[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 0.03125 similar 2.0000 g/L. The OD value reached peak level when LPS concentration was 0.2500 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.


CONCLUSIONS
(1) LPS could combine MMECs and enter into the nucleus without the presence of serum. (2) LPS could directly damage MMECs.
Wang
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Institute of Burn Research, Southwestern Hospital, The Third Military Medical University, Chongqing 400038, P.R. China.
Zhang
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Wang
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D000818:Animals; D001692:Biological Transport; D002455:Cell Division / Q000187:drug effects; D002467:Cell Nucleus / Q000187:drug effects / Q000378:metabolism; D002478:Cells, Cultured; D003331:Coronary Vessels / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D004305:Dose-Response Relationship, Drug; D004730:Endothelium, Vascular / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D016650:Fluorescein-5-isothiocyanate; D008070:Lipopolysaccharides / Q000378:metabolism / Q000494:pharmacology*; D008297:Male; D018613:Microscopy, Confocal; D051381:Rats; D017208:Rats, Wistar; D013997:Time Factors
D008070:Lipopolysaccharides; D016650:Fluorescein-5-isothiocyanate