血小板源生长因子α亚基受体结构域切除对血管平滑肌凋亡和c-sis基因表达的影响

Effect of truncated platelet-derived growth factor -α receptor on apoptosis and expression of c-sis mRNA of pulmonary artery smooth muscle cells

目的 探讨从血小板源生长因子α(platelet derivedgrowthfactor ,PDGF α)受体水平阻断对肺血管平滑肌细胞凋亡和c sis基因表达的影响。方法 采用两种方法干预肺动脉平滑肌细胞(vascularsmoothmuscularcells ,VSMC) ,一种是在培养液中分别给予不同剂量的受体结构域切除的PDGF α受体腺病毒重组体Ad5CMV PaRtr(ACP) ,另一种是加入固定的中浓度ACP液后再分别加入不同浓度的血小板源生长因子BB(PDGF BB) ,对不同组细胞的细胞周期、凋亡细胞百分率和c sis原癌基因表达进行测定。结果 中、大浓度ACP对细胞增殖有明显的抑制作用 ,且使凋亡细胞百分率明显升高。在中浓度ACP的作用下 ,加入PDGF BB不能促进VSMC的增殖 ,亦不改变细胞的凋亡水平。不同浓度的ACP可使c sismRNA的表达上调。在中浓度ACP的作用下 ,加入PDGF BB可下调c sismRNA的表达。结论 ACP作为细胞增殖的抑制剂 ,能在中、大剂量下明显增加Go/G1期细胞的数量 ,抑制肺VSMC的增殖 ,增加细胞的凋亡率 ,同时使c sismRNA的表达上调。

Objective Platelet-derived growth factor (PDGF) plays an important role during the pathophysiological changes in vascular remodeling. The study aimed to investigate the effect of truncated PDGF-α receptor on apoptosis and expression of c-sis mRNA of pulmonary artery smooth muscle cells (VSMCs). Methods Tissue mass culture was done to get vascular smooth muscle cells of pulmonary artery in newborn pigs. Two methods were used to interfere VSMCs: adding adenoviral recombined body (Ad5CMV-PαRtr, ACP) with three different concentrations of truncated PDGF-α receptor into the cultures, or adding three concentrations of PDGF-BB after the treatment with mid-concentration of ACP. VSMC apoptosis, cellular cycle and expression of c-sis were observed using flow-cytometry, and the expression of c-sis mRNA was detected by reverse transcription polymerase chain reaction (RT-PCR). Results ACP with mid- to- high concentrations could restrain the proliferation of VSMCs apparently with the increase of G 0/G 1 cells. The apoptotic rate presented an ascending tendency. The differences among the groups were of statistically significant. Affected by mid- concentration of ACP, PDGF-BB did not exhibit a significantly accelerating effect on the changes of cellular cycle and VSMC apoptosis. The expression of c-sis mRNA was up-regulated under the effect of ACP. Affected by mid-concentration of ACP and PDGF-BB, c-sis mRNA expressed was down-regulated. Conclusion Mid- to- high concentration of ACP is a powerful inhibitor of cellular proliferation for pulmonary artery VSMCs. It can significantly increase cells in number in G 0/G 1 phase, apoptosis and c-sis mRNA expression.