Gurbuz H Alper, Durukan A Baris, Sevim Handan, Ergin Ekin, Gurpinar Aylin, Yorgancioglu Cem
aDepartment of Cardiovascular Surgery, Medicana International Ankara Hospital bDepartment of Biology, Hacettepe University Faculty of Science, Ankara, Turkey.
Blood Coagul Fibrinolysis. 2013 Oct;24(7):742-5. doi: 10.1097/MBC.0b013e3283629bbc.
Heparin is a universal drug used frequently for its anticoagulant effects. The variabilities in distribution and tendency of heparin to accumulate in tissues cause increased tissue concentrations despite normal serum levels. We aimed to underline the toxic effects of heparin in cell culture make projections for clinical applications. L929 mouse fibroblastic cell line was plated in 96-well culture plates at an initial density of 5000 cells/well. Heparin was prepared in 10 different concentrations (10-300 units/well). Following 3 days of incubation, viabilities were assessed by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay for each concentration in each day and compared. The viability of cells decreased significantly with increasing doses of heparin; at least 50 units/well in the first and second days and at least 20 units/well in the third day (P < 0.05 for each). There was statistically significant difference when the viabilities of cells treated with same heparin concentration in different days were compared (P < 0.05). The authors clearly demonstrated the toxic effects of heparin in cell culture, toxic effects increased as the dose increased. To prevent the unwanted clinical side-effects of heparin further studies should be made and more accurate testing methods should be developed to determine the effective tissue concentration of heparin.
肝素是一种因其抗凝作用而经常使用的通用药物。尽管血清水平正常，但肝素在组织中的分布和蓄积趋势的变异性会导致组织浓度升高。我们旨在强调肝素在细胞培养中的毒性作用，并为临床应用做出预测。将L929小鼠成纤维细胞系以5000个细胞/孔的初始密度接种于96孔培养板中。肝素以10种不同浓度（10 - 300单位/孔）配制。孵育3天后，每天对每种浓度通过3 -（4,5 - 二甲基噻唑 - 2 - 基）- 2,5 - 二苯基四氮唑溴盐（MTT）法评估细胞活力并进行比较。随着肝素剂量增加，细胞活力显著降低；第一天和第二天至少为50单位/孔，第三天至少为20单位/孔（每种情况P < 0.05）。比较不同天数用相同肝素浓度处理的细胞活力时存在统计学显著差异（P < 0.05）。作者清楚地证明了肝素在细胞培养中的毒性作用，毒性作用随剂量增加而增强。为防止肝素产生不良临床副作用，应进一步开展研究并开发更准确的检测方法以确定肝素的有效组织浓度。
