文章摘要
李翠莹,王丽红,王婷婷,辛馥辰,刘德江.软枣猕猴桃根内生真菌Trichoderma sp.总皂苷抗肝癌初步探究[J].,2024,(3):434-441
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
投稿时间:2023-07-19  修订日期:2023-08-16
DOI:10.13241/j.cnki.pmb.2024.03.007
中文关键词: 软枣猕猴桃  内生真菌  肝癌  总皂苷
英文关键词: Actinidia arguta  Endophytic fungi  Liver cancer  Total saponins
基金项目:中央支持地方高校改革发展资金优秀青年人才项目(2020YQ09)
作者单位E-mail
李翠莹 佳木斯大学药学院 黑龙江 佳木斯 154007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 Licuiying0911@163.com 
王丽红 佳木斯大学药学院 黑龙江 佳木斯 154007  
王婷婷 佳木斯大学生物与农业学院 黑龙江 佳木斯 154007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  
辛馥辰 佳木斯大学生物与农业学院 黑龙江 佳木斯 154007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  
刘德江 佳木斯大学药学院 黑龙江 佳木斯 154007佳木斯大学生物与农业学院 黑龙江 佳木斯 154007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  
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中文摘要:
      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50~400 μg/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、400 μg/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、400 μg/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、400 μg/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、400 μg/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。
英文摘要:
      ABSTRACT 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 400 μg/mL. The cell scratch test showed that the concentration of AARTS and C15TS at 200 and 400 μg/mL could decreased the migration abilities of HepG 2 and Huh-7 cells. In Transwell assay, AARTS and C15TS at concentrations of 200 and 400 μg/mL inhibited the invasion abilities of HepG 2 and Huh-7 cells. In WB experiment, after incubation with 200 and 400 μg/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 400 μg/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.
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