Zheng Zhen-Zhong, Liu Zheng-Xiang
Department of Cardiology, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430030, China.
Acta Pharmacol Sin. 2007 Jan;28(1):66-72. doi: 10.1111/j.1745-7254.2007.00490.x.
To investigate the effects of CD151 on the activity of endothelial NO synthase (eNOS), and ECV304 migration, proliferation and tube formation.
pAAV-CD151 and pAAV-anti-CD151 were constructed and used to transiently transfect ECV304 mediated with Lipofectamine 2000. After transfection, the expression of CD151 was measured by Western blotting. Cell migration assay was performed using Boyden transwell; proliferation assay was evaluated using the 3- [4,5-dimethylthiazol-2-yl]-2,5, diphenyltetrazolium bromide (MTT) method, and tube formation test was examined on matrigel. eNOS activity was assayed by L- [3H]citrulline production from L-[3H]arginine. The involvement of eNOS was explored using an eNOS inhibitor (L-NAME) and the effects in the process were observed.
CD151 promotes cell migration, proliferation and tube formation. In addition, CD151 increases eNOS activity. Moreover, cell migration, proliferation and tube formation induced by CD151 are inhibited when L-NAME is used, which indicates that there is an involvement of eNOS in CD151-induced cell migration, cell proliferation and tube formation.
CD151 promotes ECV304 migration, proliferation and tube formation. The mechanism is that CD151 increases eNOS activity. This result also suggests that eNOS is involved in the angiogenic effects of CD151.
研究CD151对内皮型一氧化氮合酶（eNOS）活性以及人脐静脉内皮细胞株（ECV304）迁移、增殖和管腔形成的影响。
构建pAAV-CD151和pAAV-抗CD151，并用于通过脂质体2000介导瞬时转染ECV304。转染后，通过蛋白质免疫印迹法检测CD151的表达。使用Boyden小室进行细胞迁移实验；使用3-[4,5-二甲基噻唑-2-基]-2,5-二苯基四氮唑溴盐（MTT）法评估增殖实验，并在基质胶上进行管腔形成实验。通过从L-[3H]精氨酸产生L-[3H]瓜氨酸来测定eNOS活性。使用eNOS抑制剂（L-NAME）探讨eNOS的参与情况，并观察该过程中的作用。
CD151促进细胞迁移、增殖和管腔形成。此外，CD151增加eNOS活性。而且，当使用L-NAME时，CD151诱导的细胞迁移、增殖和管腔形成受到抑制，这表明eNOS参与了CD151诱导的细胞迁移、细胞增殖和管腔形成。
CD151促进ECV304迁移、增殖和管腔形成。其机制是CD151增加eNOS活性。该结果还表明eNOS参与了CD151的血管生成作用。
