Regulatory Changes of Apoptotic Factors in the Bovine Corpus Luteum After Induced Luteolysis
Regulatory Changes of Apoptotic Factors in the Bovine Corpus Luteum After Induced Luteolysis
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DOI：
10.1002/mrd.20946
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发表时间：
2009-03-01
影响因子：
2.5
通讯作者：
Schams, D.
中科院分区：
文献类型：
--
作者：
Kliem, H.;Berisha, B.;Schams, D.
黄体（CL）不仅提供了研究增殖过程的机会，而且还提供了研究退化过程的机会。在CL的黄体溶解过程中，似乎涉及黄体和内皮细胞的突然死亡（细胞凋亡）。本研究的目的是检测已知参与凋亡过程的因子的mRNA表达：单核细胞趋化蛋白-1（MCP-1），外源性和内源性凋亡途径的因子，半胱天冬酶3、-6、-7和干扰素γ（IFN γ）。在黄体中期注射500 μ g氯前列醇诱导黄体溶解。在PGF 2 α注射后0.5、2、4、12、24、48和64 h收集CL。在动物饲养室采集对照CL（第8 - 12天）。实时定量RT-PCR检测mRNA的表达。聚（ADP-核糖）聚合酶（PARP-1）和IFN γ的蛋白质印迹分析以及肿瘤坏死因子α的蛋白质测量。(TNFα）进行了EIA。MCP-1、IFN-γ和大多数外源性途径因子的mRNA水平在0.5~2 h显著升高，内源性途径因子在24-48 h显著上调。Caspase 6和3分别从2和12小时开始显著增加，而Caspase 7在24小时后显著上调。TNF α的蛋白水平在12小时显著增加至最高水平。Western印迹显示，从12至24小时，PARP-1的89 kDa片段的水平增加，这是细胞凋亡的特异性。我们认为，外源性途径是更重要的发病黄体溶解，因为它更早和更高的增加在诱导黄体溶解。
The corpus luteum (CL) offers the opportunity to study not only proliferative, but also regressive processes. During luteolysis of the CL a sudden death of luteal and endothelial cells seems to be involved (apoptosis). The aim of this study was to examen the mRNA expression of factors known to be involved in apoptotic processes: monocyte chemoattractant protein-1 (MCP-1), factors of the extrinsic and intrinsic apoptotic pathways, caspase3, -6, -7 and interferone gamma (IFN gamma). Luteolysis was induced by injection of 500 mu g Cloprostenol during mid-luteal phase. The CLs were collected at 0.5, 2, 4, 12, 24, 48, and 64 hr after PGF2 alpha-injection. Control CLs (Days 8-12) were collected at the slaugtherhouse. Real-time RT-PCR determined the mRNA expressions. Western blot analysis of poly(ADP-ribose) polymerase (PARP-1) and IFN gamma as well as protein measurement of tumor necrosis factor alpha. (TNF alpha) by EIA were performed. The mRNA levels of MCP-1, IFN gamma and most factors of the extrinsic pathway were significantly increased between 0.5 and 2 hr. The factors of the intrinsic pathway were mostly later up-regulated at 24-48 hr after PGF2 alpha. Caspase6 and 3 revealed a significant increase from 2 and 12 hr, respectively, whereas caspase7 was significantly up-regulated after 24 hr. The protein level of TNF alpha increased significantly to a maximum level at 12 hr. The Western blot revealed an increasing level of an 89 kDa fragment of PARP-1 from 12 to 24 hr, which is specific for apoptosis. We assume that the extrinsic pathway is more important for the onset of luteolysis, because of its earlier and higher increase during induced luteolysis.