肉苁蓉苯乙醇总苷对Bel-7405肝癌细胞增殖、凋亡的影响及机制研究  

Effect of phenylethanol glycoside from Cistache deserticola on proliferaion and apoptosis of Bel-7405 cells in vitro

作　　者：齐鑫鑫 赵军 何转霞 苏德奇 刘涛 QI Xin-xin;ZHAO Jun;HE Zhuan-xia;SU De-qi;LIU Tao(Department of Toxicology,College of Public Health,Xinjiang Medical University,Urumqi Xinjiang 830011,China;Xinjiang Uygur Autonomous Region Institute of materia Medicine,Urumqi Xinjiang 830004,China)

机构地区：[1]新疆医科大学公共卫生学院卫生毒理学教研室,新疆乌鲁木齐830011 [2]新疆维吾尔自治区药物研究所,新疆乌鲁木齐830004

出　　处：《毒理学杂志》2021年第3期225-230,240,共7页Journal of Toxicology

基　　金：新疆维吾尔自治区自然科学基金(2019D01C199);新疆维吾尔自治区研究生创新项目(XJ2020G183)。

摘　　要：目的探讨肉苁蓉苯乙醇总苷(CPhGs)对Bel-7405肝癌细胞增殖、凋亡的影响及其作用机制。方法以不同浓度CPhGs干预Bel-7405肝癌细胞后,计算其IC_(50),根据IC_(50)设置后续实验干预浓度分别为0、35、70和140μg/ml,采用MTT与划痕实验检测CPhGs对Bel-7405肝癌细胞活性及增殖情况、Annexin V-FITC/PI、Hochest33258及线粒体染色检测细胞凋亡变化,Western blot检测Pro-PARP、cleaved-PARP、Pro-Caspase-9、cleaved-Caspase-9及Cleaved-Caspase-3等凋亡蛋白表达情况。结果CPhGs对Bel-7405肝癌细胞的IC_(50)为298μg/ml,随着CPhGs浓度增加,Bel-7405细胞增殖抑制作用逐渐增强(P<0.01);细胞在24 h及48 h迁移率随CPhGs剂量的增加而减少(P<0.01);CPhGs各剂量组均可诱导Bel-7405肝癌细胞凋亡,且有一定的剂量依赖性,差异有统计学意义(P<0.05)。Hoechst33258及线粒体染色发现,CPhGs各剂量组均可诱导细胞凋亡;凋亡相关蛋白Pro-PARP、Cleaved-PARP、Pro-Caspase-3、Cleaved-Caspase-3、Pro-Caspase-9及Cleaved-Caspase-9表达明显增加,差异有统计学意义(P<0.01)。结论CPhGs对Bel-7405肝癌细胞具有增殖抑制作用,其机制可能与诱导细胞凋亡有关,但其具体细胞凋亡机制还需进一步探讨。Objective To investigate the effects and its mechanism of phenylethanol glycoside from Cistanche deserticola(CPhGs)on the proliferation and apoptosis of Bel-7405 cells.Methods After mediation of Bel-7405 cells with diverse concentrations of CPhGs,the IC_(50) was calculated,and the ensuing test mediation concentrations were set to 0,35,70 and 140μg/ml agreeing to the IC_(50).Influence of CPhGs on Bel-7405 cell proliferation was measured by MTT method and scratch test.Apoptosis was detected by Annexin V-FITC/PI staining,Hoechst33258 staining method and mitotracker.The protein expressions of apoptotic proteins including Pro-PARP,Cleaved-PARP,Pro-Caspase-9,cleaved-Caspase-9 and cleaved-Caspase-3 were detected by Western blot.Results The experiment result showed that CPhGs could significantly inhibit proliferation of Bel-7405(P<0.01),and IC_(50) value was 289μg/ml.The result of the scratch experiment also showed that the migration area decreased at a dose dependent relation(P<0.01).Hoechst33258 staining and motiotracker found that more and more nuclear appeared shrinking and fragmentation,the mitochondria damage became greater with the increase of CPhGs concentration.Compared with control group,CPhGs could significantly induce Bel-7405 cell apoptosis in Annexin V/PI experiment(P<0.05).In addition,Western blot result showed that CPhGs can increase the expressions of Pro-PARP,cleaved-PARP,Pro-Caspase-9,cleaved-Caspase-9 and cleaved-Caspase-3(P<0.01).Conclusion It can be seen that CPhGs can inhibit proliferation of tumor by inducing cell apoptosis,and increasing apoptosis-related proteins,but the mechanism need to be further explored.

关 键 词：肉苁蓉苯乙醇总苷 Bel-7405肝癌细胞 细胞增殖 细胞凋亡 

分 类 号：R285.5[医药卫生—中药学] R114[医药卫生—中医学]