Effects of total flavone of Cydonia oblonga on high glucose-induced oxidative damage in human lens epithelial cells and its mechanism
10.13389/j.cnki.rao.2024.0147
- VernacularTitle:榅桲总黄酮对高糖诱导的人晶状体上皮细胞氧化损伤的影响及作用机制
- Author:
Yuanyuan LUO
1
;
Jingjie CAO
;
Haiying WANG
;
Chuan FENG
;
Taofu TANG
;
Jie HU
Author Information
1. 425000 湖南省永州市,湖南省永州职业技术学院医学技术学院
- Keywords:
total flavone of Cydonia oblonga;
miR-370;
oxidative stress;
apoptosis;
human lens epithelial cells
- From:
Recent Advances in Ophthalmology
2024;44(10):774-779
- CountryChina
- Language:Chinese
-
Abstract:
Objective To investigate the effect of total flavone of Cydonia oblonga on oxidative damage of human lens epithelial cells induced by high glucose and its mechanism.Methods A cell injury model was established by inducing human lens epithelial cells with high glucose.Human lens epithelial cells were cultured in the medium containing 30 mmol·L-1 glucose for 24 h,which was recorded as the high glucose group.Cells in the control group were cultured in a medium con-tainning 5.5 mmol·L-1 glucose for 24 h.Human lens epithelial cells were inoculated into 96-well plates with 5 × 103 cells per well,and treated with mediums containing 10 mmol·L-1,20 mmol·L-1,and 40 mmol·L-1 total flavone of Cydonia ob-longa combined with 30 mmol·L-1 glucose for 24 h.They were recorded as high glucose+low total flavone of Cydonia oblonga group,high glucose+medium total flavone of Cydonia oblonga group,and high glucose+high total flavone of Cydonia oblonga group.Human lens epithelial cells were transfected with anti-miR-NC and anti-miR-370 with Lipo-fectamine2000 transfection reagent,and treated with 30 mmol·L-1 glucose for 24 h,which were recorded as high glucose+anti-miR-NC group and high glucose+anti-miR-370 group.Human lens epithelial cells were transfected with miR-NC and miR-370 mimics,and treated with medium containing 30 mmol·L-1 glucose and 40 mmol·L-1 total flavone of Cydonia oblonga for 24 h,which were labeled as high glucose+total flavone of Cydonia oblonga+miR-NC group and high glucose+total flavone of Cydonia oblonga+miR-370 group.The activities of superoxide dismutase(SOD)and cata-lase(CAT),and the content of malondialdehyde(MDA)were measured by Enzyme-linked immunosorbent assay;flow cy-tometry was applied to detect apoptosis rate;quantitative reverse transcription polymerase chain reaction was applied to detect the expression level of miR-370;Western blot was applied to detect the expression of apoptosis-related proteins.Results Compared with the control group,the activities of SOD and CAT decreased and the content of MDA increased in the human lens epithelial cells of the high glucose group,and the differences were statistically significant(all P<0.05);compared with the high glucose group,the activities of SOD and CAT significantly increased and the content of MDA signifi-cantly decreased in the high glucose+low total flavone of Cydonia oblonga group,the high glucose+medium total fla-vone of Cydonia oblonga group and the high glucose+high total flavone of Cydonia oblonga group(all P<0.05).Com-pared with the control group,apoptosis rate,and protein expressions of Caspase-3 and Caspase-9 of human lens epithelial cells in the high glucose group significantly increased(all P<0.05);compared with the high glucose group,the apoptosis rate,Caspase-3 and Caspase-9 protein expressions of human lens epithelial cells in the high glucose+low total flavone of Cydonia oblonga group,high glucose+medium total flavone of Cydonia oblonga group,and high glucose+high total fla-vone of Cydonia oblonga group significantly decreased(all P<0.05).The expression levels of miR-370 in human lens epi-thelial cells were 1.00±0.00,4.04±0.36,3.22±0.24,2.42±0.23 and 1.62±0.14 in the control group,high glucose group,high glucose+low total flavone of Cydonia oblonga group,high glucose+medium total flavone of Cydonia oblon-ga group,and high glucose+high total flavone of Cydonia oblonga group,respectively.There was a statistically different significance among the five groups(F=256.138,P<0.05).Compared with the high glucose+anti-miR-NC group,the ex-pression of miR-370 significantly decreased,the activities of SOD and CAT significantly increased,and the content of MDA significantly decreased in human lens epithelial cells of the high glucose+anti-miR-370 group(all P<0.001).Compared with the high glucose+anti-miR-NC group,apoptosis rate,and protein expressions of Caspase-3 and Caspase-9 of the hu-man lens epithelial cells in the high glucose+anti-miR-370 group significantly decreased(all P<0.001).Compared with the high glucose+total flavone of Cydonia oblonga+miR-NC group,the activities of SOD and CAT significantly de-creased,and the content of MDA,apoptosis rate,and protein expressions of Caspase-3 and Caspase-9 significantly in-creased in the human lens epithelial cells of high glucose+total flavone of Cydonia oblonga+miR-370 group(all P<0.001).Conclusion The expression of miR-370 increases in high glucose-induced human lens epithelial cells.Total fla-vone of Cydonia oblonga can inhibit the oxidative stress and apoptosis of high glucose-induced human lens epithelial cells,and the mechanism may be related to the decreased expression of miR-370.