摘要
目的优化桦褐孔菌总三萜提取物的纯化工艺,考察其纯化物的抗肿瘤活性。方法以桦褐孔菌醇为对照品,建立桦褐孔菌总三萜的含量测定方法。采用单因素实验筛选大孔吸附树脂种类、上样体积、上样液质量浓度、上样流速、洗脱剂体积分数、洗脱剂用量、洗脱流速,确定并验证该粗提物的纯化工艺。通过细胞增殖实验、迁移实验、流式细胞术和AO/EB试剂盒检测桦褐孔菌总三萜纯化物对人宫颈癌细胞HeLa增殖、迁移及凋亡的影响。结果桦褐孔菌总三萜粗提物的最佳纯化工艺为:AB-8型大孔吸附树脂,上样液质量浓度为2.0 mg/mL,上样体积为140 mL,上样流速为1.0 mL/min;洗脱时先用50%的乙醇40 mL除杂,再用95%的乙醇160 mL洗脱,洗脱流速为3.0 mL/min。纯化后桦褐孔菌总三萜的质量分数从34.36%升至73.39%。桦褐孔菌总三萜纯化物可抑制HeLa细胞增殖,半数抑制浓度为184.20μg/mL;与对照组相比,该纯化物可显著抑制HeLa细胞的迁移,促进其凋亡(P<0.05或P<0.01)。结论成功优化了桦褐孔菌总三萜提取物的纯化工艺,该纯化物可抑制HeLa细胞增殖和迁移,诱导其凋亡。
OBJECTIVE To optimite the purification technology of total triterpenoid extracts from Inonotus obliquus,and to investigate the anti-tumor activity of its purified products.METHODS Using inotodiol as control,the method was established for the content determination of total triterpenoid in I.obliquus.The type of macroporous adsorption resin,sample volume,sample concentration,sample flow rate,eluent volume,eluent dosage and elution flow rate were selected by single factor experiments.The purification technology of the crude extract was determined and verified.The effects of total triterpenoid purified from I.obliquus on the proliferation,migration and apoptosis of human cervical cancer HeLa cells were detected by cell proliferation test,migration test,flow cytometry and AO/EB kit.RESULTS The best purification technology of total triterpenoid crude extracts from I.obliquus was as follows:AB-8 macroporous adsorption resin was used;mass concentration of the sample solution was 2.0 mg/mL;sample volume was 140 mL,and the flow rate was 1.0 mL/min;the impurity was removed with 50%ethanol 40 mL,then eluted with 95%ethanol 160 mL,at the elution flow rate of 3.0 mL/min.After purification,mass concentration of total triterpenoid from I.obliquus increased from 34.36%to 73.39%.The total triterpenoid of I.obliquus could inhibit the proliferation of HeLa cells,and the 50%inhibitory concentration was 184.20μg/mL.Compared with control group,the purified products could significantly inhibit the migration and promote the apoptosis of HeLa cells(P<0.05 or P<0.01).CONCLUSIONS The purification technology of total triterpenoids extracts from I.obliquus is successfully optimited.The purified product could inhibit the proliferation and migration of HeLa cells and induce their apoptosis.
作者
邵骏菁
杨颖
马大龙
吕志强
田景振
张晓平
SHAO Junjing;YANG Ying;MA Dalong;LYU Zhiqiang;TIAN Jingzhen;ZHANG Xiaoping(School of Pharmacy,Shandong University of Traditional Chinese Medicine,Jinan 250355,China;Jining Medical University,Shandong Jining 272067,China;Dept.of Drug Dispensing,the Affiliated Hospital of Qingdao University,Shandong Qingdao 266000,China;Qingdao Academy of Chinese Medicine,Shandong University of Chinese Medicine,Shandong Qingdao 266112,China)
出处
《中国药房》
CAS
北大核心
2022年第18期2198-2203,共6页
China Pharmacy
基金
山东省重点研发计划(重大科技创新工程)(No.2020CXGC010505,No.2021CXGC010511)。
关键词
桦褐孔菌总三萜
纯化
细胞增殖
细胞迁移
细胞凋亡
total triterpenoid of Inonotus obliquus
purification
cell proliferation
cell migration
cell apoptosis
