Aloperin improves liver non-alcoholic steatohepatitis in vitro and in vivo

Zhongsheng Guo; Ruyu Liu; Liwei Guo; Sumei Lu; Min Yang; Shaofei Wu; Yong An

doi:10.5114/aoms.2020.95629

Aloperin improves liver non-alcoholic steatohepatitis in vitro and in vivo

Arch Med Sci. 2020 May 25;21(1):285-297. doi: 10.5114/aoms.2020.95629. eCollection 2025.

Authors

Zhongsheng Guo¹, Ruyu Liu², Liwei Guo¹, Sumei Lu³, Min Yang⁴, Shaofei Wu¹, Yong An¹

Affiliations

¹ Department of Infection Diseases and Liver Diseases, The First Affiliated Hospital of Shandong First Medical University (Shandong Provincial Qianfoshan Hospital), Jinan, China.
² Department of Hepatology, Beijing Ditan Hospital Capital Medical University, Beijing, China.
³ PCR laboratory, The First Affiliated Hospital of Shandong First Medical University (Shandong Provincial Qianfoshan Hospital), Jinan, China.
⁴ Department of Clinical Pathology, The First Affiliated Hospital of Shandong First Medical University (Shandong Provincial Qianfoshan Hospital), Jinan, China.

Abstract

Introduction: The aim of this work is to investigate the inhibitory effect of aloperin (Alo) on hepatocyte apoptosis in non-alcoholic fatty liver disease, and the underlying mechanism.

Material and methods: Rats in the Alo groups were fed a high-fat + high-sugar diet for 8 weeks and then treated with low-, moderate-, and high-dose Alo for another 8 weeks via gavage. Oxidative stress indices were tested by a colourimetric method, and pathological changes were observed by haematoxylin-eosin staining. Apoptosis was detected by TUNEL staining. TLR4, TRIF, and NF-κB(p65) mRNA and protein expressions were detected by RT-qPCR, Western blot assay and immunohistochemistry. In the in vitro study, L02 cells were treated with FFA (free fatty acid) for 24 h to establish a non-alcoholic steatohepatitis (NASH) model. Inhibition of cell proliferation was measured by the MTT method, and cell apoptosis was evaluated by flow cytometry. Finally, the nuclear import volume of NF-κB(p65) was evaluated by cellular immunofluorescence.

Results: Cell apoptosis significantly decreased in the Alo-treatment groups in a dose-dependent manner (p < 0.05). TLR4, TRIF, and NF-κB(p65) expression in the Alo-treatment groups was significantly downregulated compared with model group (p < 0.05). The cell proliferation rate significantly increased, cell apoptosis significantly decreased (p < 0.05), and the TLR4/TRIF/NF-κB pathway was significantly inhibited (p < 0.05) in the Alo-treatment groups. The nuclear import volume of NF-κB(p65) in the Alo-treatment groups was significantly decreased compared with that in the model group in a dose-dependent manger (p < 0.05).

Conclusions: Alo could improve NASH via the TLR4/TRIF/NF-κB pathway.

Keywords: TLR4/TRIF/NF-κB; aloperin; cell apoptosis; non-alcoholic steatohepatitis.

Grants and funding

Funding This work was funded by the 12^th 5-year S&T Program of Shandong Province P.R. China. (Grant no. 2014ZZCX02201) and Key R&D Project of Shandong Province P.R. China (grant no. 2019GSF108129).