Effects of sodium arsenite on oxidative stress, apoptosis and Hippo signaling pathway in AML12 hepatocytes

doi:10.12307/2022.822

Abstract

Abstract: BACKGROUND: As the main target organ of arsenic toxicity, the liver has become the focus of research on the mechanism of arsenic toxicity.
OBJECTIVE: To investigate the effects of sodium arsenite (NaAsO2) on oxidative stress and apoptosis of AML12 hepatocytes, as well as the influence on the expression of key molecules in the Hippo signaling pathway.
METHODS: AML12 hepatocytes were cultured in medium containing 0, 10, 15, 20, 25, 30 μmol/L NaAsO2 for 24 hours. The cell morphology was observed by phase-contrast microscopy and cell viability was assessed by cell counting kit-8 assay. The content of intracellular reactive oxygen species was detected by fluorescent probe technique combined with flow cytometry. The activity of caspase-3 was measured by colorimetry. The apoptotic rate of hepatocytes was detected by annexin V/propidium iodide double staining combined with flow cytometry. The mRNA expression level of Yes-associated protein was detected by real-time fluorescence quantitative PCR. The protein expression level of phosphorylated mammalian sterile20-like kinases1/2, phosphorylated Yes-associated protein and Yes-associated protein were detected by western blot assay.
RESULTS AND CONCLUSION: Compared with the control group, the viability of AML12 hepatocytes decreased gradually along with the increase of NaAsO2 concentration. With the increase of NaAsO2 concentration, the reactive oxygen species contents, the activity of caspase-3 and the apoptotic rate increased gradually in AML12 hepatocytes. There was no significant change in the mRNA expression of Yes-associated protein in NaAsO2 treated AML12 hepatocytes. NaAsO2 elevated the relative protein expression of phosphorylated mammalian sterile20-like kinases 1/2 and phosphorylated Yes-associated protein, while inhibited the relative protein expression of Yes-associated protein. Therefore, NaAsO2-induced oxidative stress and apoptosis injury in AML12 hepatocytes may be related to the activation of Hippo signaling pathway.

Key words: sodium arsenite, AML12 hepatocyte, oxidative stress, apoptosis, Yes-associated protein, Hippo signaling pathway

CLC Number:

Zhao Zheyi, Wang Zhengrong, Fang Xingyan, Wang Tian, Xie Tingting. Effects of sodium arsenite on oxidative stress, apoptosis and Hippo signaling pathway in AML12 hepatocytes[J]. Chinese Journal of Tissue Engineering Research, 2022, 26(26): 4186-4191.

Figures/Tables 7

References

[1] CHEN Q, COSTA M. Arsenic: A Global Environmental Challenge. Annu Rev Pharmacol Toxicol. 2021;61:47-63.
[2] BJØRKLUND G, OLIINYK P, LYSIUK R, et al. Arsenic intoxication: general aspects and chelating agents. Arch Toxicol. 2020;94(6):1879-1897.
[3] PRAKASH C, CHHIKARA S, KUMAR V. Mitochondrial Dysfunction in Arsenic-Induced Hepatotoxicity: Pathogenic and Therapeutic Implications. Biol Trace Elem Res. 2021.
[4] RENU K,SARAVANAN A,ELANGOVAN A, et al. An appraisal on molecular and biochemical signalling cascades during arsenic-induced hepatotoxicity. Life Sci. 2020;260:118438.
[5] WU Z, GUAN K. Hippo Signaling in Embryogenesis and Development. Trends Biochem Sci. 2021;46(1): 51-63.
[6] ZHENG J, YU H, ZHOU A, et al. It takes two to tango: coupling of Hippo pathway and redox signaling in biological process. Cell Cycle. 2020; 19(21):2760-2775.
[7] LI C, ZHANG S, LI L, et al. Ursodeoxycholic Acid Protects Against Arsenic Induced Hepatotoxicity by the Nrf2 Signaling Pathway. Front Pharmacol. 2020;11:594496.
[8] JAHANGIRNEJAD R, GOUDARZI M, KALANTARI H, et al. Subcellular Organelle Toxicity Caused by Arsenic Nanoparticles in Isolated Rat Hepatocytes. Int J Occup Environ Med. 2020;11(1):41-52.
[9] BUHA A, BARALIĆ K, DJUKIC-COSIC D, et al. The Role of Toxic Metals and Metalloids in Nrf2 Signaling. Antioxidants (Basel). 2021;10(5):630.
[10] 王甜,赵哲仪,穆银贵,等.亚砷酸钠所致L-02人肝细胞损伤与p14ARF表达下调及MDM2, p53表达增加有关[J].细胞与分子免疫学杂志,2020,36(6):507-512.
[11] FURTH N, AYLON Y, OREN M. p53 shades of Hippo. Cell Death Differ. 2018;25(1):81-92.
[12] 马文靓, 吕曼, 杨杰, 等. 砷致肝损伤作用机制的研究进展[J]. 中华地方病学杂志,2021,40(1):75-80.
[13] CHAZELAS P, STEICHEN C, FAVREAU F, et al. Oxidative Stress Evaluation in Ischemia Reperfusion Models: Characteristics, Limits and Perspectives. Int J Mol Sci. 2021;22(5):2366.
[14] SIES H. Oxidative Stress: Concept and Some Practical Aspects. Antioxidants (Basel). 2020;9(9):852.
[15] FU Y, YE Y, ZHU G, et al. Resveratrol induces human colorectal cancer cell apoptosis by activating the mitochondrial pathway via increasing reactive oxygen species. Mol Med Rep. 2021;23(3):170.
[16] NGUYEN-LEFEBVRE AT, SELZNER N, WRANA JL, et al. The hippo pathway: A master regulator of liver metabolism, regeneration, and disease. FASEB J. 2021;35(5):e21570.
[17] LI HL, LI QY, JIN MJ, et al. A review: hippo signaling pathway promotes tumor invasion and metastasis by regulating target gene expression. J Cancer Res Clin Oncol. 2021;147(6):1569-1585.
[18] MA S, MENG Z, CHEN R, et al. The Hippo Pathway: Biology and Pathophysiology. Annu Rev Biochem. 2019;88:577-604.
[19] LI D, NI H, RUI Q, et al. Mst1: Function and Mechanism in Brain and Myocardial Ischemia Reperfusion Injury. Curr Neuropharmacol. 2018; 16(9):1358-1364.
[20] ZHANG J, LIU J, GAO S, et al. Antiapoptosis and Antifibrosis Effects of Qishen Granules on Heart Failure Rats via Hippo Pathway. Biomed Res Int. 2019;2019:1642575.
[21] MOYA IM, HALDER G. Hippo-YAP/TAZ signalling in organ regeneration and regenerative medicine .Nat Rev Mol Cell Biol. 2019;20(4):211-226.
[22] HENG B, ZHANG X, AUBEL D, et al. An overview of signaling pathways regulating YAP/TAZ activity. Cell Mol Life Sci. 2021;78(2):497-512.
[23] ZHAO W, WANG M, CAI M, et al. Transcriptional co-activators YAP/TAZ: Potential therapeutic targets for metastatic breast cancer. Biomed Pharmacother. 2021;133:110956.
[24] JIN M, WU L, CHEN S, et al. Arsenic trioxide enhances the chemotherapeutic efficiency of cisplatin in cholangiocarcinoma cells via inhibiting the 14-3-3ε-mediated survival mechanism. Cell Death Discov. 2020;6:92.
[25] OOKI A, BEGUM A, MARCHIONNI L, et al. Arsenic promotes the COX2/PGE2-SOX2 axis to increase the malignant stemness properties of urothelial cells. Int J Cancer. 2018;143(1):113-126.
[26] LIU Y, BEYER A, AEBERSOLD R. On the Dependency of Cellular Protein Levels on mRNA Abundance. Cell. 2016;165(3):535-550.
[27] ZHOU W, LIU M, LI X, et al. Arsenic nano complex induced degradation of YAP sensitized ESCC cancer cells to radiation and chemotherapy. Cell Biosci. 2020;10(1):146.
[28] ZHAO Y, ZANG G, YIN T, et al. A novel mechanism of inhibiting in-stent restenosis with arsenic trioxide drug-eluting stent: Enhancing contractile phenotype of vascular smooth muscle cells via YAP pathway. Bioact Mater. 2021;6(2):375-385.