中华眼视光学与视觉科学杂志
中華眼視光學與視覺科學雜誌
중화안시광학여시각과학잡지
CHINESE JOURNAL OF OPTOMETRY OPHTHALMOLOGY AND VISUAL SCIENCE
2013年
8期
479-483
,共5页
蔡晓静%朱煌%赵立全%孔繁琪%冯婕妤%牛宗镇
蔡曉靜%硃煌%趙立全%孔繁琪%馮婕妤%牛宗鎮
채효정%주황%조립전%공번기%풍첩여%우종진
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
葛根素%極低頻電磁場%人胚胎眼鞏膜成纖維細胞%Ⅰ型膠原蛋白%鞏膜基質重塑
갈근소%겁저빈전자장%인배태안공막성섬유세포%Ⅰ형효원단백%공막기질중소
Puerarin%Extremely low frequency electromagnetic field%Human fetal sclera fibroblast%Collagen type Ⅰ%Sclera remodeling
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(COL1A1)表达的影响.方法 实验研究.体外培养HFSFs,根据是否暴露于0.2 mT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(0.0、0.1、1.0、10.0 μmol/L).MTT比色法检测不同暴露时间(0、12、24、48 h)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0.2 mT电磁场24 h时增殖活性;用Real-time PCR和Western-Blot检测非暴露组和暴露于0.2 mT电磁场24 h时各浓度葛根素组HFSFsmRNA转录水平和COL1A1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(F=4.560,P＜0.05),暴露于0.2 mT磁场24 h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1.0 μmol/L葛根素开始使HFSFs增殖活性增强(P,＜0.01),0.1 μmol/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COL1A1 mRNA和蛋白表达下调,可能对巩膜基质重塑具有预防作用.
目的 研究葛根素對極低頻電磁場(ELF-EMFs)作用下人胚胎眼鞏膜成纖維細胞(HFSFs)增殖活性及Ⅰ型膠原蛋白(COL1A1)錶達的影響.方法 實驗研究.體外培養HFSFs,根據是否暴露于0.2 mT電磁場分為暴露組和非暴露組,暴露組又分為不同終濃度葛根素組(0.0、0.1、1.0、10.0 μmol/L).MTT比色法檢測不同暴露時間(0、12、24、48 h)暴露組和非暴露組HFSFs增殖活性,及各濃度葛根素組HFSFs暴露于0.2 mT電磁場24 h時增殖活性;用Real-time PCR和Western-Blot檢測非暴露組和暴露于0.2 mT電磁場24 h時各濃度葛根素組HFSFsmRNA轉錄水平和COL1A1蛋白錶達.2組間比較採用獨立樣本t檢驗,各組整體比較採用單因素方差分析,組間多重比較採用LSD-t檢驗.結果 暴露組各時間點HFSFs增殖活性差異有統計學意義(F=4.560,P＜0.05),暴露于0.2 mT磁場24 h開始即齣現HFSFs增殖受抑(t=5.000,P＜0.01)；與非暴露組相比,暴露組細胞內COL1A1蛋白錶達下調(t=7.956,P＜0.01),同時mRNA轉錄也下調(t=17.364,P＜0.01),而1.0 μmol/L葛根素開始使HFSFs增殖活性增彊(P,＜0.01),0.1 μmol/L葛根素即使暴露組HFSFs內COL1A1蛋白(P,＜0.05)和mRNA錶達上調(P＜0.05),且濃度越高變化越明顯.結論 葛根素可逆嚮改變ELF-EMFs引起的HFSFs增殖活性下降、細胞內COL1A1 mRNA和蛋白錶達下調,可能對鞏膜基質重塑具有預防作用.
목적 연구갈근소대겁저빈전자장(ELF-EMFs)작용하인배태안공막성섬유세포(HFSFs)증식활성급Ⅰ형효원단백(COL1A1)표체적영향.방법 실험연구.체외배양HFSFs,근거시부폭로우0.2 mT전자장분위폭로조화비폭로조,폭로조우분위불동종농도갈근소조(0.0、0.1、1.0、10.0 μmol/L).MTT비색법검측불동폭로시간(0、12、24、48 h)폭로조화비폭로조HFSFs증식활성,급각농도갈근소조HFSFs폭로우0.2 mT전자장24 h시증식활성;용Real-time PCR화Western-Blot검측비폭로조화폭로우0.2 mT전자장24 h시각농도갈근소조HFSFsmRNA전록수평화COL1A1단백표체.2조간비교채용독립양본t검험,각조정체비교채용단인소방차분석,조간다중비교채용LSD-t검험.결과 폭로조각시간점HFSFs증식활성차이유통계학의의(F=4.560,P＜0.05),폭로우0.2 mT자장24 h개시즉출현HFSFs증식수억(t=5.000,P＜0.01)；여비폭로조상비,폭로조세포내COL1A1단백표체하조(t=7.956,P＜0.01),동시mRNA전록야하조(t=17.364,P＜0.01),이1.0 μmol/L갈근소개시사HFSFs증식활성증강(P,＜0.01),0.1 μmol/L갈근소즉사폭로조HFSFs내COL1A1단백(P,＜0.05)화mRNA표체상조(P＜0.05),차농도월고변화월명현.결론 갈근소가역향개변ELF-EMFs인기적HFSFs증식활성하강、세포내COL1A1 mRNA화단백표체하조,가능대공막기질중소구유예방작용.
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 μmol/L).The proliferation of HFSFs under different exposure time (0,12,24,48 hours) and under different final puerarin concentrations were detected by MTT 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 significant 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＜0.01) with shrinking protein expression of COL1A1 (t=7.956,P＜0.01) and decreasing mRNA expression (t=17.364,P＜0.01).However,Puerarin (1.0 μmol/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.
