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牛黄体发情周期及诱导黄体溶解后细胞外基质降解蛋白酶及其抑制剂的表达与定位

Expression and localisation of extracellular matrix degrading proteases and their inhibitors during the oestrous cycle and after induced luteolysis in the bovine corpus luteum.

作者信息

Kliem H, Welter H, Kraetzl W D, Steffl M, Meyer H H D, Schams D, Berisha B

机构信息

Physiology Weihenstephan, Technical University Munich, 85354 Freising, Germany.

出版信息

Reproduction. 2007 Sep;134(3):535-47. doi: 10.1530/REP-06-0172.

Abstract

The corpus luteum (CL) offers the opportunity to study high proliferative processes during its development and degradation processes during its regression. We examined the mRNA expression of matrix metalloproteases (MMP)-1, MMP-2, MMP-9, MMP-14, MMP-19, tissue inhibitor of MMP (TIMP)-1, TIMP-2, tissue plasminogen activator (tPA), urokinase plasminogen activator (uPA), uPA-receptor (uPAR), PA-inhibitors (PAI)-1, PAI-2 in follicles 20 h after GnRH application, CLs during days 1-2, 3-4, 5-7 and 8-12 of the oestrous cycle as well as after induced luteolysis. Cows in the mid-luteal phase were injected with Cloprostenol and the CLs were collected at 0.5, 2, 4, 12, 24, 48 and 64 h after PGF2alpha injection. Real-time RT-PCR determined mRNA expressions. Expression from 20 h after GnRH to day 12: MMP-1, MMP-2, MMP-14 and tPA showed a clear expression, but no regulation. TIMP-1 and uPAR mRNA increased when compared with the follicular phase. TIMP-2, MMP-9, MMP-19 and uPA increased from the follicular phase to days 8-12. PAI-1 and PAI-2 expression increased from days 1-7 and decreased to days 8-12. Induced luteolysis: MMP-1, MMP-2, MMP-9, MMP-14, MMP-19 and TIMP-1 all increased at different time points and intensities, whereas TIMP-2 was constantly decreased from 24 to 64 h. The plasminogen activator system and their inhibitors were up-regulated from 2 to 64 h, tPA was already increased after 0.5 h. Immunohistochemistry for MMP-1, MMP-2, MMP-14: an increased staining for MMP-1 and MMP-14 was seen in large luteal cells beginning 24 h after PGF2alpha application. MMP-2 showed a strong increase in staining in endothelial cells at 48 h.

摘要

黄体(CL)为研究其发育过程中的高增殖过程以及退化过程中的降解过程提供了机会。我们检测了促性腺激素释放激素(GnRH)应用后20小时卵泡、发情周期第1 - 2天、3 - 4天、5 - 7天和8 - 12天的黄体以及诱导黄体溶解后的基质金属蛋白酶(MMP)-1、MMP-2、MMP-9、MMP-14、MMP-19、基质金属蛋白酶组织抑制剂(TIMP)-1、TIMP-2、组织型纤溶酶原激活剂(tPA)、尿激酶型纤溶酶原激活剂(uPA)、uPA受体(uPAR)、PA抑制剂(PAI)-1、PAI-2的mRNA表达。处于黄体中期的母牛注射氯前列醇,并在注射前列腺素F2α(PGF2α)后0.5、2、4、12、24、48和64小时收集黄体。实时逆转录聚合酶链反应(RT-PCR)测定mRNA表达。从GnRH应用后20小时到第12天的表达情况:MMP-1、MMP-2、MMP-14和tPA呈现明显表达,但无调节。与卵泡期相比,TIMP-1和uPAR mRNA增加。TIMP-2、MMP-9、MMP-19和uPA从卵泡期到第8 - 12天增加。PAI-1和PAI-2表达从第1 - 7天增加,到第8 - 12天减少。诱导黄体溶解:MMP-1、MMP-2、MMP-9、MMP-14、MMP-19和TIMP-1在不同时间点和强度均增加,而TIMP-2从24小时到64小时持续下降。纤溶酶原激活剂系统及其抑制剂从2小时到64小时上调,tPA在0.5小时后即已增加。MMP-1、MMP-2、MMP-14的免疫组织化学:PGF2α应用后24小时开始,大黄体细胞中MMP-1和MMP-14染色增加。MMP-2在48小时时内皮细胞染色强烈增加。

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