Nitric oxide induces apoptosis in bovine luteal cells
Nitric oxide induces apoptosis in bovine luteal cells
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DOI：
10.1262/jrd.17092
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发表时间：
2006-06-01
影响因子：
1.8
通讯作者：
Skarzynski, Dariusz J.
中科院分区：
文献类型：
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作者：
Korzekwa, Anna J.;Okuda, Kiyoshi;Skarzynski, Dariusz J.
我们之前在体内和体外研究中表明，一氧化氮（NO）参与牛体内的黄体溶解。牛黄体（CL）局部产生的一氧化氮抑制黄体酮（P4）的合成，被认为是子宫前列腺素（PG） F-2 α诱导的黄体溶解级联反应的一个组成部分。本研究在体外培养的牛黄体细胞（发情周期第15-17天）中，研究了NO在结构黄体溶解过程中的分子机制。比较NO供体（NONOate; 10(-4) M）与PGF（2 α）对DNA片段化、细胞活力、P4生成和caspase-3活性的影响。(10 (6) M)。此外，通过半定量RT-PCR检测NONOate处理后细胞内钙[Ca2+](i)的动员和Fas- l、Fas、bcl-2、bax和caspase-3的基因表达。用量热法检测Caspase-3活性。与PGF（2 α）相反，NONOate降低细胞活力。NONOate处理后的DNA断裂比PGF（2 α）增加更多。NONOate增加[Ca2+](i)在细胞中的动员。NO供体虽然不影响Fas- l和bcl-2基因表达，但刺激了Fas和bax mRNA和caspase-3的表达。NONOate处理的细胞bcl-2与bax mRNA的比值降低。此外，NONOate比PGF（2 α）更有效地刺激caspase-3活性。综上所述，一氧化氮是一种黄体溶解因子，通过诱导黄体细胞凋亡，在黄体结构性溶解的发情周期调控中起重要作用。
We previously showed in in vivo and in vitro studies that nitric oxide (NO) is engaged in luteolysis in cattle. Nitric oxide produced locally in the bovine corpus luteum (CL) inhibits progesterone (P4) synthesis and is suggested to be a component of the luteolytic cascade induced by uterine prostaglandin (PG) F-2 alpha. In the present study, the molecular mechanisms of NO action during structural luteolysis were studied in cultured bovine luteal cells (Days 15-17 of the estrous cycle). The effects of the NO donor (NONOate; 10(-4) M) on DNA fragmentation, cell viability, P4 production and caspase-3 activity were compared with those of PGF(2 alpha). (10(-6) M). Moreover, mobilization of intracellular calcium [Ca2+](i) and gene expressions of Fas-L, Fas, bcl-2, bax, and caspase-3 in the cells were determined by semi-quantitative RT-PCR after NONOate treatment. Caspase-3 activity was examined calorimetrically. Contrary to PGF(2 alpha) NONOate decreased cell viability. DNA fragmentation after NONOate treatment increased by more than with PGF(2 alpha). NONOate increased mobilization of [Ca2+](i) in the cells. Although the NO donor did not affect Fas-L and bcl-2 gene expression, it stimulated Fas and bax mRNA and caspase-3 expression. The ratio of bcl-2 to bax mRNA level decreased in the cells treated with NONOate. Moreover, NONOate stimulated caspase-3 activity more effectively than PGF(2 alpha). The overall results suggest that NO is a luteolytic factor that plays a crucial role in regulation of the estrous cycle in structural luteolysis by inducing apoptosis of luteal cells in cattle.