目的:构建THP-1源性泡沫细胞模型,探讨血管紧张素-(1-7)[Ang-(1-7)]和腺苷酸环化酶抑制剂MDL对THP-1源性泡沫细胞胆固醇逆转运相关蛋白--ATP结合盒转运子A1(ABCA1)表达和胆固醇含量的影响.方法:将THP-1源性单核细胞诱导为巨噬细胞,加入氧化低密度脂蛋白(ox-LDL)建立泡沫细胞模型.实验分为4组:正常泡沫细胞组、MDL组、Ang-(1-7)组和MDL+Ang-(1-7)组.处理24 h后,分别通过ELISA法、荧光定量PCR法和Western blotting检测各组细胞cAMP表达水平、ABCA1 mRNA相对表达量和蛋白含量,闪烁计数法检测各组细胞胆固醇流出率,高效液相色谱法检测各组细胞胆固醇含量.结果:(1)Ang-(1-7)组cAMP表达水平、ABCA1 mRNA相对表达量和蛋白含量较泡沫细胞组有明显升高(P<0.05),胆固醇流出率较泡沫细胞组明显增高(P<0.05),胆固醇含量较泡沫细胞组显著降低(P<0.05).(2)MDL组cAMP表达水平、ABCA1 mRNA相对表达量和蛋白含量较泡沫细胞组有明显降低(P<0.05),胆固醇流出率较泡沫细胞组明显降低(P<0.05),胆固醇含量较泡沫细胞组增高(P<0.05).(3)MDL+Ang-(1-7)组数值介于Ang-(1-7)组与泡沫细胞组之间.结论:(1)Ang-(1-7)能促进泡沫细胞ABCA1的表达增加,促进胆固醇外流,减少泡沫细胞胆固醇含量,逆转泡沫细胞的形成.(2) MDL可以抑制腺苷酸环化酶的活性,减少cAMP的生成,从而部分拮抗Ang-(1-7)的上述作用,但不能完全阻断Ang-(1-7)的作用.%AIM; To establish the THP - 1 - derived foam cell formalion and Lo evaluale the effecls of angio-Lensin - ( 1 - 7 ) [ Ang - ( 1 - 7 ) ] and MDL ( an inhibitor of adenylale cyclase) on the expression of ATP - binding cassele transporter Al(ABCA1) and the conlenl of choleslerol. METHODS; THP - 1 - derived macrophages were trealed with oxidized low - densily lipoprolein( ox - LDL) lo develop inlo foam cells. The foam cells were divided inlo 4 groups; conlrol group, MDL group, Ang - ( 1 - 7 ) group and MDL + Ang - ( 1 - 7 ) group. Al 24 h after trealment, the conlenl of cAMP was measured by ELISA. The mRNA and prolein levels of ABCA1 were determined by real - lime RT - PCR and Western blotting, respectively. The conlenl of choleslerol was delecled by high performance liquid chromalography. RESULTS; The cAMP, the mRNA and prolein levels of ABCA1 in Ang - (1 - 7 ) group were significantly higher, and the conlenl of choleslerol was significantly lower than those in conlrol group (P < 0. 05). On the contrary, the cAMP, the mRNA and protein levels of ABCA1 in MDL group were significantly lower and the conlenl of choleslerol was significantly higher than those in conlrol group ( P < 0. 05 ) . The results in MDL + Ang - ( 1 - 7 ) group were belween Ang - ( 1 - 7 ) group and control group. CONCLUSION: Ang - ( 1 -7) inhibits the formalion of foam cells by promoting the expression of ABCA1 and decreasing the conlenl of choleslerol. MDL partly anlagonizes the effecl of Ang - (1 - 7) by inhibiting the adenylate cyclase and decreasing the conlenl of cAMP.
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