2022, Vol. 26 ›› Issue (26): 4186-4191

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

Zhao Zheyi1, 2, Wang Zhengrong3, 2, Fang Xingyan2, Wang Tian4, Xie Tingting1, 2   

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