基于Keap1/Nrf2/ARE信号通路探讨丹皮酚改善酒精性肝、脑损伤小鼠氧化应激损伤与炎症的作用机制

目的 探讨丹皮酚基于Kelch样环氧氯丙烷相关蛋白1(Kelch-like ECH-associated protein 1,Keap1)/核因子E2相关因子2(nuclear factor erythroid 2-related factor 2,Nrf2)/抗氧化反应元件(antioxidant response element, ARE)信号通路改善乙醇刺激所致小鼠肝、脑炎症和氧化应激损伤的作用机制。方法 将C57BL/6小鼠随机分为空白组，模型组，水飞蓟宾组(36.8 mg/kg),丹皮酚高、中、低(480、240、120 mg/kg)剂量组，每组15只；适应期各组小鼠自由饮用Liber-DeCarli对照液体饲料，模型复制期空白组小鼠自由饮用Lieber-DeCarli对照液体饲料，其他组小鼠自由饮用Lieber-DeCarli乙醇液体饲料并连续灌胃相对应的药液10 d;测定小鼠血脂[三酰甘油(triglyceride, TG)、总胆固醇(total cholesterol, TC)]、肝功能[丙氨酸氨基转移酶(alanine aminotransferase, ALT)、天...

Mechanism of Action of Paeonol in Improving Oxidative Stress Injury and Inflammation in Mice with Alcohol-induced Liver and Brain Injury:A Study Based on the Kelch-like ECH-associated Protein 1/Nuclear Factor Erythroid 2-related Factor 2/Antioxidant Response Element Signaling Pathway

Objective To investigate the mechanism of action of paeonol improving liver and brain inflammation and oxidative stress injury induced by acute alcohol stimulation in mice based on the on Kelch-like ECH-associated protein 1 (Keap1)/nuclear factor erythroid 2-related factor 2 (Nrf2)/antioxidant response element (ARE) signaling pathway.Methods C57BL/6 mice were randomly divided into blank group,model group, silybin group (36.8 mg/kg),and high-,middle-,and low-dose paeonol groups (480, 240, and 120 mg/kg),with 15 mice in each group.During the adaptive phase, the mice in all groups were given Lieber-DeCarli control liquid feed freely;during modeling, the mice in the blank group were given Lieber-DeCarli control liquid feed freely,and those in the other groups were given Lieber-DeCarli alcohol liquid feed freely and the corresponding drug by gavage for 10 days. The mice were measured in terms of blood lipids [triglyceride (TG) and total cholesterol (TC)],liver function [alanine aminotransferase (ALT) and aspartate aminotransferase (AST)],inflammatory factors [interleukin-6(IL-6), interleukin-1α(IL-1α),interleukin-1β (IL-1β),and tumor necrosis factor-α (TNF-α)],and oxidative stress indicators [catalase (CAT),glutathione(GSH),superoxide dismutase(SOD),and malondialdehyde (MDA)]; HE staining and oil red O staining were used to observe the pathological changes of the liver and brain;Western blot,immunofluorescence assay,and qRT-PCR were used to measure the expression levels of Keap1/Nrf2/ARE signaling pathway-related proteins and their mRNA expression levels in mouse liver and brain.Results Compared with the model group,the high-dose paeonol group had a significant increase in body weight (P<0.05),significant increases in blood lipids (TG and TC), liver function parameters (ALT and AST),inflammatory factors (IL-6,IL-1α,IL-1β,and TNF-α),and oxidative stress indicators (CAT,GSH,and SOD) (P<0.05),and a significant reduction in the content of MDA (P<0.05);there were significant increases in the protein and mRNA expression levels of heme oxygenase-1,NAD(P)H quinone oxidoreductase 1,and glutamate-cysteine ligase catalytic subunit and significant reductions in the protein and mRNA expression levels of Keap1 in mouse liver and brain (P<0.05);the high-dose paeonol group showed significant improvements in the pathological state of the mouse liver and brain.Conclusion Paeonol can significantly alleviate alcohol-induced liver and brain inflammation and oxidative stress injury in mice,possibly by regulating the Keap1/Nrf2/ARE signaling pathway.