临床儿科杂志 ›› 2016, Vol. 34 ›› Issue (6): 465-.doi: 10.3969 j.issn.1000-3606.2016.06.016

• 实验研究 • 上一篇    下一篇

米贝地尔对血小板衍生生长因子诱导的人肺动脉平滑肌细胞增殖的影响

李红红, 肖婷婷, 谢利剑, 黄敏, 沈捷   

  1. 上海交通大学附属儿童医院 上海市儿童医院心内科(上海 200062)
  • 收稿日期:2016-06-15 出版日期:2016-06-15 发布日期:2016-06-15
  • 通讯作者: 沈捷 E-mail:she6nt@163.com
  • 基金资助:
    上海市科委科技攻关重大项目(No.12411952400)子课题(No.12411952403)

Effect of mibefradil on proliferation of human pulmonary artery smooth muscle cells induced by platelet-derived growth factor

LI Honghong, XIAO Tingting, XIE Lijian, HUANG Min, SHEN Jie   

  1. Shanghai Children’s Hospital, Children’s Hospital Affiliated to Shanghai Jiaotong University, Shanghai 200062, China
  • Received:2016-06-15 Online:2016-06-15 Published:2016-06-15

摘要: 目的 探讨新型钙通道拮抗剂米贝地尔(mibefradil,Mib)对血小板衍生生长因子(PDGF)诱导的人肺动脉平滑肌细胞(HPASMCs)增殖的影响。方法 体外培养HPASMCs,随机分为对照组、PDGF刺激组、Mib干预组、PDGF+Mib组,PDGF刺激组细胞用25 ng/ml PDGF刺激,Mib干预组细胞加入10 μmol/L 的Mib干预,PDGF+Mib组细胞予PDGF和Mib联合干预。通过MTT法检测48 h 和72 h 各组HPASMCs增殖率,流式细胞术检测细胞周期,免疫荧光染色法观察增殖细胞核抗原(PCNA)的表达。结果 MTT法检测48 h 和72 h 各组HPASMCs增殖率的差异均具有统计学意义(P < 0.05),并以72 h时的差异更明显;其中PDGF组均为最高,与其余3组间的差异均有统计学意义(P均 < 0.05);而PDGF+Mib组与Mib组、对照组,以及Mib 组与对照组间的差异则无统计学意义(P > 0.05)。流式细胞术检测各组G0/G1 期及S期细胞比例差异均有统计学意义(P < 0.05);其中PDGF刺激组G0/G1 期细胞比例最低、S期细胞比例最高,与其余3 组比较差异均有统计学意义(P < 0.05);PDGF+Mib组与Mib 组、对照组,以及Mib 组与对照组间的差异均无统计学意义(P > 0.05)。免疫组化染色结果表明各组间PCNA表达差异有统计学意义(P < 0.05);其中PDGF组的PCNA表达最高,与其余3 组间的差异均有统计学意义(P均 < 0.05);而PDGF+Mib组与Mib组、对照组,以及Mib组与对照组间的差异则无统计学意义(P > 0.05)。结论 Mib组能显著抑制PDGF诱导的HPASMC增殖,抑制PDGF促进的细胞周期进程,对PCNA的表达具有明显的抑制作用。

Abstract: Objectives To explore the effect of mibefradil, a kind of novel calcium channel antagonists, on proliferation of human pulmonary artery smooth muscle cells (HPASMCs) induced by platelet-derived growth factor (PDGF). Methods HPASMCs were cultured in vitro, and randomly divided into control group, PDGF group, Mib group, and PDGF+Mib group.  The PDGF group was stimulated by 25 ng/ml of PDGF. Mib group was intervented by 10 μmol/L of Mib. PDGF+Mib group was treated by PDGF and Mib. The reproduction rate in 48 hours and 72 hours were detected by MMT. Cell cycle was detected by flow cytometry. The expression of proliferating cell nuclear antigen (PCNA) was observed by immunofluorescence staining (IFS). Results There were statistical differences among four groups in both 48 hours and 72 hours (P all < 0.05), especially in 72 hours. PDGF group had the highest level of HPASMC reproduction rate, and there were statistical differences as compared with the other three groups (P all < 0.05). However, the HPASMC reproduction rates were similar among PDGF+Mib group, Mib group, and control group (P all > 0.05). There were statistically differences of G0/G1 phase and S phase among four groups (P < 0.05). PDGF group had lowest G0/G1 phase cells and highest S phase cells, and there were statistically differences as compared with the other three groups (P all < 0.05). No differences were found among PDGF+Mib group, Mib group, and control group (P all < 0.05). There was obviously difference in the expression of PCNA among four groups (P < 0.05). PDGF group had the highest expression of PCNA and there were statistically differences as compared with other three groups, (P < 0.05). While the expression of PCNA was similar among PDGR+Mib group, Mib group, and control group. Conclusion Mibefradil was able to restrain remarkably the proliferation of HPASMC by inhibit the cell cycle that stimulated by PDGF, and by the expression of PCNA.