Experimental study on the regulation of migration of retinal pigment epithelial cells by bone morphogenetic protein 4
10.3760/cma.j.cn511434-20230217-00069
- VernacularTitle:骨形成蛋白4调控视网膜色素上皮细胞迁移
- Author:
Wenbo LI
1
;
Jingjing CAO
;
Tongtong ZHUANG
;
Qing WANG
;
Lijie DONG
Author Information
1. 天津医科大学眼科医院、眼视光学院、眼科研究所 国家眼耳鼻喉疾病临床医学研究中心天津市分中心 天津市视网膜功能与疾病重点实验室, 天津 300384
- Keywords:
Proliferative diabetic retinopathy;
Bone morphogenetic protein 4;
Epithelial-mesenchymal transformation;
Retinal pigment epithelium cells;
Proliferation;
M
- From:
Chinese Journal of Ocular Fundus Diseases
2024;40(3):208-214
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To observe the effect of bone forming protein 4 (BMP4) on the proliferation and migration of human retinal pigment epithelium (RPE) cells under oxidative stress, and to preliminarily explore its effect on epithelial-mesenchymal transition (EMT) of RPE cells.Methods:Human RPE cells cultured in vitro were divided into normal group, pure 4-hydroxynonenal (HNE) group (4-HNE group), 4-HNE+NC group and 4-HNE+ small interfering BMP (siBMP4) group. The effect of 4-HNE on the proliferation of RPE cells was detected by thiazole blue colorimetry. The effects of 4-HNE and BMP4 on cell migration were determined by cell scratch test. The expression of BMP4 was detected by immunofluorescence staining, Western blot and real-time quantitative polymerase chain reaction. The transfection efficiency of siBMP4 was observed by fluorescence microscopy. Mitochondrial reactive oxygen species (MitoSOX) were detected by flow cytometry. The expression of EMT markers E-cadherin and Fibronection were detected by immunofluorescence assay. t-test was used for comparison between the two groups, and one-way analysis of variance was used for comparison between the three groups. Results:Compared with normal group, cell proliferation and migration ability of 4-HNE group were significantly enhanced, with statistical significance ( t=21.619, 24.469; P<0.05). The expression of BMP4 in cells was significantly increased, and the difference was statistically significant ( t=19.441, P<0.05). The relative expression levels of BMP4 mRNA and protein were also significantly increased, with statistical significance ( t=26.163, 37.163; P<0.05). After transfection with siBMP4 for 24 h, the transfection efficiency of BMP4 in RPE cells was>90%. Compared with 4-HNE group and 4-HNE+NC group, the relative expression levels of BMP4 protein ( F=27.241), mRNA ( F=36.943), cell mobility ( F=46.723) and MitoSOX expression levels ( F=39.721) in normal group and 4-HNE+siBMP4 group were significantly decreased. The differences were statistically significant ( P<0.05). The epithelial marker E-cadherin increased significantly, while the mesenchymal marker Fibronection decreased significantly, with statistical significance ( F= 51.722, 45.153; P<0.05). Conclusions:BMP4 inhibits RPE proliferation and migration under oxidative stress. BMP4 is involved in inducing EMT in RPE cells.
