【摘要】 目的:改进一种新的硫化氢供体合成方法,在培养的HepG2细胞上观察了其细胞毒性,予ICR小鼠腹腔注射观察其组织分布及代谢途径,为硫化氢研究中合理使用该供体提供实验依据。方法:根据文献方法加以改进合成新型供体,采用台盼蓝染色鉴定细胞活力及乳酸脱氢酶(lactate dehydrogenase,LDH)活性检测细胞毒性;利用电极法测定注射供体后不同时间点心、肝、脑以及肾组织中硫化氢的浓度。结果:成功制备出新型硫化氢缓释供体。供体溶液在4℃或-20℃存储情况下基本不影响其释放能力,供体最大达到2mmol/L浓度处理HepG2细胞不影响细胞活力及LDH活性,持续给药9d也不影响细胞活力。注射供体后,可短暂提高心、肝及肾的组织硫化氢水平,心和肝维持20min以上,而肾组织持续高水平,到2h恢复正常水平,脑组织中硫化氢浓度基本不变。表明此新的硫化氢供体可以提高组织硫化氢浓度,但不能通过血脑屏障,最终通过肾进行排泌,而慢性给药(4周)则易导致肝或皮肤局部损伤。结论:新合成的化合物可作为较稳定、缓释、毒副作用相对低的硫化氢供体,可用于研究硫化氢的细胞功能调节和急性心血管调节病理生理学作用机制。更多还原

【Abstract】 Objective:To improve a method of a new hydrogen sulfide slow-releasing donor,to observe its cellular toxicity in HepG2 cells and tissue distribution and metabolic pathway after administration of the donor by intraperitoneal injection in ICR mice and to afford experimental evidences for rationally using this donor in hydrogen sulfide research.Methods:We synthesized the new chemical compound which slowly released hydrogen sulfide.After administration of the donor,the cell toxicity was evaluated for cell viability using trypan blue staining in HepG2 cells and lactate dehydrogenase (LDH) activity in culture medium.After administration of this donor by intrasperitoneal injection,we measured the tissue hydrogen sulfide content in the liver,heart,kidney and brain using sensitive-sulfur electrode assay.Results:We successfully prepared the donor which could release hydrogen sulfide.The releasing ability of the donor solution stored at 4 ℃ or 20 ℃ did not change as compared with the freshly-prepared one.Treated at various concentrations of the donor (0.062 5,0.125,0.25,0.5,1 and 2 mmol/L) for 24 hours in HepG2 cells,the cell viability and LDH leak from the cells were not different as compared with the controls.The donor (2 mmol/L) was administrated everyday and the culture medium was changed every 3 days.After 9 days,the cell viability and LDH leak did not change.Administration of the donor (200 μmol/kg) quickly increased the tissue hydrogen sulfide concentrations in the liver and heart and maintained about 20 min;the hydrogen sulfide level in the kidney elevated and maintained a longer time,then recovered after 2 hours,which implicated that the donor might exclude the kidney;the hydrogen sulfide concentration in the brain did not change in the present study,which suggested that the donor could not pass the blood-brain barrier.Long time (4 weeks) treatment with this compound might induce hepatic or cutaneous injury.Conclusion:The new chemical compound is a relative stable,slow-releasing donor of hydrogen sulfide with low cellular toxicity,which may be used to study the regulatory role of hydrogen sulfide in the cellular physiological and pathophysiological mechanism of the animal model with acute diseases.更多还原