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oxLDL对转人apoAI基因小鼠动脉平滑肌细胞胆固醇摄取和清除的影响
Effect of oxLDL on the Uptake and Clearance Rate of Cholesterol in Vascular Smooth Muscle Cells Originated from Human apoAI Transgenic Mice
【摘要】 目的 明确氧化低密度脂蛋白(oxLDL)对转人apoAI基因 C57BL/6小鼠主动脉平滑肌细胞(tr-SMC)胆固醇清除与摄取的影响;转入的apoAI基因是否有助于减轻oxLDL引起的SMC胆固醇积聚和对细胞的损害;以及oxLDL的刺激,对细胞已转入的apoAI基因表达的影响。方法 用已建立的含小鼠金属硫蛋白I(MT-I)启动子的人apoAI转基因C57BL/6 小鼠主动脉SMC,通过3H-胆固醇标记率和清除率测定,观察oxLDL对tr-SMC胆固醇摄取与清除的影响。通过RT-PCR及 Northern blot技术检测oxLDL对tr-SMC基因表达的影响。结果 30 μg/ml oxLDL明显促进小鼠SMC增生;未见tr-SMC与非转基因小鼠平滑肌细胞(n-SMC)在标记率间的差别,oxLDL刺激后标记率增加约100%;tr-SMC较n-SMC细胞内胆固醇清除率增加40% ~ 50%。oxLDL刺激后,n-SMC清除率下降28%,而 tr-SMC仅下降10%。RT-PCR 及Northern blot表明oxLDL刺激后tr-SMC 的h-apoAI基因表达明显增加。结论 转人apoAI 基因小鼠SMC的基因表达有助于减轻oxLDL引起的SMC胆固醇积聚;oxLDL的刺激促进tr-SMC 的人apo AI基因表达,表达的 h-apo AI可抑制oxLDL对平滑肌细胞的损害。
【Abstract】 Objective Study on(1)inhibition of oxidized low density lipoprotein(oxLDL)effect on the upt- ake and clearance of intra-cellular 3H-cholesterol in vascular smooth muscle cells(v-SMC)originated from the human-apoAI transgenic mice(C57BL/6);(2)change of human-apolipoprotein AI(h-apoAI)mRNA expression in v-SMC after oxLDL stimulation and the protective effect of expressed h-apoAI on v-SMC against oxLDL intoxication. Methods (1)v-SMC isolated from human apoAI transgenic mice possessing a recombined gene connected beforehand with a mouse metallothionein-I(MT-I)as the pro- moter;(2)study of h-apoAI mRNA expression from v-SMC of the transgenic mice by RT-PCR and Northern blot. Results oxLDL(30 μg/ml) strongly promoted v-SMC proliferation. No difference found on 3H-cholesterol uptake between smooth muscle from normal mouse aorta(n-SMC)and smooth muscle cells from transgenic mouse aorta(tr-SMC) of the control groups, the uptake rates of both kinds of SMC rose 100% after oxLDL stimulation. The efflux rates of 3H-cholesterol in tr-SMC were much higher than those of n-SMC(40%~ 50%). After oxLDL stimulation, the clearance rates fell by 28% and 10% respectively for n-SMC and tr-SMC. The result of RT-PCR and Northern blot showed a marked increase of h-apoAI gene expression after oxLDL stimulation Conclusions Expression of h-apoAI gene in C57BL/6 mice enables to decrease the accumulation of cholesterol in v-SMC. tr-SMC are capable to alleviate the harmful effect of oxLDL on v-SMC due to the increase of h-apoAI expression.
【Key words】 oxidized low density lipoprotein; apolipoprotein AI; transgenic mice; smooth muscle cells; cholesterol;
- 【文献出处】 中国医学科学院学报 ,Acta Academiae Medicinae Sinicae , 编辑部邮箱 ,2001年04期
- 【分类号】R543.5
- 【被引频次】1
- 【下载频次】83