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
{"title":"Resistin as a new player in the regulation of porcine corpus luteum luteolysis: in vitro effect on proliferation/viability, apoptosis and autophagy.","authors":"P Kurowska,&nbsp;K Gazdzik,&nbsp;A Jasinska,&nbsp;E Mlyczynska,&nbsp;D Wachowska,&nbsp;A Rak","doi":"10.26402/jpp.2023.1.03","DOIUrl":null,"url":null,"abstract":"<p><p>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.</p>","PeriodicalId":50089,"journal":{"name":"Journal of Physiology and Pharmacology","volume":null,"pages":null},"PeriodicalIF":2.0000,"publicationDate":"2023-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"Journal of Physiology and Pharmacology","FirstCategoryId":"3","ListUrlMain":"https://doi.org/10.26402/jpp.2023.1.03","RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q3","JCRName":"PHYSIOLOGY","Score":null,"Total":0}
引用次数: 0

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.

查看原文
分享 分享
微信好友 朋友圈 QQ好友 复制链接
本刊更多论文
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。
求助全文
约1分钟内获得全文 去求助
来源期刊
CiteScore
4.00
自引率
22.70%
发文量
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.
期刊最新文献
Adherence and acceptance of a new oral nutritional supplement in cancer patients - a pilot study in crossover design. Chromosomal analysis in pregnant women of advanced maternal age: indications for prenatal diagnosis. Determining the expression of vitamin D receptor, regulator of iron metabolism hepcidin and cathelicidin antimicrobial peptide genes for development of future diagnostic and therapeutic options in patients with inflammatory bowel disease. Expression patterns of interleukin-6 and microRNA-146A during orthodontic relapse in a rat model. Ginsenoside Rg1 induces ferroptosis by regulating the focal adhesion kinase/protein kinase B-forkhead box O3A signaling pathway and alleviates sepsis-induced myocardial damage.
×
引用
GB/T 7714-2015
复制
MLA
复制
APA
复制
导出至
BibTeX EndNote RefMan NoteFirst NoteExpress
×
×
提示
您的信息不完整,为了账户安全,请先补充。
现在去补充
×
提示
您因"违规操作"
具体请查看互助需知
我知道了
×
提示
现在去查看 取消
×
提示
确定
0
微信
客服QQ
Book学术公众号 扫码关注我们
反馈
×
意见反馈
请填写您的意见或建议
请填写您的手机或邮箱
已复制链接
已复制链接
快去分享给好友吧!
我知道了
×
扫码分享
扫码分享
Book学术官方微信
Book学术文献互助
Book学术文献互助群
群 号:481959085
Book学术
文献互助 智能选刊 最新文献 互助须知 联系我们:info@booksci.cn
Book学术提供免费学术资源搜索服务,方便国内外学者检索中英文文献。致力于提供最便捷和优质的服务体验。
Copyright © 2023 Book学术 All rights reserved.
ghs 京公网安备 11010802042870号 京ICP备2023020795号-1