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《天津医科大学学报》[ISSN:1006-8147/CN:12-1259/R]

卷:

30卷

期数:

2024年06期

页码:

514-521

栏目:

基础医学

出版日期:

2024-11-20

文章信息/Info

Title:

The role of p67phox in hepatic fibrosis and the underlying mechanism

文章编号:

1006-8147(2024)06-0514-08

作者:

马立业; 程欣欣; 徐哲龙

（天津医科大学基础医学院病理生理学系，天津300070）

Author(s):

MA Liye; CHENG Xinxin; XU Zhelong

（Department of Pathophysiology，School of Basic Medical Sciences，Tianjin Medical University，Tianjin 300070，China）

关键词:

p67phox; 信号转导及转录激活因子3; 蛋白激酶B; 细胞外调节蛋白激酶; 肝纤维化

Keywords:

p67phox; STAT3; AKT; ERK; liver fibrosis

分类号:

R363

DOI:

10.20135/j.issn.1006-8147.2024.06.0514

文献标志码:

A

摘要:

目的：探讨NADPH氧化酶的胞浆亚基p67phox在肝纤维化中的作用及其机制。方法：用四氯化碳（CCl4）腹腔注射的方法建立小鼠肝脏纤维化模型，随机将小鼠分为 5 组：野生型组（WT）、对照组（NCF2fl/fl-Control）、模型组（NCF2fl/fl-CCl4）、NCF2基因敲除加模型组（NCF2LKO- CCl4）和NCF2过表达加模型组（NCF2OE-CCl4），每组6只小鼠。分别用HE 染色、Masson染色、天狼猩红染色观察肝脏的病理学改变；免疫组化法检测肝脏平滑肌肌动蛋白（α-SMA）表达水平； Western 印迹检测肝脏组织中p67phox、纤维化相关蛋白α-SMA和信号传导及转录激活因子（STAT）3、蛋白激酶B（AKT）、细胞外调节蛋白激酶（ERK）通路蛋白的表达。结果：与NCF2fl/fl-Control组相比，注射CCl4引起小鼠肝纤维化及损伤，p67phox表达下降（t=2.96，P<0.05）。与NCF2fl/fl-CCl4组相比，NCF2LKO-CCl4组肝脏α-SMA表达增强（t=3.6， P<0.05）。此外，与NCF2fl/fl-CCl4组相比，NCF2LKO-CCl4组AKT通路相关蛋白（p-PDK1、AKT、p-AKT473、p-AKT308）表达上升（t=3.17、1.68、3.34、2.47，均P<0.05），STAT3通路相关蛋白（STAT3、p-STAT3705）表达升高（t=3.95、2.67，均P<0.05），ERK通路相关蛋白（p-c-Raf、p-ERK）表达增加（t=3.68、1.96，均P<0.05）。与NCF2fl/fl-CCl4组相比，NCF2OE-CCl4组小鼠肝脏病理损伤及纤维化程度减轻，p-PDK1、AKT、p-AKT473及p-AKT308表达下降（t=2.31、3.53、3.54、2.71，均P<0.05），STAT3及p-STAT3705表达减少（t=3.47、2.5，均P<0.05），p-c-Raf和p-ERK蛋白表达下降（t=2.5、6.27，均P<0.05）。结论：p67phox表达下调，通过激活STAT3、AKT及ERK通路来诱导肝纤维化的发生。

Abstract:

Objective： To investigate the role of cytoplasmic subunit p67phox of NOX in hepatic fibrosis and its underlying mechanism. Methods：The mouse hepatic fibrosis model was established by intraperitoneal injection of CCl4. Mice were grouped into five groups：WT group，NCF2fl/fl-Control group，NCF2fl/fl-CCl4 group，NCF2LKO-CCl4 group and NCF2OE-CCl4 group，with 6 mice in each group. HE，Masson and Sirius red staining were adopted to observe the pathological changes of liver tissue. Expression of α-smooth muscle actin（α-SMA） was detected by immunohistochemistry. Expressions of p67phox and fibrosis-related proteins including α-SMA， signal transducer and activator of transcription （STAT）3，AKT and extracellular regulated protein kinases （ERK） were detected by Western blotting. Results：CCl4 injection induced liver fibrosis and injury，compared with NCF2fl/fl-Control group，the expression of p67phox was decreased（t=2.96，P<0.05）. Compared with NCF2fl/fl-CCl4 group，NCF2LKO-CCl4 group had increased expression of α-SMA （t=3.6，P<0.05）. In addition，compared with NCF2fl/fl-CCl4 group，NCF2LKO-CCl4 group showed increased expression of the AKT signaling proteins （P-PDK1，AKT，p-AKT473，and p-AKT308） （t=3.17，1.68，3.34，2.47，all P<0.05），enhanced expression of the STAT3 signaling proteins （STAT3 and p-STAT3705） （t=3.95，2.67，both P<0.05），and upregulated expression of the ERK signaling proteins （p-c-Raf and p-ERK） （t=3.68，1.96，both P<0.05）. Compared with NCF2fl/fl-CCl4 group，NCF2OE-CCl4 group showed less pathological damages and fibrosis. Moreover，NCF2OE-CCl4 group had reduced expression of P-PDK1， AKT， p-AKT473 and p-AKT308（t=2.31，3.53，3.54，2.7，all P<0.05），decreased expression of STAT3 and p-STAT3705 （t=3.47，2.5，both P<0.05），and reduced expression of p-c-Raf and p-ERK（t=2.5，6.27，both P<0.05）. Conclusion：Down-regulation of p67phox induces hepatic fibrosis by activating STAT3， AKT and ERK signaling pathways.

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相似文献/References:

备注/Memo

备注/Memo:

基金项目 国家自然科学基金项目（81970255）
作者简介 马立业（1998-），女，硕士在读，研究方向：病理学与病理生理学；
通信作者：徐哲龙，E-mail：zxu@tmu.edu.cn。

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更新日期/Last Update: 2024-11-25