石斛酚在人晶状体上皮细胞中跨膜吸收机制研究  

Mechanism of gigantol in transmembrane transport in human lens epithelial cells

作　　者：胡小红[1] 魏小勇[1] HU Xiao-hong;WEI Xiao-yong(Fundamental Medicine School,Guangzhou University of Chinese Medicine,Guangzhou 510006,China)

机构地区：[1]广州中医药大学基础医学院,广东广州510006

出　　处：《中国中药杂志》2023年第7期1936-1942,共7页China Journal of Chinese Materia Medica

基　　金：国家自然科学基金项目(81973537);广东省自然科学基金项目(2020A1515011419)。

摘　　要：石斛酚为中药石斛的酚类成分,具有抗肿瘤、抗糖尿病性白内障等多种药理活性。该文探索其在人晶状体上皮细胞(HLECs)中跨膜转运的分子机制。体外培养永生化的HLECs,按5000个/mL细胞接种于激光共聚焦扫描显微镜(LSCM)培养皿中,通过LSCM观察荧光标记的石斛酚在HLECs内的荧光分布和荧光强度,以荧光强度表示石斛酚的吸收量,监测石斛酚在HLECs内的跨膜转运过程,比较时间、温度、浓度、转运抑制剂以及不同细胞系对石斛酚跨膜吸收和转运的影响;HLECs接种于6孔培养板的爬片上,原子力显微镜(AFM)检测非荧光标记的石斛酚跨膜吸收过程中HLECs表面超微结构的变化。结果显示,石斛酚跨膜吸收存在时间、温度、浓度依赖性,并能够专一性地靶向识别HLECs;能量及载体转运抑制剂降低HLECs对石斛酚的吸收量;在石斛酚跨膜过程中,HLECs细胞膜表面变得更粗糙,并呈现不同程度的凹坑。结果表明,石斛酚跨膜转运方式是通过能量及载体介导的胞吞作用主动吸收实现的。Gigantol is a phenolic component of precious Chinese medicine Dendrobii Caulis,which has many pharmacological activities such as prevent tumor and diabetic cataract.This paper aimed to investigate the molecular mechanism of gigantol in transmembrane transport in human lens epithelial cells(HLECs).Immortalized HLECs were cultured in vitro and inoculated in the laser scanning confocal microscopy(LSCM)medium at 5000 cells/mL.The fluorescence distribution and intensity of gigantol marked by fluorescence in HLECs were observed by LSCM,and the absorption and distribution of gigantol were expressed as fluorescence intensity.The transmembrane transport process of gigantol in HLECs were monitored.The effects of time,temperature,concentration,transport inhibitors,and different cell lines on the transmembrane absorption and transport of gigantol were compared.HLECs were inoculated on climbing plates of 6-well culture plates,and the ultrastructure of HLECs was detected by atomic force microscopy(AFM)during the transmembrane absorption of non-fluorescent labeled gigantol.The results showed that the transmembrane absorption of gigantol was in time and concentration-dependent manners,which was also able to specifically target HLECs.Energy and carrier transport inhibitors reduced gigantol absorption by HLECs.During transmembrane process of gigantol,the membrane surface of HLECs became rougher and presented different degrees of pits,indicating that the transmembrane transport of gigantol was achieved by active absorption of energy and carrier-mediated endocytosis.

关 键 词：石斛酚 跨膜转运 转运方式 人晶状体上皮细胞 

分 类 号：R285[医药卫生—中药学]