Study On The Protective Effect Of Hydrogen Sulfide On The Permeability Of PMVECs By Lipopolysaccharide

Objective:To explore the effect of different concentrations exogenous H₂S on the increasing PMVECs induced by LPS were investigated by testing PMVECs activity and the expression of VE-cad.Method:1.The primary culture of PMVECs in rats was cultured by tissue patch.Immunofluorescence and immunohistochemical were used to identify the cells.2.The different concentrations of LPS（1μg/mL,4μg/mL,7μg/mL,10μg/m L）were added to DMEM/F-12 medium for 24h.MTT assay was used to select the optimum concentration of PMVECs permeability induced by LPS.3.The experiment was randomly divided into blank control group,LPS group,LPS+NaHS 50μmol/L group,LPS+NaHS 100μmol/L group,LPS+NaHS 200μmol/L group and each group was contain of 5 compound pores.Blank control group:DMEM/F-12 medium;LPS group:the concentration of LPS was 7μg/mL;LPS+NaHS 50μmol/L group:LPS（7μg/mL）+NaHS（50μmol/L）;LPS+NaHS 100μmol/L group:LPS（7μg/mL）+aHS（100μmol/L）,LPS+NaHS 200μmol/L group:LPS（7μg/mL）+NaHS（200μmol/L）.After LPS was given for 24h,MTT assay was used to detect the activity of PMVECs and immunohistochemical was used to detect the expression of VE-cad in each group.4.SPSS19.0 and image-pro Plus（IPP）were used for statistical analysis of immunohistochemical images and measurement data.Result:1.The result of cell morphology:the primary culture of PMVECs were typical of paving stones.The cell morphology after the transmission was Long fusiform and slightly different from the original generation.2.The result of cell identification:The reagent was combined with PMVECs cytoplasm,which presented green fluorescence and brown granules,VIII antibody and vWF antibody were positive.3.The result of MTT assay Select the concentration of LPS:LPS with a concentration of 7μg/mL for 24h,the cells growth were significantly inhibited.Therefore,the results of selecting model concentration was 7μg/mL.4.The result of cell activity after NaHS intervention:Compared with blank control group,the cell activity in the LPS group decreased significantly（P<0.05）.In LPS+NaHS 50μmol/L group,LPS+NaHS 100μmol/L group,LPS+NaHS 200μmol/L group showed a higher dose dependence than the LPS group and the difference was significant（P<0.05）.5.The result of the expression of VE-cad after NaHS intervention:Compared with the blank control group,VE-cad amount decreased obviously in LPS group（P<0.05）;Compared with the LPS group,the expression of VE-cad were higher in LPS+NaHS 50μmol/L group,LPS+NaHS 100μmol/L group,LPS+NaHS 200μmol/L group.At the same time,the comparison between the NaHS groups were also statistically significant,indicating that the increase was dose-dependent（P<0.05）.6.The result of correlation analysis:PMVECs activity and VE-cad expression in PMVECs showed a significant positive correlation（P<0.001）.Conclusion:1.LPS can induce the reduction of VE-cad expression in PMVECs and establish a model of increased permeability of PMVECs.2.H₂S participated in the regulation of the damaged and increased permeability of PMVECs and improved the damaged PMVECs.By modified the expression of VE-cad to and reduced the increased permeability of PMVECs by LPS.