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苍术酮对肝癌HepG2细胞活性、凋亡的影响及其相关机制

杨雪丽 薛建华 陈天阳 平键 侯天禄 陈建杰 成扬

引用本文:
Citation:

苍术酮对肝癌HepG2细胞活性、凋亡的影响及其相关机制

DOI: 10.3969/j.issn.1001-5256.2021.11.020
基金项目: 

上海市浦东新区卫计委“中医肝病”临床中医特色学科建设项目 (PDZY-2018-0607)

上海市浦东新区名中医工作室建设项目 (PWRzm2020-14)

详细信息
    通信作者:

    成扬,drchengyang@126.com

  • 中图分类号: R735.7

Effect of atractylone on the viability and apoptosis of hepatoma HepG2 cells and related mechanism

Research funding: 

The Project of "Chinese Medicine Liver Disease" Clinical TCM Specialty Construction Project of Shanghai Pudong New Area Health Planning Commission (PDZY-2018-0607);

The project of Shanghai Pudong New Area Famous Chinese Medicine Studio Construction (PWRzm2020-14)

  • 摘要:   目的  观察苍术酮对肝癌HepG2细胞活性、凋亡的影响,并探讨其作用机制。  方法  以肝癌HepG2细胞为研究对象,分为苍术酮低、中、高剂量组(5、10、20 μmol/L),对照组加入等体积的DMSO。MMT比色法检测不同浓度苍术酮处理后HepG2细胞活性。流式细胞仪检测HepG2细胞凋亡率及线粒体膜电位。DCFH-DA荧光探针标记法检测HepG2细胞内ROS水平。Transwell实验检测苍术酮对HepG2细胞迁移能力的影响。蛋白质印迹法检测Bcl-2、Bax和Cleaved caspase-3蛋白表达水平。计量资料多组间比较采用单因素方差分析,组间的两两比较采用LSD-t检验。  结果  在苍术酮处理细胞24、48 h后,与对照组同一时间相比,苍术酮低、中、高剂量组的细胞活性呈下降趋势,且差异均有统计学意义(P值均<0.05),苍术酮处理HepG2细胞72 h的半数抑制率为26.19 μmol/L。苍术酮低、中、高剂量组的细胞凋亡率分别为14.34%、29.32%和50.12%,均高于对照组(0.32%)(P值均<0.05)。与对照组比较,苍术酮低、中、高剂量组HepG2细胞中ROS的荧光强度明显升高(P值均<0.05)。在苍术酮处理HepG2细胞48 h后,与对照组比较,苍术酮低、中、高剂量组的细胞迁移数量明显降低(132.67 ± 18.36、57.00 ± 9.26、31.00 ± 2.45 vs 258.11 ± 38.54,P值均<0.05);与对照组比较,苍术酮低、中、高剂量组能显著抑制抗凋亡因子Bcl-2的表达,促进凋亡因子Bax和Cleaved caspase-3的表达,差异均有统计学意义(P值均<0.05)。  结论  苍术酮能够诱导HepG2细胞凋亡,并抑制其迁移,为进一步开发利用苍术酮提供一定的实验基础。

     

  • 图  1  苍术酮对HepG2细胞活性的影响

    注: 与对照组同一时间比较, *P < 0.05。

    图  2  苍术酮对HepG2细胞凋亡率的影响

    图  3  苍术酮对HepG2细胞线粒体膜电位丢失的影响

    注: a, 对照组;b,5 μmol/L组;c, 10 μmol/L组;d, 20 μmol/L组。

    图  4  苍术酮对HepG2细胞中ROS水平的影响

    注: a, 对照组;b,5 μmol/L组;c, 10 μmol/L组;d, 20 μmol/L组,e,苍术酮对HepG2细胞内ROS水平定量分析。

    图  5  结晶紫染色观察苍术酮对HepG2细胞迁移的影响(×200)

    注: a, 对照组;b,5 μmol/L组;c, 10 μmol/L组;d, 20 μmol/L组。

    图  6  苍术酮对HepG2细胞中Bcl-2,Bax和Cleaved caspase-3蛋白表达水平的影响

    表  1  苍术酮对HepG2细胞迁移的影响

    组别 迁移数量
    对照组 258.11±38.54
    5 μmol/L组 132.67±18.361)
    10 μmol/L组 57.00±9.261)2)
    20 μmol/L组 31.00±2.451)2)
    注:与对照组比较,1)P<0.05;与5 μmol/L组比较,2)P<0.05。
    下载: 导出CSV
  • [1] SUNG H, FERLAY J, SIEGEL RL, et al. Global Cancer Statistics 2020: GLOBOCAN estimates of incidence and mortality worldwide for 36 cancers in 185 countries[J]. CA Cancer J Clin, 2021, 71(3): 209-249. DOI: 10.3322/caac.21660.
    [2] WU YX, SHENG QS. Advances in traditional Chinese medicine therapy for primary hepatocarcinoma[J]. Guangxi Med J, 2020, 42(4): 483-485. DOI: 10.1165/j.issn.0253-4304.2020.04.26.

    吴玉潇, 盛庆寿. 中医药治疗原发性肝癌的研究进展[J]. 广西医学, 2020, 42(4): 483-485. DOI: 10.11675/j.issn.0253-4304.2020.04.26.
    [3] LI ZW. Analysis of postoperative prognosis and risk factors in patients with primary liver cancer[J]. China's Naturopathy, 2020, 28(13): 73-75. DOI: 10.19621/j.cnki.11-3555/r.2020.1334.

    李柱威. 原发性肝癌患者术后预后及其危险因素分析[J]. 中国民间疗法, 2020, 28(13): 73-75. DOI: 10.19621/j.cnki.11-3555/r.2020.1334.
    [4] FORNER A, REIG ME, de LOPE CR, et al. Current strategy for staging and treatment: The BCLC update and future prospects[J]. Semin Liver Dis, 2010, 30(1): 61-74. DOI: 10.1055/s-0030-1247133.
    [5] HUANG JH, HUANG ZM, ZHANG TQ, et al. Progress in comprehensive treatment of advanced liver cancer based on interventional therapy[J/CD]. Electronic J Liver Tumor, 2019, 6(4): 27-31. DOI: 10.3969/j.issn.2095-7815.2019.04.007.

    黄金华, 黄职妹, 张天奇, 等. 以介入治疗为基础的中晚期肝癌综合治疗进展[J/CD]. 肝癌电子杂志, 2019, 6(4): 27-31. DOI: 10.3969/j.issn.2095-7815.2019.04.007.
    [6] YUAN SX, ZHOU WP. Progress and hot spots of comprehensive treatment for primary liver cancer[J]. Chin J Dig Surg, 2021, 20(2): 163-170. DOI: 10.3760/cma.j.cn115610-20201211-00776.

    袁声贤, 周伟平. 原发性肝癌综合治疗的进展和热点[J]. 中华消化外科杂志, 2021, 20(2): 163-170. DOI: 10.3760/cma.j.cn115610-20201211-00776.
    [7] ZHAN YP, ZHAI XF. Study on prescription compatibility law of contemporary famous doctors of TCM in treating primary liver cancer based on data mining[J]. J Liaoning Univ Tradit Chin Med, 2018, 20(3): 159-162. DOI: 10.13194/j.issn.1673-842x.2018.03.046.

    占义平, 翟笑枫. 基于数据挖掘当代名中医治疗原发性肝癌用药配伍规律研究[J]. 辽宁中医药大学学报, 2018, 20(3): 159-162. DOI: 10.13194/j.issn.1673-842x.2018.03.046.
    [8] ZHANG Y, CHEN HG, ZHAO C, et al. Research progress on anti-hepatocellular carcinoma mechanism of active ingredients of traditional Chinese medicine[J]. China J Chin Mater Med, 2020, 45(14): 3395-3406. DOI: 10.19540/j.cnki.cjcmm.20200429.601.

    张宇, 陈华国, 赵超, 等. 中药有效成分抗肝癌作用机制研究进展[J]. 中国中药杂志, 2020, 45(14): 3395-3406. DOI: 10.19540/j.cnki.cjcmm.20200429.601.
    [9] ZHAO ZJ, XIAO SN, ZHAO YX, et al. Re-evaluation of the literature on the pharmacological effects of Atractylodes macrocephala[J]. Chin J Hosp Pharm, 2011, 31(7): 607-609. https://www.cnki.com.cn/Article/CJFDTOTAL-ZGYZ201107035.htm

    赵子剑, 肖胜男, 赵永新, 等. 苍术药理作用的文献再评价[J]. 中国医院药学杂志, 2011, 31(7): 607-609. https://www.cnki.com.cn/Article/CJFDTOTAL-ZGYZ201107035.htm
    [10] LIU C. Progress of medicinal research on Atractylodes macrocephala[J]. Heilongjiang Sci Technol Inform, 2013, 12: 101. DOI: 10.3969/j.issn.1673-1328.2013.12.099.

    刘畅. 苍术药用研究进展[J]. 黑龙江科技信息, 2013, 12: 101. DOI: 10.3969/j.issn.1673-1328.2013.12.099.
    [11] CHEN TY, LIU YL, HOU TL, et al. Study on the protective effect of atractylone on acute lung injury in mice[J]. Mod J Integr Tradit Chin West Med, 2018, 27(24): 2623-2626. DOI: 10.3969/j.issn.1008-8849.2018.24.001.

    陈天阳, 刘廷亮, 侯天禄, 等. 苍术酮对急性肺损伤小鼠保护作用的研究[J]. 现代中西医结合杂志, 2018, 27(24): 2623-2626. DOI: 10.3969/j.issn.1008-8849.2018.24.001.
    [12] ATHINARAYANAN S, FAN YY, WANG X, et al. Fatty acid desaturase 1 influences hepatic lipid homeostasis by modulating the PPARα-FGF21 axis[J]. Hepatol Commun, 2021, 5(3): 461-477. DOI: 10.1002/hep4.1629.
    [13] SUN YP, CHENG Z, WU Z, et al. Research on the inheritance, innovation and high-quality development of Chinese herbal medicine Atractylodes[J]. Hubei Agricult Sci, 2020, 59(11): 203-207. DOI: 10.14088/j.cnki.issn0439-8114.2020.11.040.

    孙元鹏, 程正, 吴喆, 等. 中药材苍术的传承创新与高质量发展研究[J]. 湖北农业科学, 2020, 59(11): 203-207. DOI: 10.14088/j.cnki.issn0439-8114.2020.11.040.
    [14] ZHANG MF, SHEN YQ. Advances in studies on anti-inflammation, antitumor, and immunoregulation of Atractylodis Rhizoma[J]. Drug Eval Res, 2016, 39(5): 885-890. DOI: 10.7501/j.issn.1674-6376.2016.05.037.

    张明发, 沈雅琴. 苍术抗炎、抗肿瘤和免疫调节作用的研究进展[J]. 药物评价研究, 2016, 39(5): 885-890. DOI: 10.7501/j.issn.1674-6376.2016.05.037.
    [15] ZHOU Y, LU JT, ZHANG XD, et al. Potential target prediction and forward molecular docking verification of atractylon[J]. J Shaoyang Uni(Natural Science Edition), 2019, 16(1): 98-104. DOI: CNKI:SUN:SYXZ.0.2019-01-013.

    周域, 陆建图, 张小丁, 等. 苍术酮潜在靶点预测及正向分子对接验证[J]. 邵阳学院学报(自然科学版), 2019, 16(1): 98-104. DOI: CNKI:SUN:SYXZ.0.2019-01-013.
    [16] GENG W, LIANG W, YE ZB, et al. Mechanism of HT29 apoptosis in colorectal cancer cells by atractylone[J]. Chin Tradit Patent Med, 2018, 40(4): 937-940. DOI: 10.3969/j.issn.1001-1528.2018.04.034.

    耿玮, 梁巍, 叶智斌, 等. 苍术酮对结直肠癌细胞HT29凋亡的机制[J]. 中成药, 2018, 40(4): 937-940. DOI: 10.3969/j.issn.1001-1528.2018.04.034.
    [17] YAO XY, DOU FF. Role of apoptosis in traditional Chinese medicine treatment of liver cancer[J]. Pract J Cancer, 2020, 35(6): 1040-1044. DOI: 10.3969/j.issn.1001-5930.2020.06.046.

    姚晓云, 钭方芳. 凋亡在中医药治疗肝癌中的作用研究[J]. 实用癌症杂志, 2020, 35(6): 1040-1044. DOI: 10.3969/j.issn.1001-5930.2020.06.046.
    [18] SHAHAR N, LARISCH S. Inhibiting the inhibitors: Targeting anti-apoptotic proteins in cancer and therapy resistance[J]. Drug Resist Updat, 2020, 52: 100712. DOI: 10.1016/j.drup.2020.100712.
    [19] QIAO Z, CHENG Y, LIU S, et al. Casticin inhibits esophageal cancer cell proliferation and promotes apoptosis by regulating mitochondrial apoptotic and JNK signaling pathways[J]. Naunyn Schmiedebergs Arch Pharmacol, 2019, 392(2): 177-187. DOI: 10.1007/s00210-018-1574-5.
    [20] ZHANG Y, WANG Y, ZHAO Y, et al. Novel camphor-based pyrimidine derivatives induced cancer cell death through a ROS-mediated mitochondrial apoptosis pathway[J]. RSC Adv, 2019, 9(51): 29711-29720. DOI: 10.1039/C9RA05900H.
    [21] ZHONG FR, CHENG HL, ZHANG H, et al. Effect of kaempferol on the proliferation, migration, invasion, and apoptosis of human hepatoma Bel-7402 cells[J]. J Clin Hepatol, 2020, 36(12): 2725-2729. DOI: 10.3969/j.issn.1001-5256.2020.12.017.

    仲富瑞, 程宦立, 张浩, 等. 山萘酚对人肝癌Bel-7402细胞增殖、迁移、侵袭及凋亡的影响[J]. 临床肝胆病杂志, 2020, 36(12): 2725-2729. DOI: 10.3969/j.issn.1001-5256.2020.12.017.
    [22] LI R, ZOU X, ZHU T, et al. Destruction of neutrophil extracellular traps promotesthe apoptosis and inhibits the invasion of gastric cancer cells by regulating the expression of Bcl-2, Bax and NF-κB[J]. Onco Targets Ther, 2020, 13: 5271-5281. DOI: 10.2147/OTT.S227331.
    [23] EDLICH F. The great migration of Bax and Bak[J]. Mol Cell Oncol, 2015, 2(3): e995029. DOI: 10.4161/23723556.2014.995029.
    [24] GUO DF, WANG RT, HUANG JC, et al. Curcumin derivative C086 induces the apoptosis of hepatoma HepG2 cells via the PI3K-Akt pathway[J]. Chin J Gerontol, 2021, 41(6): 1270-1274. DOI: 10.3969/j.issn.1005-9202.2021.06.042.

    郭登方, 王若涛, 黄建成, 等. 姜黄素衍生物C086通过PI3K-Akt通路诱导肝癌HepG2细胞凋亡[J]. 中国老年学杂志, 2021, 41(6): 1270-1274. DOI: 10.3969/j.issn.1005-9202.2021.06.042.
    [25] WU XY, ZHAO YN, WANG XJ, et al. Effect of galectin-3 expression suppression on expressions of Bcl-2 and Bax in gastric cancer MGC-803 cells and its promotion on apoptosis[J]. J Jilin Univ(Med Edit), 2020, 46(2): 335-339. DOI: 10.13481/j.1671-587x.20200221.

    吴雪艳, 赵依纳, 王小杰, 等. 半乳糖凝集素3表达抑制对人胃癌MGC-803细胞中Bcl-2和Bax表达的影响及其促凋亡作用[J]. 吉林大学学报(医学版), 2020, 46(2): 335-339, 前插3. DOI: 10.13481/j.1671-587x.20200221.
    [26] FU K, SUI GC, SHI JM. Inhibitory effect of triterpenoid saponins from Dioscorea gracillima on human hepatoma HepG2 cells[J/CD]. Cardiovasc Dis J Integr Tradit Chin Western Med (Electronic), 2020, 8(1): 67-68. DOI: 10.16282/j.cnki.cn11-9336/r.2020.01.052.

    付康, 隋广超, 史金铭. 三萜纤细薯蓣皂苷对人肝癌HepG2细胞的抑制作用研究[J/CD]. 中西医结合心血管病电子杂志, 2020, 8(1): 67-68. DOI: 10.16282/j.cnki.cn11-9336/r.2020.01.052.
    [27] NIE SS, LI X, ZHAO YH, et al. Effect of modified Si Junzitang drug serum on expression of apoptosis-related molecules of gastric cancer cell SGC-7901[J]. Chin J Exp Med Formul, 2019, 25(9): 25-30. DOI: 10.13422/j.cnki.syfjx.20190824.

    聂闪闪, 李洵, 赵玉航, 等. 加味四君子汤含药血清对胃癌细胞SGC-7901凋亡相关因子表达的影响[J]. 中国实验方剂学杂志, 2019, 25(9): 25-30. DOI: 10.13422/j.cnki.syfjx.20190824.
    [28] MAO M, HUA Y, JIANG X, et al. Expression of tumor necrosis factor alpha and neuronal apoptosis in the developing rat brain after neonatal stroke[J]. Neurosci Lett, 2006, 403(3): 227-232. DOI: 10.1016/j.neulet.2006.03.078.
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