Cell lines

Cardiomyocyte cells from 1- to 2-d-old Wistar rats

Preparation Method

Cells incubated with a mixture of 100 µg/ml of Oleic acid (OA) and 400 µg/ml of bovine serum albumin for 12, 24, or 48 h at 37 °C, while the control group was treated with BSA alone for 24 or 48 h. To determine which protein kinases were activated by OA, use the

Reaction Conditions

1 µM for 24 hours

Applications

Calphostin C blocked OA-induced Cx43 Ser368 phosphorylation, showing involvement of PKC in this signaling cascade. In addition, PKCε Inhibitor Peptide, also blocked the effect, showing that PKCε was involved.

Animal models

Male FVB (H-2q) and C57BL/6J (H-2b) mice

Preparation Method

Recipient mice were treated with PKCε Inhibitor Peptide (n = 9, 20 mg/kg/day) or with TAT as a control (13 mg/kg/day; n = 8) using 0.1 mL osmotic pumps (release rate; 0.25 µL/h, 30 mM of each peptide in sterile saline) implanted subcutaneously on day 3, replaced on day 17 and left them until 30 days after transplantation.

Dosage form

osmotic pumps , 20 mg/kg/day

Applications

PKCε Inhibitor Peptide treatment significantly improved the beating score of cardiac allografts compared to TAT-peptide treatment, suggesting that adding PKCε Inhibitor Peptide treatment to CyA augmented preservation of graft function without toxic side effects. The beating score in the PKCε Inhibitor Peptide treated group at 30 days was equivalent to that after 14 days in the TAT control group

PKCε Inhibitor Peptide, also called εV1-2, is a protein kinase C ε (PKCε)-derived peptide, act as a selective PKCε inhibitor, inhibits the translocation of PKCε^[1]. PKCε Inhibitor Peptide is a peptide designed to compete with native nPKC ε to bind ε-Receptors for activated C Kinase (ε-RACK) and thereby inhibits nPKC ε catalytic activity due to decreased substrate accessibility.

PKCε Inhibitor Peptide, a selective PKCε inhibitor, the addition of PKCε Inhibitor Peptide interferes with the interaction between PKCε and its anchoring protein, and abolishes the cardioprotective effects of PKCε^[2]. ADP-induced thromboxane generation in human platelets pretreated with PKCε Inhibitor Peptide was more compared to the platelets pretreated with control peptide^[4].

PKCε Inhibitor Peptide were used to inhibit PKCε expression and activity. Apigenin-7-O-β-D-(-6"-p-coumaroyl)-glucopyranoside (APG) preconditioning-induced PKCε translocation to the mitochondria and anti-mitochondrial oxidative stress effects were attenuated by PKCε-targeted ε-V1-2 treatment in IR-injured hearts^[3].

References:
[1]. M Yedovitzky, et al. Translocation inhibitors define specificity of protein kinase C isoenzymes in pancreatic beta-cells. J Biol Chem. 1997 Jan 17;272(3):1417-20.
[2]. L. Tang, Y. Peng, T. Xu, X. Yi, Y. Liu, Y. Luo, D. Yin, M. He. The effects of quercetin protect cardiomyocytes from A/R injury is related to its capability to increasing expression and activity of PK C epsilon protein.Mol. Cell. Biochem., 382 (2013), pp. 145-152
[3]. Zhu Y, Di S, Hu W, et al.. A new flavonoid glycoside (APG) isolated from Clematis tangutica attenuates myocardial ischemia/reperfusion injury via activating PKCε signaling.Biochim Biophys Acta Mol Basis Dis. 2017; 1863:701-711. doi: 10.1016/j.bbadis.2016.12.013
[4]. Yamini Saraswathy Bynagari, B-Tech., Bela Nagy, M.D., et al. nPKC Epsilon Negatively Regulates Platelet Functional Responses 2008. Blood (2008) 112 (11): 2855.