Alipui C, Tenner T E, Ramos K
Department of Pharmacology, Texas Tech University Health Sciences Center, Lubbock 79430.
J Pharmacol Methods. 1991 Nov;26(3):211-22. doi: 10.1016/0160-5402(91)90045-7.
Atherosclerotic vascular disease is the most common complication of diabetes mellitus. Enhanced vascular smooth muscle cell proliferation plays a central role in atherosclerotic lesion formation. Studies using explant cultures have demonstrated that aortic smooth muscle cells from rats with experimental or genetic diabetes have enhanced rates of proliferation when compared to controls. However, this method of culture may select for cells with enhanced migratory potential. In the present studies, aortic smooth muscle cells were successfully cultured from control and diabetic rabbits after enzymatic and mechanical dispersion from thoracic aortic segments. The proliferative patterns of control cells were characterized and growth rates of diabetic cells were compared to controls. Primary cultures from control rabbits grew after an initial 5-day lag period to achieve threefold increases in cell number by 9 days. Subcultures of aortic smooth muscle cells entered the logarithmic phase of growth after 2 days, reaching the plateau phase of growth in 5-7 days and achieving three to fourfold increases in cell number. The final density to which cultures grew was not affected by the number of cells attached on day 1 for the range studied. Cells from diabetic rabbits displayed shorter doubling times and reached greater densities at confluence than did cells from controls. These data support the hypothesis that diabetes induces an atherogenic response. The dissociated rabbit aortic smooth muscle cell culture provides a model in which to study diabetes-induced modulation of cell proliferation that is amenable to pharmacological manipulation to investigate agonist and growth factor-induced responses.
动脉粥样硬化性血管疾病是糖尿病最常见的并发症。血管平滑肌细胞增殖增强在动脉粥样硬化病变形成中起核心作用。使用外植体培养的研究表明，与对照组相比，患有实验性或遗传性糖尿病的大鼠的主动脉平滑肌细胞增殖速率更高。然而，这种培养方法可能会选择具有增强迁移潜力的细胞。在本研究中，通过酶解和机械分散从胸主动脉段成功培养出对照兔和糖尿病兔的主动脉平滑肌细胞。对对照细胞的增殖模式进行了表征，并将糖尿病细胞的生长速率与对照细胞进行了比较。对照兔的原代培养物在最初5天的延迟期后开始生长，到第9天时细胞数量增加了三倍。主动脉平滑肌细胞的传代培养在2天后进入对数生长期，在5 - 7天达到生长平台期，细胞数量增加三到四倍。在所研究的范围内，培养物生长的最终密度不受第1天附着细胞数量的影响。糖尿病兔的细胞比对照兔的细胞具有更短的倍增时间，汇合时达到更高的密度。这些数据支持糖尿病诱导致动脉粥样硬化反应的假说。解离的兔主动脉平滑肌细胞培养提供了一个模型，可用于研究糖尿病诱导的细胞增殖调节，该模型适合进行药理操作以研究激动剂和生长因子诱导的反应。
