[Application of sensitive sulphur electrode assay to measure and analyze cystathionine-gamma-lyase/hydrogen sulfide pathway of cardiovascular tissues, cells and plasma in rats].
OBJECTIVE
To construct a method of measurement microamount hydrogen sulfide (H(2)S) using sensitive sulphur electrode.


METHODS
According to the physical and chemical characters of H(2)S, H(2)S, which in the fluids by mean of physical dissolve and chemical shape, is turned to sulphur ion (S(2-)) by chemical responses. After the microamount of S(2-) was measured by sensitive sulphur electrode, and the concentration of H(2)S was converted, a method was constructed to measure the H(2)S. It was used to analyze the concentrations of H(2)S of plasma in rats and humans, the endogenous concentration of H(2)S of cardiovascular tissue in rats, and CSE activity of cardiovascular tissues and cells.


RESULTS
The exponential regression of S(2-) in the extent including 1 to 80 micromol/L was found using sensitive sulphur electrode. The H(2)S levels of plasma in male and female rats were 40+/-4 and 41+/-5 micromol/L, respectively, and significant difference was not found; those in venous blood plasma of men and women were 33+/-4 micromol/L and 35+/-5 micromol/L respectively, without significant difference. There were not significant differences in the aortic endogenous levels of H(2)S (24+/-6 and 25+/-5 nmol/mg pro) and myocardial levels (19+/-4 and 19+/-6 nmol/mg protein) between female and male rats. There were no different results of CSE activity in aortal tissue using sensitive sulphur electrode or traditional methods, however, the CSE activity of vascular smooth muscle cells could be accurately measured using sensitive sulphur electrode, which was difficult in using traditional method.


CONCLUSIONS
The sensitive sulphur electrode assay was fit for the analysis of CSE/H(2)S pathway in cardiovascular research.
Geng
Bin
B
Institute of Cardiovascular Research, Peking University First Hospital, Beijing 100034, China.
Du
Jun-bao
JB
Tang
Chao-shu
CS
D000818:Animals; D002319:Cardiovascular System / Q000378:metabolism; D003542:Cystathionine gamma-Lyase / Q000378:metabolism*; D004566:Electrodes; D005260:Female; D006862:Hydrogen Sulfide / Q000097:blood / Q000378:metabolism*; D008297:Male; D009131:Muscle, Smooth, Vascular / Q000378:metabolism*; D009206:Myocardium / Q000378:metabolism*; D051381:Rats; D017207:Rats, Sprague-Dawley; D015398:Signal Transduction; D013455:Sulfur
D013455:Sulfur; D003542:Cystathionine gamma-Lyase; D006862:Hydrogen Sulfide