酰基辅酶A:胆固醇酰基转移酶1反义寡核苷酸对泡沫细胞形成的影响  

Effect of acyl coenzyme A:cholesteryl acyltransferase 1 antisense oligonucleotides on the formation of foam cells

作　　者：何平[1] 成蓓[1] 戚本玲[1] 

机构地区：[1]华中科技大学同济医学院协和医院老年病科,湖北武汉430022

出　　处：《中国病理生理杂志》2008年第8期1616-1619,共4页Chinese Journal of Pathophysiology

基　　金：国家自然科学基金资助项目(No.30170378)

摘　　要：目的:观察酰基辅酶A:胆固醇酰基转移酶1(ACAT1)反义寡核苷酸对泡沫细胞(FC)形成的影响。方法:体外培养人THP-1单核细胞系,由佛波酯(PMA)作用使其分化为巨噬细胞;并建立过表达ACAT1基因的THP-1单核细胞系。在脂质体的介导下将针对ACAT1基因序列的反义和错义寡核苷酸分别作用于上述细胞6h后加入Ac-LDL进行脂质负荷,以Western blotting法检测ACAT1蛋白表达,放射性同位素标记底物法检测ACAT酶活性的变化,油红O染色法观察泡沫细胞的形成情况。结果:ACAT1反义寡核苷酸可明显抑制巨噬细胞和过表达ACAT1基因的THP-1单核细胞内的ACAT酶活性,而且可明显抑制Ac-LDL负荷后泡沫细胞的形成;错义寡核苷酸则无此作用。结论:ACAT1反义寡核苷酸可抑制ACAT的生物学活性及泡沫细胞的形成。AIM ： To study the effect of acyl coenzyme A ： cholesteryl acyltransferase 1 （ ACAT1 ） antisense oligonucleotides on the formation of foam cells （ FC ）. METHODS ： THP - 1 cells were cultured and differentiated into macrophages （MP） by phorbol myfistate acetate （PMA）. Over - expressing ACAT1 gene THP - 1 cells were constructed. The ACAT1 antisense and missense oligonucleotides conducted by LipofectamineTM 2000 were incubated with above cells. Ac - LDL was added 6 h later and incubated for 24 h. The expression of ACAT1 protein was detected by Western blotting. The ACAT activity was measured by quantifying the incorporation of [ 1 - 14C] oleoyl CoA into cholesteryl esters. The formation of foam cells was detected by oil red O staining. RESULTS ： The ACAT1 antisense oligonucleotides inhibited the activity of ACAT in macrophages and over - expressing ACAT1 gene THP - 1 cells. It also inhibited the formation of foam cell in mac- rophages and over - expressing ACAT1 gene THP - 1 cells with lipid loading. The missense oligonucleotides did not show the inhibitory effects. CONCLUSION ： The ACAT1 antisense oligonucleotides inhibit the activity of ACAT and the formation of foam cells.

关 键 词：酰基辅酶A 胆固醇酰基转移酶 泡沫细胞 寡核苷酸类 反义 

分 类 号：R363[医药卫生—病理学]