天然产物研究与开发 ›› 2023, Vol. 35 ›› Issue (12): 2127-2136.doi: 10.16333/j.1001-6880.2023.12.012

• 开发研究 • 上一篇    下一篇

甘草内生菌对顺铂诱导HEK293细胞损伤的保护作用及机制研究

张   楠1，张尚龙1，连小龙3，杨志军1，马趣环1，叶礼巧1，邓   毅1,2*，杨秀娟1,2*   

  1. 1甘肃中医药大学；2甘肃省中药药理与毒理学重点实验室，兰州 730000；3青海大学，青海 810000
  • 出版日期:2023-12-28 发布日期:2023-12-27
  • 基金资助:
    国家自然科学基金（地区）项目（81960723）；甘肃省中医药管理局（GZKP-2022-37）；甘肃省自然科学基金（21JR11RA145）；兰州市科技计划（2022-3-21）

Protective effect and mechanism of endophytic bacteria of Glycyrrhiza uralensis Fisch against cisplatin-induced damage of HEK293 cells

ZHANG Nan1,ZHANG Shang-long1,LIAN Xiao-long3,YANG Zhi-jun1,MA Qu-huan1,YE Li-qiao1,DENG Yi1,2*,YANG Xiu-juan1,2*   

  1. 1Gansu University of Traditional Chinese Medicine;2Key Laboratory of Pharmacology and Toxicology of traditional Chinese Medicine in Gansu Province,Lanzhou 730000,China;3Qinghai University,Qinghai 810000,China
  • Online:2023-12-28 Published:2023-12-27

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摘要：

基于体外实验建立顺铂诱导肾细胞损伤模型，探讨甘草内生菌对顺铂诱导人胚肾293（HEK293）细胞损伤的减毒作用机制。通过MTT法检测甘草内生菌、顺铂及两者联合用药对HEK293细胞存活率的影响，流式细胞术检测不同干预下HEK293细胞凋亡率的变化，Q-PCR及Western blot法检测凋亡相关指标Bcl-2、Bax、Caspase-3、Caspase-8、Caspase-9及内质网应激通路（GRP78、CHOP）mRNA和蛋白表达的变化。结果表明，顺铂诱导HEK293细胞损伤，具有时间和剂量依赖性。甘草内生菌能够明显降低顺铂损伤模型细胞凋亡率，通过下调Bax、Caspase-3、Caspase-8、Caspase-9、GRP78、CHOP mRNA和蛋白表达，上调Bcl-2 mRNA和蛋白表达发挥对顺铂诱导肾细胞损伤的保护作用。综上，甘草内生菌能够改善顺铂诱导的HEK293细胞损伤，其作用机制可能是通过调控Bcl-2家族、Caspase家族凋亡蛋白及内质网应激通路蛋白来实现抗凋亡作用，减轻细胞损伤。

关键词: 甘草内生菌, 顺铂, HEK293细胞, 凋亡, 内质网应激

Abstract:

The model of cisplatin-induced kidney cell damage was established based on in vitro experiment to investigate the attenuated mechanism of Glycyrrhiza uralensis endophyte on cisplatin-induced human embryonic kidney 293 (HEK293) cell damage.MTT assay was used to detect the effects of Glycyrrhiza uralensis endophyte,cisplatin and the combination of two drugs on the survival rate of HEK293 cells.The apoptosis rate of HEK293 cells was detected by flow cytometry,the mRNA and protein expressions of Bcl-2,Bax,Caspase-3,Caspase-8,Caspase-9 and endoplasmic reticulum stress pathway (GRP78,CHOP) were detected by Q-PCR and Western blot.The results showed that cisplatin induced HEK293 cell damage in a time-and dose-dependent manner.Glycyrrhiza uralensis endophyte could significantly reduce the apoptosis rate of cisplatin-damaged model by down-regulating the mRNA and protein expressions of Bax,Caspase-3,Caspase-8,Caspase-9,GRP78,and CHOP,up-regulating the mRNA and protein expression of Bcl-2 exerts a protective effect on cisplatin-induced renal cell injury.In conclusion.Glycyrrhiza uralensis endophyte could significantly reduce the apoptosis rate of cisplatin-damaged model,have anti-apoptotic effects on cisplatin-induced HEK293 cell damage,and the mechanism may be through regulation of apoptotic proteins of Bcl-2 family,Caspase family and endoplasmic reticulum stress pathway proteins to achieve anti-apoptotic effects and reduce cell damage.

Key words: Glycyrrhiza uralensis endophytes, cisplatin, HEK293 cells, apoptosis, endoplasmic reticulum stress

中图分类号:  R285.5

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