一种评估氰化钾及其解毒剂毒性的稳定体外方法。
A stable in vitro method for assessing the toxicity of potassium cyanide and its antidote.
作者信息
Yamagiwa Takeshi, Inokuchi Sadaki, Saito Takeshi, Inoue Shigeaki, Morita Seiji, Kawaguchi Akira T
机构信息
Department of Emergency and Critical Care Medicine, Tokai University School of Medicine, 143 Shimokasuya, Isehara, Kanagawa 259-1193, Japan.
出版信息
Tokai J Exp Clin Med. 2013 Dec 20;38(4):114-22.
BACKGROUND
Hydrogen cyanide possesses a high acid-dissociation constant of 9.14, favoring its vaporization and depletion from the culture media at physiological pH, which may cause the cyanide toxicity unstable in vitro.
OBJECTIVE
We investigated whether adjustment of culture medium pH stabilizes cyanide concentration and decreases the effective concentration of potassium cyanide (KCN).
METHODS
Murine fibroblast cells were exposed to different concentrations of KCN in media maintained at pH 7.4 or 9.2, in the presence or absence of hydroxocobalamin. After incubation for 1 h, we evaluated medium pH, cyanide concentration, cytochrome activity, and cell viability.
RESULTS
Cyanide concentration decreased to 18.8% in pH 7.4 medium compared to 83.2% in pH 9.2 medium. A significant decrease in cytochrome activity was observed at 40 mM and 1.25 mM KCN in pH 7.4 and pH 9.2 media, respectively. In pH 9.2 medium, dose-dependent cytotoxicity of KCN and antidotal effects of hydroxocobalamin were observed.
CONCLUSION
Adjustment of culture medium pH to 9.2 could stabilize cyanide concentration and decrease the effective concentration of KCN, allowing stable evaluation of KCN toxicity and antidotal efficacy.
背景
氰化氢具有较高的酸解离常数9.14，这有利于其在生理pH值下从培养基中蒸发和消耗，这可能导致体外氰化物毒性不稳定。
目的
我们研究了调节培养基pH值是否能稳定氰化物浓度并降低氰化钾（KCN）的有效浓度。
方法
在存在或不存在羟钴胺素的情况下，将小鼠成纤维细胞暴露于pH值为7.4或9.2的培养基中不同浓度的KCN。孵育1小时后，我们评估了培养基pH值、氰化物浓度、细胞色素活性和细胞活力。
结果
与pH值为9.2的培养基中83.2%相比，pH值为7.4的培养基中氰化物浓度降至18.8%。在pH值为7.4和9.2的培养基中，分别在40 mM和1.25 mM KCN时观察到细胞色素活性显著降低。在pH值为9.2的培养基中，观察到KCN的剂量依赖性细胞毒性和羟钴胺素的解毒作用。
结论
将培养基pH值调节至9.2可稳定氰化物浓度并降低KCN的有效浓度，从而能够稳定评估KCN的毒性和解毒效果。
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