姜黄素抑制人晶状体上皮细胞增生过程中蛋白激酶通路的信号转导变化  

Changes of protein kinase regulatory pathway in inhibition of curcumin on human lens epithelial cells proliferation

作　　者：胡艳红[1] 黄秀榕[2] 祁明信[1] 陈胜[1] 冯春燕[1] 柯发杰[1] 

机构地区：[1]福建中医药大学附属第二人民医院眼科,福州350003 [2]福建中医药大学中西医结合学院,福州350003

出　　处：《中华实验眼科杂志》2015年第6期507-511,共5页Chinese Journal Of Experimental Ophthalmology

基　　金：福建省科技厅青年人才基金项目（2008F3050）;福建省卫生厅重点课题项目（wzzb0605）

摘　　要：背景我们先前的研究证实姜黄素可抑制体外培养的人晶状体上皮细胞（LECs）的增生并影响细胞中环磷酸腺苷（cAMP）和环磷鸟嘌呤核苷（cGMP）的信号转导过程，但实际上体内LECs生物学行为的调控机制非常复杂。目的探讨姜黄素抑制体外培养的HLE-B3细胞增生过程中蛋白激酶（PK）通路的信号转导变化。方法HLE-B3经培养和传代后分为空白对照组、重组人碱性成纤维细胞生长因子（rhbFGF）组和rhbFGF＋姜黄素组。rhbFGF组在培养液中添加10ng／mlrhbFGF，rhbFGF＋姜黄素组在rhbFGF培养的基础上添加20mg／L姜黄素作用于细胞24h，利用流式细胞术检测各组细胞内PKA、PKC、PKG和钙调蛋白（CaM）的表达率。结果空白对照组、rhbFGF组和rhbFGF＋姜黄素组HLE-B3细胞中PKA蛋白的表达率分别为（46．8474±1．673）％、（33．250±2．242）％和（71．645±2．432）％，rhbFGF组较空白对照组PKA蛋白的表达率明显下降，而rhbFGF＋姜黄素组较rhbFGF组明显升高，差异均有统计学意义（t=11．904、28．430，均P〈0．01）。空白对照组、rhbFGF组和rhbFGF＋姜黄素组细胞中PKC蛋白的表达率分别为（35．5754-1．937）％、（50．652±2．068）％和（27．662±4．481）％，PKG蛋白的表达率分别为（63．838±0．486）％,（86．562±1．325）％和（40．733±2．175）％，CaM蛋白的表达率分别为（67．408±1．391）％、（83．887±3．499）％和（53．785±1．942）％，其中rhbFGF组细胞中PKC、PKG和CaM蛋白的表达率均明显低于空白对照组，差异均有统计学意义（均P〈0．01），而rhbFGF＋姜黄素组细胞中PKC、PKG和CaM蛋白的表达率均明显低于rhbFGF组，差异均有统计学意义（均P〈0．01）。结论姜黄素对HLE-B3细胞增生的抑制作用可能与其上调细胞中PKA的表达和下调PKC、PKG和CaM的表达有关。Background Our previous study showed that curcumin suppresses the proliferation of human lens epithelial cells （LECs） in vitro and has an influence on the signal transduction of cyclic adenosine monophosphate （cAMP） and cyclic guanosinc monophosphate （cGMP）. Actually, the regulation for biological behavior of LECs in vivo is complex. Objective This study was to investigate the changes of signal transduction of protein kinase （PK） in inhibition of curcumin on human LECs proliferation. Methods HLE-B3 was cultured and then divided into the blank control group, recombinant human basic fibroblast growth factor （rhbFGF） group and rhbFGF＋ curcumin group, rbbFGF of 10 ng/ml was added in the medium in the rhbFGF group, and 20 mg/L curcumin was added into rhbFGF-induced cell medium for 24 hours in the rhbFGF＋ curcumin group. The expression rates of PKA,PKC,PKG and calmodulin （CAM） in the cells were assayed using flow cytometry. Results The expression rates of PKA protein were （46. 847 ± 1. 673 ） % , （ 33. 250 ± 2. 242 ） % and （ 71. 645 ± 2.432 ） % in the blank control group,rhbFGF group and the rhbFGF＋ eurcumin group, respectively, and the expression rate of PKA protein was significantly reduced in the rhbFGF group compared with the blank control group （t = 11. 904, P〈0. 01 ） , but the expression rate of PKA protein in the rhbFGF＋ curcumin group was significnatly higher than that in the rhbFGF group （t=28. 430, P〈O. 01 ）. In the blank control group, rhbFGF group and the rhbFGF＋ curcumin group, the expression rates of PKC protein in the cells were （ 35. 575 ± 1. 937 ） % , （50. 652±2.068 ） % and （27.662±4.481 ） % ,those of PKG protein were （63. 838 ±0. 486） %, （ 86. 562 ± 1. 325 ） % and （40. 733 ± 2. 175 ） %, while those of CaM protein were （ 67. 408 ± 1.391 ） % , （ 83. 887±3. 499 ） % and （53. 785 ± 1. 942 ） % , the expression rates of PKC, PKG and CaM were significantly lower in the rhbFGF group in comparison with the b

关 键 词：姜黄素 蛋白激酶 信号转导 晶状体上皮细胞 

分 类 号：R285[医药卫生—中药学]