Ziyin Mingmu Formula improve retinitis pigmentosa through the GRP78/IRE1/ATF6 pathway in vitro study
10.3969/j.issn.1006-2157.2024.06.006
- VernacularTitle:滋阴明目方调控GRP78/IRE1/ATF6通路改善视网膜色素变性的体外研究
- Author:
Chen OU
1
,
2
;
Houpan SONG
;
Wei XIE
;
Jun PENG
;
Meiyan ZENG
;
Qinghua PENG
Author Information
1. 湖南中医药大学第一附属医院 长沙 410007
2. 湖南中医药大学
- Keywords:
Ziyin Mingmu Formula;
adult retinal pigment epithelial cell line-19;
endoplasmic reticulum stress;
78 kDa glucose regulated protein/inositol-requiring enzyme 1/activating transcription factor 6 pathway;
apoptosis
- From:
Journal of Beijing University of Traditional Chinese Medicine
2024;47(6):773-781
- CountryChina
- Language:Chinese
-
Abstract:
Objective We aimed to explore the mechanism of Ziyin Mingmu Formula in improving retinitis pigmentosa.Methods Twenty SD rats were randomly divided into the blank serum group and the Ziyin Mingmu Formula containing serum group using a random number table method,with 10 rats in each group.The rats in the Ziyin Mingmu Formula containing serum group were given Ziyin Mingmu Formula(46.875 g/kg),while the rats in the blank serum group were given distilled water.The Ziyin Mingmu Formula containing serum and blank serum were prepared.Analysis of chemical components in Ziyin Mingmu Formula containing serum using liquid chromatography-mass spectrometry technology.Adult retinal pigment epithelial cell line-19(ARPE-19)cells were treated with tunicamycin to induce endoplasmic reticulum stress injury model.The optimal volume fraction of Ziyin Mingmu Formula containing serum was screened by CCK-8.ARPE-19 cells were divided into the blank group,the model group,the blank serum group,the Ziyin Mingmu Formula containing serum group and the tauroursodeoxycholic acid group,each group is intervened separately.After 24 hours of intervention,the morphological observation of cells was performed using a multi-time dynamic cell function analysis system,cell survival rate was detected by CCK-8,cell apoptosis rate was detected by Annexin V-FITC/PI,and protein and mRNA expressions of 78 kDa glucose regulated protein(GRP78),inositol-requiring enzyme 1(IRE1)and activating transcription factor 6(ATF6)were detected by Western blotting and ddPCR.Results The main chemical components in Ziyin Mingmu Formula containing serum were obtained by liquid chromatography-mass spectrometry analysis,such as arbutin,salicylic acid,luteolin,Salvianolic acid A,erysovine,taurine,quercetin,maltol,baicalin,and danshensu.Compared with the model group,the number of cells in Ziyin Mingmu Formula containing serum group increased,the growth was more uniform,and the floating dead ARPE-19 cells and fragments decreased.The cell survival rate of Ziyin Mingmu Formula containing serum group and taursodeoxycholic acid group increased,and the cell apoptosis rate decreased(P<0.05).The protein expressions of GRP78,IRE1,and ATF6 in Ziyin Mingmu Formula containing serum group were decreased,and IRE1 and ATF6 in taursodeoxycholic acid group were decreased(P<0.05).The mRNA expressions of GRP78,IRE1,and ATF6 in Ziyin Mingmu Formula containing serum group and tauroursodeoxycholic acid group were decreased(P<0.05).Conclusion Ziyin Mingmu Formula can reduce the cell apoptosis of ARPE-19 cells induced by endoplasmic reticulum stress,and its molecular mechanism is related to down-regulating the expression of GRP78,IRE1,ATF6 and inhibiting the endoplasmic reticulum stress response.
