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Regulation of mouse blastocyst adhesion, outgrowth and secretion of matrix metalloproteinase-2 by cGMP and nitric oxide in vitro

机译:cGMP和一氧化氮对小鼠胚泡黏附,生长和基质金属蛋白酶-2分泌的调节

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摘要

Nitric oxide (NO) is a multifunctional messenger molecule produced through oxidation of L-arginine to L-citrulIine by enzyme NO synthase (NOS). In the current study, mouse blastocysts were cultured in the different media, and the implantation capacity of blastocyst was evaluated by evaluating the percentage of embryos adhesion and outgrowth after culture for 12, 24 or 48 h. Matrix metalloproteinase-2 (MMP-2) mRNA was detected by RT-PCR, and MMP-2 protein was detected by gelatin zymography. Inhibition of blastocyst adhesion and outgrowth was observed in embryo cultured with 500 (mu)mol/L NOS inhibitor NG-mono-methyl-L-arginine (L-NMMA) alone; however, 100 (mu)mol/L S-nitroso-N-acetylpenicillamine (SNAP), a NO donor, and 20 (mu)mol/L cGMP analogue, 8-Br-cGMP could block this inhibition. The expression and production of MMP-2 in the blastocysts were suppressed by L-NMMA, and SNAP or 8-br-cGMP could reverse this suppression. These results suggest that NO induces embryo implantation by cGMP signaling pathway.
机译:一氧化氮(NO)是通过NO合酶(NOS)将L-精氨酸氧化为L-瓜氨酸而产生的多功能信使分子。在当前的研究中,小鼠胚泡在不同的培养基中培养,并通过评估培养12、24或48 h后胚胎粘附和生长的百分比来评估胚泡的植入能力。 RT-PCR检测基质金属蛋白酶-2(MMP-2)mRNA,明胶酶谱检测MMP-2。在单独用500μmol/ L NOS抑制剂NG-单甲基-L-精氨酸(L-NMMA)培养的胚胎中观察到了胚泡粘附和生长抑制。但是,NO供体100μmol/ L S-亚硝基-N-乙酰青霉胺(SNAP)和20μmol/ L cGMP类似物8-Br-cGMP可以阻止这种抑制作用。 L-NMMA抑制了胚泡中MMP-2的表达和产生,SNAP或8-br-cGMP可以逆转这种抑制作用。这些结果表明NO通过cGMP信号通路诱导胚胎着床。

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