Basic Study
Copyright ©The Author(s) 2023. Published by Baishideng Publishing Group Inc. All rights reserved.
World J Gastrointest Oncol. Mar 15, 2023; 15(3): 504-522
Published online Mar 15, 2023. doi: 10.4251/wjgo.v15.i3.504
Possible mechanisms associated with immune escape and apoptosis on anti-hepatocellular carcinoma effect of Mu Ji Fang granules
Yi-Bing Zhang, Yong-Rui Bao, Shuai Wang, Tian-Jiao Li, He Tai, Jia-Peng Leng, Xin-Xin Yang, Bo-Cai Wang, Xian-Sheng Meng
Yi-Bing Zhang, Yong-Rui Bao, Shuai Wang, Tian-Jiao Li, He Tai, Jia-Peng Leng, Xin-Xin Yang, Bo-Cai Wang, Xian-Sheng Meng, College of Pharmacy, Liaoning University of Traditional Chinese Medicine, Dalian 116600, Liaoning Province, China
Yi-Bing Zhang, Department of Clinical Trail Institution Office, Dalian Municipal Central Hospital, Dalian 116033, Liaoning Province, China
Yong-Rui Bao, Shuai Wang, Tian-Jiao Li, Xian-Sheng Meng, Technical Innovation Center of Multidimensional Analysis of Traditional Chinese Medicine of Liaoning Province, Dalian 116600, Liaoning Province, China
Yong-Rui Bao, Shuai Wang, Tian-Jiao Li, Xian-Sheng Meng, Engineering Laboratory of Modern Chinese Medicine Research of Liaoning Province, Dalian 116600, Liaoning Province, China
Author contributions: Zhang YB, Bao YR, Wang S, Wang BC and Meng XS contributed to conception and design of the study; Zhang YB, Wang S, Li TJ, Tai H, Leng JP, Wang BC and Yang XX conducted the experiments, organized the database, carried out the statistical analysis and drafted the manuscript; All authors contributed to manuscript revision, and approved the submitted version.
Supported by National Natural Science Foundation of China, No. 81874342; and Natural Science Foundation of Liaoning Province, No. 2020-MZLH-35.
Institutional animal care and use committee statement: All experimental procedures were performed with the approval of the animal guidelines and protocols established by the Medicine Ethics Review Committee of Animal Experiments of Liaoning University of Traditional Chinese Medicine. The study was reviewed and approved by the Animal Ethics Committee of The Affiliated Hospital of Liaoning University of TCM. [Approval No. 2019YS(DW)-033-01].
Conflict-of-interest statement: All the authors report no relevant conflicts of interest for this article.
Data sharing statement: Data are available from the corresponding authors upon request (mxsvvv@163.com).
ARRIVE guidelines statement: The authors have read the ARRIVE Guidelines, and the manuscript was prepared and revised according to the ARRIVE Guidelines.
Open-Access: This article is an open-access article that was selected by an in-house editor and fully peer-reviewed by external reviewers. It is distributed in accordance with the Creative Commons Attribution NonCommercial (CC BY-NC 4.0) license, which permits others to distribute, remix, adapt, build upon this work non-commercially, and license their derivative works on different terms, provided the original work is properly cited and the use is non-commercial. See: https://creativecommons.org/Licenses/by-nc/4.0/
Corresponding author: Xian-Sheng Meng, PhD, Dean, Professor, College of Pharmacy, Liaoning University of Traditional Chinese Medicine, No. 77 Life one Road, DD Port, Jinpu District, Dalian 116600, Liaoning Province, China. mxsvvv@163.com
Received: December 20, 2022
Peer-review started: December 20, 2022
First decision: January 9, 2023
Revised: January 19, 2023
Accepted: March 2, 2023
Article in press: March 2, 2023
Published online: March 15, 2023
Processing time: 84 Days and 16 Hours
Abstract
BACKGROUND

Hepatocellular carcinoma (HCC) is one of the most common digestive system cancers with high mortality rates worldwide. The main ingredients in Mu Ji Fang Granules (MJF) are alkaloids, flavonoids, and polysaccharides. MJF has been used in the clinical treatment of hepatitis, cirrhosis and HCC for more than 30 years. Few previous studies have focused on the mechanism of MJF on tumor immu-nology in the treatment of HCC.

AIM

To explore the mechanism of action of MJF on tumor immunology in the treatment of HCC.

METHODS

The absorbable ingredients of MJF were identified using Molecule Network related to High Performance Liquid Chromatography-Electron Spray Ionization-Time of Flight- Mass Spectrometry, and hub potential anti-HCC targets were screened using network pharmacology and pathway enrichment analysis. Forty male mice were randomly divided into the Blank, Model, and MJF groups (1.8, 5.4, and 10.8 g/kg/d) following 7 d of oral administration. Average body weight gain, spleen and thymus indices were calculated, tumor tissues were stained with hematoxylin and eosin, and Interferon gamma (IFN-γ), Tumor necrosis factor α (TNF-α), Interleukin-2, aspartate aminotransferase, alanine aminotransferase, alpha-fetoprotein (AFP), Fas, and FasL were measured by Enzyme-linked Immunosorbent Assay. Relevant mRNA expression of Bax and Bcl2 was evaluated by Real Time Quantitative PCR (RT-qPCR) and protein expression of Transforming growth factor β1 (TGF-β1) and Mothers against decapentaplegic homolog (SMAD) 4 was assessed by Western blotting. The HepG2 cell line was treated with 10 mg/mL, 20 mg/mL, 30 mg/mL, 40 mg/mL of MJF, and another 3 groups were treated with TGF-β1 inhibitor (LY364947) and different doses of MJF. Relevant mRNA expression of TNF-α, IFN-γ, Bax and Bcl2 was evaluated by RT-qPCR and protein expression of TGF-β1, SMAD2, p-SMAD2, SMAD4, and SMAD7 was assessed by Western blotting.

RESULTS

It was shown that MJF improved body weight gain and tumor inhibition rate in H22 tumor-bearing mice, protected immune organs and liver function, reduced the HCC indicator AFP, affected immunity and apoptosis, and up-regulated the TGF-β1/SMAD signaling pathway, by increasing the relative expression of TGF-β1, SMAD2, p-SMAD2 and SMAD4 and decreasing SMAD7, reducing immune factors TNF-α and IFN-γ, decreasing apoptosis cytokines Fas, FasL and Bcl2/Bax, and inhibiting the effect of LY364947 in HepG2 cells.

CONCLUSION

MJF inhibits HCC by activating the TGF-β1/SMAD signaling pathway, and affecting immune and apoptotic cytokines, which may be due to MJF adjusting immune escape and apoptosis.

Keywords: Mu Ji Fang granules; Hepatocellular carcinoma; Transforming growth factor β1/Mothers against decapentaplegic homolog; Immune escape; H22 tumor-bearing mice; HepG2 cells

Core Tip: Mu Ji Fang Granules (MJF), a Chinese patent medicine, has been used in hepatitis, cirrhosis and hepatocellular carcinoma (HCC) for more than 30 years. MJF was identified with Molecule Network related to High Performance Liquid Chromatography-Electron Spray Ionization-Time of Flight- Mass Spectrometry, and hub potential anti-HCC targets were screened using network pharmacology and pathway enrichment analysis in H22 tumor-bearing mice and HepG2 cells. It was shown that MJF improved body weight gain and tumor inhibition rate, protected immune organs and liver function, affected immunity and apoptosis, up-regulated the Transforming growth factor β1(TGF-β1)/ Mothers against decapentaplegic homolog(SMAD) signaling pathway, and inhibited the effect of TGF-β1 inhibitor (LY364947).