Effects Of Hes130/0.4 On The Inflammatory Injury Of Rat Pulmonary Microvascular Endothelial Cells Induced By Lipopolysaccharide

Objective:Acute lung injury(ALI) is an acute progress respiratory failure which induced by various reasons except cardiogenic factors. Theserious stage of ALI is acute respiratory distress syndrome(ARDS). At present,there is still no effective therapeutic method. The mortality of ALI is 30-60%. Among he enormous lung parenchymal cells, pulmonary microvascular endothelial cells(PMVEC) are important cells. They are not only the target cells of inflammatory cells and mediators,but are active inflammatory cells and effector cells as well. PMVEC are different to culture,so pulmonary vascular endothelial cells and umbilical vein endothelial cells,easy for culturing, often substitute PMVEC in the past rese-arch. Nevertheless PMVEC and pulmonary vascular endothelial cells are similar in shapes,their biological characteristics are not the same.So ot-her endothelial cells can not substitute PMVEC. It is basal to culture PMVEC in vitro successfully for investigating the cells in ALI pathogenesis. HES130/0.4, the lastest hydroxyethyl starch, is used for various kinds of patients' capability substitution therapy.It is reported that HES130/0.4 has antiinflammatory effect besides the role in capability substitution therapy. The purpose of this study is to investigate in vitro, HES130/0.4 wheather has the effects on the inflammatory injury of rat pulmonary microvascular endothelial cells induced by lipopolysaccharide.Methods:1 PMVEC cultureThe SD rat weighing 30-50g were disinfected with 75% alcohol after anesthetized with pentobarbital sodium.Their chest cavities were opened and lungs were taken out and then were put into PBS liquid to was hed away the residual blood,carefully cut the lung tissues into 1mm3 sections,which would be put into the labeled sterile culture bottle and were left for solidification for 2 hours in culture apparatus.After this process, add DMEM into culture bottle.When the cells converged into mono-layer,continued the passage culture after digested by trypsin.Third generation of PMVEC was used in this study.2 The PMVECs from cell suspension were adjusted their number to 1×105/ml,and then inoculated into a 48-pore culture plate.There were 36 pors needed.1 ml cell suspension was put into each pore.The 36 pores were randomly divided into nine groups (n=4). With different treatments, the LPS group,the HES+LPS group and the control group were divided. In each group above, according to the different PMVEC culturing time,2,6,12h, 3 deuto-groups were divided. The LPS group was added 1μg/ml LPS.The HES+LPS group ,cells were received both 1μg/ml LPS and 30 mg/ml HES130/0.4.PBS liquid was put to the contronl group with the same volumes of the two groups above. The cell culture plate was discarded after cultured for 2,6 and 12 hours in the CO2 incubator (37℃,5%CO2 ).After collecting the supernatant,test the release of proinflammatory mediators of TNF-αand IL-6 by ELISA method.Results:1 the primary culture of PMVECThe cultured PMVECs of rats showed fusiform shape or polygon,and the monolayer cultures displayed a typical pavingstone morphology and were inhibited when contacted each other.The PMVECs expressed factorⅧassociated antigen.2 Effects of HES130/0.4 on the inflammatory injury of rat pulmonary microvascular endothelial cells induced by lipopolysaccharide2.1 The proinflammatory mediators of TNF-αand IL-6 were significantly increased in the LPS group.At 2,6 and 12 hours ,the 3 time points, TNF-αand IL-6 was increased apparentely.2.2 HES130/0.4 can decrease the amout of proinflammatory mediators o -f TNF-αand IL-6 indued by LPS at every time point.Conclusions:1 in this study, the primary culture of PMVEC was established successfully.2 HES130/0.4 can decrease the amout of proinflammatory mediators of TNF-αand IL-6 indued by LPS.It indicates that HES130/0.4 has the advantage of anti-inflammatory effect and the protective effect on acute lung injury.