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手术准备在人隐静脉培养物中诱导损伤并促进平滑肌细胞增殖。

Surgical preparation induces injury and promotes smooth muscle cell proliferation in a culture of human saphenous vein.

作者信息

Soyombo A A, Angelini G D, Bryan A J, Newby A C

机构信息

Department of Cardiology, University of Wales College of Medicine, Cardiff, United Kingdom.

出版信息

Cardiovasc Res. 1993 Nov;27(11):1961-7. doi: 10.1093/cvr/27.11.1961.

Abstract

OBJECTIVE

The aim was to investigate the influence of vessel wall injury, incurred during routine vein preparation, on smooth muscle cell proliferation.

METHODS

A newly developed quantitative organ culture was used, in which segments of human saphenous vein were cultured in medium containing 30% fetal bovine serum and 1 microCi.ml-1 of [3H]thymidine for up to 14 d. Endothelial integrity was measured by scanning electron microscopy and medial cell viability by adenine nucleotide concentrations. Cell proliferation was measured by DNA concentration, global incorporation of [3H]thymidine, and by counting labelled cells in autoradiographs of transverse sections.

RESULTS

Surgical preparation led to endothelial injury and reduced adenine triphosphate concentration by 60%. Surgically prepared veins also suffered a significant decline in DNA concentration during culture, which implied that injury led to cell necrosis. Surgically prepared veins showed 2.1- and 2.7-fold greater global incorporation of [3H]thymidine than freshly isolated veins after 7 and 14 d in culture, respectively, which corresponded with a 23-fold and 11-fold greater abundance of thymidine labelled cells in the medial layer. Intimal thickening and the numbers of total and thymidine labelled cells in the intimal layer were similar.

CONCLUSIONS

The data show that injury incurred during routine surgical preparation is associated with enhanced medial smooth muscle cell proliferation. The effect of injury was most probably to permit an increased response of medial smooth muscle cells to serum derived mitogens.

摘要

目的

研究常规静脉制备过程中发生的血管壁损伤对平滑肌细胞增殖的影响。

方法

采用一种新开发的定量器官培养方法,将人隐静脉段在含有30%胎牛血清和1微居里/毫升[3H]胸腺嘧啶核苷的培养基中培养长达14天。通过扫描电子显微镜测量内皮完整性,通过腺嘌呤核苷酸浓度测量中层细胞活力。通过DNA浓度、[3H]胸腺嘧啶核苷的整体掺入以及在横切面放射自显影片中计数标记细胞来测量细胞增殖。

结果

手术制备导致内皮损伤,三磷酸腺苷浓度降低60%。手术制备的静脉在培养过程中DNA浓度也显著下降,这表明损伤导致细胞坏死。手术制备的静脉在培养7天和14天后,[3H]胸腺嘧啶核苷的整体掺入分别比新鲜分离的静脉高2.1倍和2.7倍,这与中层中胸腺嘧啶核苷标记细胞的丰度分别高23倍和11倍相对应。内膜增厚以及内膜层中总细胞和胸腺嘧啶核苷标记细胞的数量相似。

结论

数据表明,常规手术制备过程中发生的损伤与中层平滑肌细胞增殖增强有关。损伤的影响很可能是使中层平滑肌细胞对血清源性促有丝分裂原的反应增加。

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