Voltage-independent calcium channels mediate lipopolysaccharide-induced hyporeactivity to endothelin-1 in the rat aorta.

The roles of intracellular calcium concentration ([Ca(2+)](i)) and Ca(2+) sensitization in lipopolysaccharide (LPS)-induced vascular smooth muscle (VSM) hyporesponsiveness are incompletely understood. To investigate these roles, contraction responses to endothelin-1 (ET-1) and 80 mM KCl; relaxation responses to nifedipine; the expression levels of mRNAs of ET-1 and its receptors (ET(A) or ET(B)); the expression levels of protein kinase C (PKC) and phosphorylation of Rho kinase (ROKalpha), CPI-17, and myosin phosphatase target subunit-1 (MYPT1); and changes in aortic VSM cell [Ca(2+)](i) were measured in LPS-treated aortic rings from male Wistar rats (250-300 g). LPS (10 mug/ml, 20 h) decreased contraction induced by ET-1 (0.3-100 nM) or 80 mM KCl. LPS-induced hypocontractility was not observed in the absence of external Ca(2+), but LPS-treated aorta remained hypocontractile on subsequent stepwise restoration of extracellular Ca(2+) (0.01-10 mM). Vascular relaxation to nifedipine; mRNA expression levels of ET-1, ET(A), or ET(B); protein expression levels of PKC; and phosphorylation levels of ROKalpha, CPI-17, and MYPT1 were not affected by LPS. In isolated aortic VSM cells, ET-1 caused a transient initial increase in [Ca(2+)](i), followed by a maintained tonic increase in [Ca(2+)](i), which was decreased by LPS pretreatment and was dependent on external Ca(2+). Subsequent restoration of extracellular Ca(2+) increased [Ca(2+)](i), but this increase was lower in the LPS-treated group. This difference in response to extracellular Ca(2+) addition was not affected by diltiazem, but was abolished by SKF-96365. Therefore, LPS induces hyporeactivity to ET-1 in rat aorta that depends on external Ca(2+) influx through non-voltage-operated Ca(2+) channels, but not on ET-1 receptor expression or Ca(2+) sensitization.