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目的:探讨白细胞介素-1β(interleukin-1β,IL-1β)对HepG2细胞羟甲基戊二酸单酰辅酶A(3-hydroxy-3-methyl-glutaryl-coenzyme A,HMGCo A)还原酶及脂质积聚的影响。方法:200μg/m L低密度脂蛋白(low density lipoprotein,LDL)负荷下的HepG2细胞分别给予0 ng/m L(对照组)、20 ng/m L(炎症组)IL-1β处理24 h。荧光定量PCR(RT-PCR)、Western blot分别测定HMGCo A还原酶mRNA及蛋白表达水平;薄层层析法检测HMGCo A还原酶活性;同位素标记法评估胆固醇合成情况;油红O染色及酶法定量检测细胞内脂质。结果:IL-1β处理下,HepG2细胞HMGCo A还原酶mRNA表达水平为对照组的(1.54±0.04)倍(t=5.619,P=0.001)、蛋白表达水平为对照组的(1.92±0.12)倍(t=7.745,P=0.002),且HMGCo A还原酶的酶活性为对照组的(1.73±0.25)倍(t=2.476,P=0.048)。IL-1β处理使HepG2细胞的胆固醇合成增多为对照组的(1.32±0.11)倍(t=2.316,P=0.036)。IL-1β处理的HepG2细胞内红染的脂滴显著多于对照组,且IL-1β处理组细胞内总胆固醇含量(22.43±1.62)为对照组(14.90±1.11)的1.51倍(t=3.845,P=0.018),胆固醇酯的含量(3.84±0.20)为对照组(1.41±0.05)的2.72倍(t=11.730,P=0.000)。结论:IL-1β可以上调HepG2细胞HMGCo A还原酶的mRNA和蛋白表达水平,并增强其酶活性,促进HepG2细胞内胆固醇合成和脂质积聚。
AIM: To investigate the effect of interleukin-1β (IL-1β) on HepG2 cells induced by 3-hydroxy-3-methyl-glutaryl coenzyme A Effect of lipid accumulation. Methods: HepG2 cells were treated with 200 ng / ml L-1 (low density lipoprotein) and 20 ng / ml IL-1β (inflammation group) for 24 h. The mRNA and protein levels of HMGCo A reductase were determined by RT-PCR and Western blot respectively. The activity of HMGCo A reductase was determined by TLC. The cholesterol synthesis was evaluated by isotope labeling. Quantitative detection of intracellular lipid. Results: The mRNA expression level of HMGCo A reductase in HepG2 cells was (1.54 ± 0.04) times higher than that in control group (t = 5.619, P = 0.001) and the protein expression level was (1.92 ± 0.12) times (t = 7.745, P = 0.002), and the enzyme activity of HMGCo A reductase was (1.73 ± 0.25) times that of the control group (t = 2.476, P = 0.048). IL-1β treatment increased the cholesterol synthesis in HepG2 cells to 1.32 ± 0.11 times of the control group (t = 2.316, P = 0.036). The lipid droplets of red stained HepG2 cells treated with IL-1β were significantly more than those of the control group, and the intracellular total cholesterol (IL-1β) was 1.51 times (22.43 ± 1.62) in the control group (14.90 ± 1.11) (t = 3.845 , P = 0.018). The cholesterol ester content (3.84 ± 0.20) was 2.72 times of that of the control group (1.41 ± 0.05) (t = 11.730, P = 0.000). CONCLUSION: IL-1β can up-regulate the mRNA and protein expression of HMGCo A reductase in HepG2 cells and enhance its enzymatic activity and promote the synthesis of cholesterol and lipid accumulation in HepG2 cells.