紫杉醇对TNF-α诱导的人脐静脉内皮细胞Cat K表达的影响

Effect of Paclitaxel on TNF-α Induced Human Umbilical Vein Endothelial Cells Expression of Cat K

目的观察紫杉醇对TNF-α诱导的人脐静脉内皮细胞中Cat K及NF-κB表达的影响,探讨紫杉醇在防治内膜增生的作用机制。方法采用HUVEC细胞株传代第3~6代的细胞作为研究对象。不同浓度的紫杉醇干预1h再用10μg/L TNF-a刺激人脐静脉内皮细胞24 h,并用RT-PCR方法检测细胞中Cat K mRNA、NF-κB mRNA的表达;用Western blot方法检测细胞中的Cat K和NF-κB蛋白的表达。结果紫杉醇可以呈剂量依赖性地抑制TNF-α所致的HUVEC中Cat K mRNA和Cat K蛋白的表达;紫杉醇可以呈剂量依赖性地抑制TNF-α所致的HUVEC中Cat K蛋白的表达,紫杉醇可以呈剂量依赖性地抑制TNF-α所致的HUVEC中NF-κB蛋白的表达。结论紫杉醇可以呈剂量依赖性地抑制TNF-α所致的HUVEC中Cat K和NF-κB的表达,紫杉醇可能通过抑制NF-κB使Cat K表达减少。

Aim To investigate the effect of Paclitaxel on TNF-α induced human umbilical vein endothelial cells expression of Cat K and NF-κB and explore the mechanism of paclitaxel inhibiting intimal hyperplasia. Methods HUVEC in the third to sixth generation were used in the present study. The cells were treated with different concentrations of paclitaxel for 1 hour and then with 10 μg / L TNF-α were added into it for 24 hours,mRNA and protein level of Cathepsin K and NF-κB were measured by reverse transcription polymerase chain reaction（ RT-PCR） and Western blot respectively in these HUVEC. Results Paclitaxel could inhibit TNF-α induced Cathepsin K mRNA and Cathepsin K protein expression in a dose-dependent manner. There was no significant difference between 1 nmol / L paclitaxel group and TNF-α group; Paclitaxel could inhibit TNF-α induced Cathepsin K protein expression in a dose-dependent manner,comparing 1 nmol / L paclitaxel group and TNF-α group. Paclitaxel could inhibit TNF-α induced NF-κB mRNA and NF-κB protein expression in a dose-dependent manner. Conclusion Paclitaxel could decrease the expression of Cathepsin K and NF-κB stimulated with TNF-α in a dose-dependent manner,paclitaxel may inhibit the activation of NF-κB to reduce Cat K expression.