Introduction
Calpain is a family of cysteine proteases that has an important role in the initiation,
regulation, and execution of cell death. Our recent studies using a hypercholesterolemic
swine model demonstrated that in the setting of the metabolic syndrome, calpain inhibition
(CI) improved collateral-dependent perfusion and increased expression of proteins
implicated in angiogenesis and vasodilation. In this study, we hypothesized that CI
(by MLD28170) would decrease myocardial apoptosis in the same model.
Methods
Yorkshire swine, all fed a high-cholesterol diet for 4 weeks underwent placement of
an ameroid constrictor on the left circumflex coronary artery. Three weeks later,
animals received either no drug, termed the high-cholesterol control group (HCC; n = 8); low-dose CI (0.12 mg/kg; LCI, n = 9); or high-dose CI (0.25 mg/kg; HCI, n = 8). The high-cholesterol diet and the CI were continued for 5 weeks, after which
the pig was humanely killed and the left ventricular myocardium was harvested and
analyzed via terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL)
staining, oxyblot analysis, and Western blots. Data were analyzed using the Kruskal–Wallis
test.
Results
The percentage of apoptotic cells to total cells in ischemic myocardial territory
was decreased in the LCI and HCI groups compared with the HCC group as shown by TUNEL
staining (P = .018). There was a decrease in proapoptotic proteins, including cleaved caspase
3, caspase 9, cleaved caspase 9, Bax, BAD, p-BAD, and Erk 1/2 (P ≤ .049 each), but no decrease in caspase 3 (P = .737). There was also an increase in antiapoptotic proteins, including BCL-2 and
p-BCL2 (P ≤ .025 each). In the ischemic myocardium, several proangiogenic proteins were increased
in the LCI and HCI groups compared with the HCC group, including p-AKT, p-eNOS, and
eNOS (P ≤ .006 each) but there was no increase in AKT (P = .311). CI decreased tissue oxidative stress in both the LCI and HCI groups compared
to the HCC group as shown by oxyblot analysis (P = .021).
Conclusion
In the setting of hypercholesterolemia, CI decreases apoptosis and the expression
of proteins in proapoptotic signaling pathways. CI also increased expression of proteins
implicated in anti apoptotic pathways and improves oxidative stress in ischemic myocardial
tissue.
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Article info
Publication history
Published online: May 16, 2015
Accepted:
March 18,
2015
Identification
Copyright
© 2015 Elsevier Inc. Published by Elsevier Inc. All rights reserved.
