摘要
目的研究丹参酮ⅡA (tanshinoneⅡA,TSN)对转化生长因子β1 (transforming growth factor-β1,TGF-β1)诱导的人角膜基质细胞(human keratocyte,HK)纤维化的作用,探索该药物是否具有防治角膜瘢痕的潜能。方法采用胰蛋白酶消化培养HK,并传代至4-7代用于实验。通过TGF-β1诱导建立HK纤维化模型,并分别用5.0μg·L^(-1)TGF-β1和不同浓度的TSN联合给药方法对HK进行处理。采用Western blot检测α-平滑肌肌动蛋白(α-smooth muscle actin,α-SMA)与Vimentin的表达,qPCR检测纤连蛋白(fibronectin,FN)和I型胶原(collgen I,COL I) mRNA的表达;利用细胞免疫荧光染色观察HK细胞形态的改变氉。结果 5.0μg·L^(-1)TGF-β1可以显著上调α-SMA与Vimentin蛋白的表达和胞外基质成分FN mRNA(2.238±0.227)、COL I mRNA(3.554±0.526)的表达。2.5μmol·L^(-1)和5.0μmol·L^(-1)TSN均能抑制α-SMA蛋白的表达和FN mRNA(0.619±0.017、0.263±0. 006)、COL I mRNA(0.631±0.011、0.275±0.081)的表达。5.0μg·L^(-1)TGF-β1诱导组HK形态呈长梭形,具有两极,细胞呈极性分布; 5.0μmol·L^(-1)TSN联合5.0μg·L^(-1)TGF-β1培养HK形态同原始细胞形态,细胞呈三角形或星形。结论 TSN可抑制TGF-β1诱导的HK纤维化,具有抗角膜瘢痕的潜力。
Objective To investigate the effects of tanshinoneⅡA(TSN)on the fibrosis of human keratocyte(HK)induced by transforming growth factor-β1(TGF-β1),and to test whether tanshinoneⅡA has the potential to prevent corneal scarring.Methods Human corneal stromal cells were harvested by trypsin and cultured to 4-7 passages.The establishment of keratocyte fibrosis model was induced by TGF-β1,and treated with 5.0μg·L^-1 TGF-β1 and different concentrations of TSN.The expression ofα-SMA,Vimentin protein were determined by Western blot,and the expression of FN and collagen Ⅰ(COL Ⅰ)gene were detected by Real-time Quantitative PCR Detection System,and immunofluorescence staining for cell morphology,to examine the inhibitory effect of TSN on HK fibrosis.Results The expression ofα-SMA and Vimentin protein and the transcription of extracellular matrix components FN(2.238±0.227)and COL I(3.554±0.526)genes were up-regulated significantly after the treatment with 5.0μg·L^-1 TGF-β1.Both 2.5μmol·L-1 and 5.0μmol·L^-1 TSN could inhibit the expression ofα-SMA protein and the transcription of FN(0.619±0.017,0.263±0.006)and COL Ⅰ(0.631±0.011,0.275±0.081)genes.The morphology of the cells induced by 5.0μg·L^-1 TGF-β1 was long fusiform with polarities and the cells were polar.The cells cultured with 5.0μmol·L^-1 TSN combined with 5.0μg·L^-1 TGF-β1 showed the same morphology as the original cells,and the cells were triangular or star-shaped.Conclusion TSN can inhibit the differentiation of human corneal fibroblasts induced by TGF-β1 and has the potential to resist corneal scarring.
作者
胡红利
肖中举
HU Hong-Li;XIAO Zhong-Ju(Department of Physiology,School of Basic Medical Sciences,Southern Medical University,Guangzhou 510515,Guangdong Province,China)
出处
《眼科新进展》
CAS
北大核心
2018年第12期1114-1118,共5页
Recent Advances in Ophthalmology
基金
国家自然科学基金资助(编号:31671083)~~
