Protective effect of cyclin-dependent kinase inhibitor AT7519 on carbon tetrachloride induced acute liver injury in mice

Abstract

Abstract: Objective To study on the hepatoprotective effect AT7519, a cyclin-dependent kinase (CDK) inhibitor, on carbon tetrachloride (CCl4) induced acute liver injury in mice.Methods Lipopolysaccharide (LPS)-induced RAW264.7 cell inflammation model was used for the in vitro study. RAW264.7 cells were pretreated with different concentrations of AT7519 (20uM, 10uM, and 5uM) for 1h followed by LPS stimulation for 4 h. The effect of AT7519 on the expression of proinflammatory cytokines were then evaluated with quantitative reverse transcription PCR (RT-qPCR) analysis. For the in vivo study, CCl₄^- induced acute liver injury model in mice was employed. Thirty C57BL/6 mice were randomly divided into three groups including a normal control group, a model group, and an AT7519 treated group. The levels of serum transaminases were investigated, along with hepatic histopathological evaluation with H&E and immunohistochemistry staining. The levels of proinflammatory cytokine expression were tested by RT-qPCR. Results RT-qPCR results of the in vitro experiment revealed that the mRNA expression levels of IL-6, IL-1β and TNF-? were markedly increased after LPS stimulation (299.8±6.1, 8198±360.1, 172.0±5.9, respectively) when compared with the normal control group (1.1±0.1, 1.1±0.0, 1.0±0.0, respectively), of which the changes could be reversed by AT7519 treatment. The relative expression of IL-6, IL-1β and TNF-α mRNA in 20uM AT7519 treated RAW264.7 cells were 6.5±0.4, 275.2±10.4, 41.5±1.0, in 10uM AT7519 treated RAW264.7 cells were 36.7±5.4, 1585±26.0, 106.2±12.2, and in 5uM AT7519 treated RAW264.7 cells were 71.7±7.1, 1615±77.9, 147.8±11.5, respectively, which were dose-dependently lower than those in the model group (P<0.05). In the murine model of CCl4-induced acute liver injury, the serum levels of ALT and AST presented a sharp increase in the model group [ (1116±197.0) U/L and (966±157.3) U/L], while AT7519 administration [ (230.6±103.1) U/L and (247.8±110.7) U/L] brought about a significant reduction in serum transaminase levels (P<0.05). As for liver histopathology, AT7519 treatment alleviated the pathologic changes in the livers of mice in model group, including massive hepatocytes necrosis and inflammatory cells infiltration. Moreover, the mRNA expression levels of IL-6, IL-1β and TNF-α in the livers were dramatically reduced after AT7519 treatment (1.6±0.7, 1.6±0.5, and 2.1±0.9, respectively) when compared with those of the model group (10.7±5.3, 4.6±1.5 and 16.4±3.0, respectively) (P<0.05).Conclusion AT7519 demonstrated a promising hepatoprotective effect against CCl4-induced acute liver injury in mice. The underlying mechanisms may correlate with a reduction of inflammatory cells infiltration and an inhibition of inflammatory cytokines expression.

Key words: Carbon tetrachloride, Acute liver injury, Cyclin-dependent kinase inhibitor, Inflammatory cells, Cytokine

LIU Yan-jun, HUANG He-ming, FU Rong, SHI Cui-cui, FAN Jian-gao, ZHANG Yuan-yuan, LUO Cheng, LI Guang-ming. Protective effect of cyclin-dependent kinase inhibitor AT7519 on carbon tetrachloride induced acute liver injury in mice[J]. Chinese Hepatolgy, 2021, 26(4): 375-378.

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