软枣猕猴桃根内生真菌Trichoderma sp.总皂苷抗肝癌初步探究  

Study on Total Saponins of Endophytic Fungi in Actinidia Arguta Root Against Hepatocellular Carcinoma

作　　者：李翠莹 王丽红[1] 王婷婷[2,3] 辛馥辰 刘德江 LI Cui-ying;WANG Li-hong;WANG Ting-ting;XIN Fu-chen;LIU De-jiang(School of Pharmacy,Jiamusi University,Jiamusi,Heilongjiang,154007,China;College of Biology and Agriculture,Jiamusi University,Jiamusi,Heilongjiang,154007,China;China-Ukraine Agriculture&Forestry Technology Development And Application International Cooperation Joint Lab,Jiamusi,Heilongjiang,154007,China)

机构地区：[1]佳木斯大学药学院,黑龙江佳木斯154007 [2]佳木斯大学生物与农业学院,黑龙江佳木斯154007 [3]中-乌农林技术开发与应用国际合作联合实验室,黑龙江佳木斯154007

出　　处：《现代生物医学进展》2024年第3期434-441,552,共9页Progress in Modern Biomedicine

基　　金：中央支持地方高校改革发展资金优秀青年人才项目(2020YQ09)。

摘　　要：目的:研究软枣猕猴桃根内生真菌C15(Trichoderma sp.)发酵提取物中的总皂苷成分对2种肝癌细胞HepG 2和Huh-7细胞的凋亡作用与初步作用机制。方法:采用CCK-8细胞计数(Cell Counting Kit-8,CCK-8)法检测软枣猕猴桃根总皂苷(AARTS)和内生真菌C15总皂苷(C15TS)对HepG 2和Huh-7细胞的细胞活力的影响;采用细胞划痕实验和Transwell小室实验对细胞的迁移和侵袭能力进行检测;采用蛋白质印记法(Western Blot,WB)检测P53信号通路中3种凋亡相关蛋白(Bax、Bcl-2、Caspase-3)的表达水平;采用Annexin V-FITC/PI染色检测HepG 2和Huh-7细胞的凋亡时期。结果:根据细胞活力测试得出浓度为50~400μg/mL时AARTS和C15TS对HepG 2和Huh-7细胞的活力有抑制作用,且呈现剂量依赖性;细胞划痕实验表明浓度为200、400μg/mL时AARTS和C15TS可以降低HepG 2和Huh-7细胞的迁移能力;在Transwell小室实验中,浓度为200、400μg/mL时AARTS和C15TS均可抑制对HepG 2细胞和Huh-7细胞的侵袭能力;在WB实验中,AARTS和C15TS经200、400μg/mL孵育24 h后,HepG 2和Huh-7细胞胞内Bax和Caspase-3蛋白表达量增加,Bcl-2蛋白表达量减少;在Annexin V-FITC/PI染色实验中,浓度为200、400μg/mL时AARTS组和C15TS组均具有促进HepG 2和Huh-7细胞凋亡的能力,且多为早期凋亡。结论:C15TS与AARTS的作用效果相似,也可抑制HepG 2和Huh-7细胞的增殖、迁移和侵袭,促进细胞凋亡,其机制可能与P53信号通路有关。Objective:To study the total saponins in the extract of endophytic fungus C15 from Actinidia arguta root and their effects on the apoptosis of hepatoma HepG 2 and Huh-7 cells,and to explore the possible mechanism.Methods:The effects of total saponins of AARTS(Total Saponins from the roots of Actinidia arguta,AARTS)and C15TS(C15 Total Saponins,C15TS)on the cell viabilities of HepG 2 and Huh-7 hepatoma cells were determined by CCK-8(Cell Counting Kit-8)method;The cell migration and invasion abilities were detected by cell scratch assay and Transwell assay;Western Blot(WB)was used to detect the expression levels of three apoptosis-related proteins(Bax,Bcl-2,Caspase-3)in HepG 2 and Huh-7 hepatoma cells;Annexin V-FITC/PI staining was used to detect the apoptotic period of HepG 2 and Huh-7 cells.Results:According to cell viability tests,AARTS and C15TS inhibited the viabilities of HepG 2 and Huh-7 cells in a dose-dependent manner at concentrations ranging from 50 to 400μg/mL.The cell scratch test showed that the concentration of AARTS and C15TS at 200 and 400μg/mL could decreased the migration abilities of HepG 2 and Huh-7 cells.In Transwell assay,AARTS and C15TS at concentrations of 200 and 400μg/mL inhibited the invasion abilities of HepG 2 and Huh-7 cells.In WB experiment,after incubation with 200 and 400μg/mL AARTS and C15TS for 24 h,the expression of Bax and Caspase-3 protein increased in HepG 2 and Huh-7 cells,while the expression of Bcl-2 protein decreased.In the Annexin V-FITC/PI staining experiment,both the AARTS groups and the C15TS groups with concentrations of 200 and 400μg/mL had the abilities to promote the apoptosis of HepG 2 and Huh-7 cells,and the apoptosis was mostly early.Conclusion:Similar to AARTS,the C15TS can also inhibit the proliferation,migration and invasion of HepG 2 and Huh-7 cells,and promote cell apoptosis,which may be related to P53 signaling pathway.

关 键 词：软枣猕猴桃 内生真菌 肝癌 总皂苷 

分 类 号：S663.4[农业科学—果树学] R285[农业科学—园艺学] R735.7[医药卫生—中药学]