金星散水提液诱导ECA-109细胞凋亡和氧化应激的作用及机制研究  

Aqueous extract of Jinxing powder induces the apoptosis and oxidative stress in ECA-109 cells

作　　者：李博 雷琨 刘家邑 刘军锋 张珍 LI Bo;LEI Kun;LIU Jia-yi;LIU Jun-feng;ZHANG Zhen(Shaanxi Hospital of Traditional Chinese Medicine,Xi’an 710003;Shaanxi Collaborative Innovation Center of Chinese Medicinal Resources Industrialization,Shaanxi University of Chinese Medicine,Xianyang Shaanxi 712046;College of Clinical Medical,Beijing University of Chinese Medicine,Beijing 100029)

机构地区：[1]陕西省中医医院,西安710003 [2]陕西中医药大学陕西省中药资源产业化协同创新中心,陕西咸阳712046 [3]北京中医药大学临床医学院,北京100029

出　　处：《中南药学》2024年第1期100-106,共7页Central South Pharmacy

基　　金：陕西省自然科学基础研究计划项目(No.2016ZDJC-18)。

摘　　要：目的研究金星散(JXS)水提液抗人食管癌ECA-109细胞的作用及机制。方法体外培养ECA-109细胞,用JXS水提液刺激,采用CCK-8法检测JXS水提液对ECA-109细胞活性的影响;Hoechst染色和流式细胞术检测JXS水提液对ECA-109细胞凋亡和周期的影响;ROS/Superoxide染色揭示JXS水提液对ECA-109细胞ROS和Superoxide水平的影响;Western blot分析JXS水提液对ECA-109细胞抗氧化及凋亡相关蛋白表达的影响。结果JXS水提液能抑制ECA-109细胞活性(3.9~1000μg·mL^(-1));Hoechst染色观察到JXS水提液在不同时间(6、12、24、48 h),以不同浓度(31.25、62.5、125、250μg·mL^(-1))刺激ECA-109细胞后,细胞核均发生不同程度的固缩、变形和碎裂,流式细胞术结果表明JXS水提液可诱导ECA-109细胞凋亡并将其周期阻滞在G2/M期;ROS/Superoxide结果显示JXS水提液刺激可引起ROS和Superoxide显著升高;Western blot分析证明JXS水提液可降低Caspase-3表达,升高Cleaved-Caspase-3水平,并明显下调Nrf2蛋白的表达。结论JXS水提液能诱导ECA-109细胞凋亡和氧化应激。Objective To determine the effect of Jinxing powder aqueous extract on human esophageal cancer ECA-109 cells and related mechanism.Methods ECA-109 cells were cultured in vitro and stimulated by Jinxing powder aqueous extract.CCK-8 assay was used to detect the effect of Jinxing powder aqueous extract on ECA-109 cell viability.Hoechst staining and flow cytometry were used to check the effect of Jinxing powder aqueous extract on ECA-109 cell apoptosis and cell cycle.The ROS/Superoxide assay was used to uncover the effect of Jinxing powder aqueous extract on ECA-109cell ROS and Superoxide level.Western blot was utilized to examine the impact of Jinxing powder aqueous extract on the oxidative resistance and expression of apoptosis-related proteins in ECA-109cells.Results Jinxing powder aqueous extract inhibited the viability of ECA-109 cells at 3.9 ~ 1000μg·mL~(-1).Hoechst staining revealed that the nucleus underwent various degrees of contractions,deformation,and fragmentation after ECA-109 cells were stimulated with Jinxing powder aqueous extract for different time(6,12,24,and 48 h) and concentrations(31.25,62.5,125,and 250 μg·mL^(-1)).Jinxing powder aqueous extract induced the apoptosis of ECA-109 cells and arrested their cell cycles at G2/M phase,as confirmed by flow cytometry.The ROS and Superoxide levels were significantly increased,as indicated by ROS/Superoxide assay.Caspase-3 was downregulated and CleavedCaspase-3 was upregulated by Jinxing powder aqueous extract,while Nrf2 expression was noticeably decreased,as demonstrated by Western blot analysis.Conclusion Jinxing powder aqueous extract can induce ECA-109 cell apoptosis and oxidative stress.

关 键 词：金星散 食管癌 凋亡 氧化应激:Caspase-3 

分 类 号：R286[医药卫生—中药学]