β-arrestin2 通过 AKT 和 TLR4 通路抑制缺血再灌注诱导的小鼠细胞凋亡和肝脏炎症。

β-arrestin2 Inhibits Apoptosis and Liver Inflamation Induced by Ischemia-reperfusion in Mice via AKT and TLR4 Pathway.

机构信息

Department of General Surgery, The First Affiliated Hospital of Soochow University, Suzhou, China.

出版信息

Arch Med Res. 2019 Oct;50(7):413-422. doi: 10.1016/j.arcmed.2019.10.012. Epub 2019 Nov 21.

DOI:10.1016/j.arcmed.2019.10.012

PMID:31760331

Abstract

BACKGROUND

Liver ischemia and reperfusion (I/R) is a common but severe clinical problem. Previous studies have revealed that the expression level of β-arrestin2 affects serum deprivation (SD)-induced cell apoptosis and was involved in lipopolysaccharide (LPS) stimulated TLR4 signaling pathway. However, little is known about β-arrestin2 in liver apoptosis and immune response induced by I/R.

METHODS

A non-lethal model of segmental (70%) hepatic ischemia was utilized. Histology examination, cell apoptosis and cytokine levels were measured using H&E staining, TUNEL assay, and ELISA, respectively. Apoptosis-related protein and gene level of cytokines were respectively detected using Western blot and Real-time PCR.

RESULTS

Our data showed that knockout (KO) of β-arrestin2 gene significantly deteriorated the injury of liver caused by I/R according to liver histology, higher serum liver enzyme, and increased level of cell apoptosis. β-arrestin2 KO could result in increased level of apoptosis related protein and decreased level of Akt phosphorylation. Furthermore, decreased levels of Bcl-2 and Bad phosphorylation, but increased level of Bax were found in β-arrestin2 KO group. In addition, the levels of p-ERK1/2, p-p38MAPKs, and p-NF-κB in β-arrestin2 KO group were significantly higher than that in WT group.

CONCLUSIONS

β-arrestin2 protected liver from I/R injury and this effect may be due to the regulating of Akt pathway, Bcl-2/Bax ratio, MAPKs and NF-κB pathway.

摘要

背景

肝脏缺血再灌注（I/R）是一种常见但严重的临床问题。先前的研究表明，β-arrestin2 的表达水平影响血清剥夺（SD）诱导的细胞凋亡，并参与脂多糖（LPS）刺激的 TLR4 信号通路。然而，关于β-arrestin2 在 I/R 诱导的肝凋亡和免疫反应中的作用知之甚少。

方法

利用非致死性 70%肝节段缺血模型。通过 H&E 染色、TUNEL 检测和 ELISA 分别检测组织学检查、细胞凋亡和细胞因子水平。使用 Western blot 和 Real-time PCR 分别检测凋亡相关蛋白和细胞因子基因水平。

结果

我们的数据显示，β-arrestin2 基因敲除（KO）显著加重了 I/R 引起的肝损伤，表现为肝酶血清水平升高和细胞凋亡增加。β-arrestin2 KO 可导致凋亡相关蛋白水平升高和 Akt 磷酸化水平降低。此外，β-arrestin2 KO 组中 Bcl-2 和 Bad 磷酸化减少，而 Bax 水平升高。此外，β-arrestin2 KO 组中 p-ERK1/2、p-p38MAPKs 和 p-NF-κB 的水平明显高于 WT 组。