5-HT inhibits actinomycin D induced hepatocyte apoptosis by upregulating 2B receptor

Abstract

Abstract: Objective To investigate the role and mechanism of 5-hydroxytryptamine (5-HT) in hepatocyte apoptosis, and to provide evidence for the application of 5-HT in liver diseases.Methods Apoptosis of HepG2 and 7702 cells was induced by different concentrations of actinomycin-D (AcD). After AcD pretreatment of HepG2 cells, 5-HT, 5-HT 2A receptor agonist DOI, 5-HT 2B receptor agonist M110, and 5-HT 2B receptor antagonist SB204 with 5-HT were added respectively. Then, apoptotic rates and survival rates were detected by Annexin V-FITC/PI flow cytometry and MTT assay, expressions of aspartate specific protease 3 (caspase-3), serine/threonine protein kinase (AKT) and phosphorylated serine/threonine protein kinase (p-AKT) were detected by Western blot method. Results AcD induced apoptosis of HepG2 cells and 7702 cells in a concentration-dependent manner. The apoptotic rates of cells treated with 50 ng/ml AcD were higher than those of serum-free control group (HepG2 cells t=1.71, 7702 cells t=1.98, P<0.05). Compared with the AcD group, 5-HT decreased the apoptosis rate of HepG2 cells from 31.96% ± 2.13% to 10.24% ± 2.59% (t=1.62, P<0.05), and the 5-HT 2B receptor agonist decreased the apoptosis rate to 8.41% ± 0.96% (t=1.75, P<0.05). On the contrary, the 5-HT 2B receptor antagonist antagonize the anti-apoptotic effect of 5-HT, making the apoptotic rate 25.92% ± 1.33% (t=1.45, P>0.05). Both 5-HT and 5-HT 2B receptor agonists reduced the activation of caspase3, increased the phosphorylation of AKT, and decreased the apoptosis.Conclusion 5-HT promotes AKT phosphorylation by up-regulating 5-HT 2B receptor, thereby inhibiting AcD-induced hepatocyte apoptosis.

Key words: Actinomycin-D, 5-hydroxtryptamine, Hepatocyte apoptosis, AKT phosphorylation, 5-HT 2B receptor

MA Li-xia, GAO Yu-juan, LIU Xiao-hui, ZHANG Jing. 5-HT inhibits actinomycin D induced hepatocyte apoptosis by upregulating 2B receptor[J]. Chinese Hepatolgy, 2019, 24(12): 1387-1392.

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