PPARγ对猪血管内皮细胞增殖、迁移及小管形成的影响

Effect of PPARγ on Proliferation,Migration and Tube Formation of Swine Vein Endothelial Cells

试验旨在研究过氧化物酶体增殖物激活受体γ(peroxisome proliferator-activated receptorsγ,PPARγ)对猪血管内皮细胞增殖、迁移及小管形成的影响,并探讨PPARγ在猪体外血管生成中的作用。设置PPARγ激动剂组(5、10、15、20μmol/L罗格列酮)、抑制剂组(5、10、15、20μmol/L T0070907)及对照组,通过iCelligence细胞功能分析、划痕试验和Matrigel基质胶三维培养,构建猪体外血管生成的模型,模拟猪体内血管生成的环境,分别对猪血管内皮细胞的增殖、迁移、小管形成能力进行测定,同时根据NCBI已有的相关序列,应用Primer Premier 5.0软件设计PPARγ基因特异性引物,利用SYBR GreenⅠ实时荧光定量PCR检测PPARγ基因mRNA相对表达量,对PPARγ的体外作用效果进行验证。结果显示,5、10μmol/L罗格列酮能促进猪血管内皮细胞增殖、迁移及小管形成,T0070907的抑制效果在试验浓度区间内(5~15μmol/L)随浓度升高而加强,较高浓度(20μmol/L)的两种药物均由于药物毒性的影响对细胞活动产生干扰。此外,5~20μmol/L罗格列酮和5~20μmol/L T0070907能分别提高和降低PPARγ基因mRNA相对表达量,且浓度趋势与增殖、迁移、小管形成的试验结果一致。综上所述,通过激活PPARγ可以对猪血管内皮细胞的增殖、迁移及小管形成产生促进效果,提示其在猪体外血管生成中具有积极作用,可为研究PPARγ对猪胎盘血管发生的影响提供参考依据。

In order to investigate the effect of peroxisome proliferator-activated receptors γ（PPARγ）on proliferation,migration and tube formation of swine vein endothelial cells,and explore its role in swine placental angiogenesis,PPARγagonist group（5,10,15 and 20μmol/L rosiglitazone）,inhibitor group（5,10,15 and 20μmol/L T0070907）and control group were set in this study.Through iCelligence cell function analyzer,scratch test,Matrigel matrix three dimensional cell culture,a model of angiogenesis of swine vein endothelial cells in vitro was established for simulating the environment of pig angiogenesis in vivo and detecting the proliferation,migration,tube formation ability of cells.A pair of specific primer was designed by Primer Premier 5.0software according to PPARγsequence from NCBI.SYBR GreenⅠ Real-time PCR were performed to detect relative quantity of PPARγgene mRNA and verify the effect of PPARγon cells in vitro.The results showed that 5and 10μmol/L rosiglitazone could promote proliferation,migration and tube formation of swine vein endothelial cells,T0070907 had inhibition on cells andnegatively correlated with concentration（5to 15μmol/L）,both of rosiglitazone and T0070907 at high concentration（20μmol/L）interfere in cells activity because of toxic effect.In addition,rosiglitazone（5to 20μmol/L）and T0070907（5to 20μmol/L）increased and reduced relative quantity of PPARγgene mRNA,respectively,and its trends of concentration were similar with the former three experiment.The results indicated that the activation of PPARγpromoted proliferation,migration and tube formation of swine vein endothelial cells,and it might play apositive role in swine placental angiogenesis.