Injury Of Cultured Endothelial Cells By Thrombin-Stimulated Platelets
Injury Of Cultured Endothelial Cells By Thrombin-Stimulated Platelets
复制标题
凝血酶刺激血小板对培养内皮细胞的损伤
DOI：
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发表时间：
1986
影响因子：
6.7
通讯作者：
J. Mustard
中科院分区：
文献类型：
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作者：
L. Jørgensen;A. Grøthe;T. Larsen;R. Kinlough;J. Mustard
体内刺激的血小板可能会导致内皮损伤。问题是血小板刺激是否可以在体外显示出类似的效果。内皮细胞是从人脐静脉在含有 20% 热灭活血清的 199 培养基中培养的。半汇合培养物，每皿 100-400.000 个细胞，用 Na2Cr51O4 标记。 24 小时后，将人血小板悬浮液（最终浓度为 200.000 个/mm3）和凝血酶（最终浓度为 4 U/ml）添加到培养基中，并将培养皿摇动 15 分钟。测定摇动过程中从培养皿脱离的细胞百分比和从内皮细胞释放到环境液体中的 Cr51 百分比，并将其用作细胞损伤的参数。与没有血小板悬浮液和/或凝血酶的对照相比，使用血小板悬浮液和凝血酶观察到松散细胞和Cr51进入环境液体的百分比增加。凝血酶与血小板悬浮液反应后的不含血小板的上清液也具有类似的效果。凝血酶单独引起 Cr51 的适度释放，但没有增加细胞的松弛。当血小板计数保持恒定时，这两个参数都随着凝血酶剂量的增加而增加。当凝血酶剂量恒定时，松散细胞的百分比随着血小板数量的增加而增加。 Cr51 的显着释放仅发生在高浓度的血小板中。对暴露于凝血酶刺激的血小板的内皮细胞进行扫描和透射电子显微镜检查，证实存在与血小板相关的松动和损伤的细胞。内皮细胞膨出，细胞间连接打开，管腔侧质膜出现绒毛并破裂，细胞质电子密度增加。因此，凝血酶刺激的血小板在体外也可能引起内皮细胞损伤。
Platelets stimulated in vivo may cause endothelial injury. The question is whether a similar effect of platelet stimulation may be shown in vitro. Endothelial cells were cultured from human umbilical veins in Medium 199 with 20 per cent heat-inactivated serum. Semiconfluent cultures, 100-400.000 cells per dish, were labelled with Na2Cr51O4. Twenty-four hours later human platelet suspension (final cone. 200.000 per mm3) and thrombin (final cone. 4 U/ml) were added to the medium and the culture dish shaken for 15 min. The percentage of cells detached from the culture dish and the percentage of Cr51 released from the endothelial cells into the ambient fluid during the shaking were determined and used as parameters of cell injury. Increased percentages of loosened cells and Cr51 into the ambient fluid were observed with platelet suspension and thrombin compared to controls without platelet suspension and/or thrombin. Platelet-free supernatant after reaction of thrombin with platelet suspension had a similar effect. Thrombin alone caused a moderat release of Cr51, but no increased loosening of cells. Both parameters increased with increasing dose of thrombin when the platelet count was kept constant. The percentage of loosened cells increased with increasing platelet number when the dose of thrombin was constant. A significant release of Cr51 occurred only at high concentrations of platelets. Scanning and transmission electron microscopy of endothelial cells exposed to thrombin-stimulated platelets confirmed the presence of loosening and injured cells in association with platelets. The endothelial cells were bulging, intercellular junctions opened up, the plasma membrane on the luminal side showed villi and breaks and the cytoplasm had increased electron density. Thus, platelets stimulated by thrombin may cause injury of endothelial cells also in vitro.