血小板源性生长因子和血小板源性内皮细胞生长因子在内皮细胞和血管平滑肌细胞中的作用研究  

Role of PDGF and PD- ECGF in Endothelial Cells and Vascular Smooth Muscle Cells

作　　者：朱晋坤[1] 毛华[1] 尹扬光[2] 董文[1] 杜峰[1] 鲁玉明[1] 熊宗华[1] 邓梦扬[2] 

机构地区：[1]贵州省贵阳市第一人民医院心血管内科,550002 [2]第三军医大学附属新桥医院心血管内科

出　　处：《中国全科医学》2015年第9期1023-1028,共6页Chinese General Practice

基　　金：贵州省科技厅社会发展项目(黔科合SY[2010]3087号)

摘　　要：目的通过体外转染血小板源性生长因子(PDGF)和血小板源性内皮细胞生长因子(PD-ECGF)质粒,观察两种生长因子对人脐静脉内皮细胞EAHY926和主动脉血管平滑肌细胞T/G HA-VSMC的影响,评估两种生长因子对血管损伤治疗的可行性。方法构建人pc DNA 3.1(+)空载(空载组)、pc DNA 3.1(+)-PDGF-透明质酸(HA)(PPH组)和pc DNA 3.1(+)-PD-ECGF-HA(PPEH组)质粒,分别转染入EAHY926和T/G HA-VSMC中,四甲基偶氮唑盐微量酶反应比色法(MTT法)检测内源性PD-ECGF和外源性PD-ECGF对EAHY926和T/G HA-VSMC细胞增殖的影响;细胞伤痕实验观察内源性PD-ECGF和外源性PD-ECGF对EAHY926和T/G HA-VSMC细胞迁移速度的影响。结果 MTT法检测3组EAHY926吸光度值比较,差异有统计学意义(F=235.18,P<0.001),其中PPH组高于空载组和PPEH组(P<0.05);空载组与PPEH组比较,差异无统计学意义(P>0.05)。MTT法检测3组T/G HA-VSMC吸光度值比较,差异有统计学意义(F=82.89,P<0.001),其中PPH组高于空载组(P<0.05);PPEH组低于空载组(P<0.05)。MTT法检测转染PPEH质粒的培养基(L-PPEH)对PPH诱导的EAHY926和T/G HA-VSMC增殖的影响,空载组、PPH组、PPH+L-PPEH组EAHY926吸光度值比较,差异无统计学意义(F=512.89,P=0.183)。空载组、PPH组、PPH+L-PPEH组T/G HA-VSMC吸光度值比较,差异有统计学意义(F=317.40,P<0.001);其中PPH组高于空载组和PPH+L-PPEH组(P<0.05)。细胞伤痕实验证实,转染PPEH质粒的EAHY926、T/G HA-VSMC细胞迁移能力增强;PPEH组EAHY926平均细胞迁移距离百分比(56.1%±2.2%)较空载组(27.8%±3.4%)升高(t=-15.08,P<0.001);PPEH组T/G HA-VSMC平均细胞迁移距离百分比(69.1%±2.3%)较空载组(43.6%±5.8%)升高(t=-33.64,P<0.001)。用转染PPEH质粒的培养基培养EAHY926、T/G HA-VSMC发现,外源性PD-ECGF处理的EAHY926、T/G HA-VSMC细胞迁移能力亦明显增强。结论通过转染PDGF质粒,模拟血管损伤后内皮细胞和平滑肌细胞受到PDGF和/或PD-ECGF刺激,发现PDGF能明显上调内皮细胞�Objective To investigate the influence of platelet derived growth factor（ PDGF） and platelet- derived endothelial cell growth factor（ PD- ECGF） on human umbilical endothelial cells EAHY926 and human- aorta vascular smooth muscle cells T / G HA- VSMC by transfecting the plasmids of PDGF and PD- ECGF in vitro and to evaluate the possibility of treating vascular injury by the two kinds of plasmids. Methods pc DNA 3. 1（ ＋） no- load（ no- load group）,pc DNA 3. 1（ ＋）- PDGF- HA（ PPH group） and pc DNA 3. 1（ ＋）- PD- ECGF- HA（ PPEH group） plasmids were constructed and transfected into EAHY926 and T / G HA- VSMC. MTT method was used to evaluate the influence of endogenous and exogenous PD- ECGF on the proliferation of EAHY926 and T / G HA- VSMC before and after transfection. Cell wound healing assay was used to evaluate the influence of endogenous and exogenous PD- ECGF on the migration speed of EAHY926 and T / G HA- VSMC after transfection. Results MTT method was used to detect the absorbance value of EAHY926 among the three groups,and the differences were statistically significant（ F = 235. 18,P 0. 001）,with the PPH group significantly higher than the no- load group and PPEH group（ P〈0. 05）. The no- load group and PPEH group showed no statistically significant differences（ P〉0. 05）. MTT method was also used to detect the absorbance value of T / G HA- VSMC among the three groups,and the differences were statistically significant（ F = 82. 89,P 0. 001）,with the PPH group significantly higher than the no-load group（ P〈0. 05）,and the PPEH group significantly lower than the no- load group（ P〈0. 05）. MTT method detecting the influence of medium transfected with PPEH on the proliferation of EAHY926 and T / G HA- VSMC induced by PPH. There was no significant difference in the absorbance value of EAHY926 among no- load group,PPH group and PPH ＋ L- PPEH group（ F = 512. 89,P = 0. 183）. There was significant difference in the absorbance value of T/G HA- VSMC am

关 键 词：血小板源性生长因子 血小板源性内皮细胞生成因子 血管成形术 气囊 冠状动脉 肌细胞 平滑肌 内皮细胞 细胞增殖 

分 类 号：R349.51[医药卫生—基础医学]