胰岛素对高糖诱导HepG2细胞ACAT2表达的影响

Influence of insulin on acyltransferase-2 expression induced by high glucose in HepG2 cells

目的探讨胰岛素对高糖诱导人肝癌组织(HepG2)细胞中酰基辅酶A:胆固醇酰基转移酶2(ACAT2)表达的影响。方法将培养好的HepG2细胞以0.7×106L-1密度接种于6孔培养板内过夜培养,加入无血清培养基继续培养24 h后分别用正常培养基(对照组)、含终浓度为25 mmol/L甘露醇溶液(甘露醇组)和25 mmol/L葡萄糖溶液(高糖组)培养基分别与细胞孵育12 h;胰岛素组用含总浓度25 mmol/L葡萄糖培养基与细胞孵育4 h后,加入终浓度为30 mU/L胰岛素再孵育8 h。采用反转录-聚合酶链反应(RT-PCR)和蛋白免疫印迹(Western blot)法检测ACAT2的表达水平。结果高糖组HepG2细胞ACAT2表达明显高于对照组(P<0.01),胰岛素组ACAT2表达显著低于高糖组(P<0.01)。结论胰岛素可下调高糖诱导的ACAT2的表达。

Objective To study the effects of insulin on acyl-CoA： cholesterol acyltransferase-2 （ACAT2） expression in- duced by high glucose in HepG2 cells. Methods Cultivated HepG2 cells with density of 0.7×10^6 L^-1 were inoculated in six well culture plates for overnight culture, and continued cultivation in serum-free medium for 24 h,then the HepG2 cells were incubat- ed for 12 h in normal culture medium （control group）, 25 mmol/L mannitol solution （mannitol group） and 25 mmol/L glucose solu- tion （high glucose group ） respectively. In the insulin group, HepG2 cells were incubated by 25mmol/L glucose for 4 h, then re-in- cubated by 30 mU/I, insulin for 8 h. The expression level of ACAT2 was detected by RT-PCR and Western blot. Results The expression of ACAT2 in the high glucose group was significant higher than that in the control group（P〈0.01 ） ,the expression of A CAT2 in the insulin group was significant lower than that in the high glucose group（P〈0.01 ）. Conclusion Insulin can down-regu- late the expression of ACAT2 induced by high glucose.