Application of sensitive sulphur electrode assay to measure and analyze cystathionine-?-lyase/hydrogen sulfide pathway of cardiovascular tissues, cells and plasma in rats / 北京大学学报(医学版)

ABSTRACT

Objective:To constract a method of measurement microamount hydrogen sulfide (H_2S) using sensitive sulphur electrode. Methods: According to the physical and chemical characters of H_2S, H_2S, 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_2S was converted, a method was constructed to measure the H_2S. It was used to analyze the concentrations of H_2S of plasma in rats and humans, the endogenous concentration of H_2S 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 ?mol/L was found using sensitive sulphur electrod. The H_2S levels of plasma in male and female rats were 40?4 and 41?5 ?mol/L, respectively, and significant difference was not found; those in venous blood plasma of men and women were 33?4 ?mol/L and 35?5 ?mol/L respectively, without significant difference. There were not significant differences in the aortic endogenous levels of H_2S (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. Conclusion: The sensitive sulphur electrode assay was fit for the analysis of CSE/H_2S pathway in cardiovascular research.