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Articles in PresS, published online ahead of print June 27, 2002
Am J Physiol Lung Cell Mol Physiol, 10.1152/ajplung.00389.2001
Submitted on October 2, 2001
Accepted on May 30, 2002
1 Department of Physiology, University of Manitoba, Winnipeg, Manitoba, Canada
2 Unite de recherche en pheumologie, Universite Laval, Sainte-Foy, Quebec, Canada
* To whom correspondence should be addressed. E-mail: Newman_Stephens{at}UManitoba.ca.
Whether contractility of bronchial smooth muscle cells(BSMC) from asthmatic subjects is significantly altered has never been validated. Therefore we tested the hypothesis that BSMC from asthmatic subjects show increased contractility compared to normal cells. The cells were isolated by enzymatic methods from endobronchial biopsies. Electrically elicited shortening properties of BSMC were measured under an inverted microscope. Statistically significant increases in maximum shortening capacity (
LMAX) and velocity (VO) were found in asthmatic BSMC compared to normal cells (p<0.05, n = 5 in each group). Mean LMAX in asthmatic BSMC was 39.05% ±1.99 (SE) of resting cell length compared to 28.6% ±1.1 in normal cells; mean VO was 7.2% ±0.8% of resting cell length per second in asthmatic cells and 5.23% ±0.46 in normal cells. These values are of sufficient magnitude to represent a major component of asthmatic bronchospasm if they occurred in vivo. To investigate the mechanism of the observed increase in BSMC contractility, we measured mRNA abundance of smooth muscle types of myosin light chain kinase and myosin heavy chain. Analysis of RT-PCR data revealed that the content of message for myosin light chain kinase was higher in asthmatic BSMC (0.106 ±0.021 SE arbitrary densitometric units, n = 7) than in control cells (0.04 ±0.008, n =11); p < 0.05. Content and isoform distributions of the messages for myosin heavy chain isoforms showed no difference. Increased kinase message content could account for the increased velocity and capacity of shortening we report, on the assumption that the message is an indicator of protein content, and that the myosin Mg2-ATPase activity of smooth muscle is linear.
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