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[吉非替尼在人晶状体上皮细胞跨膜转运中的作用机制]

[Mechanism of gigantol in transmembrane transport in human lens epithelial cells].

作者信息

Hu Xiao-Hong, Wei Xiao-Yong

机构信息

Fundamental Medicine School, Guangzhou University of Chinese Medicine Guangzhou 510006, China.

出版信息

Zhongguo Zhong Yao Za Zhi. 2023 Apr;48(7):1936-1942. doi: 10.19540/j.cnki.cjcmm.20230118.501.

Abstract

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 5 000 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.

摘要

石斛酚是名贵中药材石斛茎中的一种酚类成分,具有多种药理活性,如预防肿瘤和糖尿病性白内障。本文旨在研究石斛酚在人晶状体上皮细胞(HLECs)跨膜转运中的分子机制。体外培养永生化HLECs,并以5000个细胞/mL接种于激光扫描共聚焦显微镜(LSCM)培养基中。通过LSCM观察HLECs中荧光标记的石斛酚的荧光分布和强度,以荧光强度表示石斛酚的吸收和分布情况。监测石斛酚在HLECs中的跨膜转运过程。比较时间、温度、浓度、转运抑制剂和不同细胞系对石斛酚跨膜吸收和转运的影响。将HLECs接种于6孔培养板的爬片上,在非荧光标记的石斛酚跨膜吸收过程中,用原子力显微镜(AFM)检测HLECs的超微结构。结果表明,石斛酚的跨膜吸收呈时间和浓度依赖性,且能特异性靶向HLECs。能量和载体转运抑制剂降低了HLECs对石斛酚的吸收。在石斛酚跨膜过程中,HLECs膜表面变得更粗糙,并呈现出不同程度的凹陷,表明石斛酚的跨膜转运是通过能量主动吸收和载体介导的内吞作用实现的。

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