雷帕霉素对血管平滑肌细胞增殖影响的实验研究  

Experimental Study of Effect of Rapamycin on Proliferation of Rat Aortic Smooth Muscle Cells

作　　者：王建波[1] 杨建勇[1] 陈伟[1] 庄文权[1] 张珑涓[2] 

机构地区：[1]中山大学附属第一医院放射介入科,广东广州510080 [2]中山大学附属第一医院外科实验室,广东广州510080

出　　处：《中山大学学报（医学科学版）》2006年第3期254-257,共4页Journal of Sun Yat-Sen University:Medical Sciences

基　　金：国家自然科学基金资助项目(30270417)

摘　　要：目的观察雷帕霉素(rapamycin)对体外培养的大鼠胸主动脉平滑肌细胞(SMC)增殖的作用。方法组织贴块法体外原代培养大鼠胸主动脉平滑肌细胞,取对数生长期细胞(第6￣10代)用作实验,MTT法检测不同浓度雷帕霉素对平滑肌细胞增殖的效果,并用RT-PCR法检测其对平滑肌细胞增殖细胞核抗原(proliferatingcellnuclearantigen,PCNA)mRNA表达情况的影响。结果①MTT法检测显示雷帕霉素呈剂量依赖性抑制体外培养的大鼠平滑肌细胞增殖,各组吸光值(A)分别为:对照组0.902±0.106;0.1ng/mL组0.873±0.079;1ng/mL组0.797±0.046;10ng/mL组0.692±0.061;100ng/mL组0.624±0.075。方差分析差异有显著性,F=28.85,P<0.001;两两比较1ng/mL以上浓度组与对照组间差异均有显著性,不同浓度间差异显著。0.1ng/mL组与对照组相比差异无显著性。②RT-PCR显示雷帕霉素以浓度依赖模式抑制大鼠平滑肌细胞PCNA-mRNA合成。各组校正后光密度值分别为:对照组219.35±7.20;0.1ng/mL组212.39±6.56;1ng/mL组113.15±10.09;10ng/mL组97.17±12.25;100ng/mL组84.38±8.66。方差分析F=195.49,P<0.001。两两比较与MTT检测相同。结论雷帕霉素呈剂量依赖性显著抑制大鼠平滑肌细胞的增殖,1ng/mL时即可明显起效。雷帕霉素有望成为治疗内支架术后再狭窄的理想药物之一。[Objective] To evaluate the proliferative effect of rapamycin on the cultured rat thoracic aortic smooth muscle cells （SMC） in vitro. [Methods] The rat thoracic aortic SMC were harvested and cultured for six to ten passages. Cultured SMC were synchronized, then restimulated to enter the cell cycle, and treated with incremental concentrations of rapamycin or without rapamycin as contrast. Forty-eight hours after managed, the proliferative assay was performed with MTr method and RT-PCR was performed to assay the level of proliferating cell nuclear antigen （PCNA） mRNA. [Result]（1） Rapamycin progressively inhibits rat aortic SMC proliferation in a concentration-dependent fashion. The absorption （A） value of each group was statistically significant for different concentrations （F=28.85,P〈 0.001） and the results were： control, 0.902±0.106; 0.1 ng/mL, 0.873±0.079; 1 ng/mL, 0.797±0.046; 10 ng/mL, 0.692±0.061; 100 ng/mL, 0.624±0.075. Compared to control group or among the different concentration, the difference of A value was statistically significant when the concentration of rapamycin was greater than 1 ng/mL. The difference was not statistically significant when the concentration was 0.1 ng/mL. （2） RT-PCR suggested rapamycin significantly decreased rat aortic SMC PCNA-mRNA transcription in a concentrationdependent fashion. The results of statistical analysis were similar with MTr assay （ F=195.491, P〈 0.001）. The optic density values of each group were： control, 219.35±7.20; 0.1 ng/mL, 212.39±6.56; 1 ng/mL, 113.15±10.09; 10 ng/ mL, 97.17±12.25; 100 ng/mL, 84.38±8.66. [Conclusion] Rapamycin inhibits rat aortic SMC proliferation in a concentration-dependent fashion. It is suggested the action concentration was 1 ng/mL. Rapamycin maybe one of the best drugs in the treatment of the in-stent restenosis after vascular interventional operation.

关 键 词：雷帕霉素 血管平滑肌细胞 细胞增殖 大鼠 

分 类 号：R363.11[医药卫生—病理学]