mTORC2-mediated signaling pathway affects protective role of alveolar epithelial sodium channel in mice of acute lung injury

钟曦; 赵燕; 王导新

您当前的位置：

首页 >

文章列表页 >

mTORC2-mediated signaling pathway affects protective role of alveolar epithelial sodium channel in mice of acute lung injury

更新时间：2026-01-09

- mTORC2-mediated signaling pathway affects protective role of alveolar epithelial sodium channel in mice of acute lung injury
- Journal of Army Medical University Vol. 37, Issue 24, Pages: 2438-2443(2015)
- 作者机构：
- 作者简介：
- 基金信息：
- DOI：
  CLC： R322.35;R363;R563
- Published：2015
- 稿件说明：
移动端阅览
钟曦,赵燕,王导新.mTORC2介导的信号通路影响肺泡上皮钠通道对急性肺损伤肺组织的保护作用[J].陆军军医大学学报(原第三军医大学学报),2015,37(24):2438-2443.

　Zhong Xi,Zhao Yan,Wang Daoxin.mTORC2-mediated signaling pathway affects protective role of alveolar epithelial sodium channel in mice of acute lung injury[J].J Amry Med Univ (J Third Mil Med Univ),2015,37(24):2438-2443.
钟曦,赵燕,王导新.mTORC2介导的信号通路影响肺泡上皮钠通道对急性肺损伤肺组织的保护作用[J].陆军军医大学学报(原第三军医大学学报),2015,37(24):2438-2443. DOI：

　Zhong Xi,Zhao Yan,Wang Daoxin.mTORC2-mediated signaling pathway affects protective role of alveolar epithelial sodium channel in mice of acute lung injury[J].J Amry Med Univ (J Third Mil Med Univ),2015,37(24):2438-2443. DOI：

摘要

目的通过调控肺泡上皮细胞α钠离子通道（epithelial sodium channel α，ENaC-α）上游信号通路，探讨mTORC2/SGK1途径在急性肺损伤（acute lung injury，ALI）情况下对肺水清除方面的作用机制。方法将50只BALB/c雄性小鼠按随机数表法分成对照组、模型组、胰岛素组、雷帕霉素组及PP242抑制剂组，每组10只。滴鼻给予5 mg/kg LPS构建ALI动物模型。建模2 h后，分别腹腔注射0.8 U/kg胰岛素、4 mg/kg雷帕霉素、60 mg/kg PP242干预，8 h后处死小鼠。A549细胞分别以培养基、10 mmol/mL胰岛素、5 mmol/mL 雷帕霉素、5 mmol/mL PP242处理。采用ELISA检测各组肺泡灌洗液（BALF）中IL-6及TNF-α浓度，并测量各组右上肺组织的湿/干质量比（W/D）评估肺水清除（alveolar fluid clearance，AFC），HE染色观察各组肺组织病理改变。采用Real-time PCR、Western blot、免疫荧光法检测A549细胞rictor、糖皮质激素调节蛋白激酶1(serum and glucocorticoid-regulated protein kinase1

SGK1)、磷酸泛素化连接酶（E3 ubiquitin protein ligase，Nedd4-2）及ENaC-α的表达。结果体内实验发现，与模型组比较，胰岛素组小鼠BALF中IL-6 [(77.48±6.04） vs （49.17±8.49) pg/mL]和TNF-α[（116.2±13.5） vs （92.4±8.37） pg/mL]水平显著降低，并伴有湿/干质量比减低，病检示肺损伤程度减轻（P0.05）。PP242抑制剂组BALF中IL-6[（77.48±6.04） vs （100.6±14.69） pg/mL]和TNF-α[（116.2±13.5） vs （192.5±16.01） pg/mL]水平上升，湿/干质量比显著升高，肺组织损伤明显，程度较模型组更重（P0.05）。而雷帕霉素组与模型组中各项指标及病理学改变差异无统计学意义（P0.05）。体外实验中，A549细胞中rictor、SGK1、Nedd4-2及ENaC-α mRNA和蛋白表达在胰岛素组明显增高，在PP242抑制剂组中明显降低（P0.05），而在雷帕霉素组中无明显改变（P0.05）。结论调控 mTORC2/SGK1信号通路可影响ENaC-α的表达，从而发挥其在ALI情况下对肺组织的保护作用。

Abstract

Objective To explore the role and mechanism of mTORC2/SGK1 signaling pathway in alveolar fluid clearance (AFC) of acute lung injury (ALI) by regulating alveolar epithelial alpha sodium channel (ENaC-α) upstream signaling pathway. Methods Fifty male BALB/c mice were randomly divided into blank group

lipopolysaccharide (LPS) group

insulin group

LPS+rapamycin (mTORC1 inhibitor) group

and LPS+PP242 (mTORC1 and mTORC2 double inhibitor) group (n=10). ALI animal models were constructed by intranasal administration of 5 mg/kg LPS. After 2 h

the models were treated by intraperitoneal injection of 0.8 U/kg insulin

4 mg/kg rapamycin

and 60 mg/kg PP242

respectively

and the mice were put to death after 8 h. A549 cells were divided into control group

insulin group

rapamycin group and PP242 group

which were treated with culture medium

10 mmol/L insulin

5 mmol/L rapamycin

and 5 mmol/L PP242

respectively. The bronchoalveolar lavage fluid (BALF) IL-6 and TNF-α were measured by ELISA

and wet/dry (W/D) weight ratio was determined to evaluate AFC. HE staining was adopted to observe the lung tissue pathological changes. Real-time PCR

Western blotting and immunofluorescence assay were used to detect the expressions of rictor

SGK1

Nedd4-2 and ENaC-α in A549 cells. Results In vivo experiment showed that

compared with the LPS group

BALF IL-6 (77.48±6.04 vs 49.17±8.49)

TNF-α (116.2±13.5 vs 92.4±8.37) and W/D weight ratio were significantly decreased

AFC was increased

and lung injury was alleviated in the insulin group （all P0.05）. Compared with the LPS group

BALF IL-6 (77.48±6.04 vs 100.6±14.69)

TNF-α (116.2±13.5 vs 192.5±16.01) and W/D weight ratio were significantly increased

AFC was decreased

and lung injury was significantly worse in the LPS+PP242 group（all P0.05）. The inflammation indexes and pathological change had no significantly statistical differences between the LPS group and LPS+rapamycin group（all P0.05）. In vitro experiment showed that

compared with the control group

the expression of rictor

SGK1

Nedd4-2 and ENaC-α in A549 cells were obviously increased in the insulin group

and obviously decreased in the PP242 group

but no obvious changes were found between the rapamycin group and LPS group（all P0.05）. Conclusion Activated mTORC2/SGK1 pathway signaling can increase the expression of ENaC-α

and improve the AFC

thereby playing a protective role in lung tissue.

关键词

Keywords

references

Views

下载量

CSCD

Alert me when the article has been cited

提交

Tools

Publicity Resources

Role and mechanism of SIRT3 in oxidative stress and mitochondrial biogenesis in alveolar type II cells

Dexmedetomidine pretreatment enhances protective effect efficacy of pericytes agaist acute lung injury in septic mice

Time-course changes of remote apoptosis and necroptosis of pulmonary cells induced by acute kidney injury in mice

Effect of geraninn on acute lung injury induced by lipopolysaccharide in mice

Effect and mechanism of cryptotanshinone on early pulmonary fibrosis in LPS-induced acute respiratory distress syndrome in mice

Related Author

ZHOU Jun

LIU Yaojun

LIU Jing

GAO Ruobing

LI Changhong

FENG Xifeng

SHEN Hao

HUANG Hong

Related Institution

Department of Geriatrics and Special Service Medicine， First Affiliated Hospital（Southwest Hospital）， Army Medical University （Third Military Medical University）

Institute of Field Surgery， Daping Hospital， Army Medical University

Laboratory of Stem Cell and Tissue Engineering， Chongqing Medical University

Department of Human Anatomy， Anhui Medical University

Department of Anesthesiology, Department of War Wound Shock and Transfusion， Army Medical Center of PLA/Daping Hospital of Third Military Medical University

⁰