TY - JOUR
T1 - Role of the TRPM4 channel in mitochondrial function, calcium release, and ROS generation in oxidative stress
AU - Wang, Chen
AU - Chen, Jian
AU - Wang, Mengxue
AU - Naruse, Keiji
AU - Takahashi, Ken
N1 - Funding Information:
This research was funded by JSPS KAKENHI , Fund for the Promotion of Joint International Research (Fostering Joint International Research), 17KK0168 and JSPS KAKENHI Grant-In-Aid for Scientific Research (B), 20H04518 .
Publisher Copyright:
© 2021 The Author(s)
PY - 2021/8/20
Y1 - 2021/8/20
N2 - Ischemic heart disease is one of the most common causes of death worldwide. Mitochondrial dysfunction, excessive reactive oxygen species (ROS) generation, and calcium (Ca2+) overload are three key factors leading to myocardial death during ischemia-reperfusion (I/R) injury. Inhibition of TRPM4, a Ca2+-activated nonselective cation channel, protects the rat heart from I/R injury, but the specific mechanism underlying this effect is unclear. In this study, we investigated the mechanism of cardioprotection against I/R injury via TRPM4 using hydrogen peroxide (H2O2), a major contributor to oxidative stress, as an I/R injury model. We knocked out the TRPM4 gene in the rat cardiomyocyte cell line H9c2 using CRISPR/Cas9. Upon H2O2 treatment, intracellular Ca2+ level and ROS production increased in wild type (WT) cells but not in TRPM4 knockout (TRPM4KO) cells. With this treatment, two indicators of mitochondrial function, mitochondrial membrane potential (ΔΨm) and intracellular ATP levels, decreased in WT but not in TRPM4KO cells. Taken together, these findings suggest that blockade of the TRPM4 channel might protect the myocardium from oxidative stress by maintaining the mitochondrial membrane potential and intracellular ATP levels, possibly through preventing aberrant increases in intracellular Ca2+ and ROS.
AB - Ischemic heart disease is one of the most common causes of death worldwide. Mitochondrial dysfunction, excessive reactive oxygen species (ROS) generation, and calcium (Ca2+) overload are three key factors leading to myocardial death during ischemia-reperfusion (I/R) injury. Inhibition of TRPM4, a Ca2+-activated nonselective cation channel, protects the rat heart from I/R injury, but the specific mechanism underlying this effect is unclear. In this study, we investigated the mechanism of cardioprotection against I/R injury via TRPM4 using hydrogen peroxide (H2O2), a major contributor to oxidative stress, as an I/R injury model. We knocked out the TRPM4 gene in the rat cardiomyocyte cell line H9c2 using CRISPR/Cas9. Upon H2O2 treatment, intracellular Ca2+ level and ROS production increased in wild type (WT) cells but not in TRPM4 knockout (TRPM4KO) cells. With this treatment, two indicators of mitochondrial function, mitochondrial membrane potential (ΔΨm) and intracellular ATP levels, decreased in WT but not in TRPM4KO cells. Taken together, these findings suggest that blockade of the TRPM4 channel might protect the myocardium from oxidative stress by maintaining the mitochondrial membrane potential and intracellular ATP levels, possibly through preventing aberrant increases in intracellular Ca2+ and ROS.
KW - CRISPR/Cas9
KW - Calcium
KW - Mitochondrial function
KW - Myocardial ischemia-reperfusion injury
KW - Reactive oxygen species
KW - TRPM4
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U2 - 10.1016/j.bbrc.2021.03.077
DO - 10.1016/j.bbrc.2021.03.077
M3 - Article
C2 - 34144257
AN - SCOPUS:85107265746
VL - 566
SP - 190
EP - 196
JO - Biochemical and Biophysical Research Communications
JF - Biochemical and Biophysical Research Communications
SN - 0006-291X
ER -