Enhanced phospholipase C activity in the cultured skin fibroblast obtained from patients with coronary spastic angina: possible role for enhanced vasoconstrictor response

Abstract

OBJECTIVESWe measured phospholipase C (PLC) activity in the cultured skin fibroblasts obtained from patients with and without coronary spasm and examined its correlation with coronary artery vasomotility.BACKGROUNDCoronary artery vasomotility is enhanced in coronary spastic angina (CSA), but no information is available for the intracellular signaling. In spontaneously hypertensive rats, PLC activity in the skin fibroblasts has been shown to be enhanced.METHODSSkin fibroblasts obtained from 24 patients with CSA—14 with organic coronary artery disease (CAD) and 12 control subjects—were cultured by the explant method. Activity of PLC was determined by incubating the membrane fraction with 3H-phosphatidyl inositol bisphosphate and by quantifying 3H-inositol trisphosphate. In patients with CSA and control subjects, the relations between PLC activity and coronary artery basal tone and constrictor response to intracoronary acetylcholine (ACh) were examined.RESULTSActivity of PLC (pmol/protein [mg] per min) was 1.74 ± 0.19 in patients with CSA; 0.90 ± 0.12 in patients with CAD; and 0.65 ± 0.07 in control subjects (p < 0.001, patients with CSA vs. patients with CAD and control subjects; p = NS, patients with CAD vs. control subjects). According to the Lineweaver-Burk plot, Michaelis constant (μmol/liter) of PLC was 28 ± 4 in patients with CSA; 49 ± 14 in patients with CAD; and 56 ± 10 in control subjects (p < 0.05, patients with CSA vs. control subjects), whereas the maximal velocity was not different between the three groups. There were significant positive correlations between PLC activity and both basal tone (p = 0.0108) and response to ACh (p = 0.0053). Western blot analysis using membrane fraction demonstrated that 89% of PLC isoenzymes detected was of the δ1 isoform.CONCLUSIONSBecause the PLC activity measured was genetically defined and was positively correlated with coronary artery vasomotility, enhanced PLC activity may be involved in the pathogenesis of coronary spasm.