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目的观察白细胞介素1β(IL-1β)能否干扰美伐他汀降低HepG2细胞内胆固醇含量的作用。方法给予不同浓度的美伐他汀(0,1,10,50,100μmol·L~(-1))单独处理或联合20 ng·mL~(-1)IL-1β处理HepG2细胞24 h。用酶学反应法检测3-羟基3-甲基戊二酸单酰辅酶A还原酶(HMG-CoA-R)的酶活性,用实时荧光定量聚合酶链式反应(Real-Time PCR)方法测定低密度脂蛋白胆固醇(LDL)受体中mRNA的表达水平。然后选取100μmol·L~(-1)美伐他汀单独处理或联合20 ng·mL~(-1)IL-1β处理HepG2细胞24 h,用酶学反应法测定细胞内胆固醇含量,用Real-Time PCR测定细胞内酰基辅酶A-胆固醇酰基转移酶(ACAT)酶表达。结果美伐他汀能显著抑制HepG2细胞的HMG-CoA还原酶活性。IL-1β能有效地削弱美伐他汀对酶活性的抑制作用,同时进一步上调LDL受体的mRNA表达。IL-1β还能促进美伐他汀处理的HepG2细胞中ACAT mRNA的表达,增加细胞内胆固醇酯的含量。结论 IL-1β能增强HepG2细胞的HMG-CoA还原酶活性,促进ACAT酶和LDL受体表达,导致细胞脂质摄入和酯化增加。
Objective To investigate whether Interleukin-1β (IL-1β) can interfere with the effect of mevastatin on reducing the cholesterol content in HepG2 cells. Methods HepG2 cells were treated with different concentrations of mevastatin (0, 1, 10, 50 and 100 μmol·L -1) alone or in combination with 20 ng · mL -1 IL-1β for 24 h. The enzymatic activity of 3-hydroxy-3-methylglutaryl coenzyme A reductase (HMG-CoA-R) was detected by enzymatic reaction method and was determined by Real-Time PCR Low density lipoprotein cholesterol (LDL) receptor mRNA expression levels. Then HepG2 cells were treated with 100 μmol·L -1 mevastatin alone or in combination with 20 ng · mL -1 IL-1β for 24 h. The intracellular cholesterol content was determined by Real Time PCR determination of intracellular acyl-CoA-cholesterol acyltransferase (ACAT) enzyme expression. Results Mevastatin significantly inhibited the HMG-CoA reductase activity of HepG2 cells. IL-1β can effectively weaken the inhibitory effect of mevastatin on the enzyme activity, and further up-regulate the mRNA expression of LDL receptor. IL-1β can promote ACAT mRNA expression in mevastatin-treated HepG2 cells and increase intracellular cholesterol ester content. Conclusion IL-1β can enhance the HMG-CoA reductase activity of HepG2 cells and promote the expression of ACAT and LDL receptors, resulting in increased lipid uptake and esterification.