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| Effect of AMPK on Starvation-induced Autophagy and Apoptosis of Sheep (Ovis aries) Leydig Cells |
| WANG Qi*, PANG Jing*, CAI Yu, WANG Feng, ZHANG Yan-Li** |
| College of Animal Science and Technology/Jiangsu Livestock Embryo Engineering Laboratory, Nanjing Agricultural University, Nanjing 210095, China |
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Abstract Nutrition and energy metabolism affect the reproductive ability of male animals, and AMP-activated protein kinase (AMPK) plays an important role in its reproductive function as an energy receptor molecule. This study aims to study the effect of AMPK on autophagy and apoptosis of sheep (Ovis aries) leydig cells (LCs) induced by starvation. Sheep LCs were used as experimental materials and cultivated to the fourth generation. After 0, 3, 6, 12, and 24 h in serum-free medium, used cell counting kit-8 to detect cell viability and selected appropriate treatment time. After starving for 12 h, the autophagy and apoptosis of LC were observed by transmission electron microscopy and flow cytometry, qRT-PCR and Western blot were used to detect the changes in gene and protein expression related to autophagy and apoptosis. The results showed that compared with the control group, there were more autophagosomes in starvation-induced LCs, and the apoptosis rate was significantly higher (P<0.05), programmed cell death 1 (Beclin1) and microtubule-associated protein 1 light chain 3 (LC3) and other genes expression level were significantly increased (P<0.05), Becline1 protein expression and LC3Ⅱ/LC3Ⅰ ratio were significantly increased (P<0.05), pro-apoptosis related genes Bcl-2 related X protein (BAX) and other genes expression level were significantly increased (P<0.05), anti-apoptosis related genes B-cell lymphoma 2 (Bcl-2) gene expression level was significantly reduced (P<0.01), and the ratio of BAX/Bcl-2 protein was increased; AMPK/mammalian target of rapamycin (mTOR)/UNC-51-like kinase 1 (ULK1) pathway key genes expression level were increased extremely significantly (P<0.01). In summary, starvation-induced sheep LCs could activate the AMPK/mTOR/ULK1 pathway and promote the occurrence of autophagy and apoptosis. This research provides a theoretical reference for further exploring the impact of AMPK on testicular development.
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Received: 24 June 2020
Published: 01 April 2021
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Corresponding Authors:
** zhangyanli@njau.edu.cn
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| About author:: * The authors who contribute equally |
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