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机构地区:[1]华中科技大学同济医学院基础医学院生物化学与分子生物学系,武汉430030
出 处:《华中科技大学学报(医学版)》2006年第1期14-17,共4页Acta Medicinae Universitatis Scientiae et Technologiae Huazhong
基 金:国家自然科学基金资助项目(No.39970307)
摘 要:目的探讨氧化型低密度脂蛋白(oxLDL)对巨噬细胞内ABCA1表达及功能的影响。方法分别以100μg/ml乙酰化低密度脂蛋白(acLDL)和oxLDL温育RAW264.7细胞24 h,应用胆固醇外排实验、半定量RT-PCR、Western blot检测oxLDL及acLDL对小鼠巨噬细胞内ABCA1功能、mRNA及蛋白质表达的影响。结果加入载脂蛋白apoA-I 24 h后,acLDL负荷的胆固醇约(37.65±2.29)%被排出细胞,而oxLDL组仅排出(9.78±2.16)%,差异有显著性意义(P<0.05);oxLDL及acLDL均可使ABCA1 mRNA水平升高3倍左右,并分别使其蛋白质表达水平提高(1.57±0.18)倍和(1.26±0.09)倍,两组相比,差异无显著性意义(P>0.05)。结论oxLDL胆固醇负荷对ABCA1表达及功能的影响不一致,推测还存在其他调控ABCA1功能的机制使oxLDL负荷的胆固醇外排障碍。Objective To study the in vitro effect of oxLDL, on the expression and function of ATP binding cassette transporter A1 ATP binding cassette transporter A1 (ABCAI). Methods RAW264.7 cells were incubated with 100μg/ml acLDL or oxLDL for 24 h. The function of ABCA1 was examined by cholesterol efflux experiment, and the expression levels of ABCA1 were detected by semi-quantitative RT -PCR and Western blot. Results After the RAW264.7 cells were incubated with apoA-Ⅰ for 24 h. about (37.65±2.29) %cholesterol loaded by acLDL, was expelled from RAW264.7 cells, and only (9.78± 2. 16) % cholesterol was expelled in oxLDL, group with the difference being significant (P〈0. 05). Incubation with 100μg/ml acLDL or oxLDL both increased the level of ABCA1 mRNA to about three times, and elevated the level of ABCA1 protein to (1.57±0. 18) times in acLDL, group and to (1.26±0.09) times in oxLDL, group respectively (P〉0.05). Conclusion There is difference between the effect of oxLDL, on the expression and function of ABCA1. It was speculated that oxLDL decreased the cholesterol efflux mediated by ABCA1 through other unknown mechanisms.
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