oxLDL 对转人apo A1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 v-SMC originated from human apoA1 transgenic mice
目的探讨①oxLDL对转人apoAI C57BL/6小鼠主动脉SMC(trSMC)胆固醇清除与摄取的影响,以衡量转入的apoAI基因是否有助于减轻oxLDL引起的SMC胆固醇积聚;②oxLDL刺激下,转入的apoAI基因的表达状况,以探知在致病的环境下,所转入的基因是否能增加表达,提高抗病能力.方法①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3 H-胆固醇标记率和清除率测定,观察oxLDL 对trSMC胆固醇摄取与清除的影响.②通过RT-PCR及 Northern blot技术检测oxLDL对trSMC基因表达的影响.结果 30μg/ml oxLDL明显促进小鼠SMC增生;未见trSMC与nSMC在标记率间的差别,oxLDL刺激后标记率增加约100%;trSMC清除率与nSMC相比,增加40%-50%; oxLDL刺激后,nSMC清除率下降28%,而 trSMC下降10%;RT-PCR 及Northern blot表明oxLDL刺激后trSMC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;在oxLDL刺激下,trSMC 的人apo A1基因表达增加,减轻了oxLDL的致病作用.
更多Objective To study the inhibition effect of oxLDL on the uptake and clearance of intra-cellular 3 H-cholesterol in v-SMC from the human-apoA1 transgenic mice (C57BL/6) and the changes in human-apoA1 mRNA expression in v-SMC from human apoA1 transgenic mice after oxLDL stimulation. Methods v-SMC originally isolated from human apoA1 transgenic mice connected with a recombined mouse metallothionein-Ⅰ (MT-Ⅰ) promoter was used, and the effect of oxLDL on the uptake and clearance of intracellular 3 H-cholesterol was studied in v-SMC of the transgenic and control mice respectively, the study of h-apoAⅠ mRNA expression from v-SMC of the transgenic mice were done by RT-PCR and Northern blot. Results oxLDL (30μg/ml) strongly promoted the SMC proliferation. No difference was found in 3 H-cholesterol uptake between nSMC and trSMC, and the uptake rates of both kinds of SMC rose 100% after oxLDL stimulation. The efflux rates of 3 H-cholesterol in trSMC were much higher than those of nSMC (40%-50%). After oxLDL stimulation, the clearance rates fell by 28% and 10%, respectively, for nSMC and trSMC. The result of RT-PCR and Northern blot showed that h-apoA1 gene expression was markedly increased by the stimulation of oxLDL. Conclusion Expression of the h-apoA1 gene in C57BL/6 mice enables them to reduce the accumulation of cholesterol in v-SMC. The trSMC can alleviate the harmful effect of oxLDL due to the increase of h-apoA1 expression.
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