山豆根主要成分苦参碱诱导人正常肝细胞L-02凋亡及其对细胞周期的影响
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摘要
目的:探索山豆根主要成分苦参碱诱导人肝(实质)细胞L-02凋亡的作用及其对细胞周期的影响。方法:以人肝细胞株L-02为模型,应用CCK8法以及流式细胞仪检测山豆根提取物(water extracts of sophorae tonkinensis radix et rhizoma,WE-SRR)及苦参碱(matrine,MT)在不同时间与浓度对L-02细胞增殖的影响以及作用15 h后对L-02凋亡及细胞周期的影响。结果:WE-SRR及MT均能时间及剂量依赖性抑制L-02细胞增殖。12和18 mg·m L-1WE-SRR及1. 67和2 mg·m L-1MT可诱导早期凋亡,而20 mg·m L-1WESRR组则表现为诱导晚期凋亡。所有浓度WE-SRR均可引起G2/M期细胞比例升高,且18和20 mg·m L-1WE-SRR组S期细胞比例升高; MT组低浓度时G0/G1期细胞比例明显升高,高浓度(2 mg·m L-1)时G2/M期细胞、S期细胞比例均明显升高。结论:WE-SRR及MT对人正常肝细胞L-02具有细胞毒性,其机制包括抑制增殖、诱导凋亡和细胞周期阻滞。MT和WE-SRR对细胞的影响变化趋势一致,推测MT是WE-SRR的主要毒性物质成分之一。
Objective: To investigate the effect of matrine(MT),the main component of Radix Sophorae Tonkinensis on cell proliferation,apoptosis and cell cycle of human liver cells L-02. Methods: CCK8 was used to measure the effect of water extracts of sophorae tonkinensis radix et rhizoma(WE-SRR) and MT on L-02 proliferation at various concentrations and time points and flow cytometry was used to examine their effects on the apoptosis and cell cycles of L-02 cells when treated with WE-SRR and MT after 15 h. Results: WE-SRR and MT both can inhibit proliferation of L-02 cells in a time and dose-dependent manner. 12,18 mg·m L-1 WE-SRR and 1. 67,2 mg·m L-1 MT could induce early apoptosis; while 20 mg·m L-1 WE-SRR group mainly showed late apoptosis. All concentrations of WE-SRR could significantly increase the proportions of G2/M phase cells,and the proportions of S phase cells in18 and 20 mg·m L-1 WE-SRR group were increased. For MT,in low concentration group,the proportions of G0/G1 phase cells were significantly increased; while in high concentration(2 mg·m L-1) group,the proportions of G2/M and S phase cells were both significantly increased. Conclusion: WE-SRR and MT both are cytotoxic tohuman normal hepatocytes L-02,and the mechanisms include inhibition of proliferation,induction of apoptosis,and cell cycle arrest. It is speculated that MT maybe one of the most important toxic components of WE-SRR as the change trend of the effects of MT and WE-SRR on cells was consistent.
Objective: To investigate the effect of matrine(MT),the main component of Radix Sophorae Tonkinensis on cell proliferation,apoptosis and cell cycle of human liver cells L-02. Methods: CCK8 was used to measure the effect of water extracts of sophorae tonkinensis radix et rhizoma(WE-SRR) and MT on L-02 proliferation at various concentrations and time points and flow cytometry was used to examine their effects on the apoptosis and cell cycles of L-02 cells when treated with WE-SRR and MT after 15 h. Results: WE-SRR and MT both can inhibit proliferation of L-02 cells in a time and dose-dependent manner. 12,18 mg·m L-1 WE-SRR and 1. 67,2 mg·m L-1 MT could induce early apoptosis; while 20 mg·m L-1 WE-SRR group mainly showed late apoptosis. All concentrations of WE-SRR could significantly increase the proportions of G2/M phase cells,and the proportions of S phase cells in18 and 20 mg·m L-1 WE-SRR group were increased. For MT,in low concentration group,the proportions of G0/G1 phase cells were significantly increased; while in high concentration(2 mg·m L-1) group,the proportions of G2/M and S phase cells were both significantly increased. Conclusion: WE-SRR and MT both are cytotoxic tohuman normal hepatocytes L-02,and the mechanisms include inhibition of proliferation,induction of apoptosis,and cell cycle arrest. It is speculated that MT maybe one of the most important toxic components of WE-SRR as the change trend of the effects of MT and WE-SRR on cells was consistent.
引文
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[16] XU GP,ZHAO W,ZHUANG JP,et al. Matrine inhibits the growth and induces apoptosis of osteosarcoma cells in vitro by inactivating the Akt pathway[J]. Tumour Biol,2015,36(3):1653-1659.
[2]李厚聪,袁德培,刘圆.山豆根属植物化学成分与生物活性研究进展[J].中草药,2014,45(23):3486-3493.
[3]李妃,李成平,付晖,等.山豆根研究进展及毒性成分检测方法补充报道[J].药物分析杂志,2013,33(8):1453-1463.
[4]陈影,陈两绵,仝燕,等.山豆根药理毒理研究进展[J].中国中药杂志,2017,42(13):2439-2442.
[5]曹洛云,李天娇,孟宪生,等.山豆根生物碱对DEN诱发肝癌大鼠的作用及机制研究[J].中国现代应用药学,2018,35(3):370-374.
[6]田雪松.山豆根中生物碱的毒性研究进展[J].中国实验方剂学杂志,2016,22(6):230-234.
[7]蔡丽云,吴莉莉,于小明,等.氧化苦参碱在大鼠肠道的吸收与代谢机制[J].药学学报,2015,50(10):1336-1341.
[8]董玲娜.(氧化)苦参碱与去甲乌药碱药代动力学特征与代谢机理[D].广州:南方医科大学,2013.
[9]戴五好,钱利武,王丽丽,等.苦参碱、氧化苦参碱对小鼠的毒性研究[J].安徽医药,2012,16(7):904-905.
[10]陈龙,吴谦,耿娅,等.山豆根水煎液致大鼠亚急性肝脏毒性研究[J].中国实验方剂学杂志,2013,19(18):293-296.
[11]耿娅,郭胜亚,邓中平,等.基于斑马鱼模型的不同产地山豆根急性毒性比较研究[C].西安:2015毒性测试替代方法与转化毒理学学术研讨会,2015.
[12]贾歌,刘畅,庞晶瑶,等.何首乌及其成分对人正常肝细胞凋亡的影响[J].中国中国医药信息杂志,2015,22(11):46-49.
[13]张茜,李峰杰,金若敏,等.苦参碱和氧化苦参碱致HL7702细胞毒性研究[J].中华中医药学刊,2011,29(6):1222-1224.
[14]杨春秀,陈建斌.苦参碱抑制RPMI8226细胞增殖及对Notch通路的影响[J].中成药,2017,39(10):2167-2170.
[15] AN Q,HAN C,ZHOU Y,et al. Matrine induces cell cycle arrest and apoptosis with recovery of the expression of miR-126 in the A549 non-small cell lung cancer cell line[J]. Mol Med Rep,2016,14(5):4042-4048.
[16] XU GP,ZHAO W,ZHUANG JP,et al. Matrine inhibits the growth and induces apoptosis of osteosarcoma cells in vitro by inactivating the Akt pathway[J]. Tumour Biol,2015,36(3):1653-1659.