基于HK-2细胞的左金丸中生物碱类组分体外毒性筛选及机制初探  

Nephrotoxicity screening of main active components in Zuojin Pills

作　　者：杨春启 连闻雨 林毅 谢光辉 马增春 谭洪玲 肖成荣 王宇光 高月 YANG Chun-qi;LIAN Wen-yu;LIN Yi;XIE Guang-hui;MA Zeng-chun;TAN Hong-ling;XIAO Cheng-rong;WANG Yu-guang;GAO Yue(Anhui Medical University,Hefei 230032,China;Dept of Pharmaceutical Sciences,Beijing Institute of Radiation Medicine,Beijing 100850,China;Guangdong Pharmaceutical University,Guangzhou 510006)

机构地区：[1]安徽医科大学,安徽合肥230032 [2]军事科学院军事医学研究院辐射医学研究所,北京100850 [3]广东药科大学,广东广州510006

出　　处：《中国药理学通报》2022年第1期110-118,共9页Chinese Pharmacological Bulletin

基　　金：国家重点研发计划(No 2018YFC1704504)。

摘　　要：目的基于肾小管上皮细胞HK-2筛选左金丸中主要生物碱类组分对肾细胞损伤。方法采用CCK-8法和高内涵筛选技术筛选左金丸主要生物碱中潜在肾细胞损伤成分。通过细胞形态、乳酸脱氢酶和细胞色素C释放率及凋亡相关蛋白表达等进行确证,初步确定造成细胞损伤作用的生物碱,为组分中药配伍组方和配伍减毒提供体外毒理学依据。结果采用CCK-8法和高内涵技术初步筛选,表明吴茱萸碱(EVO)能够显著降低细胞数目,升高细胞膜通透性,降低线粒体膜电位。此外,细胞形态、细胞凋亡和细胞色素C表达与高内涵筛选结果一致,Western blot实验表明EVO能够诱导细胞凋亡造成肾细胞损伤。结论 EVO可明显导致肾细胞损伤,可能是通过影响线粒体、细胞色素C及细胞膜通透性诱导细胞凋亡。Aim To study the nephrotoxicity effects of the main monomers in Zuojin Pills. Methods CCK-8 and high-content toxicity screening were used to preliminarily screen the main alkaloids in Zuojin Pills that may cause renal cell damage. Further, by confirmation of cell morphology, release rate of lactate dehydrogenase and cytochrome C, and expression of apoptosis-related proteins, the alkaloids causing cell damage were preliminarily identified, providing in vitro toxicological evidence for the compatibility of components of traditional Chinese medicine and compatibility attenuation. Results Preliminary screening using CCK-8 method and high-content technology showed that evodiamine(EVO) could significantly reduce cell number, increase cell membrane permeability, and reduce mitochondrial membrane potential. In addition, cell morphology, apoptosis and cytochrome C expression were consistent with the results of high-content screening. Western blot experiments indicate that EVO could induce apoptosis and cause renal cell damage. Conclusions EVO can obviously cause renal cell damage, and may induce apoptosis by affecting mitochondria, cytochrome C and cell membrane permeability.

关 键 词：左金丸 吴茱萸碱 高内涵 肾毒性 凋亡 HK-2细胞 

分 类 号：R284.1[医药卫生—中药学] R289.5[医药卫生—中医学]