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目的观察高表达脂肪分化相关蛋白对酰基辅酶A∶胆固醇酰基转移酶1表达及脂质蓄积的影响。方法构建表达载体pcDNA3.1-HA-脂肪分化相关蛋白,使用THP-1巨噬细胞,通过瞬时转染使之高表达脂肪分化相关蛋白,依次用氧化型低密度脂蛋白和(或)丙泮尼地处理。逆转录聚合酶链反应和Western blot检测酰基辅酶A∶胆固醇酰基转移酶1及脂肪分化相关蛋白的表达,油红O染色和高效液相色谱检测细胞内脂质的蓄积。结果随着氧化型低密度脂蛋白浓度的增加,巨噬细胞脂肪分化相关蛋白及酰基辅酶A∶胆固醇酰基转移酶1表达明显增强,两者呈伴行关系。丙泮尼地能抑制酰基辅酶A∶胆固醇酰基转移酶1表达上调,且随处理时间延长,其表达逐渐减少,并且能减少细胞内脂滴生成。与对照组相比,瞬时转染pcDNA3.1-HA-脂肪分化相关蛋白能使脂肪分化相关蛋白表达明显升高。高表达脂肪分化相关蛋白的巨噬细胞能使酰基辅酶A∶胆固醇酰基转移酶1表达增加,促进细胞内胆固醇酯蓄积,并协同增强氧化型低密度脂蛋白的作用。加入丙泮尼地后,高表达脂肪分化相关蛋白的作用被减弱。结论高表达脂肪分化相关蛋白能上调THP-1巨噬细胞酰基辅酶A∶胆固醇酰基转移酶1表达,促进细胞内脂质蓄积。脂肪分化相关蛋白可能通过酰基辅酶A∶胆固醇酰基转移酶1促进细胞内胆固醇酯的蓄积。
Objective To observe the effect of overexpression of fat differentiation-related proteins on the expression of acyl-CoA: cholesterol acyltransferase 1 and lipid accumulation. Methods The expression vector pcDNA3.1-HA-lipofiltration was constructed. THP-1 macrophages were transiently transfected to express lipofectin-related proteins, followed by oxidized low density lipoprotein and / or diazepam Nigeria to deal with. Reverse transcriptase polymerase chain reaction and Western blot were used to detect the expression of acyl-CoA: cholesterol acyltransferase 1 and adipogenic differentiation related proteins. Oil red O staining and high performance liquid chromatography were used to detect the accumulation of intracellular lipid. Results With the increase of the concentration of oxidized low density lipoprotein, the expression of adipogenic differentiation related protein and acyl coenzyme A: cholesterol acyltransferase 1 in macrophages increased significantly. Pripnodipine inhibited the up-regulation of acyl-CoA: cholesterol acyltransferase 1 expression, and gradually decreased with the prolongation of treatment time, and decreased the intracellular lipid droplets formation. Compared with the control group, transient transfection of pcDNA3.1-HA-fat differentiation-related protein expression of fat differentiation-related protein was significantly increased. Macrophages highly expressing adipogenic differentiation-related proteins can increase the expression of acyl-CoA: cholesterol acyltransferase 1, promote the accumulation of intracellular cholesterol ester and synergistically enhance the action of oxidized low-density lipoprotein. The effect of increasing expression of proteins related to adipogenesis was diminished by the addition of clavulanic acid. Conclusion Highly expressed lipofunctional proteins can up-regulate the expression of acyl-CoA: cholesterol acyltransferase 1 in THP-1 macrophages and promote intracellular lipid accumulation. Lipid differentiation-related proteins may promote the accumulation of intracellular cholesterol esters by acyl-CoA: cholesterol acyltransferase 1.