In vitro study on injury effect of IL-18 activating NF-κB cell signal pathway on human umbilical vein endothelial cells / 重庆医学

Objective To investigate the influence of proinflammatory factor interleukin-18(IL-18) on vein endothelial cell function by activating NF-κB mediated cell signal pathway and its association with deep vein thrombosis(DVT).Methods Recombinant human IL-18 was used to act on in vitro cultured human umbilical vein endothelial cell(HUVECs).The NF-κB activation inhibitor was used to conduct interference.The detection measures of real time fluorescence quantitative PCR,Western blot,immunofluorescence and flow cytometry were used to verify whether IL-18 affect the expression of endothelial cellular function markers such as HUVECs normal statusand vWF,P-selectin and tissue plasminogen activator(t-PA) by activating NF-κB mediated cell signal pathway.Moreover the mechanism of IL-18 participating in the DVT was performed the comprehensive analysis by combining with previous study.Results IL-18 could activate NF-κB in endothelial cell,increased the p65 expression in nucleus,decreased the intracellular IκBα expression and significantly increased early apoptosis cells in HUVECs;adding QNZ(EVP4593) could significantly inhibit the activation effect of IL-18 on NF-κB,the occurrence of cellular injury and apoptosis was significantly reduced;IL-18 could promote the abnormal expression of DVT related endothelial cell markers vWF,P-selectin and t-PA (P<0.05).But various markers could recover conventional expression after inhibiting NF-κB activation.Conclusion The interaction between Il-18 and NF-κB causes the abnormality of HUVECs growth status and function,which may be the DVT onset related pathogenic mechanism.

Correlation between interleukin-18 and deep venous thrombosis disease / 重庆医学

Objective To investigate the correlation between IL‐18 and deep venous thrombosis disease and its clinical significa‐tion .Methods To detect the expression of IL‐18 by ELISA ,we collected the blood samples of DVT patients as the experimental group(n=40) compared to the control group(n=40) and normal group(n=20) .IL‐18 over expression/interference vectors were constructed and transfected human vein endothelial cells ,analyzed by microarray and KEGG Pathway as biology information tech‐nology .Then discuss the association between IL‐18 and DVT .Results Results of ELISA showed that compared with control group and normal group ,the expression of IL‐18 gene in DVT patient were up‐regulated(F=11 .248 ,P0 .05) .Immunofluorescence detected IL‐18 gene expression in cytoplasm of human umbilical vein endothelial cells (HUVECs) .According to the microarray analysis we found in the IL18‐pCDH‐GFP transfected cells 17 signaling pathways were down‐expressed while 16 signaling pathways were up‐expressed .Compared with normal group cells ,in the IL18‐LMP‐shRNAmir1 transfected cells 23 signaling pathways were down‐ex‐pressed and 9 signaling pathways were up‐expressed .Conclusion Based on the above experimental data ,it is very clear that IL‐18 influenced HUVECs and plays an important role in DVT ,it is possible to predict the diagnosis of DVT and act as candidate molecu‐lar markers .

The Study of Signaling Pathways in MCP-1 Over-Expression/Interference of HUVECs / 天津医药

Objective To investigate the association between the signaling pathways of MCP-1-pCDH-GFP-trans?fected cells and deep venous thrombosis (DVT). Methods The cultured human umbilical vein endothelial cells (HUVECs) were tested by immunofluorescence and co-immunoprecipitation methods. The constructed MCP-1 over-expression/interfer?ence vector, and the change of transcription profile were detected by microarray assay and biological information technology analysis. Results MCP-1 over-expression/interference vector MCP-1-pCDH-GFP/MCP-1-LMP shRNAmir1 was con?structed and HUVECs were transfected. According to the microarray analysis we found that there were 18 down-expressed signaling pathways and 7 up-expressed signaling pathways in MCP-1-pCDH-GFP-transfected cells. There were 60 down-expressed signaling pathways and 15 up-expressed signaling pathways in the MCP-1-LMP shRNAmir1 transfected cells. Conclusion Signaling pathways of MCP-1 plays an important role in DVT formation,which may provide us a new way to study molecular mechanism of DVT.

Changes of arginase Ⅰ expression in rat deep venous thrombosis models / 中国组织工程研究

BACKGROUND:Studies in recent years have demonstrated that arginase Ⅰ contribute to the process of numerous cardiovascular diseases,however,most of studies focus on arteries,few regarding venous diseases.OBJECTIVE:To explore the changes of arginase Ⅰ expression in rat traumatic deep venous thrombosis models,and to analyze the possible function of arginase Ⅰ in deep venous thrombosis formation.METHODS:Totally 100 Sprague Dawley rats were randomly divided into the control and model groups.Traumatic deep venous thrombosis models were established by clamping the femoral vein and immobilizing the bilateral hind limbs (hip spica cast fixation),and assigned into initial thrombosis,peak thrombosis and non-thrombosis groups according to different observing time points and pathophysiological situations of thrombosis.Whole blood RNA of each group was extracted,and the change of arginase I expression in blood cells of each group was detected by real-time PCR.RESULTS AND CONCLUSUON:Expression of arginase Ⅰ in the peak thrombosis group was significantly increased compared with other 3 groups (P < 0.01).There were no significances among control,initial thrombosis and non-thrombosis groups (P > 0.05).The finding demonstrated that arginase Ⅰ is closely related to deep vein thrombosis formation.