白细胞介素-18与深静脉血栓疾病的相关性研究

目的：探讨白细胞介素‐18（IL‐18）的表达变化与深静脉血栓疾病（DVT）发生、发展的相关性及临床意义。方法收集临床DVT病例（DVT组，n＝40）、实验对照组（n＝40）、健康对照组（n＝20）的血液，以ELISA法检测其中IL‐18的表达并分析差异；培养原代培养的人脐静脉内皮细胞（HUVECs）行目标蛋白免疫荧光检测明确其定位；构建人IL‐18过表达和干扰病毒载体（IL‐18‐pCDH‐GFP、IL‐18‐LMP‐shRNAmir1）感染HUVECs并进行基因表达谱芯片检测和KEGG Pathway 等生物信息技术，分析IL‐18与疾病的相关性。结果人血 ELISA 检测发现在 DVT 组和两对照组中 IL‐18的表达差异均有统计学意义（F＝11．248，P＜0．01）；实验对照组与健康对照组之间IL‐18的表达差异无统计学意义（ P＞0．05）；免疫荧光检测显示目标蛋白定位于细胞质；人IL‐18过表达和干扰载体经病毒转染后感染 HUVECs ；基因芯片检测发现与正常细胞比较，过表达载体 IL‐18‐pC‐DH‐GFP感染细胞有17条信号通路下调、16条上调；干扰载体IL‐18‐LMP‐shRNAmir1感染细胞有23条信号通路下调、9条上调。结论 IL‐18对HUVECs有确切影响，同时与DVT疾病密切相关，有希望作为深静脉血栓疾病临床预测诊断的候选分子标记物。

Correlation between interleukin-18 and deep venous thrombosis disease

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 ,P<0 .01) .Compared with normal group ,the IL‐18 expression in control group have not been significantly up‐regulated(P>0 .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 .