摘要
目的:通过生物活性追踪法对牛蒡根氯仿提取物进行分离提取,研究其抗肿瘤有效组分及可能的作用机制。方法:牛蒡根氯仿提取物经硅胶柱洗脱分离得到不同组分。用S180细胞和K562细胞经MTT法筛选活性组分;流式细胞术检测活性组分对K562细胞周期及凋亡的影响;小鼠淋转实验验证活性组分对小鼠免疫功能的影响;MTT法观察活性组分对耐药细胞K562/O2的逆转作用。结果:牛蒡根氯仿提取物分离鉴定后得到甾体类组分(L-1)、三萜类组分(L-2)、L-3和L-44个组分。各组分对S180细胞均显示很强的细胞毒作用,其IC50值均在10μg/mL以下。而K562细胞对甾体类组分和三萜类组分敏感度较高,IC50值分别为204.50μg/mL和117.56μg/mL;三萜类组分可使K562细胞G0/G1期细胞比例较对照组的45.52%增加至53.82%,S期细胞比例由对照组的45.68%减少到37.11%(均P<0.05);甾体类组分和三萜类组分分别将K562细胞凋亡率由5.1%增加至21.8%和38.0%(均P<0.01)。三萜类组分对小鼠淋巴细胞存在一定促进增殖作用,而甾体类组分与阿霉素合用后K562/O2细胞耐药指数由1165降低至468(P<0.01)。结论:牛蒡根氯仿提取物中的甾体类组分和三萜类组分影响细胞周期、诱导细胞凋亡并在免疫调节和肿瘤耐药性方面发挥抗肿瘤作用。
Objective:To explore the extraction and isolation of the roots of Arctiumlappa and the research of antitumoractivities.Methods:Different components of the roots of Arctiumlappa chloroform extract were separated by silica gel column elution.MTT assay was applied to determine the antitumor active componentsin S180 and K562 cells.Flow cytometry was used to detect the effects of antitumor active components on cell cycle and apoptosis of K562 cells.And lymphocyte transformation was applied to explore the effect of active components on immune function in mice.MTT assay was used to observe the effect of active components on reversing drug resistance of K562/O2 cells.Results:Four components including Steroidal components(L-1),Triperpenoids components(L-2),L-3 and L-4 were separated by column chromatography of the roots of Arctiumlappa chloroform extract.Four components showed strong inhibitory effect on cell proliferation of S180 cell,and the IC50 value were below 10μg/mL.Steroidal components and Triperpenoids components were identified to have stronger inhibition effect on K562 cell by MTT assay,the IC50 value were 204.50μg/mL and 117.56μg/mL.Triperpenoids components significantly increased the proportion of G0/G1 phase cells from 45.52%to 53.82%and decreased the proportion of S phase cells from 45.68%to 37.11%in K562 cells(both P<0.05).Steroidal components and Triperpenoids components could significantly increase the apoptotic rate of K562 cells from 5.1%to 21.8%and 38.0%,respectively.Triperpenoids components promoted lymphocyte proliferation of mice.Steroidal components reversed the drug resistance of K562/O2 cells,and the resistance index decreased from Arctiumlappa 1165 to 468(P<0.01).Conclusion:Steroidal components and Triperpenoids components of chloroform extracts from the roots of Arctiumlappa exert antitumor effect by blocking cycle arrest,inducing apoptosis,regulating immunity and reversing multidrug resistance.
作者
沈洪昇
何景华
赵秀梅
顾娜
史鹏程
胡人杰
SHEN Hong-sheng;HE Jing-hua;ZHAO Xiu-mei;GU Na;SHI Peng-cheng;HU Ren-jie(Department of Pharmacology,College of Basic Medicine,Tianjin Medical University,Tianjin 300070,China;Tianjin Institute of Medical and Pharmaceutical Sciences,Tianjin 300020,China)
出处
《天津医科大学学报》
2020年第5期476-481,共6页
Journal of Tianjin Medical University
基金
天津市卫生计生委中医中西医结合课题(13114)。
关键词
牛蒡根
细胞增殖
周期
凋亡
免疫
逆转耐药
roots of Arctiumlappa
proliferation
cell cycle
apoptosis
immunity
multidrug resistance
