葛根素对工频电磁场下人胚胎眼巩膜成纤维细胞增殖活性和Ⅰ型胶原及MMP-2表达的影响  

Influence of puerarin on proliferative activity of human fetal scleral fibroblasts and expression of collagen type Ⅰ and matrix metalloproteinase-2 in power frequency electromagnetic field

作　　者：苏强[1] 杨璐[2] 

机构地区：[1]山西医科大学第一医院,太原030001 [2]山西省眼科医院杨璐

出　　处：《中华劳动卫生职业病杂志》2016年第6期447-451,共5页Chinese Journal of Industrial Hygiene and Occupational Diseases

摘　　要：目的探讨葛根素对工频电磁场下人胚胎眼巩膜成纤维细胞（HFSFs）的增殖活性和细胞内基质金属蛋白酶-2（MMP-2）及Ⅰ型胶原表达的影响。方法培养HFSFs细胞，分为4个辐射组（0．2mT、50Hz）和对照组，辐射组分别加入0．0、1．0、5．0、10．0μmol／L葛根素；采用3-（4，5-二甲基噻唑-2）-2，5-二苯基四氮唑溴盐（MTT）比色法测定HFSFs的增殖活性；采用蛋白质印迹（Western blot）法和实时定量荧光聚合酶链式反应（RT-qPCR）法分别测定葛根素对工频电磁场引起HFSFs Ⅰ型胶原和MMP-2蛋白和mRNA表达的变化。结果随着培养时间的延长，辐射组和对照组HFSFs增殖活性明显增大，差异有统计学意义（F值分别为959．472、279．468，均P〈0．01）；辐射组HFSFs24和48h增殖活性分别为0．432±0．038、0．591±0．017，明显低于对照组（0．536±0．034、0．801±0．020），差异均有统计学意义（均P〈0．01）。HFSFs在0．0、1．0、5．0、10．0μmol／L浓度葛根素的干预下增殖活性逐渐增强，A值分别为0．598±0．031、0．809±0．041、0．910±0．037、0．983±0．054，差异均有统计学意义（P〈0．05）。暴露24h后，辐射组Ⅰ型胶原蛋白表达明显低于对照组，MMP-2蛋白表达明显高于对照组，差异有统计学意义（t=7．917、7．831，均P〈0．01）；与0．0μmol／L葛根素组比较，1．0、5．0、10．0μmol／L葛根素组Ⅰ型胶原蛋白表达明显增高，MMP-2蛋白表达明显降低，差异均有统计学意义（均P〈0．01）。与对照组相比，辐射组HFSFs细胞Ⅰ型胶原mRNA表达下降，MMP-2mRNA表达升高，差异有统计学意义（t=17．293、16．378，P〈0．01）；与0．0μmol／L葛根素组比较，1．0、5．0、10．0μmol／L葛根素组Ⅰ型胶原mRNA表达明显增高，MMP-2mRNA表达明显降低，差异均有统计学意义（P〈0．01）。结论葛根素能够抑制工频电磁场下HFSFs的增�Objective To investigate the influence of puerarin on the proliferative activity of human fetal scleral fibroblasts （HFSFs） and the expression of collagen type I and matrix metalloproteinase-2 （MMP-2） in HFSFs in the power frequency electromagnetic field. Methods HFSFs were cultured and divided into four radiation groups （0.2 mT, 50 Hz） and control group. Puerarin at concentrations of 0.0, 1.0, 5.0, and 10.0 μmol/L was added to the four radiation groups, respectively. The methyl thiazolyl tetrazolium colorimetry was used to measure the influence on the proliferative activity of HFSFs, Western blot and RT-qPCR were used to measure the protein and mRNA expression of collagen type Ⅰ and MMP-2 induced by puerarin in the power frequency electromagnetic field. Results The radiation groups and control group showed significant increases in the proliferative activity of HFSFs over the culture time （F= 959.472 and 279.468, both P〈0.01）. At 24 and 48 hours, the radiation groups showed significantly lower proliferative activity than the control group （0.432±0.038/ 0.591±0.017 vs 0.536±0.034/0.801 ±0.020, both P〈0.01 ）. With the increasing concentration of puerarin （0.0, 1.0, 5.0, and 10.0 μmol/L）, the proliferative activity of HFSFs significantly increased, with A values of 0.598± 0.031, 0.809±0.041, 0.910±0.037, and 0.983±0.054, respectively （P〈0.05）. After exposure for 24 hours, the radiation groups showed significantly lower protein expression of collagen type Ⅰ and significantly higher protein expression of MMP-2（t=7.917 and 7.831, both P〈0.01 ） ; compared with the 0.0 μmol/L puerarin group, the 1.0, 5.0, and 10.0 μmol/L puerarin groups showed significant increases in the protein expression of collagen type I and significant reductions in the protein expression of MMP-2 （all P〈0.01 ）. Compared with the control group, the radiation groups showed significant reductions in the mRNA expression of collagen type Ⅰ and MMP-2 （t=17.293 and 16.378, both P〈0.01）

关 键 词：葛根素 电磁场 人胚胎眼巩膜成纤维细胞 Ⅰ型胶原 基质金属蛋白酶-2 

分 类 号：R285[医药卫生—中药学]