摘要
目的:建立一种测定组织样品中腺苷酸含量的 RP-HPLC 法,并观察异甘草素(isoliquiritigenin,ISL)对脑缺血再灌注小鼠能量代谢的改善作用。方法:采用反复阻断小鼠双侧颈总动脉造成脑缺血再灌注模型,采用 RP-HPLC 法,Hypersil C_(18)色谱柱(4.6mm×250mm,5μm),流动相为100mmol-L^(-1)磷酸二氢钾-磷酸氢二钾缓冲液(pH 6.0),流速为0.7mL·min^(-1),柱温为25℃,检测波长为254nm,测定脑组织 ATP 含量、腺苷酸池(TAN)水平和能量负荷值(EC)。结果:ATP、ADP、AMP 浓度分别在1.7-212.0μg·mL^(-1)(r=0.9999),1.6～208.0μg·mL^(-1)(r=0.9999),1.3～164.0μg·mL^(-1)(r=0.9999)范围内线性关系良好,平均回收率(n=6)分别为101.3%(RSD=1.0%),99.0%(RSD=1.4%),91.6%(RSD=1.6%);ISL(10,20,40mg·kg^(-1)·d^(-1))剂量依赖性提高脑组织中 ATP 含量、TAN 水平和 EC 值。结论:该分析方法简便、准确、可靠;ISL 对脑缺血再灌注小鼠脑能量代谢水平有明显的改善作用。
Objective:To establish an RP-HPLC method for determining the contents of adenine nucleotides in tissue samples and to study the effects of isoliquiritigenin(ISL)on brain energy metabolism by this RP-HPLC method in repeated cerebral ischemia-reperfusion mice.Methods:The repeated cerebral ischimia-reperfusion models in mice were made by repeated occlusion of bilateral common carotid arteries and reperfusion.The levels of ATP,energy charge(EC)and total adenine nucleotides(TAN)of the brain energy state were analyzed by RP- HPLC.The Hypersil C_(18)(4.6mm×250mm,5μm)column was used,the mobile phase was 100 mmol·L^(-1) KH_2PO_4-K_2HPO_4 buffer solution(pH 6.0),the flow rate was 0.7 mL·min^(-1),the column temperature was 25℃,the detection wavelength was 254nm.Results:The linear range was 1.7-212.0μg·mL^(-1)(r=0.9999)for ATP,1.6～208.0μg·mL^(-1)(r=0.9999)for ADP,1.3～164.0μg·mL^(-1)(r=0.9999)for AMP.The average recoveries(n=6)were 101.3%(RSD=1.0%)for ATP,99.0%(RSD=1.4%)for ADP,97.6%(RSD=1.6%) for AMP.ISL(10,20,40mg·kg^(-1)·d^(-1))increased the levels of ATP,EC and TAN in a dose-dependent manner in repeated cerebral ischemia-reperfusion mice brain.Conclusion:This analytical method is simple,accurate and reliable.ISL can significantly improve the brain energy metabolism in repeated cerebral ischemia-reperfusion mice.
出处
《药物分析杂志》
CAS
CSCD
北大核心
2005年第6期639-642,共4页
Chinese Journal of Pharmaceutical Analysis
