青藤碱抑制视网膜母细胞瘤细胞生长的作用机制研究  

Study on the mechanism of Sinomenine inhibiting the growth of Retinoblastoma cells

作　　者：郑虔[1] 李翠萍 潘玉 李华林[1] 耿红丽[1] 朱芹 郝爽 杨淑颖 Zheng Qian;Li Cuiping;Pan Yu;Li Hualin;Geng Hongli;Zhu Qin;Hao Shuang;Yang Shuying(Department of ophthalmology,Zibo maternal and Child Health Hospital,Shandong Province 255000,China)

机构地区：[1]淄博市妇幼保健院,255000

出　　处：《临床眼科杂志》2024年第5期444-449,共6页Journal of Clinical Ophthalmology

基　　金：山东省中医药科技项目(2020Q078);山东省医药卫生科技发展计划项目(202007020268)。

摘　　要：目的探讨青藤碱抑制视网膜母细胞瘤系WERI⁃RB⁃1生长的作用机制。方法采用免疫荧光法、CCK⁃8法和流式细胞仪分析青藤碱对WERI⁃RB⁃1细胞活力和凋亡的影响,Western blot检测相关蛋白表达。结果不同浓度青藤碱处理的WERI⁃RB⁃1细胞中自噬相关蛋白Beclin1和LC3B表达上调,p62表达下调。WER⁃RBv1细胞中加入50μmol/L青藤碱和自噬抑制剂CQ刺激24 h后,由青藤碱刺激增加的Beclin1和LC3B恢复,降低的p62蛋白也恢复;由青藤碱刺激增加的细胞自噬减少(免疫荧光法检测LC3B标记的阳性细胞);部分减轻了青藤碱诱导的细胞活力降低(CCK⁃8法);由青藤碱刺激增加的细胞凋亡蛋白Bax、Bcl⁃2蛋白恢复。青藤碱抑制PI3K/Akt/mTOR通路,激活PI3K/Akt/mTOR通路后,青藤碱对细胞生长和自噬的作用被抵消。结论青藤碱通过体外PI3K/Akt/mTOR轴失活促进自噬来抑制视网膜母细胞瘤细胞的生长。Objective To explore the mechanism of Sinomenine inhibiting the growth of Retinoblastoma line WERI-RB-1.Methods Immunofluorescence,CCK-8 and flow cytometry were used to analyze the effect of Sinomenine on the viability and apoptosis of WERI-RB-1 cells,and Western blot was used to detect the expression of related proteins.Results The expression of autophagy related proteins Beclin1 and LC3B was up-regulated and p62 was down regulated in WERI-RB-1 cells treated with different concentrations of Sinomenine.After 24 h stimulation with 50μmol/L Sinomenine and autophagy inhibitor CQ,the increased Beclin1 and LC3B stimulated by Sinomenine recovered,and the decreased p62 protein also recovered;The autophagy stimulated by Sinomenine was reduced(LC3B labeled positive cells were detected by immunofluorescence);It partially alleviated the cell viability reduction induced by Sinomenine(CCK-8 method);The increased apoptosis protein Bax and Bcl-2 stimulated by Sinomenine recovered.Sinomenine inhibits the PI3K/Akt/mTOR pathway,and after activating the PI3K/Akt/mTOR pathway,the effect of Sinomenine on cell growth and autophagy is offset.Conclusions Sinomenine can inhibits retinoblastoma cell growth by promoting autophagy via inactivation of PI3K/Akt/mTOR axis in vitro.

关 键 词：青藤碱 视网膜母细胞瘤 作用机制 

分 类 号：R285[医药卫生—中药学]