Effects of salidroside pretreatment on high glucose-induced epithelial-mesen-chymal transition in ARPE-19 cells and its mechanism
10.13389/j.cnki.rao.2024.0116
- VernacularTitle:红景天苷预处理对高糖诱导的ARPE-19细胞上皮-间充质转化的作用与机制研究
- Author:
Miao HAN
1
;
Haijuan SUI
;
Dongbo PANG
Author Information
1. 121000 辽宁省锦州市,锦州医科大学附属第一医院
- Keywords:
salidroside;
high glucose;
human retinal pigment epithelium;
epithelial-mesenchymal transition;
transfor-ming growth factor-β1
- From:
Recent Advances in Ophthalmology
2024;44(8):607-612
- CountryChina
- Language:Chinese
-
Abstract:
Objective To explore the effects of salidroside(SAL)on high glucose-induced epithelial-mesenchymal transition in ARPE-19 cells and its mechanism.Methods The appropriate glucose concentration for stimulating ARPE-19 cell proliferation was determined to be 50 mmol·L-1 using the MTT method.ARPE-19 cells were divided into the normal control group(Control group)(cultured with 5 mmol·L-1 D-glucose and 45 mmol·L-1 mannitol),high glucose group(HG group)(cultured with 50 mmol·L-1 glucose),HG+low SAL group,HG+medium SAL group,and HG+high SAL group,with low,medium,and high SAL concentrations of 20 μmol·L-1,80 μmol·L-1,and 320 μmol·L-1,respective-ly.After 4 hours of SAL pretreatment,50 mmol·L-1 glucose was added to treat cells for 48 hours.The cell proliferation rate was detected using the MTT method;the cell migration activity was detected using the scratch experiment;the expres-sion levels of α-smooth muscle actin(α-SMA)and Vimentin in cells were measured using the immunofluorescence;the protein expression levels of α-SMA,Vimentin,fibronectin(FN),collagen type Ⅰ(Col Ⅰ),E-Cadherin,transforming growth factor-β1(TGF-β1),p-Smad2 and p-Smad3 in cells were measured using Western blot.Results Compared with the Control group,the proliferation rate and wound healing percentage of ARPE-19 cells in the HG group increased;the relative expression levels of α-SMA,Vimentin,FN,and Col Ⅰ proteins in the cells increased;the relative expression level of E-cad-herin protein in the cells decreased;the relative expression levels of TGF-β1,p-Smad2,and p-Smad3 proteins in the cells increased,and the differences were statistically significant(all P＜0.01).Compared with the HG group,the proliferation rate and wound healing percentage of ARPE-19 cells in the HG+low SAL group,HG+medium SAL group,and HG+high SAL group decreased,the relative expression levels of α-SMA,Vimentin,FN,Col Ⅰ,TGF-β1,p-Smad2,and p-Smad3 pro-teins in the cells decreased in a concentration dependent manner,while the relative expression level of E-cadherin protein increased in a concentration dependent manner,and the differences were statistically significant(all P＜0.05).Conclu-sion SAL can reduce the proliferation and migration ability of high glucose-induced ARPE-19 cells and intervene in their epithelial-mesenchymal transition process.This may be related to SAL's inhibition of the TGF-β/Smads signaling pathway in ARPE-19 cells.
