Mechanisms of aortic smooth muscle hyporeactivity after prolonged hypoxia in rats.
The aim of this study was to determine whether the effects of hypoxia on aortic contractility reflect a decrease in smooth muscle activation [phosphorylation of the 20-kDa myosin regulatory light chain (LC(20))], the capacity for myofibrillar ATP hydrolysis (mATPase activity), or both. Our results indicate that, in endothelium-denuded aortic rings from rats exposed to hypoxia for 48 h (inspired O(2) concentration = 10%), contractions to phenylephrine and potassium chloride (KCl) are impaired compared with rings from normoxic rats. The proportion of phosphorylated to total LC(20) during aortic contraction induced by 10(-5) M phenylephrine was reduced after hypoxia (51.4 +/- 5.4% in normoxic control rats vs. 32.5 +/- 4.7% in hypoxic rats, P < 0.01). Aortic mATPase activity was also decreased (maximum ATPase rate = 29.6 +/- 3.4 and 20.7 +/- 3.7 nmol. min(-1). mg protein(-1) in control and hypoxic rats, respectively, P < 0.05). Neither proliferation nor dedifferentiation of aortic smooth muscle was evident in this model; immunostaining for smooth muscle expression of the proliferating cell nuclear antigen was negative and smooth muscle-specific isoforms of myosin heavy chains, h-caldesmon, and calponin were increased, not decreased, after hypoxic exposure. Decreased aortic reactivity after hypoxia is associated with both impairment of smooth muscle activation and diminished capacity of the actomyosin complex, once activated, to hydrolyze ATP. These changes cannot be attributed to smooth muscle dedifferentiation or to reduced contractile protein expression.
Zacour
Mary E
ME
Meakins-Christie Laboratories, McGill University, Montreal H3A 2T5, Canada R3T 2N2.
Teoh
Hwee
H
Halayko
Andrew J
AJ
Ward
Michael E
ME
D000251:Adenosine Triphosphatases / Q000378:metabolism; D000818:Animals; D001011:Aorta / Q000378:metabolism / Q000503:physiopathology*; D003285:Contractile Proteins / Q000378:metabolism; D000860:Hypoxia / Q000503:physiopathology*; D007150:Immunohistochemistry; D066298:In Vitro Techniques; D019394:Ki-67 Antigen / Q000378:metabolism; D008297:Male; D008970:Molecular Weight; D009131:Muscle, Smooth, Vascular / Q000378:metabolism / Q000503:physiopathology*; D009210:Myofibrils / Q000201:enzymology; D018994:Myosin Light Chains / Q000737:chemistry / Q000378:metabolism; D010766:Phosphorylation; D051381:Rats; D017207:Rats, Sprague-Dawley; D013997:Time Factors; D014661:Vasoconstriction
D003285:Contractile Proteins; D019394:Ki-67 Antigen; D018994:Myosin Light Chains; D000251:Adenosine Triphosphatases