FOXO1对急性肺损伤小鼠肺泡上皮钠通道的调控作用及其机制

doi:10.13481/j.1671-587x.20200608

Abstract

Abstract: Objective

To investigate the regulatory effect of forkhead box O1（FOXO1） on the alveolar epithelial sodium channel （ENac） in the lipopolysaccharide （LPS）-induced acute lung injury mice， and to illustrate the mechanism.

Methods

Twenty C57BL/6J mice were randomly divided into control group and LPS group， and there were 10 mice in each group. After treatment， the mouse samples were obtained. HE staining was used to observe the pathomorphology of lung tissue of the mice in two groups， RT-PCR method and Western blotting method were used to detect the expression levels of α-subunit ENaC（αENaC） mRNA and pFOXO1 （Ser256） protein in lung tissue of the mice in two groups.The alveolar epithelial A549 cells were cultured， and then infected with ADV-FOXO1 or shRNA-FOXO1， respectively.RT-PCR method was used to determine the expression of αENaC mRNA. The A549 cells were divided into control group， insulin group， insulin+PI3K inhibitor group， and insulin+AKT inhibitor group. After corresponding intervention， Western blotting method was used to detect the expression levels of pFOXO1 （Ser256） protein，RT-PCR method was used to determine the expression levels of αENaC mRNA， and immunofluorescence method was used to detect the intracellular localization of FOXO1.

Results

Compared with control group， the lung injury score of mice in LPS group was increased significantly （P<0.05）， the expression levels of pFOXO1 （Ser256） and αENaC proteins in the lung tissue were significantly decreased（P<0.05） and they showed positively correlated relationship（r=0.703， P<0.05）.Compared with control group； the expression level of αENaC mRNA in A549 cells in over-expression FOXO1 group was significantly decreased （P<0.05），and the expression level of αENaC mRNA in silencing FOXO1 group was significantly increased（P<0.05）； compared with control group， the expression levels of pFOXO1 protein and αENaC mRNA in the A549 cells in insulin group were increased significantly （P<0.05）， and FOXO1 was mainly located in the cytoplasm. The expression levels of αENaC mRNA and pFOXO1 protein in A549 cells in insulin+PI3K inhibitor group and insulin+AKT inhibitor group were lower than those in insulin group （P<0.05）.

Conclusion

After inactiviation of FOXO1 phosphorylation，FOXO1 can translocate from nucleus to cytoplasm to weaken the inhibitory effect of αENaC and upregulate the expression of αENaC.

Key words: forkhead box O1, epithelial sodium channel, insulin, acute lung injury, acute respiratory distress syndrome

CLC Number:

R563

Jing HE,Daoxin WANG,Wang DENG. Regulatory effect of FOXO1 on alveolar epithelial sodium channel in acute lung injury mice and its mechanism[J].Journal of Jilin University(Medicine Edition), 2020, 46(6): 1155-1161.

Figures/Tables 4

References 20

1	FAN E， BRODIE D， SLUTSKY A S. Acute respiratory distress syndrome： advances in diagnosis and treatment［J］. JAMA， 2018， 319（7）： 698-710.
2	MATTHAY M A， ZEMANS R L， ZIMMERMAN G A， et al. Acute respiratory distress syndrome ［J］. Nat Rev Dis Primers， 2019， 5（1）： 18.
3	YADAV H， THOMPSON B T， GAJIC O. Fifty years of research in ARDS. Is acute respiratory distress syndrome a preventable disease？［J］. Am J Respir Crit Care Med， 2017， 195（6）： 725-736.
4	NIU F F， XU X T， ZHANG R， et al. Ursodeoxycholic acid stimulates alveolar fluid clearance in LPS-induced pulmonary edema via ALX/cAMP/PI₃K pathway［J］. J Cell Physiol， 2019， 234（11）： 20057-20065.
5	WARE L B， MATTHAY M A. Alveolar fluid clearance is impaired in the majority of patients with acute lung injury and the acute respiratory distress syndrome［J］. Am J Respir Crit Care Med， 2001， 163（6）： 1376-1383.
6	HE J， QI D， WANG D X， et al. Insulin upregulates the expression of epithelial sodium channel in vitro and in a mouse model of acute lung injury： role of mTORC2/SGK1 pathway［J］. Exp Cell Res， 2015， 331（1）： 164-175.
7	DENG W， LI C Y， TONG J， et al. Regulation of ENaC-mediated alveolar fluid clearance by insulin via PI₃K/Akt pathway in LPS-induced acute lung injury［J］. Respir Res， 2012， 13： 29.
8	SZYDŁOWSKI M， JABŁOŃSKA E， JUSZCZYŃSKI P. FOXO1 transcription factor： a critical effector of the PI₃K-AKT axis in B-cell development［J］. Int Rev Immunol， 2014， 33（2）： 146-157.
9	PENG S M， LI W， HOU N N， et al. A review of FoxO1-regulated metabolic diseases and related drug discoveries［J］. Cells， 2020， 9（1）： E184.
10	MARUNAKA Y. Characteristics and pharmacological regulation of epithelial Na⁺ channel （ENaC） and epithelial Na⁺ transport［J］. J Pharmacol Sci， 2014， 126（1）： 21-36.
11	HUMMLER E， BARKER P， GATZY J， et al. Early death due to defective neonatal lung liquid clearance in alpha-ENaC-deficient mice［J］. Nat Genet， 1996， 12（3）： 325-328.
12	TIA N， SINGH A K， PANDEY P， et al. Role of Forkhead Box O （FOXO） transcription factor in aging and diseases［J］. Gene， 2018， 648： 97-105.
13	BACH D H， LONG N P， LUU T T， et al. The dominant role of forkhead box proteins in cancer［J］. Int J Mol Sci， 2018， 19（10）： E3279.
14	ZHOU M， ZHANG Y D， CHEN X L， et al. PTEN-Foxo1 signaling triggers HMGB1-mediated innate immune responses in acute lung injury［J］. Immunol Res， 2015， 62（1）： 95-105.
15	SHEN Y C， XU W F， YOU H， et al. FoxO1 inhibits transcription and membrane trafficking of epithelial Na⁺ channel［J］. J Cell Sci， 2015， 128（19）： 3621-3630.
16	SCHWEIZER-GROYER G， FALLOT G， CADEPOND F， et al. The cAMP-responsive unit of the human insulin-like growth factor-binding protein-1 coinstitutes a functional insulin-response element［J］. Ann N Y Acad Sci， 2006， 1091： 296-309.
17	MOUNIER C， POSNER B I. Transcriptional regulation by insulin： from the receptor to the gene［J］. Can J Physiol Pharmacol， 2006， 84（7）： 713-724.
18	WILHELM K， HAPPEL K， EELEN G， et al. FOXO1 couples metabolic activity and growth state in the vascular endothelium［J］. Nature， 2016， 529（7585）： 216-220.
19	ARRIOLA D J， MAYO S L， SKARRA D V， et al. FOXO1 transcription factor inhibits luteinizing hormone β gene expression in pituitary gonadotrope cells［J］. J Biol Chem， 2012， 287（40）： 33424-33435.
20	TZIVION G， DOBSON M， RAMAKRISHNAN G. FoxO transcription factors； Regulation by AKT and 14-3-3 proteins［J］. Biochim Biophys Acta， 2011， 1813（11）： 1938-1945.

[1]	Lihua ZHOU,Ying HU,Hui ZOU. Expression of miR-27a in villi tissue of patients with recurrent abortion and its effects on trophoblast cell proliferation and apoptosis and their mechanisms [J]. Journal of Jilin University(Medicine Edition), 2022, 48(4): 1018-1027.
[2]	Zhaohui WAN,Liang ZENG,Hui ZHOU,Jing LI,Changcheng LEI. Protective effect of polyphyllin Ⅶ on acute lung injury in rats with severe acute pancreatitis by inhibiting NF-κB signaling pathway [J]. Journal of Jilin University(Medicine Edition), 2022, 48(3): 668-675.
[3]	Haixin QU,Erwei YUAN,Weiping GUO,Yajing ZHANG,Wenxia MA,Dan WU. Improvement effect of luteolin on acute respiratory distress syndrome by inhibiting ROS/TXNIP/NLRP3 signaling pathway activation in mice [J]. Journal of Jilin University(Medicine Edition), 2022, 48(3): 676-683.
[4]	Qiao LI,Zhenzhong SU,Zhaoyong WANG,Yanling XU,Qun ZHANG,Peng GAO,Jie ZHANG. Bortezomib-induced acute lung injury in patient with multiple myeloma：A case report and literature review [J]. Journal of Jilin University(Medicine Edition), 2021, 47(4): 1022-1027.
[5]	Guangwen LONG, Qian ZHANG, Xiulin YANG, Chunling JI, Yukang DONG. Regulation effect of miR-146b on expression of intercellular adhesion molecule-1 in lung tissue of rats with acute respiratory distress syndrome [J]. Journal of Jilin University(Medicine Edition), 2021, 47(3): 587-594.
[6]	Lei XIE, Xiaoli LIU, Jing GE, Yawei LI. Effects of baicalin combined with metformin on endocrine, inflammatory factors and insulin resistance in rats with polycystic ovary syndrome and their mechanisms [J]. Journal of Jilin University(Medicine Edition), 2021, 47(3): 623-629.
[7]	Hao HUANG,Hong JIA,Xiaoshuang WANG,Lu ZHANG,Yating DUAN. Expressions of miRNA-508-3p and HGF in placenta tissue of patients with gestational diabetes mellitus and their effects on trophoblast insulin resistance [J]. Journal of Jilin University(Medicine Edition), 2021, 47(1): 187-195.
[8]	ZHANG Yezhuo, YANG Yuefeng, YANG Yetong, ZHANG Ying, LIU Yajuan. Effects of poria cocos compound extract on blood glucose and lipid levels in rats with type 2 diabetes mellitus [J]. Journal of Jilin University(Medicine Edition), 2020, 46(05): 937-941.
[9]	BU Yi, ZHANG Shuo, QIAN Xudong, WANG Hongmei, DOU Zhijie. Expression of insulin-like growth factor binding protein-3 in ischemic brain tissue of cerebral infarction model rats and its relationship with angiogenesis [J]. Journal of Jilin University(Medicine Edition), 2020, 46(04): 759-764.
[10]	LIU Donghui, FU Wenliang, FU Xiumei, SONG Chengjun, CHEN Zhihong. Regulatory effect of sericin on liver insulin PI3K/Akt signaling pathway in rats of type 2 diabetes mellitus and its mechanism [J]. Journal of Jilin University(Medicine Edition), 2020, 46(02): 228-232.
[11]	LI Ruiyang, LIN Zhiyi, LI Jing, FEI Jing, XU Dan, GONG Ping. Changes of IGF-1 and mTOR expressions in peripheral blood of patients with non-small cell lung cancer before and after chemotherapy combined with metformin and analysis on their therapeutic effects [J]. Journal of Jilin University(Medicine Edition), 2020, 46(01): 108-115.
[12]	SHI Yan, SUN Yan, SHI Guang, YU Qiong, TANG Yan. Systemic lupus erythematosus complicated with autoimmune hypoglycemia with pancytopenia as first manifestation: A case report and literature review [J]. Journal of Jilin University(Medicine Edition), 2020, 46(01): 164-168.
[13]	LIAO Xin, DENG Fanqu, SONG Meihui, GAO Lin, ZHANG Han, ZHANG Lin, WANG Xuemei, ZHANG Ying, ZHAO Yu. Relationship between serum levels of CTRP3, 25(OH)D and insulin resistance in individuals with different glucose metabolism states [J]. Journal of Jilin University(Medicine Edition), 2019, 45(04): 887-892.
[14]	JIN Aihong, ZHOU Xiaping, YU Zhiying. Expressions of Versican and ADAMTS-1 in polycysticovary syndrome patients and their clincal significances [J]. Journal of Jilin University(Medicine Edition), 2019, 45(02): 400-404.
[15]	LU Mingyue, LI Jing. Relationships between expression of IGF-1 in cancer tissue of gastric cancer patients and clinicopathological parameters in China: A Meta-analysis [J]. Journal of Jilin University(Medicine Edition), 2019, 45(01): 137-142.

Regulatory effect of FOXO1 on alveolar epithelial sodium channel in acute lung injury mice and its mechanism

RichHTML

PDF (PC)

Like

Knowledge

Abstract

Cite this article

share this article

Figures/Tables 4

References 20

Related Articles 15

Metrics

Comments

Recommended 0