肿瘤坏死因子-α对人脐静脉内皮细胞中组织蛋白酶K表达的影响

摘要

目的：观察不同浓度肿瘤坏死因子-α（ TNF-α）和不同TNF-α作用时间对人脐静脉内皮细胞（ HUVEC ）中组织蛋白酶K（Cat K）表达的影响。方法取对数生长期HUVEC进行实验。将HUVEC分为正常对照组、0．1 ng／ml TNF-α组、1 ng／ml TNF-α组、10 ng／ml TNF-α组、100 ng／ml TNF-α组，分别采用DMEM培养基、0．1 ng／ml TNF-α、1 ng／ml TNF-α、10 ng／ml TNF-α、100 ng／ml TNF-α作用24 h。另取HUVEC分为对照组、TNF-α6 h组、TNF-α12 h组、TNF-α24 h组、TNF-α48 h组，对照组DMEM培养基作用24 h，其他组采用10 ng／ml TNF-α作用相应时间。检测各组HUVEC 中的Cat K mRNA和蛋白的表达情况。结果与正常对照组比较，不同TNF-α浓度组的Cat K mRNA和Cat K蛋白表达水平均升高（P＜0．05）。0．1 ng／ml TNF-α组、1 ng／ml TNF-α组、10 ng／ml TNF-α组 Cat K mRNA 和 Cat K 蛋白的表达水平依次增高（P＜0．05），100 ng／ml TNF-α组Cat K mRNA和Cat K蛋白表达水平较10 ng／ml TNF-α组下降（P＜0．05）。与对照组比较，TNF-α各作用时间组Cat K mRNA和Cat K蛋白的表达水平增高（P＜0．05）；TNF-α6 h组、TNF-α12 h组、TNF-α24 h组Cat K mRNA和Cat K蛋白的表达水平依次增高（P＜0．05）；TNF-α48 h组Cat K mRNA和Cat K蛋白的表达水平较TNF-α24 h组明显下降（P＜0．05）。结论在一定作用浓度及作用时间内，TNF-α可以呈剂量和时间依赖性刺激HUVEC中Cat K表达。 TNF-α可能通过促进Cat K的表达促进动脉硬化的发生。%Objective To observe the effect of tumor necrosis factor-α( TNF-α) with different concentrations and different actuation durations on the expression of cathepsin K (Cat K) in human umbilical vein endothelial cells (HUVECs).Methods HUVECs in logarithmic growth phase were used in the experiment .The HUVECs were divided into normal control group ,0.1 ng/ml TNF-αgroup,1 ng/ml TNF-αgroup,10 ng/ml TNF-αgroup and 100 ng/ml TNF-αgroup treated with DMEM culture medium,0.1 ng/ml TNF-α,1ng/ml TNF-α,10 ng/ml TNF-αand 100 ng/m for 24 hours respectively .Other HUVECs were divided into control group ,TNF-α6 h group,TNF-α12 h group, TNF-α24 h group and TNF-α48 h group.The control group was treated with DMEM culture medium for 24 hours,and the other groups were treated with 10 ng/ml TNF-αfor the corresponding time .The expressions of Cat K mRNA and protein were detected in HUVECs of each group .Results The expression levels of Cat K mRNA and protein in the groups with different TNF-αconcentrations increased compared to the normal control group(P<0.05).The expression levels of Cat K mRNA and protein increased in the order of 0.1 ng/ml TNF-αgroup,1 ng/ml TNF-αgroup and 10 ng/ml TNF-αgroup(P<0.05).The expression levels of Cat K mRNA and protein in 100 ng/ml TNF-αgroup were lower than those in 10 ng/ml TNF-αgroup(P<0.05).Compared to the control group,the expression levels of Cat K mRNA and protein in the groups treated with TNF-αfor different times increased(P<0.05).The expression levels of Cat K mRNA and protein increased in the order of TNF-α6 h group,TNF-α12 h group and TNF-α24 h group(P<0.05).The expression levels of Cat K mRNA and protein in TNF-α48 h group were significantly lower than those in TNF-α24 h group(P<0.05).Conclusion Within certain action concentration and actuation duration , TNF-αcan stimulate the expression of Cat K in HUVEC in a time-and dose-dependent manner. TNF-αmight promote the occurrence of arteriosclerosis by promoting Cat K expression .