Role of Notch1 and autophagy on high glucose-induced human retinal pig-ment epithelial cells
10.13389/j.cnki.rao.2024.0148
- VernacularTitle:Notch1和自噬对高糖诱导下的人视网膜色素上皮细胞的作用
- Author:
Tingting QIN
1
,
2
;
Suhan WANG
;
Leying ZHANG
;
Jiaojiao WANG
;
Zongming SONG
Author Information
1. 453003 河南省新乡市,新乡医学院研究生院
2. 450003 河南省郑州市,河南省人民医院眼科,河南省立眼科医院
- Keywords:
autophagy;
Notch signaling pathway;
human retinal pigment epithelial cells;
high glucose
- From:
Recent Advances in Ophthalmology
2024;44(10):780-785
- CountryChina
- Language:Chinese
-
Abstract:
Objective To investigate the relationship between Notch1 and autophagy under high glucose conditions and to explore the effects of Notch1 inhibitor DAPT and autophagy inhibitor 3-MA on human retinal pigment epithelial cells cultured in high glucose conditions.Methods Via preliminary experiment,25 mmol·L-1 glucose was used as the high glucose culture medium of adult retinal pigment epithelial(ARPE)-19 cells,and 5 mmol·L-1 3-MA was adopted as the au-tophagy inhibitor.ARPE-19 cells cultured in vitro were randomly divided into four groups:control group(treated with 5 mmol·L-1 glucose for 48 h),high glucose group(treated with 25 mmol·L-1 glucose for 48 h),high glucose+DAPT group(treated with 40 μmol·L-1 DAPT for 2 h and then 25 mmol·L-1 glucose for 48 h),and high glucose+3-MA group(treated with 5 mmol·L-1 3-MA for 2 h and then 25 mmol·L-1 glucose for 48 h).A transmission electron microscope was used to observe the ultrastructure of cells in each group.Cell proliferation and migration were observed using Cell Counting Kit-8 and scratch assays.Western blot was used to detect the protein expression levels of Notch1 and autophagy-related proteins LC3 and Beclin1.Reverse transcription-polymerase chain reaction was used to measure the relative messenger ri-bonucleic acid(mRNA)expression levels of Notch1,LC3 and Beclin1 of cells in each group.Results Transmission elec-tron microscope showed that cells in the control group had normal structures,with round or oval nuclei and a few autopha-gosomes.In the high glucose group,cells exhibited slightly obvious injury,with uneven cytoplasm and numerous autolyso-somes.Compared to the control group,ARPE-19 cells in the high glucose group had increased proliferation and migration abilities,and higher mRNA and protein expression levels of Notch1,LC3 and Beclin1(all P＜0.05).Compared to the high glucose group,ARPE-19 cells in the high glucose+DAPT group showed decreased proliferation and migration abilities,and lower mRNA and protein expression levels of Notch1,LC3 and Beclin1(all P＜0.05).The high glucose+3-MA group showed reduced proliferation and migration abilities,as well as decreased mRNA and protein expression levels of LC3 and Beclin1(all P＜0.05)compared to the high glucose group.Conclusion High glucose can activate Notch1 and the auto-phagy process,promoting the proliferation of ARPE-19 cells.In the high glucose+DAPT group and high glucose+3-MA group,the autophagy process is inhibited to a certain extent,thereby restraining cell proliferation.
