allocated into four groups: negative control group (50 μg/ml LDL for 48 h) ; positive control group (50 μg/ml ox-LDL for48 h) ; C. pn infection group (50 μg/ml LDL plus 1× 105,4×105,5×105and 1×106 IFU C.pn for 48 h or 1×106 IFU C.pn for 0,24,48 and 72h); ACAT inhibitor 58-035 plus C. pn infection group (1, 5, 10 μg/ml ACAT inhibitor 58-035 pretreatment for 1 h,50 μg/ml LDL and 1 × 106 IFU C. pn for 48 h). The mRNA and protein expressions of ACATI were determined by RT-PCR and Western blot, respectively. Lipid droplets in cytoplasm were observed by oil red 0 staining. The contents of intracellular cholesteryl esters were detected by enzyme-fluorescence. Results The mRNA and protein expressions of ACATI were significantly up-regulated in positive control cells compared those in negative control cells and further upregulated by C. pn infection in a time-dependent and concentration-dependent manner (all P ＜ 0.05). There were significantly increases in the accumulation of lipid droplets and the ratio of cholesteryl ester to total cholesterol in positive control cells as compared with negative control cells and these were further aggravated by C. pn (at the concentrations of 5× 105 and 1×106IFU for 48 h) and C. pn infection induced increases in the accumulation of lipid droplets and the ratio of cholesteryl ester to total cholesterol could he significantly attenuated by ACAT inhibitor 58-035 (all P ＜ 0.05). Conclusion Chlamydia pneumoniae induces THP-1-derived foam cell formation by up-regulating the expression of ACATI.     

肺炎衣原体通过上调酰基辅酶A:胆固醇酰基转移酶1诱导人单核细胞株THP-1源性泡沫细胞形成的实验研究

Objective To investigate the expression changes of acyl-coenzyme A: cholesterol acyhransferase 1 (ACAT1) on Chlamydia pneumoniae (C. pn) induced foam cell formation. Methods Human monocytic cell line (THP-1) was induced into macrophages by 160 nmol/L phorbol myristate acetate (PMA)for 48 h, and were randomly allocated into four groups: negative control group (50 μg/ml LDL for 48 h) ; positive control group (50 μg/ml ox-LDL for48 h) ; C. pn infection group (50 μg/ml LDL plus 1× 105,4×105,5×105and 1×106 IFU C.pn for 48 h or 1×106 IFU C.pn for 0,24,48 and 72h); ACAT inhibitor 58-035 plus C. pn infection group (1, 5, 10 μg/ml ACAT inhibitor 58-035 pretreatment for 1 h,50 μg/ml LDL and 1 × 106 IFU C. pn for 48 h). The mRNA and protein expressions of ACATI were determined by RT-PCR and Western blot, respectively. Lipid droplets in cytoplasm were observed by oil red 0 staining. The contents of intracellular cholesteryl esters were detected by enzyme-fluorescence. Results The mRNA and protein expressions of ACATI were significantly up-regulated in positive control cells compared those in negative control cells and further upregulated by C. pn infection in a time-dependent and concentration-dependent manner (all P ＜ 0.05). There were significantly increases in the accumulation of lipid droplets and the ratio of cholesteryl ester to total cholesterol in positive control cells as compared with negative control cells and these were further aggravated by C. pn (at the concentrations of 5× 105 and 1×106IFU for 48 h) and C. pn infection induced increases in the accumulation of lipid droplets and the ratio of cholesteryl ester to total cholesterol could he significantly attenuated by ACAT inhibitor 58-035 (all P ＜ 0.05). Conclusion Chlamydia pneumoniae induces THP-1-derived foam cell formation by up-regulating the expression of ACATI.     

肺炎衣原体通过上调酰基辅酶A:胆固醇酰基转移酶1诱导人单核细胞株THP-1源性泡沫细胞形成的实验研究

Objective To investigate the expression changes of acyl-coenzyme A: cholesterol acyhransferase 1 (ACAT1) on Chlamydia pneumoniae (C. pn) induced foam cell formation. Methods Human monocytic cell line (THP-1) was induced into macrophages by 160 nmol/L phorbol myristate acetate (PMA)for 48 h, and were randomly allocated into four groups: negative control group (50 μg/ml LDL for 48 h) ; positive control group (50 μg/ml ox-LDL for48 h) ; C. pn infection group (50 μg/ml LDL plus 1× 105,4×105,5×105and 1×106 IFU C.pn for 48 h or 1×106 IFU C.pn for 0,24,48 and 72h); ACAT inhibitor 58-035 plus C. pn infection group (1, 5, 10 μg/ml ACAT inhibitor 58-035 pretreatment for 1 h,50 μg/ml LDL and 1 × 106 IFU C. pn for 48 h). The mRNA and protein expressions of ACATI were determined by RT-PCR and Western blot, respectively. Lipid droplets in cytoplasm were observed by oil red 0 staining. The contents of intracellular cholesteryl esters were detected by enzyme-fluorescence. Results The mRNA and protein expressions of ACATI were significantly up-regulated in positive control cells compared those in negative control cells and further upregulated by C. pn infection in a time-dependent and concentration-dependent manner (all P ＜ 0.05). There were significantly increases in the accumulation of lipid droplets and the ratio of cholesteryl ester to total cholesterol in positive control cells as compared with negative control cells and these were further aggravated by C. pn (at the concentrations of 5× 105 and 1×106IFU for 48 h) and C. pn infection induced increases in the accumulation of lipid droplets and the ratio of cholesteryl ester to total cholesterol could he significantly attenuated by ACAT inhibitor 58-035 (all P ＜ 0.05). Conclusion Chlamydia pneumoniae induces THP-1-derived foam cell formation by up-regulating the expression of ACATI.