Prokineticins (endocrine gland-derived vascular endothelial growth factor and BV8) in the bovine ovary: Expression and role as mitogens and survival factors for corpus luteum-derived endothelial cells
Prokineticins (endocrine gland-derived vascular endothelial growth factor and BV8) in the bovine ovary: Expression and role as mitogens and survival factors for corpus luteum-derived endothelial cells
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DOI：
10.1210/en.2005-0297
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发表时间：
2005-09-01
期刊：
影响因子：
4.8
通讯作者：
Meidan, R
中科院分区：
文献类型：
--
作者：
Kisliouk, T;Podlovni, H;Meidan, R
黄体是一种高度血管化的内分泌腺，是研究血管生成因子的良好模型。原激动素(Prokineticins，PK-1)和原激动素(Prokineticins，PK-2)又称内分泌腺源性血管内皮细胞生长因子(VEGF)和BV8，是新发现的选择性血管生成有丝分裂原蛋白。我们以前在人和牛卵巢细胞中发现了两个同源的G蛋白偶联受体PK结合位点(PK-R1和PK-R2)，但它们的功能尚不清楚。在这项研究中，我们研究了PK在CL来源的内皮细胞和类固醇生成细胞类型(分别为LEC和LSC)中的存在和作用。使用牛PK-1序列的特异性引物(检索自牛全基因组猎枪数据库)，用实时荧光定量聚合酶链式反应(Real-time PCR)鉴定了CL和卵泡中的PK-1mRNA。PK是一种有效的促血管生成有丝分裂原，可促进LEC增殖，增加[H-3]胸腺嘧啶核苷掺入、MAPK活性和c-jun/fos基因表达。通过细胞核形态(4‘，6-二氨基-2-苯基吲哚二盐酸盐染色)、DNA片段化(dUTP末端标记法)和caspase-3裂解等方法检测PK蛋白对细胞存活的影响。结果表明，PK对血清饥饿诱导的LEC凋亡有保护作用。应激条件，如血清停用、肿瘤坏死因子-α和低氧显著增加PK-R2的表达，而PK-R1mRNA水平保持不变。提示PK-1对LEC的抗凋亡作用可能是通过PK-R2介导的。PK-1通过LSC促进血管内皮生长因子的表达，提示PK-1还可能通过血管内皮生长因子间接影响黄体血管生成。综上所述，这些发现提示PK-1作为LEC的有丝分裂原和生存因子在黄体功能中发挥重要作用。
A highly vascular endocrine gland, the corpus luteum ( CL) is an excellent model for the study of angiogenic factors. Prokineticins (PK-1 and -2), also termed endocrine-gland-derived vascular endothelial growth factor ( VEGF) and BV8 are newly identified proteins described as selective angiogenic mitogens. We previously identified PK binding sites, two closely homologous G protein-coupled receptors (PK-R1 and PK-R2) in human and bovine ovarian cells, but their function remained unknown. In this study we examined the presence and effects of PK in CL-derived endothelial and steroidogenic cell types (LEC and LSC, respectively). PK-1 mRNA was identified in CL and follicles by real-time PCR, using primers specific for the bovine PK-1 sequence ( retrieved from Bos taurus whole genome shotgun database). PK were potent angiogenic mitogens for LEC; they enhanced cell proliferation, elevated [H-3] thymidine incorporation, MAPK activation, and c-jun/fos mRNA expression. The effects of PK proteins on cell survival were examined by nuclear morphology (4', 6-diamidino-2-phenylindole dihydrochloride staining), measurement of DNA fragmentation (terminal dUTP nucleotide end labeling assay), and caspase-3 cleavage. Results obtained by these techniques demonstrated that PK protected LEC from serum starvation-induced apoptosis. Stress conditions such as serum withdrawal, TNF-alpha, and hypoxia markedly increased PK-R2 expression, whereas mRNA levels of PK-R1 remained unchanged. These suggest that the antiapoptotic effect of PK-1 on LEC may be mediated via PK-R2. PK-1 increased VEGF mRNA expression by LSC, implying that it could also indirectly, via VEGF, affect luteal angiogenesis. Together, these findings suggest an important role for PK-1 in luteal function by acting as a mitogen and survival factor in LEC.