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目的:建立测定微量硫化氢(Hydrogensulfide,H2S)的敏感硫电极法。方法:根据硫化氢的理化特性,应用化学反应将溶液中物理溶解和化学形式存在的硫化氢转变成硫离子(S2-),应用敏感硫电极检测微量S2-,换算出溶液中H2S,构建了敏感硫电极检测H2S的方法。并检测了大鼠及人血浆中H2S的浓度、大鼠心血管组织中内源性H2S的含量以及大鼠心血管组织和细胞胱硫醚γ裂解酶(cystathionineγlyase,CSE)的活性。结果:敏感硫电极检测1～80μmol/L的S2-有较好的指数相关关系,应用该方法检测到雄性和雌性大鼠血浆H2S的浓度分别为(40±4)和(41±5)μmol/L,差异无统计学意义,人类男性和女性静脉血血浆H2S浓度分别为(33±4)μmol/L和(35±5)μmol/L,差异无统计学意义。雌、雄大鼠主动脉组织H2S的含量分别为每毫克蛋白(24±6)和(25±5)nmol,心肌组织含量分别为每毫克蛋白(19±4)和(19±6)nmol,差异无统计学意义。采用敏感硫电极法测量主动脉组织CSE活性与传统方法测量结果差异无统计学意义,但可精确测量出血管平滑肌细胞CSE的活性。结论:敏感硫电极法可以应用于CSE/H2S信号通路的检测。
Objective: To establish a sensitive sulfur electrode method for the determination of hydrogen sulfide (H2S). Methods: According to the physical and chemical characteristics of H2S, the chemical reaction was used to convert the H2S dissolved physically and chemically into sulfide ion (S2-). The sensitive sulfur electrode was used to detect the trace amount of S2- and the H2S in solution was calculated. Sensitive sulfur electrode detection H2S method. The concentration of H2S in rat and human plasma, the content of endogenous H2S in rat cardiovascular tissue and the activity of rat cardiovascular tissue and cystathionine gammalyase (CSE) were also measured. Results: There was a good exponential relationship between the sensitivity of sulfur electrode and the concentration of S2- (1 ~ 80μmol / L). The concentrations of H2S in male and female rats were (40 ± 4) and (41 ± 5) μmol / L, the difference was not statistically significant. There was no significant difference in plasma H2S concentration between male and female patients (33 ± 4) μmol / L and (35 ± 5) μmol / L, respectively. The contents of H2S in the aorta of female and male rats were (24 ± 6) and (25 ± 5) nmol per mg respectively, and the contents of myocardial tissue were (19 ± 4) and (19 ± 6) nmol per mg respectively No statistical significance. There was no significant difference between the CSE activity measured by the sensitive sulfur electrode method and the traditional method, but the CSE activity of vascular smooth muscle cells could be accurately measured. Conclusion: The sensitive sulfur electrode method can be applied to the detection of CSE / H2S signaling pathway.