RESUMO
Objective To explore the protective effect and mechanism of endoplasmic reticulum stress(ERS)signaling pathway PERK/eIF2α/ATF4/CHOP on BRL-3A cell injury by serum containing Zhige oral solution.Methods ①Preparation of drug-containing serum:SD male rats were randomly divided into 3 groups:Zhige oral solution group,Metadoxine group and normal group,and were gavaged with Zhige oral solution,Metadoxine and saline respectively,and the drug-containing serum was prepared by blood sampling from the abdominal aorta for 5 consecutive days of intervention.②Normal BRL-3A cells were cultured,and the cell survival rate of each group was determined by CCK-8 after 24h of cell intervention with different concentrations of ethanol(1.0%,1.5%,2.0%,2.5%,3.0%,5.0%,7.0%,10.0%).③Cultured normal BRL-3A cells were divided into normal group,model group,metadoxine group,and Zhige oral solution low,medium and high dose group(later referred to as low dose group,medium dose group and high dose group),After 24 h,the levels of γ-glutamyl transpeptidase(GGT)and lactate dehydrogenase(LDH)in the supernatant of each group were measured,and the survival rate of BRL-3A cells was measured by CCK-8.The expression levels of PERK/eIF2α/ATF4/CHOP pathway-related mRNA and protein in each group were measured by Real-time PCR and Western blot.Results ①After 24 h of cell intervention with the same concentration of ethanol(1.0%,1.5%,2.0%,2.5%,3.0%,5.0%,7.0%,10.0%),the cell survival rate gradually decreased with the increase of ethanol concentration,and when the ethanol concentration was 5%,the cell survival rate decreased significantly(P<0.05),so the ethanol concentrations of 1.0%,1.5%,2.0%,2.5%,3.0%,5.0%were chosen for the subsequent experiments.②Compared with the normal group,the GGT and LDH contents in the supernatant of the model group were significantly higher(P<0.05),and the expression of PERK/eIF2α/ATF4/CHOP pathway-related factors mRNA and protein were significantly higher(P<0.05),showing a significant state of liver injury.③Compared with the model group,the Zhige oral solution group and the metadoxine group showed different degrees of reduction in the above indexes,with the most significant effect in the Zhige oral solution high-dose group(P<0.05).Conclusion Serum containing Zhige oral solution ameliorated ethanol-induced hepatocyte injury in BRL-3A rats,and the mechanism may be related to the inhibition of endoplasmic reticulum stress PERK/eIF2α/ATF4/CHOP signaling pathway.