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组织培养中的猪主动脉平滑肌。纯化猪脂蛋白对细胞生长和形态的一些影响。

Swine aortic smooth muscle in tissue culture. Some effects of purified swine lipoproteins on cell growth and morphology.

作者信息

Brown B G, Mahley R, Assmann G

出版信息

Circ Res. 1976 Sep;39(3):415-24. doi: 10.1161/01.res.39.3.415.

Abstract

Smooth muscle cells (SMC) were grown from inner media explants of swine aorta and used as a model for studying the role of lipoproteins in atherogenesis. These cultured cells retain the characteristics of SMC through multiple passages. Cell growth curves, in time, were obtained by using standard counting techniques, SMC grew slowly (0.019 cycle/day) in modified Dulbecco-Vogt medium supplemented with 1.5% swine serum. Purified lipoproteins were prepared from three normolipidemic and two hyperlipidemic (cholesterol-fed) swine. When the medium of 84 growth experiments was supplemented with these lipoproteins, SMC growth rate increased linearly with lipoprotein cholesterol concentration up to 10 mg/dl. At 10 mg/dl of lipoprotein cholesterol, very low density lipoproteins (VLDL) increased growth rate 7.2-fold (P less than 0.01); low density lipoproteins (LDL) 5.7-fold (P less than 0.01); high density lipoproteins (HDL2) 3,4-fold (P less than 0.02); and HDLc, and lipoprotein appearing in the hyperlipidemic swine, 3.0-fold (P less than 0.01). Addition of 10% lipoprotein-free serum stimulated growth rate 6.0-fold (P less than 0.01). There was no difference between normo- and hyperlipidemic lipoproteins with respect to cell growth rate. Factors present in the ultracentrifugal bottom, and factors appearing during the platelet release reaction, were shown to contribute to the SMC growth response. Morphological alterations characteristic of intimal foam cells occurred in SMC grown in VLDL at triglyceride levels in excess of 15 mg per 100 ml. Thus there are distinct parallels between SMC response in this model in vitro and atherogenesis in vivo.

摘要

平滑肌细胞(SMC)取自猪主动脉的内膜培养基外植体并进行培养,用作研究脂蛋白在动脉粥样硬化形成中作用的模型。这些培养的细胞经过多次传代仍保留平滑肌细胞的特征。通过标准计数技术获得细胞生长曲线,在补充有1.5%猪血清的改良杜尔贝科 - 沃格特培养基中,平滑肌细胞生长缓慢(0.019个周期/天)。从三只血脂正常和两只高脂血症(喂胆固醇)猪中制备纯化的脂蛋白。当在84次生长实验的培养基中添加这些脂蛋白时,平滑肌细胞生长速率随脂蛋白胆固醇浓度线性增加,直至10mg/dl。在脂蛋白胆固醇浓度为10mg/dl时,极低密度脂蛋白(VLDL)使生长速率提高7.2倍(P小于0.01);低密度脂蛋白(LDL)提高5.7倍(P小于0.01);高密度脂蛋白2(HDL2)提高3.4倍(P小于0.02);以及高脂血症猪中出现的HDLc和脂蛋白提高3.0倍(P小于0.01)。添加10%无脂蛋白血清使生长速率提高6.0倍(P小于0.01)。正常血脂和高脂血症脂蛋白在细胞生长速率方面没有差异。超速离心底部存在的因子以及血小板释放反应期间出现的因子被证明有助于平滑肌细胞的生长反应。当平滑肌细胞在甘油三酯水平超过每100ml 15mg的VLDL中生长时,会出现内膜泡沫细胞特有的形态学改变。因此,该体外模型中平滑肌细胞的反应与体内动脉粥样硬化形成之间存在明显的相似之处。

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