葛根素对极低频电磁场下人胚胎眼巩膜成纤维细胞增殖活性及I型胶原蛋白表达的影响  

Effect of Puerarin on human fetal scleral fibroblasts＇ expression of collagen type I following exposure to extremely low frequency electromagnetic field

作　　者：蔡晓静[1] 朱煌[1] 赵立全[1] 孔繁琪[1] 冯婕妤[1] 牛宗镇[1] 

机构地区：[1]上海交通大学医学院附属新华医院眼科,200092

出　　处：《中华眼视光学与视觉科学杂志》2013年第8期479-483,共5页Chinese Journal Of Optometry Ophthalmology And Visual Science

基　　金：上海市卫生局中药科研基金（2009S023）

摘　　要：目的研究葛根素对极低频电磁场（ELF—EMFs）作用下人胚胎眼巩膜成纤维细胞（HFSFs）增殖活性及I型胶原蛋白（COLlAl）表达的影响。方法实验研究。体外培养HFSFs．根据是否暴露于0．2mT电磁场分为暴露组和非暴露组，暴露组又分为不同终浓度葛根素组（O．0、0．1、1．0、10．0μmol／L）。MTT比色法检测不同暴露时间（0、12、24、48h）暴露组和非暴露组HFSFs增殖活性，及各浓度葛根素组HFSFs暴露于0．2mT电磁场24h时增殖活性；用Real．timePCR和Western—Blot检测非暴露组和暴露于0．2mT电磁场24h时各浓度葛根素组HFSFsmRNA转录水平和COLlAl蛋白表达。2组间比较采用独立样本t检验，各组整体比较采用单因素方差分析，组间多重比较采用LSD—t检验。结果暴露组各时间点HFSFs增殖活性差异有统计学意义（t=4．560，P〈0．05）．暴露于0．2mT磁场24h开始即出现HFSFs增殖受抑（t=5．000，P〈0．01）；与非暴露组相比，暴露组细胞内COL1A1蛋白表达下调（t=7．956，P〈0．01），同时mRNA转录也下调（t=17．364，P〈0．01），而1．0μmol／L葛根素开始使HFSFs增殖活性增强（P〈0．01），0．1μmol／L葛根素即使暴露组HFSFs内COL1A1蛋白（P〈0．05）和mRNA表达上调（P〈0．05），且浓度越高变化越明显。结论葛根素可逆向改变ELF．EMFs引起的HFSFs增殖活性下降、细胞内COL1A1mRNA和蛋白表达下调，可能对巩膜基质重塑具有预防作用。[Abstract] Objective To observe the effect of Puerarin on human fetal scleral fibroblasts＇ （HFSFs） proliferation and expression of collagen type I （COL1A1） following exposure to extremely low frequency electromagnetic field （ELF-EMFs）. Methods Experiment study. HFSFs were cultured and divided into control groups and radiation groups, which were exposed to 0.2 mT ELF-EMFs. The radiation groups were divided into groups of different puerarin concentrations （0.0, 0.1, 1.0, 10.0 ~mol/L）. The proliferation of HFSFs under different exposure time （0, 12, 24, 48 hours） and under different final puerarin concentrations were detected by MT1~ skills. The expressions of COL1A1 mRNA and protein of all these groups were detected by real-time PCR and Western-Blot. Single factor analysis of variance was first used to determine whether there was difference among these groups, LSD-t was then used to figure out which groups had significant difference. And the difference between two groups are analysed by independent sample t test. Results There was signifcant difference between different radiating time in radiations groups （F--4.560, P〈0.05）, and the proliferation of HFSFs （A490） of radiation groups was inhibited after exposed to 0.2 mT ELF-EMFs for 24 hours （t=5.000, P〈0.O1） with shrinking protein expression of COL1A1 （t=7.956, P〈0.01） and decreasing mRNA expression （t=17.364, P〈O.01）. However, Puerarin （1.0μmol/L） enhanced HFSFs＇ proliferation activity （P〈0.01） and 0.1 μmol/L increased the protein expression of COL1A1 （P〈0.05） and its mRNA expression （P〈0.05）. The effects were more obvious under higher-concentration of puerarin （1.0, 10.0 μmol/L）. Conclusion ELF-EMFs inhibited the proliferation of HFSFs and expression of COL1A1, but puerarin can reverse these changes, suggesting that puerarin play a postive role in sclera remodeling.

关 键 词：葛根素 极低频电磁场 人胚胎眼巩膜成纤维细胞 I型胶原蛋白 巩膜基质重塑 

分 类 号：R285.5[医药卫生—中药学]