Intracellular mechanisms involved in the responses of cerebrovascular smooth-muscle cells to hemoglobin

Abstract

✓ An investigation was undertaken of the mechanism by which oxyhemoglobin and its analog methemoglobin might cause cerebrovascular spasm. The effect of these compounds on the levels of intracellular inositol triphosphate and calcium in cultured primate cerebrovascular smooth-muscle cells and the contractile action of oxyhemoglobin on isolated rings of primate cerebral arteries were also examined. Oxyhemoglobin, but not methemoglobin, produces a transient but highly significant increase in the intracellular levels of inositol triphosphate. Intracellular calcium levels in these cells are increased by thrombin, aluminum tetrafluoride, and oxyhemoglobin, and the sustained elevation in intracellular calcium is prevented by ethyleneglycol tetra-acetic acid and the phospholipase C inhibitor neomycin. Removal of the oxyhemoglobin after as long as 48 hours' incubation with this compound allowed cells to rapidly reduce their intracellular calcium levels to near normal. Oxyhemoglobin produced contractions of isolated rings of both normal and spastic cerebral arteries, although the response of spastic vessels was significantly smaller. This effect was inhibited by neomycin. The addition of neomycin relaxed arteries that were contracted with oxyhemoglobin, 5-hydroxytryptamine, or potassium chloride. It is thus likely that activation of phospholipase C is a critical step in the development of vasospasm, but the transient nature of the response to inositol triphosphate suggests that the sustained contraction may arise from other phospholipase C-dependent mechanisms.