内毒素与内皮细胞直接结合和损伤特性的研究

目的 观察内毒素（LPS）和内皮细胞的结合及损伤特性。方法 分离培养大鼠心肌微血管内皮细胞（MMECs），选择不同浓度的LPS及同一浓度不同时相点，作用于贴壁状态的MMECs，应用流式细胞仪，激光共聚焦荧光扫描显微镜（laser confocal scanning microscope,LCSM）进行观察和图象分析。结果 （1）随孵育时间延长，LPS作用后MMECs的OD值逐渐增加，30min为最高点，有时间依赖性。（2）LPS和MMECs共培养2h ,LPS浓度从0.03125-2.00000g/L，都可以使MMECs的阳性细胞数及OD值增加，0.2500g/L时是OD值的峰值，提示该浓度是LPS与MMECs的最佳结合浓度，有浓度的饱和性。（3）LPS可以进入MMECs胞浆和胞核。（4）LPS可导致MMECs的核移位和脱核。结论 （1）LPS在无血清状况下可以与内皮细胞结合，并进入细胞核。（2）LPS可导致内皮细胞的直接损伤。

A study on the direct binding of LPS to endothelium and the characteristics of endothelial injury

Objective To explore the characteristics of the combination of LPS to the endothelium and endothelial injury. Methods Myocardial microvascular endothelial cells (MMECs) were isolated and cultured to adhering state.Flow cytometry,laser confocal scanning microscope (LCSM) and image analysis were employed to evaluate the effects of different concentrations of LPS and different culturing time with the same concentration of LPS on the adhered MMECs Results (1)The OD values of MMECs increased progressively along with the prolongation of culturing time of MMECs with LPS and reached top level at 30 min.In addition,the increase of OD values was dependent on time.(2)The number of positive MMECs and the OD values increased after MMECs was cultured for 2 hours with LPS at concentrations ranging from 0 031 25~2.000 0 g/L.The OD value reached peak level when LPS concentration was 0 250 0 g/L,which implied that this concentration of LPS (0.25 g/L) was the optimal one for the combination of LPS with MMECs ,and there was concentration saturation.(3)LPS could enter into the cytoplasm and nucleus of MMECs.(4)LPS could induce nucleic translocation and denucleation. Conclusion (1)LPS could combine MMECs and enter into the nucleus without the presence of serum.(2)LPS could directly damage MMECs.