甘草干姜汤的抗胃癌作用及其机制研究

doi:10.3969/j.issn.1004-616x.2022.03.009

摘要/Abstract

摘要： 目的: 采用血清药理学方法探究甘草干姜汤(LDGD)的体外抗胃癌作用,并基于网络药理学和分子对接法探讨其抗胃癌机制。方法: 以不同配比的甘草干姜汤灌胃大鼠后以腹主动脉取血方式提取含药血清,利用四甲基噻唑蓝(MTT)法观察甘草、干姜不同配比LDGD大鼠含药血清对人胃腺癌细胞AGS以及人脐静脉血管内皮细胞HUVEC活力的影响;基于网络药理学构建LDGD抗胃癌的“成分-靶点”网络,通过构建蛋白-蛋白相互作用网络(PPI)筛选出关键靶点,通过GO功能和KEGG通路富集分析探究LDGD抗胃癌可能涉及的生物学功能和信号通路;通过分子对接分析LDGD中文献报道的8种主要成分甘草苷、异甘草苷、甘草素、异甘草素、甘草酸、6-姜酚、6-姜烯酚、8-姜酚和抗胃癌关键靶点的结合情况。结果: 血清药理学实验中不同配比LDGD灌胃大鼠后提取的30%含药血清对人胃腺癌细胞AGS的细胞活力均产生了显著的抑制作用,而对HUVEC无明显抑制作用;通过网络药理学分析得到LDGD有效化学成分101个,抗胃癌关键靶点为VEGFA、TNF-α、CASP3、MYC;富集分析结果表明,LDGD抗胃癌可能主要通过对凋亡信号、氧化应激、活性氧反应等途径的调节,以及对TNF、p53等信号通路发挥作用;分子对接结果表明,甘草酸、甘草苷与VEGFA对接良好,6-姜酚与TNF-α对接良好,且这3种成分在LDGD中含量相对较高。结论: 甘草干姜汤具有一定抗胃癌活性,其机制可能是通过调节氧化应激,作用于TNF、p53等信号通路,其中甘草酸、甘草苷、6-姜酚可能为关键药效成分。

关键词: 甘草干姜汤, 胃癌, 血清药理学, 网络药理学, 分子对接, 分子机制

Abstract: OBJECTIVE: Serum pharmacology was used to explore anti-gastric cancer effects of licorice and dried ginger decoction (LDGD) in vitro,and its anti-gastric cancer mechanism was discussed based on network pharmacology and molecular docking. METHODS: After intragastric administrations of LDGD in different proportions to mice,serum samples were extracted by taking blood from abdominal aorta.Effects of the drug containing serum on activities of human gastric cancer cells AGS and human umbilical vein endothelial cells HUVEC in vitro were observed using the MTT method.The "component target" network of LDGD against the gastric cancer cells was constructed by network pharmacology.Key targets of LDGD against the cancer cells were selected by constructing protein-protein interaction network (PPI).Possible biological functions and signal pathways of LDGD against the cancer cells were explored by the GO function and KEGG pathway enrichment analyses.The binding of 8 pharmacodynamic components in LDGD and key anti-gastric cancer targets reported in the literature were analyzed by molecular docking. RESULTS: In the serum pharmacology experiment,30% medicated serum with different proportions of LDGD had significant inhibitory effects on cell viability of AGS but had no significant inhibitory effect on HUVEC.A total of 101 active components of LDGD were obtained,and the key targets of LDGD in the gastric cancer cells were VEGFA,TNF-α,CASP3 and MYC.Bioinformatics enrichment analyses show that LDGD anti-gastric cancer effects may mainly regulate apoptosis signal,oxidative stress and reactive oxygen species response,as well as influence on TNF,p53 and other signal pathways.The results of molecular docking show that glycyrrhizic acid and liquiritin had strong affinity with VEGFA,and 6-gingerol had strong affinity with TNF-α. CONCLUSION: LDGD showed certain antitumor activities on gastric cancer cells.Through network pharmacology and molecular docking,the antitumor activities of LDGD involved regulating oxidative stress and acting on TNF,p53 and other signal pathways.In addition,glycyrrhizic OBJECTIVE: To study effects of Fuzi (aconite) extracts on proliferation and radiosensitization of human lung cancer A549 cells. METHODS: The cells were cultured and were divided into 4 groups:control,Fuzi extract,X-ray-irradiated,and Fuzi plus X-ray-irradiated groups. The control group was given culture medium,the Fuzi extract group was treated with IC₂₀30 μg/mL of Fuzi extract,the radiation group was given X-ray-irradiated,and the Fuzi plus X-ray-irradiated group were first treated with the Fuzi extract and then X-ray. After treatments,cell proliferation was detected by CCK-8 assay and colony formation assay was used to measure cell survival. Single-hit multi-target model was used to fit the survival curve and to calculate the sensitive enhancement ratio (SER). Apoptosis rates were detected by flow cytometry,and protein expression levels of Bax and Bcl-2 were detected by Western blot. RESULTS: Fuzi extract concentrations(<20 μg/mL) enhanced viability of A549 cells but increasing concentrations (>30 μg/mL) inhibited growth of the cells in a dose-dependent manner. The shoulder at the beginning of the survival curve in the Fuzi plus X-ray-irradiated group was slightly narrower, and the D₀values in the X-ray-irradiated and Fuzi plus X-ray-irradiated group were 1.83 Gy,1.48 Gy,and the SER was 1.24,respectively. The apoptosis rates were significantly increased in the Fuzi plus X-ray-irradiated group and were higher than that in both the Fuzi and X-ray irradiation groups (P<0.05). The results of Western blot show that protein expressions of Bax were increased but were decreased for Bcl-2 in the Fuzi extract versus the X-ray-irradiated groups. Bax and Bcl-2 protein levels were changed more significantly in the Fuzi plus X-ray-irradiated than the other groups (P<0.01). CONCLUSION: Fuzi extract at 30 μg/mL inhibited proliferation and enhanced radiosensitizing effects on human lung cancer A549 cells,and the mechanism may be related to the induction of apoptosis.

Key words: licorice and dried ginger decoction, gastric cancer, serum pharmacology, network pharmacology, molecular docking, molecular mechanism

中图分类号:

R285.5

余悦华, 梁桓熙, 辛雨濛, 孙震晓. 甘草干姜汤的抗胃癌作用及其机制研究[J]. 癌变·畸变·突变, 2022, 34(3): 219-226.

YU Yuehua, LIANG Huanxi, XIN Yumeng, SUN Zhenxiao. Investigations on anti-gastric cancer effects of licorice and dried ginger decoction[J]. Carcinogenesis, Teratogenesis & Mutagenesis, 2022, 34(3): 219-226.

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