Atherogenic L5 LDL induces cardiomyocyte apoptosis and inhibits KATP channels through CaMKII activation.
BACKGROUND
Cardiac Ca2+/calmodulin-dependent protein kinase II (CaMKII) activation plays a critical role in cardiomyocyte (CM) apoptosis and arrhythmia. Functional ATP-sensitive potassium (KATP) channels are essential for cardiac protection during ischemia. In cultured CMs, L5 low-density lipoprotein (LDL) induces apoptosis and QTc prolongation. L5 is a highly electronegative and atherogenic aberrant form of LDL, and its levels are significantly higher in patients with cardiovascular-related diseases. Here, the role of L5 in cardiac injury was studied by evaluating the effects of L5 on CaMKII activity and KATP channel physiology in CMs.


METHODS
Cultured neonatal rat CMs (NRCMs) were treated with a moderate concentration (ie, 7.5 μg/mL) of L5 or L1 (the least electronegative LDL subfraction). NRCMs were examined for apoptosis and viability, CaMKII activity, and the expression of phosphorylated CaMKIIδ and NOX2/gp91phox. The function of KATP and action potentials (APs) was analyzed by using the patch-clamp technique.


RESULTS
In NRCMs, L5 but not L1 significantly induced cell apoptosis and reduced cell viability. Furthermore, L5 decreased Kir6.2 expression by more than 50%. Patch-clamp analysis showed that L5 reduced the KATP current (IKATP) density induced by pinacidil, a KATP opener. The partial recovery of the inward potassium current during pinacidil washout was susceptible to subsequent inhibition by the IKATP blocker glibenclamide. Suppression of IKATP by L5 significantly prolonged the AP duration. L5 also significantly increased the activity of CaMKII, the phosphorylation of CaMKIIδ, and the expression of NOX2/gp91phox. L5-induced apoptosis was prevented by the addition of the CaMKII inhibitor KN93 and the reactive oxygen species scavenger Mn (III)TBAP.


CONCLUSIONS
L5 but not L1 induces CM damage through the activation of the CaMKII pathway and increases arrhythmogenicity in CMs by modulating the AP duration. These results help to explain the harmful effects of L5 in cardiovascular-related disease.
['Ma|Yanzhuo|Y|', 'Cheng|Nancy|N|', 'Sun|Junping|J|', 'Lu|Jonathan Xuhai|JX|', 'Abbasi|Shahrzad|S|', 'Wu|Geru|G|', 'Lee|An-Sheng|AS|', 'Sawamura|Tatsuya|T|', 'Cheng|Jie|J|', 'Chen|Chu-Huang|CH|', 'Xi|Yutao|Y|http://orcid.org/0000-0002-2537-2527']
D000200:Action Potentials D000818:Animals D017209:Apoptosis D015153:Blotting, Western D054732:Calcium-Calmodulin-Dependent Protein Kinase Type 2 D002470:Cell Survival D004594:Electrophysiology D054086:KATP Channels D008077:Lipoproteins, LDL D032383:Myocytes, Cardiac D018408:Patch-Clamp Techniques D010766:Phosphorylation D024661:Potassium Channels, Inwardly Rectifying D051381:Rats D017207:Rats, Sprague-Dawley D017382:Reactive Oxygen Species D015398:Signal Transduction
Cardiomyocytes Ca2+/calmodulin-dependent protein kinase II ATP-sensitive potassium Action potential Electronegative low-density lipoprotein