Astragaloside IV alleviates hypoxia/reoxygenation-induced neonatal rat cardiomyocyte injury via the protein kinase a pathway

Da Wei Zhang, Zhi Ping Bian, Jin Dan Xu, Heng Fang Wu, Chun Rong Gu, Bin Zhou, Xiang Jian Chen, Di Yang

Research output: Contribution to journalArticle

7 Citations (Scopus)

Abstract

Background: Astragaloside IV (As-IV) exerts beneficial effects on hypoxia/reoxygenation (H/R)-induced cardiomyocyte injury, possibly through normalization of sarcoplasmic reticulum Ca2+ ATPase (SERCA2a) function. The exact mechanisms remain unknown. This study was designed to investigate the role of protein kinase A (PKA) in the protective effect of As-IV on SERCA2a function. Methods: Cultured cardiomyocytes from neonatal rats were exposed to 6 h of hypoxia followed by 3 h of reoxygenation (H/R) with or without As-IV treatment. Myocyte injury was determined by the creatine kinase (CK)-MB fraction in supernatant. Myocardial SERCA2a activity and PKA kinase activity were assessed. PKA subunit mRNA expression and Ser16 phosphorylated phospholamban (Ser16-PLN) protein expression were detected by real-time PCR and Western blot, respectively. Results: The administration of As-IV significantly decreased CK-MB release and restored SERCA2a activity in H/R cardiomyocytes. The mRNAs of PKA subunits, PKA-RIα, PKA-RIIα, PKA-RIIβ, PKA-Cα and PKA-Cβ, were downregulated in H/R cardiomyocytes. However, PKA-Cα mRNA expression was significantly increased after As-IV treatment. Meanwhile, there was a tendency to recovery of the H/R-induced PKA kinase activity decrease after As-IV treatment. The expression of Ser16-PLN protein, which is specifically phosphorylated by PKA, was upregulated in As-IV-treated H/R cardiomyocytes. Conclusions: These results suggest that the cardioprotection of As-IV may be through the upregulation of PKA and Ser16-PLN, thereby restoring SERCA2a function in H/R injury.

Original languageEnglish (US)
Pages (from-to)95-101
Number of pages7
JournalPharmacology
Volume90
Issue number1-2
DOIs
StatePublished - Aug 2012

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Cyclic AMP-Dependent Protein Kinases
Cardiac Myocytes
Protein Kinases
Wounds and Injuries
MB Form Creatine Kinase
Hypoxia
astragaloside A
Messenger RNA
Protein Kinase C
Phosphotransferases
Calcium-Transporting ATPases
Sarcoplasmic Reticulum
Muscle Cells
Real-Time Polymerase Chain Reaction
Proteins
Up-Regulation
Down-Regulation
Western Blotting

Keywords

  • Astragaloside IV
  • Cardioprotection
  • Hypoxia/Reoxygenation
  • Phospholamban
  • Protein kinase A
  • Sarcoplasmic reticulum calcium ATPase

ASJC Scopus subject areas

  • Pharmacology

Cite this

Astragaloside IV alleviates hypoxia/reoxygenation-induced neonatal rat cardiomyocyte injury via the protein kinase a pathway. / Zhang, Da Wei; Bian, Zhi Ping; Xu, Jin Dan; Wu, Heng Fang; Gu, Chun Rong; Zhou, Bin; Chen, Xiang Jian; Yang, Di.

In: Pharmacology, Vol. 90, No. 1-2, 08.2012, p. 95-101.

Research output: Contribution to journalArticle

Zhang, Da Wei ; Bian, Zhi Ping ; Xu, Jin Dan ; Wu, Heng Fang ; Gu, Chun Rong ; Zhou, Bin ; Chen, Xiang Jian ; Yang, Di. / Astragaloside IV alleviates hypoxia/reoxygenation-induced neonatal rat cardiomyocyte injury via the protein kinase a pathway. In: Pharmacology. 2012 ; Vol. 90, No. 1-2. pp. 95-101.
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abstract = "Background: Astragaloside IV (As-IV) exerts beneficial effects on hypoxia/reoxygenation (H/R)-induced cardiomyocyte injury, possibly through normalization of sarcoplasmic reticulum Ca2+ ATPase (SERCA2a) function. The exact mechanisms remain unknown. This study was designed to investigate the role of protein kinase A (PKA) in the protective effect of As-IV on SERCA2a function. Methods: Cultured cardiomyocytes from neonatal rats were exposed to 6 h of hypoxia followed by 3 h of reoxygenation (H/R) with or without As-IV treatment. Myocyte injury was determined by the creatine kinase (CK)-MB fraction in supernatant. Myocardial SERCA2a activity and PKA kinase activity were assessed. PKA subunit mRNA expression and Ser16 phosphorylated phospholamban (Ser16-PLN) protein expression were detected by real-time PCR and Western blot, respectively. Results: The administration of As-IV significantly decreased CK-MB release and restored SERCA2a activity in H/R cardiomyocytes. The mRNAs of PKA subunits, PKA-RIα, PKA-RIIα, PKA-RIIβ, PKA-Cα and PKA-Cβ, were downregulated in H/R cardiomyocytes. However, PKA-Cα mRNA expression was significantly increased after As-IV treatment. Meanwhile, there was a tendency to recovery of the H/R-induced PKA kinase activity decrease after As-IV treatment. The expression of Ser16-PLN protein, which is specifically phosphorylated by PKA, was upregulated in As-IV-treated H/R cardiomyocytes. Conclusions: These results suggest that the cardioprotection of As-IV may be through the upregulation of PKA and Ser16-PLN, thereby restoring SERCA2a function in H/R injury.",
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T1 - Astragaloside IV alleviates hypoxia/reoxygenation-induced neonatal rat cardiomyocyte injury via the protein kinase a pathway

AU - Zhang, Da Wei

AU - Bian, Zhi Ping

AU - Xu, Jin Dan

AU - Wu, Heng Fang

AU - Gu, Chun Rong

AU - Zhou, Bin

AU - Chen, Xiang Jian

AU - Yang, Di

PY - 2012/8

Y1 - 2012/8

N2 - Background: Astragaloside IV (As-IV) exerts beneficial effects on hypoxia/reoxygenation (H/R)-induced cardiomyocyte injury, possibly through normalization of sarcoplasmic reticulum Ca2+ ATPase (SERCA2a) function. The exact mechanisms remain unknown. This study was designed to investigate the role of protein kinase A (PKA) in the protective effect of As-IV on SERCA2a function. Methods: Cultured cardiomyocytes from neonatal rats were exposed to 6 h of hypoxia followed by 3 h of reoxygenation (H/R) with or without As-IV treatment. Myocyte injury was determined by the creatine kinase (CK)-MB fraction in supernatant. Myocardial SERCA2a activity and PKA kinase activity were assessed. PKA subunit mRNA expression and Ser16 phosphorylated phospholamban (Ser16-PLN) protein expression were detected by real-time PCR and Western blot, respectively. Results: The administration of As-IV significantly decreased CK-MB release and restored SERCA2a activity in H/R cardiomyocytes. The mRNAs of PKA subunits, PKA-RIα, PKA-RIIα, PKA-RIIβ, PKA-Cα and PKA-Cβ, were downregulated in H/R cardiomyocytes. However, PKA-Cα mRNA expression was significantly increased after As-IV treatment. Meanwhile, there was a tendency to recovery of the H/R-induced PKA kinase activity decrease after As-IV treatment. The expression of Ser16-PLN protein, which is specifically phosphorylated by PKA, was upregulated in As-IV-treated H/R cardiomyocytes. Conclusions: These results suggest that the cardioprotection of As-IV may be through the upregulation of PKA and Ser16-PLN, thereby restoring SERCA2a function in H/R injury.

AB - Background: Astragaloside IV (As-IV) exerts beneficial effects on hypoxia/reoxygenation (H/R)-induced cardiomyocyte injury, possibly through normalization of sarcoplasmic reticulum Ca2+ ATPase (SERCA2a) function. The exact mechanisms remain unknown. This study was designed to investigate the role of protein kinase A (PKA) in the protective effect of As-IV on SERCA2a function. Methods: Cultured cardiomyocytes from neonatal rats were exposed to 6 h of hypoxia followed by 3 h of reoxygenation (H/R) with or without As-IV treatment. Myocyte injury was determined by the creatine kinase (CK)-MB fraction in supernatant. Myocardial SERCA2a activity and PKA kinase activity were assessed. PKA subunit mRNA expression and Ser16 phosphorylated phospholamban (Ser16-PLN) protein expression were detected by real-time PCR and Western blot, respectively. Results: The administration of As-IV significantly decreased CK-MB release and restored SERCA2a activity in H/R cardiomyocytes. The mRNAs of PKA subunits, PKA-RIα, PKA-RIIα, PKA-RIIβ, PKA-Cα and PKA-Cβ, were downregulated in H/R cardiomyocytes. However, PKA-Cα mRNA expression was significantly increased after As-IV treatment. Meanwhile, there was a tendency to recovery of the H/R-induced PKA kinase activity decrease after As-IV treatment. The expression of Ser16-PLN protein, which is specifically phosphorylated by PKA, was upregulated in As-IV-treated H/R cardiomyocytes. Conclusions: These results suggest that the cardioprotection of As-IV may be through the upregulation of PKA and Ser16-PLN, thereby restoring SERCA2a function in H/R injury.

KW - Astragaloside IV

KW - Cardioprotection

KW - Hypoxia/Reoxygenation

KW - Phospholamban

KW - Protein kinase A

KW - Sarcoplasmic reticulum calcium ATPase

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