Jiang Qi, Liu Yu, Duan Dandan, Gou Meng, Wang Hao, Wang Jihong, Li Qingwei, Xiao Rong
School of Life Sciences, Liaoning Normal University, Dalian 116081, PR China; Lamprey Research Center, Liaoning Normal University, Dalian 116081, PR China.
School of Life Sciences, Liaoning Normal University, Dalian 116081, PR China; Lamprey Research Center, Liaoning Normal University, Dalian 116081, PR China.
Biochimie. 2016 Apr;123:7-19. doi: 10.1016/j.biochi.2015.11.020. Epub 2015 Nov 23.
Cysteine-rich secretory proteins (CRISPs), characterized by 16 conserved cysteines, are distributed in a wide range of organisms, such as secernenteas, amphibians, reptiles and mammals. In the previous studies, a novel CRISP family member (cysteine-rich buccal gland protein, CRBGP) was separated from the buccal gland of lampreys (Lampetra japonica, L. japonica). Lamprey CRBGP could not only suppress depolarization-induced contraction of rat tail arterial smooth muscle, but also block voltage-gated sodium channels (VGSCs). In the present study, the anti-angiogenic activities of lamprey CRBGP were investigated using endothelial cells and chick chorioallantoic membrane (CAM) models. In vitro assays, lamprey CRBGP is able to induce human umbilical vein endothelial cells (HUVECs) apoptosis by disturbing the calcium homeostasis and mitochondria functions. In addition, lamprey CRBGP could inhibit proliferation, adhesion, migration, invasion and tube formation of HUVECs by affecting the organization of F-actin and expression level of matrix metallo-proteinase 2 (MMP-2), matrix metallo-proteinase 9 (MMP-9) and vascular endothelial growth factor A (VEGFA) which are related to angiogenesis. In vivo assays, lamprey CRBGP could suppress the blood vessel formation in CAM models. Therefore, lamprey CRBGP is an important protein present in the buccal gland of lampreys and might help lampreys suppress the contraction of blood vessels, nociceptive responses and wound healing of host fishes during their feeding time. In addition, lamprey CRBGP might have the potential to act as an effective anti-angiogenic factor for the treatment of abnormal angiogenesis induced diseases.
富含半胱氨酸的分泌蛋白（CRISPs）以16个保守的半胱氨酸为特征，广泛分布于多种生物中，如线虫、两栖动物、爬行动物和哺乳动物。在先前的研究中，一种新型的CRISP家族成员（富含半胱氨酸的颊腺蛋白，CRBGP）从七鳃鳗（日本七鳃鳗）的颊腺中分离出来。七鳃鳗CRBGP不仅可以抑制大鼠尾动脉平滑肌的去极化诱导收缩，还能阻断电压门控钠通道（VGSCs）。在本研究中，利用内皮细胞和鸡胚绒毛尿囊膜（CAM）模型研究了七鳃鳗CRBGP的抗血管生成活性。体外实验表明，七鳃鳗CRBGP能够通过扰乱钙稳态和线粒体功能诱导人脐静脉内皮细胞（HUVECs）凋亡。此外，七鳃鳗CRBGP可以通过影响与血管生成相关的F-肌动蛋白的组织以及基质金属蛋白酶2（MMP-2）、基质金属蛋白酶9（MMP-9）和血管内皮生长因子A（VEGFA）的表达水平，抑制HUVECs的增殖、黏附、迁移、侵袭和管腔形成。体内实验表明，七鳃鳗CRBGP可以抑制CAM模型中的血管形成。因此，七鳃鳗CRBGP是七鳃鳗颊腺中存在的一种重要蛋白质，可能有助于七鳃鳗在摄食期间抑制宿主鱼类的血管收缩、伤害性反应和伤口愈合。此外，七鳃鳗CRBGP可能有潜力作为一种有效的抗血管生成因子，用于治疗异常血管生成引起的疾病。
