Jin Hong-fang, Du Jun-bao, Li Xiao-hui, Wang Yan-fei, Liang Yin-fang, Tang Chao-shu
Department of Pediatrics, Peking University First Hospital, Beijing 100034, China.
Acta Pharmacol Sin. 2006 Dec;27(12):1561-6. doi: 10.1111/j.1745-7254.2006.00425.x.
To investigate the interaction between hydrogen sulfide (H2S)/cystathionine gamma-lyase (CSE) and carbon monoxide (CO)/heme oxygenase (HO) pathways in aortic smooth muscle cells (ASMC).
The ASMCs were divided into the following groups: (1) the control group; (2) the zinc protoporphyrin (ZnPP) 20 micromol/L group; (3) the propargylglycine (PPG) 2 mmol/L, 4 mmol/L and 10 mmol/L groups; and (4) the sodium hydrosulfide (NaHS) 1 x 10(-5) mol/L, 1 x 10(-4) mol/L and 1 x 10(-3) mol/L groups. Each of the groups was further divided into 6 h, 12 h, 18 h and 24 h subgroups. The CO level, represented by carboxyhemoglobin (HbCO) content was measured using a spectrophotometric method and H2S content was detected by a sensitive electrode method. CSE and HO-1 expressions were detected by Western blotting.
The H2S content in the medium and CSE expression by ASMC were markedly increased by ZnPP compared with the control group. HbCO content in the medium and HO-1 expression by the ASMC started strengthening following 24 h treatment with PPG at 2 mmol/L, but were further strengthened following 18 h and 24 h treatment with PPG at 4 mmol/L compared with the controls (P < 0.01). PPG at 10 mmol/L increased the HbCO level in the medium following 18 h treatment and increased HO-1 expression by the ASMC following 12 h treatment. Moreover, NaHS at 1 x 10(-5) mol/L and 1 x 10(-4) mol/L decreased the HbCO level in the medium and HO-1 expression by the ASMC after 6 h and 12 h treatment, while NaHS at 1 x 10(-3) mol/L decreased them at all time points of the treatments.
The results suggested that endogenous CO/HO and H2S/CSE pathways inhibited each other in ASMC under physiological conditions.
研究主动脉平滑肌细胞（ASMC）中硫化氢（H2S）/胱硫醚γ-裂解酶（CSE）与一氧化碳（CO）/血红素加氧酶（HO）途径之间的相互作用。
将ASMC分为以下几组：（1）对照组；（2）20 μmol/L锌原卟啉（ZnPP）组；（3）2 mmol/L、4 mmol/L和10 mmol/L炔丙基甘氨酸（PPG）组；（4）1×10⁻⁵ mol/L、1×10⁻⁴ mol/L和1×10⁻³ mol/L硫氢化钠（NaHS）组。每组再进一步分为6 h、12 h、18 h和24 h亚组。采用分光光度法测定以碳氧血红蛋白（HbCO）含量表示的CO水平，并用灵敏电极法检测H2S含量。通过蛋白质印迹法检测CSE和HO-1的表达。
与对照组相比，ZnPP显著增加了培养基中H2S含量及ASMC的CSE表达。在2 mmol/L PPG处理24 h后，培养基中HbCO含量及ASMC的HO-1表达开始增强，但与对照组相比，在4 mmol/L PPG处理18 h和24 h后进一步增强（P < 0.01）。10 mmol/L PPG在处理18 h后增加了培养基中HbCO水平，在处理12 h后增加了ASMC的HO-1表达。此外，1×10⁻⁵ mol/L和1×10⁻⁴ mol/L NaHS在处理6 h和12 h后降低了培养基中HbCO水平及ASMC的HO-1表达，而1×10⁻³ mol/L NaHS在处理的所有时间点均降低了它们。
结果表明，在生理条件下，内源性CO/HO和H2S/CSE途径在ASMC中相互抑制。
