[1]刘畅,蔡雯婷,苏途,等.槲皮素对转化生长因子β1介导的视网膜色素上皮细胞上皮-间质转化过程的影响[J].眼科新进展,2021,41(7):638-642.[doi:10.13389/j.cnki.rao.2021.0131]
　LIU Chang,CAI Wenting,SU Tu,et al.Effect of quercetin on transforming growth factor β1-induced epithelial-mesenchymal transition process in retinal pigment epithelial cells[J].Recent Advances in Ophthalmology,2021,41(7):638-642.[doi:10.13389/j.cnki.rao.2021.0131]
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槲皮素对转化生长因子β1介导的视网膜色素上皮细胞上皮-间质转化过程的影响/HTML
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《眼科新进展》[ISSN:1003-5141/CN:41-1105/R]

卷:
41卷
期数:
2021年7期
页码:
638-642
栏目:
实验研究
出版日期:
2021-07-05

文章信息/Info

Title:
Effect of quercetin on transforming growth factor β1-induced epithelial-mesenchymal transition process in retinal pigment epithelial cells
作者:
刘畅; 蔡雯婷; 苏途; 于靖
211166　江苏省南京市，南京医科大学（刘畅，于靖）；200072　上海市，同济大学附属上海市第十人民医院眼科（蔡雯婷，苏途）
Author(s):
LIU Chang1; CAI Wenting2; SU Tu2; YU Jing1
1.Nanjing Medical University, Nanjing 211166,Jiangsu Province,China
2.Department of Ophthalmology, Shanghai Tenth People’s Hospital Affiliated to Tongji University, Shanghai 200072, China
关键词:
增生性玻璃体视网膜病变; 槲皮素; 上皮-间质转化; 转化生长因子-β1; 视网膜色素上皮细胞
Keywords:
proliferative vitreoretinopathy;  quercetin;  epithelial-mesenchymal transition;  transforming growth factor-β1;  retinal pigment epithelial cell
分类号:
R774
DOI:
10.13389/j.cnki.rao.2021.0131
文献标志码:
A
摘要:
目的　探讨槲皮素对转化生长因子β1（TGF-β1）介导的视网膜色素上皮(RPE)细胞增殖、迁移、I型胶原蛋白含量、上皮-间质转化相关标志物表达以及Smad信号通路的影响。方法　取人RPE细胞系（ARPE-19细胞）进行培养，依据干预药物浓度筛选结果设置4个组：对照组、10.0 mg·L-1TGF-β1组、10.0 mg·L-1TGF-β1+25 μmol·L-1槲皮素组、10.0 mg·L-1TGF-β1+50 μmol·L-1槲皮素组，ARPE-19细胞在4种不同的条件下培养24 h和48 h。采用CCK-8法测定各组细胞活性，Transwell实验检测细胞迁移情况，ELISA检测细胞中I型胶原蛋白含量，Western blot 检测各组细胞上皮-间质转化标志物表达情况及Smad2/3磷酸化情况。结果　处理细胞24 h或48 h，10.0 mg·L-1TGF-β1组细胞存活率均高于其他各组（均为P<0.05）；10.0 mg·L-1TGF-β1+25 μmol·L-1槲皮素组细胞存活率高于10.0 mg·L-1TGF-β1+50 μmol·L-1槲皮素组（P<0.05）。处理细胞24 h或48 h，10.0 mg·L-1TGF-β1组细胞迁移数均多于其他各组（均为P<0.05）；10.0 mg·L-1TGF-β1+25 μmol·L-1槲皮素组细胞迁移数多于10.0 mg·L-1TGF-β1+50 μmol·L-1槲皮素组（P<0.05）。10 mg·L-1 TGF-β1处理细胞48 h后，培养基上清液I型胶原蛋白含量升高（P<0.05），而加入不同浓度槲皮素均能够有效抑制I型胶原蛋白含量的升高（均为P<0.05）。槲皮素以浓度依赖的方式显著降低了TGF-β1所引起的N-钙黏合素和α平滑肌肌动蛋白的表达增高作用；槲皮素可以使紧密连接蛋白ZO-1和E-钙黏合素的表达增高。10 mg·L-1 TGF-β1处理细胞48 h后，Smad2/3磷酸化显著增加，而槲皮素可以抑制Smad2/3磷酸化。结论　槲皮素能够显著抑制TGF-β1介导的RPE细胞增殖、迁移和胶原分泌，通过调节Smad2/3磷酸化来抑制TGF-β1介导的RPE细胞的上皮-间质转化过程。
Abstract:
Objective　To investigate the effect of quercetin on transforming growth factor-β1 (TGF-β1)-induced retinal pigment epithelial (RPE) cell proliferation, migration, collagen I content, expression of epithelial-mesenchymal transition markers and Smad pathway.Methods　ARPE-19 cells were cultured and divided into 4 groups according to the intervention concentration:the normal control group, the 10.0 mg·L-1 TGF-β1 group, the 10.0 mg·L-1 TGF-β1+25 μmol·L-1 quercetin group, the 10.0 mg·L-1 TGF-β1+50 μmol·L-1 quercetin group. Cells were cultured for 24 hours and 48 hours under four different conditions. Cell counting kit-8 (CCK-8) assay was used to assess each group’s cell viability, Transwell assay was used to assess migration, ELISA was used to detect collagen I content, and Western blot analysis was employed to detect expression of epithelial-mesenchymal transition markers and condition of Smad2/3 phosphorylation. Results　Whether incubating for 24 hours or 48 hours, the cell viability of the 10.0 mg·L-1 TGF-β1 group was higher than that in other groups (all P<0.05); the cell viability of the 10.0 mg·L-1 TGF-β1+25 μmol·L-1 quercetin group was higher than that in the 10.0 mg·L-1 TGF-β1+50 μmol·L-1 quercetin group (P<0.05). Whether incubating for 24 hours or 48 hours, the number of cell migrating of the 10.0 mg·L-1 TGF-β1 group were more than that in other groups (all P<0.05); the numbers of cell migrating of the 10.0 mg·L-1 TGF-β1+25 μmol·L-1 quercetin group were more than that in the 10.0 mg·L-1 TGF-β1+50 μmol·L-1 quercetin group (P<0.05). The content of collagen I in the culture supernatant increased after cells were treated with 10 mg·L-1 TGF-β1 for 48 h (P<0.05), and various concentrations of quercetin effectively suppressed the rise of TGF-β1-induced cell collagen I content (all P<0.05). The increased expression level of N-cadherin and alpha-smooth muscle actin caused by TGF-β1 were significantly inhibited by quercetin in a concentration-dependent manner. Quercetin unregulated the expression of E-cadherin and tight junction protein ZO-1. The level of Smad2/3 phosphorylation obviously increased after cells were treated with 10 mg·L-1 TGF-β1 for 48 h, of which the process was restrained by quercetin.Conclusion　Quercetin can significantly inhibit the proliferation, migration and collagen secretion induced by TGF-β1 in RPE cells, thereby inhibiting the TGF-β1-induced epithelial-mesenchymal transition process in RPE cells by regulating Smad2/3 phosphorylation.

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更新日期/Last Update: 2021-07-05