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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 oftransporter 1,ABCA-1)表达的影响,探讨罗格列酮对泡沫细胞形成的影响及可能的作用机制。方法在DMEM高糖培养基中培养RAW264.7巨噬细胞,按完全随机分组方法分为:①空白对照组(常规培养基培养巨噬细胞);②氧化低密度脂蛋白(oxidized low density lipoprotein,oxLDL)组(用终浓度为30 mg/L的oxLDL孵育48 h);③oxLDL+罗格列酮组(分别用终浓度5、10、20μmol/L的罗格列酮+30 mg/L oxLDL共同孵育48 h)(n=10)。采用油红O染色观察泡沫细胞,胆固醇检测试剂盒测定各组细胞内总胆固醇(total cholesterol,TC)和游离胆固醇(free cholesterol,FC)的含量,Western blot法检测各组细胞ACAT-1和ABCA1的表达水平。结果 oxLDL组中大量泡沫细胞的胞质被油红O染色,oxLDL+罗格列酮组泡沫细胞胞质染色明显浅于oxLDL组的细胞;与oxLDL组相比,oxLDL+罗格列酮组TC及FC显著降低(P<0.05),且呈浓度依赖性;Western blot检测结果表明,不同浓度(5、10、20μmol/L)oxLDL+罗格列酮组ACAT-1蛋白表达分别为(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ABCA1蛋白表达分别为(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。结论罗格列酮可能通过抑制ACAT-1的表达及促进ABCA-1的表达减少泡沫细胞形成,从而发挥其抗动脉粥样硬化的作用。
Objective To observe the effect of rosiglitazone on the cholesterol content and the expression of ACAT-1, adenosine triphosphate binding cassette in RAW264.7 macrophage-derived foam cells To study the effect of rosiglitazone on the formation of foam cells and its possible mechanism. Methods RAW264.7 macrophages were cultured in DMEM high glucose medium and divided into two groups according to a completely randomized method: ① blank control group (macrophages cultured in conventional medium); ② oxidized low density lipoprotein (oxLDL) group (oxLDL at a final concentration of 30 mg / L for 48 h); ③oxLDL plus rosiglitazone group (co-incubated with 5,10 and 20 μmol / L of rosiglitazone and 30 mg / 48 h) (n = 10). Foam cells were observed by oil red O staining. The content of total cholesterol (TC) and free cholesterol (FC) in each group were determined by the cholesterol detection kit. The expressions of ACAT-1 and ABCA1 The level of expression. Results The cytoplasm of a large number of foam cells in oxLDL group was stained with oil red O, and the cytoplasm of oxLDL + rosiglitazone group was significantly lighter than that in oxLDL group. Compared with oxLDL group, the levels of TC and FC in oxLDL + rosiglitazone group (P <0.05), and in a concentration-dependent manner. The results of Western blot showed that the expressions of ACAT-1 protein in oxLDL + rosiglitazone group were (0.94 ± 0.11), (0.86 ± 0.13) and (0.58 ± 0.12), respectively, compared with oxLDL group (1.19 ± 0.12) (P <0.05). The ABCA1 protein expression in oxLDL plus rosiglitazone group was (0.72 ± 0.08) (0.91 ± 0.15) and (1.15 ± 0.11) respectively, which were significantly different from those in oxLDL group (0.63 ± 0.05) (P <0.05). Conclusion Rosiglitazone may play an anti-atherogenic role by inhibiting the expression of ACAT-1 and promoting the expression of ABCA-1, thereby reducing foam cell formation.