Sustained Stimulation of β2AR Inhibits Insulin Signaling in H9C2 Cardiomyoblast Cells Through the PKA-Dependent Signaling Pathway
Jinli Pei,1,2 Zhengpan Xiao,1,2 Ziyi Guo,1,2 Yechun Pei,1,2 Shuangshuang Wei,1,2 Hao Wu,1,2 Dayong Wang1,2 1Key Laboratory of Ministry of Education for Tropical Bioresources, Hainan University, Haikou, Hainan 570228, People’s Republic of China; 2Laboratory of Biotechnology and Molecular Ph...
Guardado en:
| Autores principales: | , , , , , , |
|---|---|
| Formato: | article |
| Lenguaje: | EN |
| Publicado: |
Dove Medical Press
2020
|
| Materias: | |
| Acceso en línea: | https://doaj.org/article/65f163039fda47399073dff413d6f10d |
| Etiquetas: |
Agregar Etiqueta
Sin Etiquetas, Sea el primero en etiquetar este registro!
|
| id |
oai:doaj.org-article:65f163039fda47399073dff413d6f10d |
|---|---|
| record_format |
dspace |
| spelling |
oai:doaj.org-article:65f163039fda47399073dff413d6f10d2021-12-02T12:07:53ZSustained Stimulation of β2AR Inhibits Insulin Signaling in H9C2 Cardiomyoblast Cells Through the PKA-Dependent Signaling Pathway1178-7007https://doaj.org/article/65f163039fda47399073dff413d6f10d2020-10-01T00:00:00Zhttps://www.dovepress.com/sustained-stimulation-of-beta2ar-inhibits-insulin-signaling-in-h9c2-ca-peer-reviewed-article-DMSOhttps://doaj.org/toc/1178-7007Jinli Pei,1,2 Zhengpan Xiao,1,2 Ziyi Guo,1,2 Yechun Pei,1,2 Shuangshuang Wei,1,2 Hao Wu,1,2 Dayong Wang1,2 1Key Laboratory of Ministry of Education for Tropical Bioresources, Hainan University, Haikou, Hainan 570228, People’s Republic of China; 2Laboratory of Biotechnology and Molecular Pharmacology, School of Life and Pharmaceutical Sciences, Hainan University, Haikou, Hainan 570228, People’s Republic of ChinaCorrespondence: Dayong WangLaboratory of Biotechnology and Molecular Pharmacology, School of Life and Pharmaceutical Sciences, Hainan University, 58 People’s Road, Haikou, Hainan 570228, People’s Republic of ChinaTel +86-187-8955-6728Email wangdy@hainu.edu.cnIntroduction: This study aimed to investigate the role of β2 adrenergic receptor (β2AR) in insulin signaling transduction in H9C2 cardiomyoblast cells to understand the formation of the β2AR-insulin receptor (IR) protein complex and its role in insulin-induced Glut4 expression.Methods: H9C2 cells were treated with various protein inhibitors (CGP, β1AR inhibitor CGP20712; ICI, β2AR inhibitor ICI 118,551; PKI, PKA inhibitor myristoylated PKI; PD 0325901, MEK inhibitor; SP600125, JNK inhibitor) with or without insulin or isoproterenol (ISO) before RNA-sequencing (RNA-Seq) and quantitative-PCR (Q-PCR). Yeast two-hybrid, co-immunoprecipitation and His-tag pull-down assay were carried out to investigate the formation of the β2AR-IR protein complex. The intracellular concentrations of cAMP in H9C2 cells were tested by high performance liquid chromatography (HPLC) and the phosphorylation of JNK was tested by Western blot.Results: Gene Ontology (GO) analysis revealed that the most significantly enriched processes in the domain of molecular function (MF) were catalytic activity and binding, whereas in the domain of biological processes (BP) were metabolic process and cellular process. Furthermore, the enriched processes in the domain of cellular components (CC) were cell and cell parts. The Kyoto encyclopedia of genes and genomes (KEGG) pathway analysis showed that the most significant pathways that have been altered included the PI3K-Akt and MAPK signaling pathways. Q-PCR, which was performed to verify the gene expression levels exhibited consistent results. In evaluating the signaling pathways, the sustained stimulation of β2AR by ISO inhibited insulin signalling, and the effect was primarily through the cAMP-PKA-JNK pathway and MEK/JNK signaling pathway. Yeast two-hybrid, co-immunoprecipitation and His-tag pull-down assay revealed that β2AR, IR, insulin receptor substrate 1 (IRS1), Grb2-associated binding protein 1 (GAB1) and Grb2 existed in the same protein complex.Conclusion: The sustained stimulation of β2AR might inhibit insulin signaling transduction through the cAMP-PKA-JNK and MEK/JNK pathways in H9C2 cells.Keywords: RNA sequencing, beta adrenergic receptor, insulin receptor, protein interaction, insulin resistancePei JXiao ZGuo ZPei YWei SWu HWang DDove Medical Pressarticlerna sequencingbeta adrenergic receptorinsulin receptorprotein interactioninsulin resistanceSpecialties of internal medicineRC581-951ENDiabetes, Metabolic Syndrome and Obesity: Targets and Therapy, Vol Volume 13, Pp 3887-3898 (2020) |
| institution |
DOAJ |
| collection |
DOAJ |
| language |
EN |
| topic |
rna sequencing beta adrenergic receptor insulin receptor protein interaction insulin resistance Specialties of internal medicine RC581-951 |
| spellingShingle |
rna sequencing beta adrenergic receptor insulin receptor protein interaction insulin resistance Specialties of internal medicine RC581-951 Pei J Xiao Z Guo Z Pei Y Wei S Wu H Wang D Sustained Stimulation of β2AR Inhibits Insulin Signaling in H9C2 Cardiomyoblast Cells Through the PKA-Dependent Signaling Pathway |
| description |
Jinli Pei,1,2 Zhengpan Xiao,1,2 Ziyi Guo,1,2 Yechun Pei,1,2 Shuangshuang Wei,1,2 Hao Wu,1,2 Dayong Wang1,2 1Key Laboratory of Ministry of Education for Tropical Bioresources, Hainan University, Haikou, Hainan 570228, People’s Republic of China; 2Laboratory of Biotechnology and Molecular Pharmacology, School of Life and Pharmaceutical Sciences, Hainan University, Haikou, Hainan 570228, People’s Republic of ChinaCorrespondence: Dayong WangLaboratory of Biotechnology and Molecular Pharmacology, School of Life and Pharmaceutical Sciences, Hainan University, 58 People’s Road, Haikou, Hainan 570228, People’s Republic of ChinaTel +86-187-8955-6728Email wangdy@hainu.edu.cnIntroduction: This study aimed to investigate the role of β2 adrenergic receptor (β2AR) in insulin signaling transduction in H9C2 cardiomyoblast cells to understand the formation of the β2AR-insulin receptor (IR) protein complex and its role in insulin-induced Glut4 expression.Methods: H9C2 cells were treated with various protein inhibitors (CGP, β1AR inhibitor CGP20712; ICI, β2AR inhibitor ICI 118,551; PKI, PKA inhibitor myristoylated PKI; PD 0325901, MEK inhibitor; SP600125, JNK inhibitor) with or without insulin or isoproterenol (ISO) before RNA-sequencing (RNA-Seq) and quantitative-PCR (Q-PCR). Yeast two-hybrid, co-immunoprecipitation and His-tag pull-down assay were carried out to investigate the formation of the β2AR-IR protein complex. The intracellular concentrations of cAMP in H9C2 cells were tested by high performance liquid chromatography (HPLC) and the phosphorylation of JNK was tested by Western blot.Results: Gene Ontology (GO) analysis revealed that the most significantly enriched processes in the domain of molecular function (MF) were catalytic activity and binding, whereas in the domain of biological processes (BP) were metabolic process and cellular process. Furthermore, the enriched processes in the domain of cellular components (CC) were cell and cell parts. The Kyoto encyclopedia of genes and genomes (KEGG) pathway analysis showed that the most significant pathways that have been altered included the PI3K-Akt and MAPK signaling pathways. Q-PCR, which was performed to verify the gene expression levels exhibited consistent results. In evaluating the signaling pathways, the sustained stimulation of β2AR by ISO inhibited insulin signalling, and the effect was primarily through the cAMP-PKA-JNK pathway and MEK/JNK signaling pathway. Yeast two-hybrid, co-immunoprecipitation and His-tag pull-down assay revealed that β2AR, IR, insulin receptor substrate 1 (IRS1), Grb2-associated binding protein 1 (GAB1) and Grb2 existed in the same protein complex.Conclusion: The sustained stimulation of β2AR might inhibit insulin signaling transduction through the cAMP-PKA-JNK and MEK/JNK pathways in H9C2 cells.Keywords: RNA sequencing, beta adrenergic receptor, insulin receptor, protein interaction, insulin resistance |
| format |
article |
| author |
Pei J Xiao Z Guo Z Pei Y Wei S Wu H Wang D |
| author_facet |
Pei J Xiao Z Guo Z Pei Y Wei S Wu H Wang D |
| author_sort |
Pei J |
| title |
Sustained Stimulation of β2AR Inhibits Insulin Signaling in H9C2 Cardiomyoblast Cells Through the PKA-Dependent Signaling Pathway |
| title_short |
Sustained Stimulation of β2AR Inhibits Insulin Signaling in H9C2 Cardiomyoblast Cells Through the PKA-Dependent Signaling Pathway |
| title_full |
Sustained Stimulation of β2AR Inhibits Insulin Signaling in H9C2 Cardiomyoblast Cells Through the PKA-Dependent Signaling Pathway |
| title_fullStr |
Sustained Stimulation of β2AR Inhibits Insulin Signaling in H9C2 Cardiomyoblast Cells Through the PKA-Dependent Signaling Pathway |
| title_full_unstemmed |
Sustained Stimulation of β2AR Inhibits Insulin Signaling in H9C2 Cardiomyoblast Cells Through the PKA-Dependent Signaling Pathway |
| title_sort |
sustained stimulation of β2ar inhibits insulin signaling in h9c2 cardiomyoblast cells through the pka-dependent signaling pathway |
| publisher |
Dove Medical Press |
| publishDate |
2020 |
| url |
https://doaj.org/article/65f163039fda47399073dff413d6f10d |
| work_keys_str_mv |
AT peij sustainedstimulationofbeta2arinhibitsinsulinsignalinginh9c2cardiomyoblastcellsthroughthepkadependentsignalingpathway AT xiaoz sustainedstimulationofbeta2arinhibitsinsulinsignalinginh9c2cardiomyoblastcellsthroughthepkadependentsignalingpathway AT guoz sustainedstimulationofbeta2arinhibitsinsulinsignalinginh9c2cardiomyoblastcellsthroughthepkadependentsignalingpathway AT peiy sustainedstimulationofbeta2arinhibitsinsulinsignalinginh9c2cardiomyoblastcellsthroughthepkadependentsignalingpathway AT weis sustainedstimulationofbeta2arinhibitsinsulinsignalinginh9c2cardiomyoblastcellsthroughthepkadependentsignalingpathway AT wuh sustainedstimulationofbeta2arinhibitsinsulinsignalinginh9c2cardiomyoblastcellsthroughthepkadependentsignalingpathway AT wangd sustainedstimulationofbeta2arinhibitsinsulinsignalinginh9c2cardiomyoblastcellsthroughthepkadependentsignalingpathway |
| _version_ |
1718394693998346240 |