核转录 因 子 kappaB 通路在血小板源性生长因子诱导大鼠肺动脉平滑肌细胞增殖中的作用  

Effect of nuclear transcription factor kappaB pathway on proliferation in rat pulmonary artery smooth muscle cells induced by platelet-derived growth factor

作　　者：刘忠强[1] 王晓琴[1] 王慧卿[1] 

机构地区：[1]四川大学华西第二医院PICU,成都610041

出　　处：《中华实用儿科临床杂志》2013年第22期1739-1742,共4页Chinese Journal of Applied Clinical Pediatrics

基　　金：依耀科技基金（12H0454）

摘　　要：目的 探讨核转录因子kappaB（NF-κB）通路在血小板源性生长因子（PDGF）诱导大鼠肺动脉平滑肌细胞（PASMC）增殖中的作用。方法 体外培养大鼠 PASMC，随机分为3组：对照组（M199培养基培养）、PDGF刺激组（M199培养基与PDGF联合培养）、PDGF＋小白菊内酯干预组（在M199培养基培养的基础上进行PDGF诱导，并予NF-κB抑制剂小白菊内酯干预）。通过四唑盐（MTT）比色法和流式细胞术检测PASMC的增殖及细胞周期；通过免疫组织化学法检测NF-κB及其下游因子环氧化酶（COX-2）蛋白的表达并进行图像分析；通过荧光定量反转录-PCR检测NF-κB及COX-2mRNA的表达；采用单因素方差分析进行统计处理，两两比较采用LSD法。结果 PDGF刺激组各时间点细胞MTT值均较对照组高（P均 〈0.05），予 NF-κB抑制剂小白菊内酯干预后，各时间点细胞 MTT值均较 PDGF刺激组明显降低（P均 〈0.05）；流式细胞术检测结果显示：PDGF刺激组S期和G2＋M 期细胞比例较对照组增高（P 〈0.05），予小白菊内酯干预后，S期和G2＋M 期细胞比例较PDGF刺激组降低（P 〈0.05）；PDGF刺激组NF-κB和COX-2蛋白及mRNA表达水平均明显高于对照组（P均〈0.05），予小白菊内酯干预后，NF-κB 和 COX-2的蛋白及 mRNA 表达水平较 PDGF刺激组均下降（P均〈0.05）。结论 PDGF能诱导PASMC增殖，促进细胞周期进程，增强NF-κB和 COX-2蛋白及 mRNA 表达；NF-κB 通路参与PDGF诱导PASMC的增殖过程。Objective To explore the effect of nuclear transcription factor kappaB （NF-KB） pathway on proliferation of rat pulmonary artery smooth muscle cells （ PASMC ） induced by platelet-derived growth factor （PDGF）. Methods PASMC isolated from rats and cultured in vitro were divided into 3 groups according to the randomization principle：control group（cultured by M199）, PDGF treatment group （cultured by M199 and stimulated by PDGF） , PDGF ＋ parthenolide treatment group （cultured by M199 and stimulated by PDGF, and intervented by the NF-KB inhibitors parthenolide）. MTT colorimetric assay and flow cytometry were performed to detect cell proliferation and cell cycle distribution. Immunohistochemistry was performed to detect the expressions of NF-KB and COX-2 protein. Fluorescence quantitative RT-PCR was performed to detect NF-KB and COX-2 mRNA expressions. One-way ANOVA was used for statistical analysis, multiple comparisons were analyzed by LSD. Results Compared with the control group, MTT value of PASMC was increased significantly when induced by PDGF at each time points（ all P 〈0.05 ）. MTT value decreased dramatically after the intervention of NF-KB inhibitor parthenolide （P 〈 0. 05 ）. Data from flow cytometry detection showed that cell proportion of S phase and G2 ＋ M phase increased significantly in PDGF treatment group, which had statistical difference compared with control group （ all P 〈 0. 05 ）. Compared with PDGF induced group, after the in- tervention of parthenolide, cell proportion of S phase and G2 ＋ M phase ratio decreased dramatically （ P 〈 0.05 ）. The expressions of NF-KB and COX-2 protein and mRNA were promoted in the PDGF induced group compared with the control group（ all P 〈 0.05 ）. Compared with PDGF induced group, after the intervention of parthenolide, the expressions of NF-KB and COX-2 protein and mRNA decreased dramatically （ all P 〈 0. 05 ）. Conclusions PDGF can induce proliferation of PASMC, promote cell cycle process and enhance t

关 键 词：肺动脉平滑肌细胞 核转录因子KAPPAB 血小板源性生长因子 

分 类 号：R363[医药卫生—病理学]