Increased bax and interleukin-1beta-converting enzyme messenger ribonucleic acid levels coincide with apoptosis in the bovine corpus luteum during structural regression.
Increased bax and interleukin-1beta-converting enzyme messenger ribonucleic acid levels coincide with apoptosis in the bovine corpus luteum during structural regression.
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DOI：
10.1095/biolreprod56.1.186
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发表时间：
1997
影响因子：
3.6
通讯作者：
Bo R. Rueda;K. I. Tilly;I. Botros;Peter D. Jolly;Thomas R. Hansen;Patricia B. Hoyer;Jonathan L. Tilly-Jonathan
中科院分区：
文献类型：
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作者：
Bo R. Rueda;K. I. Tilly;I. Botros;Peter D. Jolly;Thomas R. Hansen;Patricia B. Hoyer;Jonathan L. Tilly-Jonathan
最近的报告表明，前列腺素F2 α诱导的活性氧或其中间产物的产生抑制孕酮的合成，也可能作为一个触发器的黄体（CL）细胞凋亡。BCL-2是一种广泛存在于多种细胞类型中的细胞凋亡抑制剂，据报道可防止氧化应激诱导的细胞死亡。因此，进行本研究以确定编码BCL-2和该基因家族的相关成员（BAX和BCL-Xshort，其诱导细胞凋亡; BCL-Xlong，BCL-2同源物，其防止细胞凋亡）的mRNA水平是否在牛卵巢中的功能性（妊娠第21天）与退化性（发情周期第21天）CL中不同。还评估了编码p53（bcl-2和bax基因的转录调节因子）和白细胞介素-1 β转化酶（ICE）（一种最近参与诱导细胞凋亡的蛋白质，其表达可通过氧化应激增强）的mRNA水平。使用逆转录-聚合酶链反应（RT-PCR）技术与总RNA制备的功能或回归CL的编码牛bax，bcl-x，p53，和冰的部分cDNA克隆分离。牛bcl-2的cDNA不能从黄体组织RNA中分离，尽管使用几个引物对扩增。然后从功能性或回归CL中提取总RNA，并通过北方印迹分析进行分析。与功能性CL中bax和Ice表达水平相比，退化CL中凋亡的发生（如核小体间DNA切割的存在所证明的）与bax和Ice mRNA水平的显著增加相关（p < 0.05，n = 3）。功能性CL与消退性CL中bcl-x或p53 mRNA水平无显著差异。通过RT-PCR对bcl-x mRNA的分析表明，在功能性和退化的牛黄体组织中，如果不是唯一的话，长型是主要的mRNA表达。Bax的表达增加与卵巢颗粒细胞和生殖细胞的凋亡相关，并且可能是卵巢颗粒细胞和生殖细胞凋亡所必需的，基于这些数据，我们提出BAX可能在黄体退化期间的凋亡诱导中发挥类似的作用。此外，在退化的CL中增加的Ice mRNA水平提供了死亡蛋白酶的ICE家族可能参与黄体溶解的第一个证据。
Recent reports have demonstrated that prostaglandin F2alpha-induced generation of reactive oxygen species or their intermediates inhibits progesterone synthesis and may also serve as a trigger for apoptosis in the corpus luteum (CL). BCL-2, an inhibitor of apoptosis in a wide variety of cell types, has been reported to prevent oxidative stress-induced cell death. Thus, the present studies were conducted to determine whether levels of mRNA encoding BCL-2 and related members of this gene family (BAX and BCL-Xshort, which induce apoptosis; BCL-Xlong, a BCL-2 homologue that prevents apoptosis) differed in functional (Day 21 of pregnancy) versus regressed (Day 21 of the estrous cycle) CL in the bovine ovary. Levels of mRNAs encoding p53, a transcriptional regulator of the bcl-2 and bax genes, and interleukin-1beta-converting enzyme (ICE), a protein recently implicated in the induction of apoptosis whose expression may be enhanced by oxidative stress, were also assessed. Partial cDNA clones encoding bovine bax, bcl-x, p53, and Ice were isolated using the reverse transcriptase-polymerase chain reaction (RT-PCR) technique with total RNA prepared from functional or regressed CL. A bovine bcl-2 cDNA could not be isolated from luteal tissue RNA despite the use of several primer pairs for amplification. Total RNA was then extracted from functional or regressed CL and analyzed by Northern blot analysis. The occurrence of apoptosis in regressed CL, as evidenced by the presence of internucleosomal DNA cleavage, was associated with a significant increase in both bax and Ice mRNA levels as compared with levels of bax and Ice expression in functional CL (p < 0.05, n = 3). There were no significant differences in bcl-x or p53 mRNA levels in functional versus regressed CL. Analysis of bcl-x mRNA by RT-PCR revealed that the long form was the primary, if not only, mRNA expressed in functional and regressed bovine luteal tissue. On the basis of data that increased expression of bax is associated with, and may be required for, apoptosis in ovarian granulosa cells and germ cells, we propose that BAX may play a similar role in apoptosis induction during luteal regression. Moreover, the increased Ice mRNA levels in regressed CL provides the first evidence that the ICE family of death proteases may be involved in luteolysis.