加减理冲汤提取物抗胃癌活性及机制研究  

The activities of modified Lichong Decoction extracts against gastric cancer and the underlying mechanism

作　　者：施国善 陈哲[2] 姚宏玉 王玉静 马立威 SHI Guoshan;CHEN Zhe;YAO Hongyu;WANG Yujing;MA Liwei(School of Basic Medicine,Guizhou University of Traditional Chinese Medicine,Guizhou Guiyang 550025,China;Public Health College,Qiqihar Medical University,Heilongjiang Qiqihar 161006,China;Research Institute of Medical Science and Pharmacy,Qiqihar Medical University,Heilongjiang Qiqihar 161006,China)

机构地区：[1]贵州中医药大学基础医学院,贵州贵阳550025 [2]齐齐哈尔医学院公共卫生学院,黑龙江齐齐哈尔161006 [3]齐齐哈尔医学院医药科学研究院,黑龙江齐齐哈尔161006

出　　处：《中国医院药学杂志》2025年第7期755-763,共9页Chinese Journal of Hospital Pharmacy

基　　金：贵州省科技计划项目(编号:黔科合基础-ZK[2021]一般502);贵州省普通高等学校青年科技人才成长项目(编号:黔教合KY字[2021]208);齐齐哈尔医学科学院重点培育项目(编号:2023-ZDPY-006)。

摘　　要：目的:探讨加减理冲汤(MLCD)不同提取物的抗胃癌活性并探讨其可能的分子机制。方法:分别获取加减理冲汤醇提液(MLCD-A)、水提液(MLCD-W)及醇水共提液(MLCD-AW)。选取5株胃癌细胞(MNK-45、AGS、MGC-803、SGC-7901、BGC-823)及1株胃上皮细胞(GES-1)进行细胞毒性评价,将筛选出的敏感细胞株作为实验细胞株,通过乳酸脱氢酶(LDH)试剂盒评估细胞LDH释放量,Hoechst33342-PI染色法分析细胞凋亡形态学变化,酶联免疫实验(ELISA)法评估细胞内活性氧(ROS)含量变化,JC-1染色分析细胞线粒体膜电位变化,GreenNuc^(TM)活细胞检测试剂盒结合共聚焦显微镜观察细胞Caspase-3酶活性表达,qPCR法及Western blot法检测Caspase-3/-9基因/蛋白表达。结果:制备的MLCD三种提取液中,MLCD-AW的出膏率及抗胃癌活性最佳,且对AGS细胞活性最好,能使AGS细胞LDH释放量增加(P<0.05,P<0.01),细胞凋亡形态学变化明显,细胞内ROS含量增加(P<0.05,P<0.01),线粒体膜电位变化显著,Caspase-3酶活性表达增强(P<0.01),Caspase-3/-9基因表达量增加(P<0.05,P<0.01),c-Caspase-3/-9蛋白表达量增加(P<0.05,P<0.01)。结论:MLCD-A、MLCD-W、MLCD-AW均有一定的抗胃癌细胞活性,且MLCD-AW抗胃癌活性最佳;线粒体途径可能参与了诱导胃癌细胞凋亡的调控。OBJECTIVE To explore the anti-gastric cancer activities of modified Lichong Decoction(MLCD)extracts and their possible molecule mechanisms.METHODS Alcohol extracts(MLCD-A),water extracts(MLCD-W)and alcohol&water co-extracts(MLCD-AW)of MLCD were obtained.The gastric cancer cell lines(MNK-45,AGS,MGC-803,SGC-7901,BGC-823)and gastric epithelial cell line(GES-1)were selected for cytotoxicity evaluations.Cells sensitive to MLCD extracts were selected as experimental cells.Lactate dehydrogenase(LDH)kits were used to evaluate LDH release from cells.Morphological changes in apoptotic cells were analyzed by Hoechst 33342/PI staining.Intracellular reactive oxygen species(ROS)content was evaluated by an enzyme-linked immunosorbent assay(ELISA)kit.Changes in the mitochondrial membrane potential were analyzed by JC-1 staining.GreenNuc™Live Cell Assay Kit combined with laser confocal microscopy were used to observe the expression of Caspase-3.The mRNA and protein expressions of Caspase-3 and Caspase-9 were detected by qPCR(quantitative polymerase chain reaction)and Western blot,respectively.RESULTS Among the three forms of MLCD extracts,water&alcohol co-extract had the best extraction ratio and anti-gastric cancer activity on AGS cells.MLCD-AW significantly increased LDH release in AGS cells(P<0.05,P<0.01),showing most pronounced changes in the morphology of apoptotic cells,an increase in ROS content(P<0.05,P<0.01),changes in the mitochondrial membrane potential an increase in Caspase-3 activity(P<0.01),and increases in the protein levels of Caspase-3,Caspase-9,c-Caspase-3,and c-Caspase-9(P<0.05,P<0.01).CONCLUSION MLCD-A,MLCD-W and MLCD-AW all have certain anti-gastric cancer activities,and MLCD-WA has the best activity against gastric cancer.The mitochondrial pathway is involved in inducing apoptosis of gastric cancer cells.

关 键 词：加减理冲汤 胃癌细胞 细胞凋亡 线粒体途径 

分 类 号：R961[医药卫生—药理学]