阿托伐他汀对人脐静脉内皮细胞和大鼠胸主动脉平滑肌细胞Adropin相关基因及蛋白表达的影响
发布时间：2019-06-09 14:16
【摘要】：第一部分阿托伐他汀对人脐静脉内皮细胞Adropin相关基因及蛋白表达的影响 目的 探讨阿托伐他汀对人脐静脉内皮细胞（human umbilical endothelial cell, HUVEC）Adropin相关基因及蛋白表达的影响。 方法 将不同浓度的阿托伐他汀（0.002umol/l、0.02umol/l、0.2umol/l、2umol/l、20umol/l）与HUVEC共培养6小时（hour，h）、12h、24h。采用MTT法检测HUVEC的增殖情况。半定量逆转录聚合酶链反应法检测HUVEC Adropin mRNA的表达。ELISA法测定细胞上清液中Adropin蛋白的表达。 结果 1.不同浓度的阿托伐他汀与HUVEC共培养6h时，20umol/l浓度组增殖活力明显高于对照组（P0.05），其他各浓度组差异无统计学意义。共培养12h时，0.02umol/l、0.2umol/l、2umol/l、20umol/l浓度组细胞活力显著高于对照组（P0.05），0.2umol/l、2umol/l、20umol/l浓度组活力显著高于0.002umol/l浓度组（P0.05）。共培养24h时，各浓度组细胞活力均显著高于对照组（P0.05），，0.02umol/l、0.2umol/l、2umol/l、20umol/l浓度组活力显著高于0.002umol/l浓度组(P0.05)。 同一浓度阿托伐他汀分别与HUVEC共培养6h、12h、24h，12h和24h组活力明显高于6h组（P0.05）。 2.不同浓度组阿托伐他汀与HUVEC共培养6h、12h、24h后，Adropin mRNA的表达均显著高于对照组（P0.001），0.02umol/l、0.2umol/l、2umol/l、20umol/l浓度组Adropin mRNA的表达显著高于0.002umol/l组（P0.001）。 同一浓度阿托伐他汀分别与HUVEC共培养6h、12h、24h，12h和24h组mRNA表达量明显高于6h组（P0.01）。 3.不同浓度的阿托伐他汀与HUVEC分别共培养6h、12h、24h，各浓度组HUVEC上清中Adropin蛋白的表达显著高于对照组（P0.001），0.02umol/l、0.2umol/l、2umol/l、20umol/l浓度组Adropin蛋白的表达显著高于0.002umol/l组（P0.001）。 同一浓度阿托伐他汀分别与HUVEC共培养6h、12h、24h时，12h和24h组蛋白表达量明显高于6h组(P0.001）。 4. Adropin蛋白含量与HUVEC吸光度值（OD值）进行Pearson相关性分析显示不同时间段各浓度组adropin与OD值呈显著正相关（6h，相关系数r=0.471，P=0.049；12h，r=0.756，P=0.001；24h，r=0.725，P=0.001)。 结论 1.阿托伐他汀能够促进HUVEC的增殖，并呈剂量和时间依赖性。 2.阿托伐他汀能够促进HUVEC Adropin相关基因及蛋白的表达，且呈剂量和时间依赖性。 3. HUVEC的增殖活力与其Adropin蛋白的表达呈正相关。 第二部分阿托伐他汀对大鼠胸主动脉平滑肌细胞Adropin相关基因及蛋白表达的影响 目的 探讨阿托伐他汀对大鼠胸主动脉平滑肌细胞（rat aortic smooth muscle cell，RASMC）Adropin相关基因及蛋白表达的影响。 方法 原代培养SD大鼠的RASMC，免疫荧光和免疫细胞化学染色法进行鉴定，取4-5代细胞进行试验。.不同浓度的阿托伐他汀（0.02umol/l、0.2umol/l、2umol/l、20umol/l）与RASMC分别共培养6h、12h、24h。采用MTT法检测RASMC的增殖情况。半定量逆转录聚合酶链反应法检测RASMCAdropin mRNA的表达。ELISA法测定细胞上清液中Adropin蛋白的表达。 结果 1.不同浓度的阿托伐他汀与RASMC共培养24h时，各浓度组细胞活力显著低于对照组（P0.05），0.02umol/l、0.2umol/l阿托伐他汀组活力明显高于20umol/组（P0.05）。 同一浓度阿托伐他汀与RASMC共培养6h、12h、24h，12h和24h组活力明显低于6h组（P0.05）。 2.不同浓度的阿托伐他汀与RASMC共培养，各浓度组Adropin mRNA表达量均高于对照组（P0.05）；随着浓度增大，细胞中Adropin mRNA表达量逐渐降低（P0.05）。 同一浓度阿托伐他汀分别与RASMC共培养6h、12h、24h，12h和24h组AdropinmRNA表达量明显高于6h组（P0.01）。 3.不同浓度的阿托伐他汀与RASMC分别共培养6h、12h、24h，各浓度组RASMC上清中Adropin蛋白的表达显著高于对照组（P0.05）。随着浓度增大，细胞中Adropin蛋白的表达量逐渐降低（P0.05）。 同一浓度的阿托伐他汀分别与RASMC共培养6h、12h、24h，Adropin蛋白的表达量随着时间延长明显增加（P0.05）。 4. Adropin蛋白含量与RASMC吸光度值（OD值）进行Pearson相关性分析显示不同时间段各浓度组adropin与OD值呈显著正相关(6h，相关系数r=0.694，P=0.012)vs(12h，r=0.697，P=0.002) vs(24h，r=0.826，P=0.001)。 结论 1.阿托伐他汀能抑制RASMC的增殖并呈剂量和时间依赖性。 2.阿托伐他汀能促进RASMC Adropin相关基因及蛋白的表达，但随浓度的增加其促进作用逐渐减弱。 3. RASMC的增长活力与其Adropin蛋白的表达呈正相关。
[Abstract]:The effect of the first part of atorvastatin on the expression of the related gene and protein of human umbilical vein endothelial cell Objective To study the expression of Atorvastatin on human umbilical vein endothelial cells (HUVEC) and its expression in human umbilical vein endothelial cells (HUVEC). a shadow in response to that method, different concentrations of atorvastatin (0.002 mol/ l, 0.02 mol/ l, 0.2 mol/ l,2 umol/ l,20 umol/ l) were co-cultured with HUVEC for 6 h (hour, h), 12 h,24 h. HUV was detected by MTT method. The proliferation of EC was detected by semi-quantitative reverse transcription polymerase chain reaction (RT-PCR). Expression of mRNA. Adrotp in the supernatant of the cells by ELISA in Results 1. The proliferation of Atorvastatin and HUVEC at different concentrations was significantly higher than that in the control group (P0.05). There was no statistical significance in the concentration group. In the co-culture for 12 h, the activity of the cell in the concentration group was significantly higher than that of the control group (P0.05), the cell viability of the concentration group was significantly higher than that of the control group (P0.05), and the activity of the concentration group of 0.2 umol/ l,2 umol/ l and 20 umol/ l was significantly higher than that of the control group (P0.05). The activity of cells in each concentration group was significantly higher than that of the control group (P0.05), 0.02 mol/ l, 0.2 uml/ l,2 umol/ l and 20 umol/ l in the concentration group (P0.05), and the activity of the concentration group was significantly higher than that of the control group (P0.05). The same concentration of atorvastatin were co-cultured with HUVEC for 6 h,12 h,24 h,12 h and 24 h, respectively. After 6 h,12 h and 24 h of co-culture of Atorvastatin and HUVEC in different concentration groups, the expression of Adropin mRNA was significantly higher than that in the control group (P 0.001), 0.02 mol/ l, 0.2 umol/ l,2 umol/ l and 20 umol/ l. The same concentration of atorvastatin were co-cultured with HUVEC for 6 h,12 h,24 h,12 h and 24 h, respectively. Atorvastatin and HUVEC were co-cultured for 6 h,12 h and 24 h, respectively. The expression of Atropin in HUVEC was significantly higher than that of control group (P 0.001), 0.02 mol/ l, 0.2 uml/ l,2 umol/ l and 20 umol/ l. The same concentration of atorvastatin were co-cultured with HUVEC for 6 h,12 h,24 h,12 h and 24 h, respectively. The expression of histone was significantly higher than that of the 6 h group (P0.001).4. The Pearson correlation analysis showed that the concentration of adropin was positively correlated with the OD (r = 0.471, P = 0.049; 12h, r = 0.756, P = 0.0) for different time periods. 01 ;24 h, r = 0.725, P = 0.001). Conclusion 1. torvastatin can promote the proliferation of HUVEC and dose and time-dependent.2. Atorvastatin can promote HUVE The expression of C-Adropin-related gene and protein is dose-dependent and time-dependent. 3. The proliferation activity of HUVEC is positively related to the expression of its Adropin protein. cut him Objective To study the effect of statins on the expression of the related gene and protein of the rat thoracic aortic smooth muscle cells, and to investigate the effect of atorvastatin on the rat thoracic aortic smooth muscle cells h m The effect of the expression of the related gene and protein of the rat cell (RASMC) D-rat RASMC, immunofluorescence and immunocytochemical staining for identification,4-5 generation of cells tested.. different concentrations of atorvastatin (0.02 uml/ l, 0.2 umol /l,2umol/l,20umol/l And were co-cultured with RASMC for 6 h,12 h and 24 h, respectively. C. Detection of RASMC by semi-quantitative reverse transcription polymerase chain reaction (RT-PCR) Ad Hoc Results 1. The activity of Atorvastatin and RASMC at different concentrations was significantly lower than that in the control group (P0.05). The activity of the Atorvastatin group was significantly higher than that of 20 umol/ l (P0.05). Atorvastatin and RASMC were co-cultured for 6 h,12 h,24 h,12 h and 24 h for 6 h,12 h,24 h,12 h and 24 h, respectively. At the same concentration, Atorvastatin and RASM were compared with those in the control group (P0.05). C co-culture for 6 h,12 h,24 h,12 h and 24 h was significantly higher than that in 6 h (P0.01).3. Atorvastatin and RASMC at different concentrations 6 h,12 h and 24 h were co-cultured for 6 h,12 h and 24 h. The white expression was significantly higher in the control group than in the control group (P0.05). As the concentration increased, the expression of the Adropin in the cells decreased gradually (P0.05). After 6 h,12 h and 24 h of co-culture with RASMC, the expression of ropin protein increased significantly with the increase of time (P0.05).4. The Pearson correlation analysis showed that the concentration of adropin in each concentration group was positively correlated with the OD value for different time periods (6 h, r = 0.6). 94 ,P=0.012)vs(12h,r=0.697,P=0.0 02) vs (24 h, r = 0.826, P = 0.001). Conclusion 1. Atorvastatin can inhibit the proliferation of RASMC 2. Atorvastatin can promote RASMC Ad
【学位授予单位】：苏州大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R96
[Abstract]:The effect of the first part of atorvastatin on the expression of the related gene and protein of human umbilical vein endothelial cell Objective To study the expression of Atorvastatin on human umbilical vein endothelial cells (HUVEC) and its expression in human umbilical vein endothelial cells (HUVEC). a shadow in response to that method, different concentrations of atorvastatin (0.002 mol/ l, 0.02 mol/ l, 0.2 mol/ l,2 umol/ l,20 umol/ l) were co-cultured with HUVEC for 6 h (hour, h), 12 h,24 h. HUV was detected by MTT method. The proliferation of EC was detected by semi-quantitative reverse transcription polymerase chain reaction (RT-PCR). Expression of mRNA. Adrotp in the supernatant of the cells by ELISA in Results 1. The proliferation of Atorvastatin and HUVEC at different concentrations was significantly higher than that in the control group (P0.05). There was no statistical significance in the concentration group. In the co-culture for 12 h, the activity of the cell in the concentration group was significantly higher than that of the control group (P0.05), the cell viability of the concentration group was significantly higher than that of the control group (P0.05), and the activity of the concentration group of 0.2 umol/ l,2 umol/ l and 20 umol/ l was significantly higher than that of the control group (P0.05). The activity of cells in each concentration group was significantly higher than that of the control group (P0.05), 0.02 mol/ l, 0.2 uml/ l,2 umol/ l and 20 umol/ l in the concentration group (P0.05), and the activity of the concentration group was significantly higher than that of the control group (P0.05). The same concentration of atorvastatin were co-cultured with HUVEC for 6 h,12 h,24 h,12 h and 24 h, respectively. After 6 h,12 h and 24 h of co-culture of Atorvastatin and HUVEC in different concentration groups, the expression of Adropin mRNA was significantly higher than that in the control group (P 0.001), 0.02 mol/ l, 0.2 umol/ l,2 umol/ l and 20 umol/ l. The same concentration of atorvastatin were co-cultured with HUVEC for 6 h,12 h,24 h,12 h and 24 h, respectively. Atorvastatin and HUVEC were co-cultured for 6 h,12 h and 24 h, respectively. The expression of Atropin in HUVEC was significantly higher than that of control group (P 0.001), 0.02 mol/ l, 0.2 uml/ l,2 umol/ l and 20 umol/ l. The same concentration of atorvastatin were co-cultured with HUVEC for 6 h,12 h,24 h,12 h and 24 h, respectively. The expression of histone was significantly higher than that of the 6 h group (P0.001).4. The Pearson correlation analysis showed that the concentration of adropin was positively correlated with the OD (r = 0.471, P = 0.049; 12h, r = 0.756, P = 0.0) for different time periods. 01 ;24 h, r = 0.725, P = 0.001). Conclusion 1. torvastatin can promote the proliferation of HUVEC and dose and time-dependent.2. Atorvastatin can promote HUVE The expression of C-Adropin-related gene and protein is dose-dependent and time-dependent. 3. The proliferation activity of HUVEC is positively related to the expression of its Adropin protein. cut him Objective To study the effect of statins on the expression of the related gene and protein of the rat thoracic aortic smooth muscle cells, and to investigate the effect of atorvastatin on the rat thoracic aortic smooth muscle cells h m The effect of the expression of the related gene and protein of the rat cell (RASMC) D-rat RASMC, immunofluorescence and immunocytochemical staining for identification,4-5 generation of cells tested.. different concentrations of atorvastatin (0.02 uml/ l, 0.2 umol /l,2umol/l,20umol/l And were co-cultured with RASMC for 6 h,12 h and 24 h, respectively. C. Detection of RASMC by semi-quantitative reverse transcription polymerase chain reaction (RT-PCR) Ad Hoc Results 1. The activity of Atorvastatin and RASMC at different concentrations was significantly lower than that in the control group (P0.05). The activity of the Atorvastatin group was significantly higher than that of 20 umol/ l (P0.05). Atorvastatin and RASMC were co-cultured for 6 h,12 h,24 h,12 h and 24 h for 6 h,12 h,24 h,12 h and 24 h, respectively. At the same concentration, Atorvastatin and RASM were compared with those in the control group (P0.05). C co-culture for 6 h,12 h,24 h,12 h and 24 h was significantly higher than that in 6 h (P0.01).3. Atorvastatin and RASMC at different concentrations 6 h,12 h and 24 h were co-cultured for 6 h,12 h and 24 h. The white expression was significantly higher in the control group than in the control group (P0.05). As the concentration increased, the expression of the Adropin in the cells decreased gradually (P0.05). After 6 h,12 h and 24 h of co-culture with RASMC, the expression of ropin protein increased significantly with the increase of time (P0.05).4. The Pearson correlation analysis showed that the concentration of adropin in each concentration group was positively correlated with the OD value for different time periods (6 h, r = 0.6). 94 ,P=0.012)vs(12h,r=0.697,P=0.0 02) vs (24 h, r = 0.826, P = 0.001). Conclusion 1. Atorvastatin can inhibit the proliferation of RASMC 2. Atorvastatin can promote RASMC Ad
【学位授予单位】：苏州大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R96
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