Effects of the modified Zhujing Pill-medicated serum to modulate PINK1/Parkin-mediated mitochondrial autophagy on the pressure-induced retinal ganglion cell injury model in rats
10.13389/j.cnki.rao.2025.0060
- VernacularTitle:驻景丸加减方含药血清调控PINK1/Parkin介导的线粒体自噬对加压诱导大鼠RGCs损伤模型的影响
- Author:
Jia GAO
1
;
Hongji LIU
;
Dan ZHANG
;
Wei WANG
Author Information
1. 610000 四川省成都市,成都中医药大学
- Publication Type:Journal Article
- Keywords:
modified Zhujing Pill;
retinal ganglion cell;
medicated serum;
mitochondrial autophagy;
PINK1/Parkin
- From:
Recent Advances in Ophthalmology
2025;45(5):342-347,364
- CountryChina
- Language:Chinese
-
Abstract:
Objective To investigate the effects of modified Zhujing Pill(ZJP)-medicated serum on mitochondrial autophagy and related proteins in a pressure-induced retinal ganglion cell(RGC)injury model in rats.Methods The modified ZJP-medicated serum and blank serum were prepared by the intragastric administration of modified ZJP(10 mL·kg-1)to 10 male Sprague-Dawley(SD)rats and equivalent normal saline to another 10 male SD rats.The RGC injury models of rats were constructed under the in vitro pressure of 0 mmHg,20 mmHg,40 mmHg,60 mmHg,and 80 mmHg(1 kPa=7.5 mmHg),respectively.According to the pressurization experiment results,the RGC injury model established under the in vitro pressure of 80 mmHg was selected for subsequent experiments.The RGCs of rats were divided into the normal group(NG group,untreated RGCs),model group(MG group,injured RGCs without serum intervention),blank serum group(KB group,injured RGCs treated with blank serum),and modified ZJP-medicated serum group(LVP group,injured RGCs treated with modified ZJP-medicated serum).The apoptosis of RGCs was detected by flow cytometry;the mitochondrial ultrastructure was observed via transmission electron microscopy;the protein expression level of PINK1,Parkin,LC3 Ⅱ/Ⅰ,and p62 was measured by Western blotting.Results Transmission electron microscopy revealed the in-tact double-membrane structure of mitochondria and evenly distributed mitochondrial crista in the NG group,while the MG and KB groups exhibited disrupted mitochondrial membranes,accompanied by a large number of vacuoles in the mitochon-dria.The mitochondria of the LVP group had a double-membrane structure and the vacuoles in the mitochondria decreased compared with those of the MG and KB groups,and the mitochondrial morphology was basically the same as that of the NG group.The flow cytometry results demonstrated the apoptosis rate of RGCs was(4.25±0.55)％,(20.79±3.06)％,(30.07±3.23)％,and(11.42±0.16)％in the NG,MG,KB,and LVP groups,respectively.Compared with the NG group,the ap-optosis rate of RGCs in rats increased in the MG group,and the difference was statistically significant(P＜0.01).Com-pared with the MG and KB groups,the apoptosis rate of RGCs in rats decreased in the LVP group,and the difference was statistically significant(both P＜0.05).The Western blotting results indicated that the MG group had an increased relative protein expression level of PINK1,Parkin,and LC3 Ⅱ/Ⅰ but a decreased relative protein expression level of p62 compared with the NG group,and the difference was statistically significant(all P＜0.01).Compared with the MG and KB groups,the LVP group had a decreased relative protein expression level of PINK1,Parkin,and LC3 Ⅱ/Ⅰ but an increased relative protein expression level of p62,and the difference was statistically significant(all P＜0.05).Conclusion The modified ZJP-medicated serum may protect RGCs by suppressing pressure-induced mitochondrial autophagy in vitro of rat RGCs via modulating PINK1/Parkin-mediated mitochondrial autophagy.
