苎麻根石油醚部位体外抗肿瘤生物活性及其GC-MS分析
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摘要
目的:明确苎麻根石油醚部位体外抗胃癌活性并揭示其发挥药效的物质基础,为苎麻根的开发利用奠定基础。方法:采用噻唑蓝(MTT)法考察不同给药剂量,不同给药时间下苎麻根石油醚部位对人胃癌HGC-27细胞的增殖抑制作用及其与时间、剂量的关系;采用流式细胞仪检测苎麻根石油醚部位作用于人胃癌HGC-27细胞后细胞凋亡率和细胞周期的变化;通过GC-MS对发挥药效的苎麻根石油醚部位进行成分解析。结果:不同质量浓度的苎麻根石油醚部位作用于人胃癌HGC-27细胞24,48,72 h后,均呈现出较好的细胞增殖抑制作用,并呈现一定的时间-剂量-效应关系,差异有统计学意义(P <0. 05);苎麻根石油醚部位作用人胃癌HGC-27细胞后可诱导细胞凋亡,影响细胞周期的正常变化,G0/G1期细胞比例明显减少,S期细胞比例明显增多;通过GC-MS鉴定出苎麻根石油醚部位中26种化学成分,主要包括谷甾醇、豆甾醇等。结论:苎麻根石油醚部位为其抗胃癌活性部位,其发挥药效的主要成分为甾醇类化合物,为苎麻根在临床上的合理应用及其在抗肿瘤方面的进一步研究奠定基础。
Objective: To define the anti-gastric cancer activity in vitro of petroleum ether fraction of Boehmeria nivea root and reveal the material basis of its efficacy,so as to lay the foundation for the development and utilization of B. nivea root. Method: Methyl thiazolyl tetraolium( MTT) method was used to evaluate the inhibitory rate and time-dose relationship of petroleum ether fraction of B. nivea root with different doses and delivery times on human gastric cancer HGC-27 cells. Flow cytometry was used to detect the change of cell apoptosis and cell cycle after petroleum ether fraction of B. nivea root acted on human gastric cancer HGC-27 cells.GC-MS was used to detect the components of petroleum ether fraction of B. nivea root. Result: Experiment data showed significant cell proliferation inhibition in an obvious time-dose-effect manner,with statistically significant differences( P < 0. 05),after 24,48,72 h of incubation of human gastric cancer HGC-27 cells with different concentrations of petroleum ether fraction of B. nivea root. The effect of petroleum ether fraction of B. nivea root on human gastric cancer HGC-27 cells could induce apoptosis,which affects the normal changes of cell cycle. The percentage of cells was decreased significantly in G0/G1 phase,and that in S phase was significantly increased.GC-MS was used to identify 26 chemical constituents in petroleum ether of B. nivea root,including sitosterol and stigmasterol. Conclusion: Petroleum ether fraction of B. nivea root is the active anti-gastric cancer part,and its main effective component is sterol compounds. This lays the foundation for the rational application of B. nivea root in clinic and the further research in tis anti-tumor effect.
Objective: To define the anti-gastric cancer activity in vitro of petroleum ether fraction of Boehmeria nivea root and reveal the material basis of its efficacy,so as to lay the foundation for the development and utilization of B. nivea root. Method: Methyl thiazolyl tetraolium( MTT) method was used to evaluate the inhibitory rate and time-dose relationship of petroleum ether fraction of B. nivea root with different doses and delivery times on human gastric cancer HGC-27 cells. Flow cytometry was used to detect the change of cell apoptosis and cell cycle after petroleum ether fraction of B. nivea root acted on human gastric cancer HGC-27 cells.GC-MS was used to detect the components of petroleum ether fraction of B. nivea root. Result: Experiment data showed significant cell proliferation inhibition in an obvious time-dose-effect manner,with statistically significant differences( P < 0. 05),after 24,48,72 h of incubation of human gastric cancer HGC-27 cells with different concentrations of petroleum ether fraction of B. nivea root. The effect of petroleum ether fraction of B. nivea root on human gastric cancer HGC-27 cells could induce apoptosis,which affects the normal changes of cell cycle. The percentage of cells was decreased significantly in G0/G1 phase,and that in S phase was significantly increased.GC-MS was used to identify 26 chemical constituents in petroleum ether of B. nivea root,including sitosterol and stigmasterol. Conclusion: Petroleum ether fraction of B. nivea root is the active anti-gastric cancer part,and its main effective component is sterol compounds. This lays the foundation for the rational application of B. nivea root in clinic and the further research in tis anti-tumor effect.
引文
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[18]肖志鹏.豆甾醇衍生物合成及抗肿瘤活性的研究[D].南昌:南昌大学,2013.
[19]段春燕,代荣阳,张春燕,等.阻断PI3K/AKT通路抑制棕榈酸诱导的肝癌细胞凋亡[J].泸州医学院学报,2014,37(4):353-356.
[2]皮林君,马春林,李海龙,等.参芪抑瘤含药血清对BGC-823胃癌细胞增殖和周期的影响[J].中国实验方剂学杂志,2018,24(5):121-125.
[3]包永睿,王帅,孟宪生,等.水红花子黄酮类成分对人肝癌细胞株SMMC-7721细胞周期及细胞凋亡的影响[J].中药材,2013,36(2):255-259.
[4]吴康郁,袁伟彬,黄伟斌.星点设计-效应面法优化苎麻根总黄酮超声提取工艺[J].新中医,2015,47(8):262-265.
[5]石文静,谭佳妮,沈卫星,等.清热解毒与以毒攻毒治法在肿瘤治疗中的比较研究[J].时珍国医国药,2017,28(9):2184-2186.
[6]周滢,周梅,段恒.中医药治疗胃癌的理论研究[J].中国实验方剂学杂志,2010,16(6):284-285.
[7]南京中医药大学.中药大辞典.上册[M]. 2版.上海:上海科学技术出版社,2005:1493-1496.
[8] CAI X F, JIN X J, Lee D H, et al.Phenanthroquinolizidine alkaloids from the roots of Boehmeria pannosa potently in-hibit hypoxia-inducible factor-1 in AGS human gastric cancer cells[J]. J Nat Prod,2006,69(7):1095-1097.
[9] YAN J,LUO D,LUO Y,et al. Induction of G1arrest and differentiation in MDA-MB-231 breast cancer cell by boehmeriasin A,a novel compound from plant[J]. Int J Gynecol Cancer,2006,16(1):165-170.
[10]王佳娜,张艳,宋丽艳,等.刺齿凤尾蕨化学成分及其体外抗肿瘤活性研究[J].中国中药杂志,2017,42(21):4159-4164.
[11]姜廷良.疗效中药发展提高的核心[J].中国实验方剂学杂志,2008,14(1):1.
[12]邵立军,王健农.中药苎麻根抗乙肝病毒活性及其化学成分研究[D].北京:中国中医科学院,2010.
[13]蔡利,管翰粟.中药在抗肿瘤过程中的应用及研究进展[J].中医临床研究,2017,9(11):141-142.
[14]范越,杨云柯,马骏,等.松友饮联合化疗治疗60例胃癌患者的临床观察[J].中华中医药杂志,2007,22(7):467-469.
[15]曹玫,欧阳露.植物甾醇的抗肿瘤作用及其机制研究进展[J].实用药物与临床,2015,18(9):1104-1107.
[16]王莉.β-谷甾醇对子宫颈癌细胞株Si Ha的生长抑制作用及其机制探讨[D].上海:复旦大学,2006.
[17]王娟,刘军权,陈复兴,等.β-谷甾醇对人共刺激细胞杀伤胃癌SGC-7901细胞的影响及其机制的探讨[J].免疫学杂志,2014,30(7):578-584.
[18]肖志鹏.豆甾醇衍生物合成及抗肿瘤活性的研究[D].南昌:南昌大学,2013.
[19]段春燕,代荣阳,张春燕,等.阻断PI3K/AKT通路抑制棕榈酸诱导的肝癌细胞凋亡[J].泸州医学院学报,2014,37(4):353-356.