[1]胡红利,肖中举.丹参酮ⅡA对人角膜基质细胞纤维化的抑制作用[J].眼科新进展,2018,38(12):1114-1118.[doi:10.13389/j.cnki.rao.2018.0263]
　HU Hong-Li,XIAO Zhong-Ju.Inhibitory effect of tanshinone ⅡA on fibrosis of human corneal stromal cells[J].Recent Advances in Ophthalmology,2018,38(12):1114-1118.[doi:10.13389/j.cnki.rao.2018.0263]
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丹参酮ⅡA对人角膜基质细胞纤维化的抑制作用/HTML
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《眼科新进展》[ISSN:1003-5141/CN:41-1105/R]

卷:
38卷
期数:
2018年12期
页码:
1114-1118
栏目:
实验研究
出版日期:
2018-12-05

文章信息/Info

Title:
Inhibitory effect of tanshinone ⅡA on fibrosis of human corneal stromal cells
作者:
胡红利; 肖中举
510515　广东省广州市，南方医科大学基础医学院生理学教研室
Author(s):
HU Hong-Li; XIAO Zhong-Ju
Department of Physiology，School of Basic Medical Sciences，Southern Medical University，Guangzhou 510515，Guangdong Province，China
关键词:
丹参酮ⅡA; 转化生长因子-β1; 人角膜基质细胞纤维化
Keywords:
tanshinone ⅡA; transforming growth factor-β1; human corneal stromal cell fibrosis
分类号:
R772
DOI:
10.13389/j.cnki.rao.2018.0263
文献标志码:
A
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5.0 μg·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。结果　5.0 μg·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。2.5 μmol·L-1和5.0 μmol·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。5.0 μg·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；5.0 μmol·L-1 TSN联合5.0 μg·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。
Abstract:
Objective　To investigate the effects of tanshinone ⅡA （TSN） on the fibrosis of human keratocyte （HK） induced by transforming growth factor-β1 （TGF-β1），and to test whether tanshinone ⅡA 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 5.0 μg·L-1 TGF-β1 and different concentrations of TSN.The expression of α-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 of α-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 5.0 μg·L-1TGF-β1.Both 2.5 μmol·L-1 and 5.0 μmol·L-1 TSN could inhibit the expression of α-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 5.0 μg·L-1 TGF-β1 was long fusiform with polarities and the cells were polar.The cells cultured with 5.0 μmol·L-1 TSN combined with 5.0 μg·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.

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备注/Memo

备注/Memo:
国家自然科学基金资助（编号：31671083）
更新日期/Last Update: 2018-12-04