Resistin as a new player in the regulation of porcine corpus luteum luteolysis: in vitro effect on proliferation/viability, apoptosis and autophagy.

IF 2 4区 医学 Q3 PHYSIOLOGY Journal of Physiology and Pharmacology Pub Date : 2023-02-01 DOI:10.26402/jpp.2023.1.03
P Kurowska, K Gazdzik, A Jasinska, E Mlyczynska, D Wachowska, A Rak
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Abstract

The formation and luteolysis of the corpus luteum (CL) is strictly controlled by many factors. Imbalance between proliferation and apoptosis processes leads to deficiency of the luteal phase and infertility. Our previous study showed resistin expression in porcine luteal cells and an inhibitory effect on progesterone synthesis. Thus, the aim of the present study was to examine the in vitro effect of resistin on the proliferation/viability, apoptosis and autophagy of porcine luteal cells as well as the involvement of mitogen-activated kinase (MAP3/1), protein kinase B (AKT) and signal transducer and activator of transcription 3 (STAT3) in these processes. Porcine luteal cells were incubated with resistin (0.1-10 ng/mL) for 24-72 h and viability was assessed using the alamarBlue or 3-[4,5-dimethylthiazole-2-yl]-2,5-diphenyltetrazolium bromide (MTT) assay. Then, the time-dependent effect of resistin on mRNA and protein expression of proliferating cell nuclear antigen (PCNA), caspase 3, BCL2-like protein 4 (BAX), B-cell lymphoma 2 (BCL2), beclin1, microtubule-associated protein 1A/1B-light chain 3 (LC3) and lysosomal-associated membrane protein 1 (LAMP1) was measured by real-time polymerase chain reaction (PCR) and immunoblotting, respectively. We found that resistin enhanced luteal cell viability with no effect on caspase 3 mRNA and protein, increased the BAX/BCL2 mRNA and protein ratio and significantly stimulated the initiation of autophagy, which promotes the maintenance of CL function rather than its regression. Additionally, using pharmacological inhibitors of MAP3/1 (PD98059), AKT (LY294002) and STAT3 (AG490), we observed that the effect of resistin was reversed to the control level in viability and, by influence, MAP3/1 and STAT3 in autophagy. Taken together, our results suggest that resistin, in addition to its well-known effect on granulosa cell function has direct influence on CL luteolysis and the formation and maintenance of luteal cell function.

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抵抗素在调节猪黄体黄体溶解中的新作用:对体外增殖/活力、细胞凋亡和自噬的影响。
黄体(corpus luteum, CL)的形成和黄体溶解受多种因素的严格控制。增殖与凋亡过程的不平衡导致黄体期不足和不孕。我们之前的研究表明抵抗素在猪黄体细胞中表达,并对黄体酮的合成有抑制作用。因此,本研究的目的是研究抵抗素对猪黄体细胞增殖/活力、凋亡和自噬的体外影响,以及丝裂原活化激酶(MAP3/1)、蛋白激酶B (AKT)和转录信号传导和激活因子3 (STAT3)在这些过程中的作用。猪黄体细胞与抵抗素(0.1 ~ 10 ng/mL)孵育24 ~ 72 h,采用alamarBlue或3-[4,5-二甲基噻唑-2-基]-2,5-二苯基溴化四唑(MTT)法测定细胞活力。利用实时聚合酶链反应(PCR)和免疫印迹法分别检测抵抗素对增殖细胞核抗原(PCNA)、caspase 3、BCL2样蛋白4 (BAX)、b细胞淋巴瘤2 (BCL2)、beclin1、微管相关蛋白1A/ 1b轻链3 (LC3)和溶酶体相关膜蛋白1 (LAMP1) mRNA和蛋白表达的时间依赖性。我们发现抵抗素在不影响caspase 3 mRNA和蛋白的情况下提高了黄体细胞活力,增加了BAX/BCL2 mRNA和蛋白的比值,显著刺激了自噬的开始,促进了CL功能的维持而不是退化。此外,使用MAP3/1 (PD98059)、AKT (LY294002)和STAT3 (AG490)的药物抑制剂,我们观察到抵抗素对细胞活力的影响逆转至对照水平,并通过影响MAP3/1和STAT3对细胞自噬的影响。综上所述,我们的研究结果表明抵抗素除了其众所周知的对颗粒细胞功能的影响外,还直接影响CL黄体溶解和黄体细胞功能的形成和维持。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
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来源期刊
CiteScore
4.00
自引率
22.70%
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0
审稿时长
6-12 weeks
期刊介绍: Journal of Physiology and Pharmacology publishes papers which fall within the range of basic and applied physiology, pathophysiology and pharmacology. The papers should illustrate new physiological or pharmacological mechanisms at the level of the cell membrane, single cells, tissues or organs. Clinical studies, that are of fundamental importance and have a direct bearing on the pathophysiology will also be considered. Letters related to articles published in The Journal with topics of general professional interest are welcome.
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