Lactate and PO2 modulate superoxide anion production in bovine cardiac myocytes: potential role of NADH oxidase.

Abstract

BACKGROUND:

Lactate increases lucigenin chemiluminescence (CL)-detectable superoxide anion (O2.-) generation in bovine vascular smooth muscle and endothelium, and a microsomal flavoprotein-containing NADH oxidase whose activity is regulated by PO2 and cytosolic NAD(H) redox appears to be the detected source of O2.- production. Little is known about the importance of this O2.(-)-producing system in cardiac myocytes.

METHODS AND RESULTS:

In isolated bovine cardiac myocytes, lactate (10 mmol/L) increased lucigenin-detectable O2.- levels to approximately 1.8 times baseline, whereas pyruvate (10 mmol/L) and mitochondrial probes did not increase the detection of O2.-. A nonmitochondrial NADH oxidase activity, found in microsomes containing a cytochrome b558, was a major source of O2.- production in the homogenate of myocytes, because NADH (0.1 mmol/L) increased basal lucigenin CL >100-fold. NADPH oxidases, mitochondria, and xanthine oxidase were minor sources of detectable O2.- production. However, mitochondria released H2O2 in the presence of 5 mmol/L succinate and 30 micromol/L antimycin, based on its detection as catalase-inhibitable luminol (+horseradish peroxidase)-elicited CL. Diphenyliodonium (DPI), an inhibitor of flavoprotein-containing oxidases, significantly attenuated basal, lactate, and NADH-elicited lucigenin CL. Hypoxia eliminated myocyte lucigenin CL, and posthypoxic reoxygenation caused an 8.6-fold increase in the detection of O2.- that was potentiated by lactate and inhibited by DPI.

CONCLUSIONS:

NADH oxidase activity linked to cytosolic NAD(H) redox appears to be a key source of O2.- production in cardiac myocytes that could contribute to oxidant signaling mechanisms and injury upon exposure to changes in PO2 and metabolites produced under hypoxia, such as lactate. These processes could contribute to the previously observed potentiation of injury caused by lactate in cardiac ischemia/reperfusion.

• Authors: K M Mohazzab-H, P M Kaminski, M S Wolin
• Journal: Circulation (Circulation) Vol. 96 Issue 2 Pg. 614-20 (Jul 15 1997) ISSN: 0009-7322 [Print] United States
• PMID: 9244234 (Publication Type: Journal Article, Research Support, U.S. Gov't, P.H.S.)

Chemical References

• Free Radicals
• Multienzyme Complexes
• Reactive Oxygen Species
• Superoxides
• Lactic Acid
• NADH oxidase
• NADH, NADPH Oxidoreductases

Topics

• Animals
• Cattle
• Cells, Cultured
• Free Radicals
• Lactic Acid (pharmacology)
• Multienzyme Complexes (metabolism)
• Myocardium (metabolism)
• NADH, NADPH Oxidoreductases (metabolism)
• Reactive Oxygen Species (metabolism)
• Superoxides (metabolism)