Effect of total flavones of Dracocephalum moldavica L.on high-glucose in-duced oxidative damage to retinal ganglion cells and its mechanism
10.13389/j.cnki.rao.2024.0177
- VernacularTitle:香青兰总黄酮(TFDM)对高糖诱导的视网膜神经节细胞氧化损伤的影响及其机制
- Author:
Liying GU
1
;
Shengfu YANG
;
Qiming ZHANG
;
Shuxin WANG
Author Information
1. 430014 湖北省武汉市,江汉大学附属医院武汉市第六医院眼科
- Keywords:
total flavones of Dracocephalum moldavica L.;
miR-93-5p;
E2F transcription factor 1;
high glucose;
reti-nal ganglion cells;
cell apoptosis;
oxidative stress
- From:
Recent Advances in Ophthalmology
2024;44(12):937-942
- CountryChina
- Language:Chinese
-
Abstract:
Objective To investigate the effects of total flavones of Dracocephalum moldavica L.(TFDM)on oxida-tive damage to retinal ganglion cells(RGCs)induced by high glucose(HG)and its mechanism.Methods RGCs of mice were taken as the research subjects.RGCs were inoculated in 24-well plates(with 2.5 × 104 cells in each well)and divided into control group(cultured with medium containing 10％fetal bovine serum for 48 h),HG group(cultured with medium containing 30 mmol·L-1 glucose for 48 h),HG+TFDM-L group(cultured with medium containing 30 mmol·L-1 glucose and 25 mg·L-1 TFDM for 48 h),HG+TFDM-M group(cultured with medium containing 30 mmol·L-1 glucose and 50 mg·L-1 TFDM for 48 h),HG+TFDM-H group(cultured with medium containing 30 mmol·L-1 glucose and 100 mg·L-1 TFDM for 48 h),miR-NC group(transfected with miR-NC and then cultured with medium containing 30 mmol·L-1 glu-cose for 48 h),miR-93-5p group(transfected with miR-93-5p mimics and then cultured with medium containing 30 mmol·L-1 glucose for 48 h),anti-miR-NC group(transfected with anti-miR-NC and then cultured with medium containing 100 mg·L-1 TFDM and 30 mmol·L-1 glucose for 48 h),and anti-miR-93-5p group(transfected with anti-miR-93-5p and then cultured with medium containing 100 mg·L-1 TFDM and 30 mmol·L-1 glucose for 48 h).Levels of RGCs oxidative stress indexes in each group were detected according to the kit instructions.The thiobarbituric acid method was used to measure the level of malondialdehyde(MDA),the colorimetric method was adopted to detect the levels of catalase(CAT)and 8-hydroxydeoxyguanosine(8-OHdG),apoptosis rate was detected by flow cytometry,the messenger ribonucleic acid(mR-NA)expressions were analyzed by real-time quantitative polymerase chain reaction,the targeting regulation of miR-93-5p and E2F transcription factor 1(E2F1)were verified by dual luciferase reporter assay,and the protein expressions were de-tected by Western blot.Statistical analysis was conducted on the data of each group.Results In the HG group,the lev-els of MDA and 8-OHdG,apoptosis rate and Bax protein expression of RGCs were higher than those in the control group(all P＜0.05);the CAT level and Bcl-2 protein expression were lower than those in the control group(both P＜0.05).In the HG+TFDM-L group,HG+TFDM-M group and HG+TFDM-H group,the levels of MDA and 8-OHdG,apoptosis rate and Bax protein expression of RGCs were lower than those in the HG group(all P＜0.05);the CAT level and Bcl-2 protein expression were higher than those in the HG group,and those indexes gradually tended to those in the control group with the increase of TFDM concentrations(all P＜0.05).In the HG group,the miR-93-5p expression in RGCs was lower than that in the control group,and the mRNA and protein expressions of E2F1 were higher than those in the control group(all P＜0.05).In the HG+TFDM-L group,HG+TFDM-M group and HG+TFDM-H group,the miR-93-5p expression in RGCs was higher than that in the HG group,and the mRNA and protein expressions of E2F1 were lower than those in the HG group(all P＜0.05).The protein expression of E2F1 in RGCs of the miR-93-5p group was lower than that in the miR-NC group,and the protein expression of E2F1 in RGCs of the anti-miR-93-5p group was higher than that in the anti-miR-NC group(both P＜0.05).In the miR-93-5p group,the miR-93-5p expression was higher than that in the miR-NC group,the levels of MDA and 8-OHdG,apoptosis rate,and protein expressions of Bax and E2F1 were lower than those in the miR-NC group,and the CAT level and Bcl-2 protein expression were higher than those in the miR-NC group(all P＜0.05).Con-clusion TFDM can inhibit oxidative stress and cell apoptosis,and then reduce the damage to RGCs induced by HG.The mechanism may involve the regulation of miR-93-5p/E2Fl expression.
