Knockdown of forkhead box protein P1 alleviates hypoxia reoxygenation injury in H9c2 cells through regulating Pik3ip1/Akt/eNOS and ROS/mPTP pathway

Bioengineered. 2022 Jan;13(1):1320-1334. doi: 10.1080/21655979.2021.2016046.

Abstract

Forkhead box protein P1 (Foxp1) exerts an extensive array of physiological and pathophysiological impacts on the cardiovascular system. However, the exact function of myocardial Foxp1 in myocardial ischemic reperfusion injury (MIRI) stays largely vague. The hypoxia reoxygenation model of H9c2 cells (the rat ventricular myoblasts) closely mimics myocardial ischemia-reperfusion injury. This report intends to research the effects and mechanisms underlying Foxp1 on H9c2 cells in response to hypoxia (12 h)/reoxygenation (4 h) (HR) stimulation. Expressions of Foxp1 and Phosphatidylinositol 3-kinase interacting protein 1 (Pik3ip1) were both upregulated in ischemia/reperfusion (IR)/HR-induced injury. Stimulation through HR led to marked increases in cellular apoptosis, mitochondrial dysfunction, and superoxide generation in H9c2 cells, which were rescued with knockdown of Foxp1 by siRNA. Silence of Foxp1 depressed expression of Pik3ip1 directly activated the PI3K/Akt/eNOS pathway and promoted nitric oxide (NO) release. Moreover, the knockdown of Foxp1 blunted HR-induced enhancement of reactive oxygen species (ROS) generation, thus alleviating excessive persistence of mitochondrial permeability transition pore (mPTP) opening and decreased mitochondrial apoptosis-associated protein expressions in H9c2 cells. Meanwhile, these cardioprotective effects can be abolished by LY294002, NG-nitro-L-arginine methyl ester (L-NAME), and Atractyloside (ATR), respectively. In summary, our findings indicated that knockdown of Foxp1 prevented HR-induced encouragement of apoptosis and oxidative stress via PI3K/Akt/eNOS signaling activation by targeting Pik3ip1 and improved mitochondrial function by inhibiting ROS-mediated mPTP opening. Inhibition of Foxp1 may be a promising therapeutic avenue for MIRI.

Keywords: Foxp1; H9c2 cells; Myocardial ischemic reperfusion injury; PI3K/Akt/eNOS pathway; Pik3ip1; hypoxia/reoxygenation.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Atractyloside / pharmacology
  • Cell Line
  • Cell Survival
  • Chromones / pharmacology
  • Forkhead Transcription Factors / genetics*
  • Forkhead Transcription Factors / metabolism
  • Gene Expression Regulation / drug effects
  • Gene Knockdown Techniques
  • Intracellular Signaling Peptides and Proteins / genetics
  • Intracellular Signaling Peptides and Proteins / metabolism*
  • Mitochondrial Permeability Transition Pore / metabolism*
  • Models, Biological
  • Morpholines / pharmacology
  • Myocardial Reperfusion Injury / genetics*
  • Myocardial Reperfusion Injury / metabolism
  • Myocytes, Cardiac / cytology*
  • Myocytes, Cardiac / drug effects
  • Myocytes, Cardiac / metabolism
  • NG-Nitroarginine Methyl Ester / pharmacology
  • Nitric Oxide Synthase Type III / metabolism*
  • Proto-Oncogene Proteins c-akt / metabolism
  • Rats
  • Reactive Oxygen Species / metabolism*
  • Repressor Proteins / genetics*
  • Repressor Proteins / metabolism
  • Signal Transduction / drug effects
  • Up-Regulation / drug effects

Substances

  • Chromones
  • Forkhead Transcription Factors
  • FoxP1 protein, rat
  • Intracellular Signaling Peptides and Proteins
  • Mitochondrial Permeability Transition Pore
  • Morpholines
  • Reactive Oxygen Species
  • Repressor Proteins
  • Atractyloside
  • 2-(4-morpholinyl)-8-phenyl-4H-1-benzopyran-4-one
  • Nitric Oxide Synthase Type III
  • Nos3 protein, rat
  • Proto-Oncogene Proteins c-akt
  • NG-Nitroarginine Methyl Ester

Grants and funding

This work was supported by the State key R & D plan of the Ministry of science and technology (2016YFC1301101, 2016YFC1301102); the National Major Research Plan Training Program of the National Natural Science Foundation of China (No. 92168117; 81370314; 91849111; 81770253).