| 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） |
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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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