兰州大学机构库 >学院待认领
罗格列酮对泡沫细胞中胆固醇贮存与运输相关蛋白ACAT-1、ABCA-1表达的影响
Alternative TitleEffects of rosiglitazone on expression of acyl-coenzyme A cholesterol acyltransferase 1 and ATP-binding cassette transporter A1 in foam cells
蒙颖
Thesis Advisor王志禄
2013-05-24
Degree Grantor兰州大学
Place of Conferral兰州
Degree Name硕士
Keyword罗格列酮 泡沫细胞 胆固醇 动脉粥样硬化
Abstract目的 观察罗格列酮对RAW264.7巨噬源性泡沫细胞形成中胆固醇含量及酰基辅酶A-胆固醇酰基转移酶-1(acyl coenzyme A cholesterol acyl transfer enzyme 1, ACAT-1)、三磷酸腺苷结合盒转运蛋白A-1 (ATP combination box of transporter 1, ABCA-1)表达的影响,旨在探讨罗格列酮对泡沫细胞形成的影响及可能的作用机制。 方法 (1)建立泡沫细胞模型:用30 mg/L 氧化型低密度脂蛋白(oxidized low density lipoprotein,oxLDL)与RAW264.7细胞共同孵育48 h后用油红O染色鉴定是否转化为泡沫细胞。(2)实验分组:随机分为五组。空白对照组:正常培养基培养巨噬细胞;oxLDL组,用终浓度为30 mg/L的oxLDL孵育48 h;oxLDL+罗格列酮组,分别用终浓度为5 μmol/L、10 μmol/L、20 μmol/L的罗格列酮与30 mg/L的oxLDL共同孵育48 h。(3)使用胆固醇检测试剂盒测定细胞内胆固醇含量:分别检测细胞内总胆固醇(total cholesterol,TC)及游离胆固醇(free cholesterol,FC)含量,胆固醇酯(cholesterol ester,CE)的含量由TC减FC得出。(4)Western blot 法检测各组细胞ACAT-1和ABCA-1的蛋白表达水平。 结果 (1)油红O染色观察罗格列酮对泡沫细胞形成的影响,oxLDL组中大量泡沫细胞的胞质被着色,oxLDL+罗格列酮组中细胞胞质染色明显浅于oxLDL组的细胞。(2)用胆固醇检测试剂盒检测细胞内胆固醇含量显示,与空白对照组相比,oxLDL组细胞内的TC及FC均显著升高(P<0.05);与oxLDL组相比,oxLDL+罗格列酮组中TC及FC均显著降低(P<0.05),且呈浓度依赖性。(3)Western blot检测细胞ACAT-1及ABCA-1表达显示,不同浓度(5 μmol/L、10 μmol/L、20 μmol/L)oxLDL+罗格列酮组ACAT-1蛋白相对表达量分别为0.94±0.11、0.86±0.13、0.58±0.12,与oxLDL组(1.19±0.12)相比有显著差异(P<0.05),不同浓度oxLDL+罗格列酮组ABCA-1蛋白相对表达量分别为0.72±0.08、0.91±0.15、1.15±0.11,与oxLDL组(0.63±0.05)相比有显著差异(P<0.05),且呈浓度依赖性。 结论 (1)oxLDL可刺激RAW264.7巨噬细胞转化为泡沫细胞。(2)罗格列酮可能在抑制泡沫细胞ACAT-1表达的同时促进ABCA-1的表达,这也可能是抑制泡沫细胞形成的机理之一。
Other AbstractObjective To investigate the effects of rosiglitazone on cholesterol content and the expressions of acyl-CoA: cholesterol acyltransferase 1 (ACAT-1) and ATP binding cassette transporter A-1 (ABCA-1)in RAW264.7 macropgage-derived foam cells, and explore the effects and mechanisms of rosiglitazone on foam cells formation, so as to provide theory basis of rosiglitazone preventing atherosclerosis in clinical. Methods (1) Establishment of foam cell model:RAW264.7 cells were co-cultured with 30 mg/L oxidized low density lipoprotein (oxLDL) for 48 h, then the formation of foam cells were identified by oil red O staining technique. (2) Cultured RAW264.7 cells were divided into 5 groups randomly: blank control group, cells were cultured in normal growth medium; oxLDL group, cells were co-cultured with 30 mg/L oxLDL for 48 h; oxLDL+rosiglitazone group, cells were co-cultured with 30 mg/L oxLDL and 5 μmol/L, 10 μmol/L, 20 μmol/L rosiglitazone respectively for 48 h. (3) The concentration of cellular total cholesterol (TC) and free cholesterol (FC) was measured by using cholesterol kit, and the content of cholesterol ester (CE) was acquired by subtracting the FC from TC. (4) The protein expressions of ACAT-1 and ABCA-1 were determined by western blot. Result (1) Oil red O staining showed cells in oxLDL group were stained by oil red O, but the color of cells in oxLDL+rosiglitazone group was shallower than that in oxLDL group. (2) Compared with blank control group , the content of cellular TC and FC in oxLDL group were significantly increased (P<0.05), while compared with oxLDL group, the content of cellular TC and FC in oxLDL+rosiglitazone group were significantly decreased(P<0.05) with a concentration-dependent manner. (3) Western blot analysis showed that the expression levels of ACAT-1 in different concentration (5 μmol/L, 10 μmol/L, 20 μmol/L) of oxLDL + rosiglitazone group were 0.94 ± 0.11, 0.86 ± 0.13, 0.58 ± 0.12 respectively, the result is significantly different(P<0.05) from the oxLDL group(1.19 ± 0.12), and the expression levels of ABCA-1 expression in different concentration of oxLDL + rosiglitazone group were 0.72 ± 0.08, 0.91 ± 0.15, 1.15 ± 0.11 respectively, the result also is significantly different(P<0.05) from the oxLDL group (0.63±0.05). Conclusion (1) OxLDL can transform RAW264.7 cell into foam cell. (2) Rosiglita...
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Language中文
Document Type学位论文
Identifierhttps://ir.lzu.edu.cn/handle/262010/223018
Collection学院待认领
Affiliation临床医学院
Recommended Citation
GB/T 7714
蒙颖. 罗格列酮对泡沫细胞中胆固醇贮存与运输相关蛋白ACAT-1、ABCA-1表达的影响[D]. 兰州. 兰州大学,2013.
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