YBX1 inhibits mitochondrial-mediated apoptosis in ischemic heart through the PI3K/AKT signaling pathway

Abstract

Background

Myocardial infarction (MI) is associated with higher morbidity and mortality in the world, especially in cold weather. YBX1 is an RNA-binding protein that is required for pathological growth of cardiomyocyte by regulating cell growth and protein synthesis. But YBX1, as an individual RNA-binding protein, regulates cardiomyocytes through signaling cascades during myocardial infarction remain largely unexplored.

Methods

In vivo, the mouse MI model was induced by ligating the left anterior descending coronary artery (LAD), and randomly divided into sham operation group, MI group, MI+ YBX1 knockdown/overexpression group and MI+ negative control (NC) group. The protective effect of YBX1 was verified by echocardiography and triphenyltetrazolium chloride staining. In vitro, mitochondrial-dependent apoptosis was investigated by using CCK8, TUNEL staining, reactive oxygen species (ROS) staining and JC-1 staining in hypoxic neonatal mouse cardiomyocytes (NMCMs).

Results

YBX1 expression of cardiomyocytes was downregulated in a mouse model and a cellular model on the ischemic condition. Compared to mice induced by MI, YBX1 overexpression mediated by adeno-associated virus serotype 9 (AAV9) vector reduced the infarcted size and improved cardiac function. Knockdown of endogenous YBX1 by shRNA partially aggravated ischemia-induced cardiac dysfunction. In hypoxic cardiomyocytes, YBX1 overexpression decreased lactic dehydrogenase (LDH) release, increased cell viability, and inhibited apoptosis by affecting the expression of apoptosis related proteins, while knockdown of endogenous YBX1 by siRNA had the opposite effect. Overexpression of YBX1 restored mitochondrial dysfunction in hypoxic NMCMs by increasing mitochondrial membrane potential and ATP content and decreasing ROS. In hypoxic NMCMs, YBX1 overexpression increased the expression of phosphorylated phosphatidylinositol 3 kinase (PI3K)/AKT, and the antiapoptosis effect of YBX1 was eliminated t by LY294002, PI3K/AKT inhibitor.

Conclusion

YBX1 protected the heart from ischemic damage by inhibiting the mitochondrial-dependent apoptosis through PI3K/AKT pathway. It is anticipated that YBX1 may serve as a novel therapeutic target for MI.

Keywords

YBX1 / PI3K/AKT / apoptosis / mitochondrial function / myocardial infarction

References

[1]	Yuan X, Pan J, Wen L, et al.MiR-144-3p enhances cardiac fibrosis after myocardial infarction by targeting PTEN. Front Cell Dev Biol, 2019; 7: 249. doi: 10.3389/fcell.2019.00249
[2]	Acharya D.Predictors of outcomes in myocardial infarction and cardiogenic shock. Cardiol Rev, 2018; 26(5): 255-266. doi: 10.1097/CRD.0000000000000190
[3]	Bai X, Yang C, Jiao L, et al.LncRNA MIAT impairs cardiac contractile function by acting on mitochondrial translocator protein TSPO in a mouse model of myocardial infarction. Signal Transduct Target Ther, 2021; 6(1): 172. doi: 10.1038/s41392-021-00538-y
[4]	Luo Y, Zhang Y, Han X, et al.Akkermansia muciniphila prevents cold-related atrial fibrillation in rats by modulation of TMAO induced cardiac pyroptosis. EBioMedicine, 2022; 82: 104087. doi: 10.1016/j.ebiom.2022.104087
[5]	Lewington S, Li L, Sherliker P, et al.Seasonal variation in blood pressure and its relationship with outdoor temperature in 10 diverse regions of China: the China Kadoorie Biobank. J Hypertens, 2012; 30(7): 1383-1391. doi: 10.1097/HJH.0b013e32835465b5
[6]	Brennan P J, Greenberg G, Miall W E, et al.Seasonal variation in arterial blood pressure. Br Med J (Clin Res Ed), 1982; 285(6346): 919-923. doi: 10.1136/bmj.285.6346.919
[7]	Dong Y, Chen H, Gao J, et al.Molecular machinery and interplay of apoptosis and autophagy in coronary heart disease. J Mol Cell Cardiol, 2019; 136: 27-41. doi: 10.1016/j.yjmcc.2019.09.001
[8]	Els?sser A, Suzuki K, Lorenz-Meyer S, et al. The role of apoptosis in myocardial ischemia: a critical appraisal. Basic Res Cardiol, 2001; 96(3): 219-226. doi: 10.1007/s003950170052
[9]	Slee E A, Adrain C, Martin S J.Serial killers: ordering caspase activation events in apoptosis. Cell Death & Differentiation, 1999; 6(11): 1067-1074. doi: 10.1038/sj.cdd.4400601
[10]	Jose Corbalan J, Vatner D E, Vatner S F.Myocardial apoptosis in heart disease: does the emperor have clothes? Basic Res Cardiol, 2016; 111(3): 31.
[11]	Suresh P S, Tsutsumi R, Venkatesh T.YBX1 at the crossroads of non-coding transcriptome, exosomal, and cytoplasmic granular signaling. Eur J Cell Biol, 2018; 97(3): 163-167. doi: 10.1016/j.ejcb.2018.02.003
[12]	Cui Q, Wang C, Liu S, et al.YBX1 knockdown induces renal cell carcinoma cell apoptosis via Kindlin-2. Cell cycle (Georgetown, Tex.), 2021; 20(22): 2413-2427. doi: 10.1080/15384101.2021.1985771
[13]	Zhang H, Zheng W, Li D, et al.MiR-379-5p promotes chondrocyte proliferation via inhibition of PI3K/Akt Pathway by targeting YBX1 in osteoarthritis. Cartilage, 2022; 13(1): 19476035221074024. doi: 10.1177/19476035221074024
[14]	Hussain S A, Venkatesh T.YBX1/lncRNA SBF2-AS1 interaction regulates proliferation and tamoxifen sensitivity via PI3K/AKT/MTOR signaling in breast cancer cells. Molecular Biology Reports, 2023; 50(4): 3413-3428. doi: 10.1007/s11033-023-08308-5
[15]	Lin F, Zeng Z, Song Y, et al.YBX-1 mediated sorting of miR-133 into hypoxia/reoxygenation-induced EPC-derived exosomes to increase fibroblast angiogenesis and MEndoT. Stem Cell Res Ther, 2019; 10(1): 263. doi: 10.1186/s13287-019-1377-8
[16]	Huang S, Li X, Zheng H, et al.Loss of super-enhancer-regulated circRNA nfix induces cardiac regeneration after myocardial infarction in adult mice. Circulation, 2019; 139(25): 2857-2876. doi: 10.1161/CIRCULATIONAHA.118.038361
[17]	Pham T P, Bink D I, Stanicek L, et al.Long Non-coding RNA Aerrie Controls DNA Damage Repair via YBX1 to Maintain Endothelial Cell Function. Front Cell Dev Biol, 2020; 8: 619079. doi: 10.3389/fcell.2020.619079
[18]	Varma E, Burghaus J, Schwarzl T, et al.Translational control of Ybx1 expression regulates cardiac function in response to pressure overload in vivo. Basic Res Cardiol, 2023; 118(1): 25. doi: 10.1007/s00395-023-00996-1
[19]	Jing X, Yang F, Shao C, et al.Role of hypoxia in cancer therapy by regulating the tumor microenvironment. Mol Cancer, 2019; 18(1): 157. doi: 10.1186/s12943-019-1089-9
[20]	Zamaraev A V, Kopeina G S, Prokhorova E A, et al.Post-translational Modification of Caspases: The Other Side of Apoptosis Regulation. Trends Cell Biol, 2017; 27(5): 322-339. doi: 10.1016/j.tcb.2017.01.003
[21]	Ramachandra C J A, Hernandez-Resendiz S, Crespo-Avilan G E, et al. Mitochondria in acute myocardial infarction and cardioprotection. EBioMedicine, 2020; 57: 102884. doi: 10.1016/j.ebiom.2020.102884
[22]	Glaviano A, Foo A S C, Lam H Y, et al. PI3K/AKT/mTOR signaling transduction pathway and targeted therapies in cancer. Mol Cancer, 2023; 22(1): 138. doi: 10.1186/s12943-023-01827-6
[23]	Yang R, Li L, Hou Y, et al.Long non-coding RNA KCND1 protects hearts from hypertrophy by targeting YBX1. Cell Death Dis, 2023; 14(5): 344. doi: 10.1038/s41419-023-05852-7
[24]	Sepa-Kishi D M, Sotoudeh-Nia Y, Iqbal A, et al. Cold acclimation causes fiber type-specific responses in glucose and fat metabolism in rat skeletal muscles. Sci Rep, 2017; 7(1): 15430. doi: 10.1038/s41598-017-15842-3
[25]	Liu P, Yao R, Shi H, et al.Effects of cold-inducible RNA-binding Protein (CIRP) on liver glycolysis during acute cold exposure in C57BL/6 mice. Int J Mol Sci, 2019; 20(6): 1470. doi: 10.3390/ijms20061470
[26]	Xu B, Lang L M, Li S Z, et al.Cortisol excess-mediated mitochondrial damage induced hippocampal neuronal apoptosis in mice following cold exposure. Cells, 2019; 8(6): 612. doi: 10.3390/cells8060612
[27]	Kloetgen A, Duggimpudi S, Schuschel K, et al.YBX1 indirectly targets heterochromatin-repressed inflammatory response-related apoptosis genes through regulating CBX5 mRNA. Int J Mol Sci, 2020; 21(12): 4453. doi: 10.3390/ijms21124453
[28]	Su H, Fan G, Huang J, et al.YBX1 regulated by Runx3-miR-148a-3p axis facilitates non-small-cell lung cancer progression. Cell Signal, 2021; 85: 110049. doi: 10.1016/j.cellsig.2021.110049
[29]	David J J, Subramanian S V, Zhang A, et al.Y-box binding protein-1 implicated in translational control of fetal myocardial gene expression after cardiac transplant. Experimental Biology & Medicine, 2012; 237(5): 593-607. doi: 10.1258/ebm.2012.011137
[30]	Ma L, Singh J, Schekman R.Two RNA-binding proteins mediate the sorting of miR223 from mitochondria into exosomes. Elife, 2023; 12: e85878. doi: 10.7554/eLife.85878
[31]	Kishikawa T, Otsuka M, Yoshikawa T, et al.Satellite RNAs promote pancreatic oncogenic processes via the dysfunction of YBX1. Nat Commun, 2016; 7: 13006. doi: 10.1038/ncomms13006
[32]	Feng M, Xie X, Han G, et al.YBX1 is required for maintaining myeloid leukemia cell survival by regulating BCL2 stability in an m6A-dependent manner. Blood, 2021; 138(1): 71-85. doi: 10.1182/blood.2020009676
[33]	Chen J, Liu Z, Zhang H, et al.YBX1 promotes MSC osteogenic differentiation by activating the PI3K/AKT pathway. Curr Stem Cell Res Ther, 2023; 18(4): 513-521. doi: 10.2174/1574888X17666220805143833
[34]	Tuerxun T, Li X, Lou F, et al.YBX1 protects against apoptosis induced by oxygen-glucose deprivation/reoxygenation in PC12 cells via activation of the AKT/GSK3β pathway. Folia Biol (Praha), 2021; 67(4): 150-157. doi: 10.14712/fb2021067040150
[35]	Su W, Wang L, Zhao H, et al.LINC00857 interacting with YBX1 to regulate apoptosis and autophagy via MET and phosphor-AMPKa signaling. Mol Ther Nucleic Acids, 2020; 22: 1164-1175. doi: 10.1016/j.omtn.2020.10.025
[36]	Wang J, Hu K, Cai X, et al.Targeting PI3K/AKT signaling for treatment of idiopathic pulmonary fibrosis. Acta Pharm Sin B, 2022; 12(1): 18-32. doi: 10.1016/j.apsb.2021.07.023
[37]	Liu C, Chen K, Wang H, et al.Gastrin attenuates renal ischemia/ reperfusion injury by a PI3K/Akt/Bad-mediated anti-apoptosis signaling. Front Pharmacol, 2020; 11: 540479. doi: 10.3389/fphar.2020.540479
[38]	Feng C, Wan H, Zhang Y, et al.Neuroprotective effect of danhong injection on cerebral ischemia-reperfusion injury in rats by activation of the PI3K-Akt pathway. Front Pharmacol, 2020; 11: 298. doi: 10.3389/fphar.2020.00298
[39]	Li Y, Xia J, Jiang N, et al.Corin protects H(2)O(2)-induced apoptosis through PI3K/AKT and NF-κB pathway in cardiomyocytes. Biomed Pharmacother, 2018; 97: 594-599. doi: 10.1016/j.biopha.2017.10.090
[40]	Chen R, Chen T, Wang T, et al.Tongmai Yangxin pill reduces myocardial no-reflow by regulating apoptosis and activating PI3K/Akt/ eNOS pathway. J Ethnopharmacol, 2020; 261: 113069. doi: 10.1016/j.jep.2020.113069
	Relative Articles
[1]	Yanying Wang, Jian Huang, Han Sun, Jie Liu, Yingchun Shao, Manyu Gong, Xuewen Yang, Dongping Liu, Zhuo Wang, Haodong Li, Yanwei Zhang, Xiyang Zhang, Zhiyuan Du, Xiaoping Leng, Lei Jiao, Ying Zhang.Long non-coding RNA-AK138945 regulates myocardial ischemia-reperfusion injury via the miR-1-GRP94 signaling pathway[J]. Frigid Zone Medicine, 2024, 4(1): 31-40. doi: 10.2478/fzm-2024-0004
[2]	Tongzhu Jin, Zhen Ye, Ruonan Fang, Yue Li, Wei Su, Qianqian Wang, Tianyu Li, Hongli Shan, Yanjie Lu, Haihai Liang.Notum protects against myocardial infarction-induced heart dysfunction by alleviating cardiac fibrosis[J]. Frigid Zone Medicine, 2024, 4(1): 41-50. doi: 10.2478/fzm-2024-0005
[3]	Rui Jiang, Lingling Xu, Yue Liu, Guangna Zhao, Chun Xing, Youyuan Li, Yongchen Wang.Low ambient temperature and air pollution are associated with hospitalization incidence of coronary artery disease: Insights from a cross-sectional study in Northeast China[J]. Frigid Zone Medicine, 2023, 3(4): 232-241. doi: 10.2478/fzm-2023-0030
[4]	Han Lou, Henghui Xu, Yong Zhang.FAM210A: Implications in mitochondrial dynamics and metabolic health[J]. Frigid Zone Medicine, 2023, 3(4): 196-198. doi: 10.2478/fzm-2023-0025
[5]	Yanan Ni, Dan Liu, Xiaona Zhang, Hong Qiao.Association of point in range with β-cell function and insulin sensitivity of type 2 diabetes mellitus in cold areas[J]. Frigid Zone Medicine, 2023, 3(4): 242-252. doi: 10.2478/fzm-2023-0031
[6]	Manyu Gong, Xuewen Yang, Yaqi Wang, Yanying Wang, Dongping Liu, Haodong Li, Yunmeng Qu, Xiyang Zhang, Yanwei Zhang, Han Sun, Lei Jiao, Ying Zhang.Growth differentiation factor 11 promotes macrophage polarization towards M2 to attenuate myocardial infarction via inhibiting Notch1 signaling pathway[J]. Frigid Zone Medicine, 2023, 3(1): 53-64. doi: 10.2478/fzm-2023-0008
[7]	Zhiyu Li, Kegong Chen, Chao Cui, Yinghui Wang, Dequan Wu.Cryptotanshinone increases the sensitivity of liver cancer to sorafenib by inhibiting the STAT3/Snail/epithelial mesenchymal transition pathway[J]. Frigid Zone Medicine, 2022, 2(2): 119-128. doi: 10.2478/fzm-2022-0016
[8]	Zhenghua Li, Kenichi Yamamura.Dissecting pathophysiology of a human dominantly inherited disease, familial amyloidotic polyneuropathy, by using genetically engineered mice[J]. Frigid Zone Medicine, 2022, 2(2): 65-75. doi: 10.2478/fzm-2022-0009
[9]	Garegin Movsesyan, Edita Margaryan, Marianna Arakelyan, Mikhail Stepanov.A randomized retrospective clinical study on the choice between endodontic surgery and immediate implantation[J]. Frigid Zone Medicine, 2022, 2(2): 90-93. doi: 10.2478/fzm-2022-0012
[10]	Wenchao Ma, Di Zhu, Tong Yu, Junwen Xue, Xiang Sun, Guofang Zhang, Yumeng Hou, Jinrui Li, Yingzhun Chen, Hongli Shan, Haihai Liang.Apelin aggravates the migration and invasion of nonsmall cell lung cancer cells via YAP1[J]. Frigid Zone Medicine, 2022, 2(1): 53-64. doi: 10.2478/fzm-2022-0007
[11]	Xinluan Wang, Wenyao Yang, Ling Qin.Prevention of osteoporotic fracture: from skeletal and non-skeletal perspectives[J]. Frigid Zone Medicine, 2022, 2(4): 214-224. doi: 10.2478/fzm-2022-0029
[12]	Jinxin Liu, Shaohong Fang, Shanjie Wang, Changbin Sun, Rong Sun, Hengxuan Cai, Bo Yu.Effects of intermittent cold-exposure on culprit plaque morphology in ST-segment elevation myocardial infarction patients: a retrospective study based on optical coherence tomography[J]. Frigid Zone Medicine, 2022, 2(2): 82-89. doi: 10.2478/fzm-2022-0011
[13]	Baofeng Yang.Inaugural Editorial[J]. Frigid Zone Medicine, 2021, 1(1): 1-1. doi: 10.2478/fzm-2021-0001
[14]	Hui Yu, Yun Zhou, Yu Duan, Yunlong Gao, Ning Fang, Jiawei Zhang, Yue Li.Tranilast treats cold-related hypertension by reducing the expression of NLRP3 inflammasome[J]. Frigid Zone Medicine, 2021, 1(2): 95-101. doi: 10.2478/fzm-2021-0012
[15]	Xin Xing, Shiqiang Wang.Mammalian hibernation: a unique model for medical research[J]. Frigid Zone Medicine, 2021, 1(2): 65-68. doi: 10.2478/fzm-2021-0008
[16]	Jia Chen, Jinghua Luo, Hao Liu, Xue Du, Shan Zhang, Zizhen Wang, Liu He, Zhichun Feng.Ibuprofen treatment for patent ductus arteriosus in preterm infants: a retrospective cohort study in a leading Chinese center[J]. Frigid Zone Medicine, 2021, 1(2): 103-110. doi: 10.2478/fzm-2021-0013
[17]	Yan Feng, Ying Li, Xinling Yang, Limin Han, Luning Wang, Shan Gao, Ruixue Yin, Xue Wang, Jiayang Li, Meiming Liu, Baiyan Li.Direct evidence of VEGF-mediated neuroregulation and afferent explanation of blood pressure dysregulation during angiogenic therapy[J]. Frigid Zone Medicine, 2021, 1(2): 119-126. doi: 10.2478/fzm-2021-0015
[18]	De-an Guo, Wenlong Wei, Changliang Yao, Jianqing Zhang, Qirui Bi. Chinese herbal medicines with beneficial effects[J]. Frigid Zone Medicine, 2021, 1(2): 79-83. doi: 10.2478/fzm-2021-0010
[19]	Daba Tolessa Muleta, Xiang Huang, Timur Yagudin, Ying Yang, Jiangang Wang, Xiaoyu Fu, Yue Zhao, Haiyu Gao, Yang Zhou, Zhenwei Pan, Xiangyu Zhong.Ablation of apoptosis-stimulating of p53 protein 1 protects mice from acute hepatic injury and dysfunction via NF-кB pathway in CCl4-induced hepatotoxicity[J]. Frigid Zone Medicine, 2021, 1(1): 53-64. doi: 10.2478/fzm-2021-0007
[20]	Yanyan Liu, Yahan Yu, Xinyao Wang, Guanqun Liu, Xinda Yin, Yunlong Bai, Zhimin Du.Overexpression of microRNA-135b-5p attenuates acute myocardial infarction injury through its antioxidant and anti-apptotic properties[J]. Frigid Zone Medicine, 2021, 1(2): 85-94. doi: 10.2478/fzm-2021-0011