转化生长因子-β(TGF-β)影响豚鼠巩膜成纤维细胞增殖和α-平滑肌肌动蛋白(α-SMA)表达的研究  

Effects of TGF-β on cellular proliferation and α-SMA expression in guinea pig scleral fibroblasts

作　　者：张兰兰[1] 刘琼[2] 于健[2] 唐晓娟[2] 徐静[2] 

机构地区：[1]南方医科大学南方医院临床实验中心,广东省广州市510515 [2]南方医科大学南方医院眼科,广东省广州市510515

出　　处：《眼科新进展》2016年第11期1011-1015,共5页Recent Advances in Ophthalmology

基　　金：国家自然科学基金资助(编号:30901650);南方医院院长基金重点项目(编号:2012A002)~~

摘　　要：目的本研究探讨转化生长因子-β(transforming growth factor,TGF-β)体外刺激豚鼠巩膜成纤维细胞,观察豚鼠巩膜成纤维细胞增殖和α-平滑肌肌动蛋白(α-smooth muscle actin,α-SMA)的表达。方法原代培养豚鼠巩膜成纤维细胞并鉴定,MTT法检测豚鼠巩膜成纤维细胞的增殖,培养液分别加入20 ng·m L-1TGF-β1、TGF-β2、TGF-β3,分别于12 h、24 h、48 h、72 h,Real-time PCR检测α-SMA mRNA,Western-blotting和免疫荧光化学法检测其蛋白表达。结果与对照组相比,TGF-β1、TGF-β2和TGF-β3促进豚鼠巩膜成纤维细胞增殖,呈剂量依赖性(P<0.05),其中TGF-β1促进豚鼠巩膜成纤维细胞增殖作用最强(P<0.05);TGF-β1、TGF-β2和TGF-β3均增加α-SMA的表达(均为P<0.001),TGF-β3促进α-SMA表达作用最强(P<0.001)。结论 TGF-β通过调控细胞外基质合成参与调控巩膜组织重塑。Objective To investigate the role of transforming growth factor-β （TGF-β） in cellular proliferation and （α-smooth muscle actin （α-SMA） expression in guinea pig scleral fibroblasts （GSF）. Methods Primary culture and identification of GSF were followed by the MMT cell proliferation assay to assess proliferation. The culture was supplemented with 20 ng · mL^- 1 of TGF-β1, TGF-β2 and TGF-β3, which were removed at 12 hours,24 hours,48 hours and 72 hours. Real-time polymerase chain reaction was performed to evaluate the mRNA expression of α-SMA. Western blotting and immunofluorescent staining were also conducted to examine expression of α-SMA. Resuits TGF-β up-regulated GSF proliferation, and this proliferation was dose-depend- ent （P 〈 0.05 ）, while TGF-β1 demonstrated the strongest regulative effect （ P 〈 0.05 ）. All three TGF-β isoforms enhanced the expression of α-SMA （ all P 〈 0. 001 ）, wihile TGF-β3 demonstrated the strongest regulative effect （ P 〈 0. 001 ）. Conclusion TGF- β participates in scleral remodeling through the regulation of extracellular matrix synthesis.

关 键 词：转化生长因子-Β Α-平滑肌肌动蛋白 细胞增殖 近视 

分 类 号：R772.3[医药卫生—眼科]