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首页> 外文期刊>Cellular Physiology and Biochemistry >FSHR and LHR Expression and Signaling as Well as Maturation and Apoptosis of Cumulus-Oocyte Complexes Following Treatment with FSH Receptor Binding Inhibitor in Sheep
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FSHR and LHR Expression and Signaling as Well as Maturation and Apoptosis of Cumulus-Oocyte Complexes Following Treatment with FSH Receptor Binding Inhibitor in Sheep

机译：FSH受体结合抑制剂处理绵羊后卵丘卵母细胞复合物的FSHR和LHR表达和信号以及成熟和凋亡

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>Background/Aims: Currently, it remains unknown whether FSH receptor binding inhibitor (FRBI) influences follicular development and reproduction functions in humans and animals. The present study aimed to investigate FRBI effects on in vitro maturation (IVM) and apoptosis of cumulus-oocyte complexes (COCs) of sheep, to determine the effect of FRBI on mRNA and protein levels of FSHR and LHR in COCs, and to elucidate the signal pathway of FRBI effects. Methods: COCs were in vitro cultured for 24h in the IVM media supplemented with varying concentrations of FRBI (0, 10, 20, 30 and 40?μg/mL) and FSH (10IU/mL). The harvested COCs were observed under an inverted microscope and maturation rates of COCs were determined. Real time RT-PCR and Western blotting were utilized to detect mRNA and protein levels of FSHR and LHR. The concentrations of FSH, LH and caspase-3 were determined using especial ELISA kits for sheep, respectively. Results: Maturation rates of COCs decreased gradually as FRBI concentrations increased from 0 to 40?μg/mL, reaching a bottom value of 23.76% of the FRBI-4 group. The maximal apoptosis rate was detected in the FRBI-4 group. IP3 contents of FRBI-3 and FRBI-4 groups were reduced as compared to control group (CG) and FSH groups (P<0.05). Levels of FSHR protein of FRBI-3 and FRBI-4 groups as well as LHR protein of FRBI-4 group were significantly less than that of CG and FSH group. FSH contents of four FRBI treatment groups were gradually decreased along with the supplementation doses of FRBI. Caspase-3 contents of FRBI groups were reduced with a maximum reduction of the FRBI-2 group. Conclusion: Our results revealed supplement of FRBI into IVM media could dose-dependently decrease the maturation rate and increase apoptosis rate of sheep COCs. A lower dose of FRBI treatment slightly promoted IP3 production, but a higher dose of FRBI reduced IP3 production. FRBI suppressed the mRNA and protein expression levels of FSHR and LHR in sheep COCs. Our study will help to therapy effectively ovarian diseases, improve ovarian and follicular functions, and further to promote fertility of humans and animals.
机译：> 背景/目的： 目前，尚不知道FSH受体结合抑制剂（FRBI）是否会影响人和动物的卵泡发育和生殖功能。本研究旨在研究FRBI对绵羊卵丘卵母细胞复合物（IVC）的体外成熟（IVM）和凋亡的影响，以确定FRBI对FSHR和LHR的mRNA和蛋白水平的影响在COC中的作用，并阐明FRBI效应的信号途径。 方法： 将COCs在补充了不同浓度FRBI（0、10、20、30和40的IVM培养基）中体外培养24小时。 ？μg/ mL）和FSH（10IU / mL）。在倒置显微镜下观察收获的COC，并确定COC的成熟率。利用实时RT-PCR和蛋白质印迹法检测FSHR和LHR的mRNA和蛋白水平。分别使用绵羊专用ELISA试剂盒测定FSH，LH和caspase-3的浓度。 结果： 随着FRBI浓度从0增加到40？μg/ mL，COC的成熟率逐渐降低，达到FRBI-4组的23.76％的最低值。在FRBI-4组中检测到最大凋亡率。与对照组（CG）和FSH组相比，FRBI-3和FRBI-4组的IP3含量降低（P <0.05）。 FRBI-3和FRBI-4组的FSHR蛋白水平以及FRBI-4组的LHR蛋白水平均显着低于CG和FSH组。随着FRBI补充剂量的增加，四个FRBI治疗组的FSH含量逐渐降低。 FRBI组的Caspase-3含量降低，而FRBI-2组最大降低。 结论： 我们的研究结果表明，在IVM培养基中补充FRBI可以剂量依赖性地降低绵羊COC的成熟率并增加其凋亡率。较低剂量的FRBI处理略微促进了IP3的产生，但是较高剂量的FRBI降低了IP3的产生。 FRBI抑制绵羊COC中FSHR和LHR的mRNA和蛋白表达水平。我们的研究将有助于有效治疗卵巢疾病，改善卵巢和卵泡功能，并进一步促进人类和动物的生育能力。

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