实时定量PCR检测胰岛素和高糖对巨噬细胞ACAT-1表达的影响  

Impact of Insulin and Glucose on the ACAT-1 mRNA Expression in Macrophage Detected by Real-time Quantitative PCR

作　　者：陆菡[1] 成蓓[1] 何平[1] 王彦富[1] 曾永利[1] 葛晶[1] 

机构地区：[1]华中科技大学同济医学院附属协和医院老年病科,湖北省武汉市430022

出　　处：《中国全科医学》2007年第4期269-272,共4页Chinese General Practice

基　　金：国家自然科学基金资助项目(30471921)

摘　　要：目的利用实时定量聚合酶链反应(PCR)法检测巨噬细胞酰基辅酶A:胆固醇酰基转移酶-1(ACAT-1)mRNA的表达。探讨胰岛素和高糖对人THP-1细胞分化的巨噬细胞ACAT-1mRNA表达的影响。方法体外培养人单核细胞株THP-1细胞,由佛波醇肉豆蔻酸乙酸盐(PMA)作用使其分化为巨噬细胞,后者再由胰岛素和高糖共同干预不同时间(0、6、12、24、48h),用实时定量PCR检测ACAT-1mRNA表达。结果胰岛素和高糖共同作用不同时间后巨噬细胞ACAT-1mRNA的表达量间差别有显著性意义(P<0.01),作用12h与0、6、24、48h的ACAT-1mRNA表达量间差别均有显著性意义(P<0.01),作用48h的ACAT-1mRNA表达量与0h相比差别无显著性意义(P>0.05)。结论胰岛素和高糖共同作用下,巨噬细胞ACAT-1mRNA表达逐渐增加,12h达到高峰,然后逐渐下降,48h恢复到0h时水平。成功建立和应用实时定量PCR检测巨噬细胞ACAT-1mRNA水平是准确评价巨噬细胞功能的方法之一。Objective To establish a real - time quantitative PCR for detecting the expression of ACAT - 1 （ acyl co- enzymcA： cholesteryl acyltransferase - 1 ） gene in THP - 1 - derived macrophages, to evaluate the impact of glucose and insulin on ACAT - 1 mRNA expression in THP - 1 - derived macrophages. Methods The THP - 1 cells were in vitro incubated with PMA for 48 hours, then exposed to high concentration of glucose and insulin for different time. The ACAT - 1 mRNA level of the cells was determined by real - time PCR. Results The expression of ACAT - 1 mRNA in macrophages differed significantly in a time - dependent manner under the combined administration of insulin and high concentration glucose （ P 〈0. 01 ）. The level of ACAT - 1 mRNA was significantly higher at the 12 h point than that of 0, 6, 24, 48 h point （ P 〈0.01 ）. The level of ACAT - 1 mRNA had no difference between the 0 h point and the 48 h point （ P 〉 0. 05 ）. Conclusion The level of ACAT - 1 mRNA increases gradually under the combined administration of insulin and high concentration glucose from 0 h to 12 h point and reached the peak at 12 h, then it goes down gradually, it is at the same level asthe beginning （0 h point） at the 48 h point. The function of macrophages could be accurately evaluated by detecting the ACAT - 1 mRNA levels with the real - time PCR.

关 键 词：聚合酶链反应 酰基辅酶A 胆固醇酰基转移酶-1 胰岛素 葡萄糖 

分 类 号：R587.1[医药卫生—内分泌]