Shi H B, Shi H L, Zhang X Y, Chen D X, Duan Z P, Ren F
Beijing You'an, Hospital Affiliated to Capital Medical University, Beijing 100069, China.
Zhonghua Gan Zang Bing Za Zhi. 2017 Mar 20;25(3):211-216. doi: 10.3760/cma.j.issn.1007-3418.2017.03.010.
To investigate the role of the glycogen synthase kinase 3β (GSK3β) and the peroxisome proliferator-activated receptor alpha (PPARα) signaling pathway in acute liver failure and related mechanisms in a mouse model of acute liver failure induced by D-galactosamine/lipopolysaccharide (D-GalN/LPS). C57BL/6 mice were given intraperitoneal injection of D-GalN/LPS to establish a mouse model of acute liver failure. SB216763 was used to inhibit the activity of GSK3β and PPARα siRNA was used to inhibit the expression of PPARα. Western blotting was used to measure the expression of PPARα protein. The changes in liver pathology were observed to evaluate liver injury, and the serum levels of alanine aminotransferase (ALT) and aspartate aminotransferase (AST) were measured to assess liver function. Quantitative real-time PCR was used to measure the mRNA expression of tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β), interleukin-12p40 (IL-12p40), and PPARα. A one-way analysis of variance was used for comparison of means between multiple groups; the least significant difference test was used for data with homogeneity of variance, and the Games-Howell method was used for data with heterogeneity of variance. In the mice with liver failure induced by D-GalN/LPS, GSK3β inhibition promoted the mRNA and protein expression of PPARα ( = 13.18 and 301.36, = 0.00 and 0.00). In the mice with acute liver failure induced by D-GalN/LPS, GSK3β inhibition alleviated liver bleeding, inflammation, and necrosis and reduced the serum levels of ALT ( = 25.16, = 0.000) and AST ( = 12.96, = 0.001), as well as the mRNA expression of TNF-α ( = 32.17, = 0.00), IL-1β ( = 11.57, = 0.005), and IL-12p40 ( = 14.17, = 0.015) in liver tissue. The inhibition of PPARα expression reversed the liver-protecting effect of GSK3β inhibition, which manifested as aggravation in liver bleeding, inflammation, and necrosis, increases in the serum levels of ALT ( = 25.16, = 0.001) and AST ( = 12.96, = 0.000), and an increase in the mRNA expression of TNF-α ( = 32.17, = 0.00), IL-1β ( = 11.57, = 0.024), and IL-12p40 ( = 14.17, = 0.001) in liver tissue. In mice with acute liver failure induced by D-GalN/LPS, the GSK3β-PPARα-inflammatory factor signaling pathway may play an important role. GSK3β inhibition has a protective effect in mice with acute liver failure possibly by activating the inhibitory inflammatory factor of PPARα.
为研究糖原合酶激酶3β（GSK3β）和过氧化物酶体增殖物激活受体α（PPARα）信号通路在急性肝衰竭中的作用及相关机制，采用D-氨基半乳糖/脂多糖（D-GalN/LPS）诱导的急性肝衰竭小鼠模型进行研究。将C57BL/6小鼠腹腔注射D-GalN/LPS以建立急性肝衰竭小鼠模型。使用SB216763抑制GSK3β的活性，使用PPARα siRNA抑制PPARα的表达。采用蛋白质免疫印迹法检测PPARα蛋白的表达。观察肝脏病理变化以评估肝损伤，检测血清丙氨酸氨基转移酶（ALT）和天冬氨酸氨基转移酶（AST）水平以评估肝功能。采用定量实时PCR检测肿瘤坏死因子-α（TNF-α）、白细胞介素-1β（IL-1β）、白细胞介素-12p40（IL-12p40）和PPARα的mRNA表达。多组间均值比较采用单因素方差分析；方差齐性数据采用最小显著差检验，方差不齐性数据采用Games-Howell法。在D-GalN/LPS诱导的肝衰竭小鼠中，抑制GSK3β可促进PPARα的mRNA和蛋白表达（F = 13.18和301.36，P = 0.00和0.00）。在D-GalN/LPS诱导的急性肝衰竭小鼠中，抑制GSK3β可减轻肝脏出血、炎症和坏死，降低血清ALT水平（F = 25.16，P = 0.000）和AST水平（F = 12.96，P = 0.001），以及肝脏组织中TNF-α（F = 32.17，P = 0.00）、IL-1β（F = 11.57，P = 0.005）和IL-12p40（F = 14.17，P = 0.015）的mRNA表达。抑制PPARα表达可逆转抑制GSK3β的肝脏保护作用，表现为肝脏出血、炎症和坏死加重，血清ALT水平（F = 25.16，P = 0.001）和AST水平（F = 12.96，P = 0.000）升高，以及肝脏组织中TNF-α（F = 32.17，P = 0.00）、IL-1β（F = 11.57，P = 0.024）和IL-12p40（F = 14.17，P = 0.001）的mRNA表达增加。在D-GalN/LPS诱导的急性肝衰竭小鼠中，GSK3β-PPARα-炎症因子信号通路可能起重要作用。抑制GSK3β对急性肝衰竭小鼠具有保护作用，可能是通过激活PPARα的抑制性炎症因子实现的。
