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目的探讨胆固醇酯转运蛋白(CETP)对脂肪细胞脂代谢的影响。方法构建稳定表达CETP的单克隆小鼠3T3-L1脂肪前体细胞株,并通过有限稀释法和流式细胞术分选法获得不同CETP表达水平的3T3-L1细胞株加以鉴定,测定细胞内胆固醇含量。结果 pcDNA3.1-CETP-GFP重组质粒构建成功,建立并获得12株稳定表达不同CETP水平的3T3-L1细胞株,挑选出高克隆(克隆编号1)、中克隆(克隆编号6)、低克隆(克隆编号8)CETP表达水平的3T3-L1细胞株。胆固醇测定显示,表达CETP的3T3-L1细胞总胆固醇及游离胆固醇含量增加,并且胆固醇含量随着CETP的水平升高而升高(P<0.05或P<0.01)。结论 CETP可能通过调节脂肪细胞内胆固醇的含量从而影响葡萄糖代谢。
Objective To investigate the effect of cholesteryl ester transfer protein (CETP) on lipid metabolism in adipocytes. Methods Monoclonal mouse 3T3-L1 adipose precursor cell line stably expressing CETP was constructed and 3T3-L1 cell lines with different expression levels of CETP were identified by limiting dilution and flow cytometry. The intracellular cholesterol content. Results The recombinant plasmid pcDNA3.1-CETP-GFP was successfully constructed and 12 stable 3T3-L1 cell lines stably expressing different levels of CETP were established. The clones (clone No. 1), middle clone (clone No. 6) (Clone number 8) CETP expression level 3T3-L1 cell line. Cholesterol assay showed that 3T3-L1 cells expressing CETP increased total cholesterol and free cholesterol levels, and cholesterol levels increased as CETP levels increased (P <0.05 or P <0.01). Conclusion CETP may affect glucose metabolism by regulating the content of cholesterol in adipocytes.