摘要
目的:建立测定心肌组织样品中腺苷酸含量的RP-HPLC法,并观察薯蓣皂苷(D)对缺血再灌注大鼠心肌能量代谢的改善作用。方法:建立大鼠心肌缺血再灌注模型,以薯蓣皂苷进行干预,大鼠随机分为4组:空白对照组(C组),心肌缺血再灌组(IR组),薯蓣皂苷高剂量组(DH组),薯蓣皂苷低剂量组(DL组)。采用反相高效液相色谱法,C18色谱柱(4.6 mm×250 mm,5μm),流动相为100 mmol.L-1磷酸二氢钾-磷酸氢二钾(10∶1)缓冲液(pH=6.0),流速为1.0 ml.min-1,柱温为35℃,检测波长254 nm,测定心肌组织ATP、ADP、AMP含量、腺苷酸池(TAN)水平。结果:ATP、ADP、AMP在浓度2.5～320μg.ml-1(r=0.999 9)范围内线性关系良好,平均回收率(n=6)分别为ATP 97.5%、ADP 105%、AMP 99.5%;批内RSD分别为0.9%、1.0%、0.8%,批间RSD分别为3.1%、3.3%、3.2%。结论:该方法可简便、快速、准确地测定大鼠心肌腺苷酸含量;薯蓣皂苷对大鼠缺血再灌注心肌能量代谢有明显改善作用。
Objective： To establish an RP-HPLC method for determining the contents of adenosine phosphates in myocardium tissue samples and study the effects of dioscin （D） on myocardium energy metabolism in rat myocardium with ischemia-reperfusion（IR）. Methods： The rats were randomly divided into four groups： Control group; IR group; dioscin H, L group. The levels of ATP, ADP, AMP and total adenine nucleotides （TAN） of the myocardium energy state were analyzed by RP-HPLC.The Hypersil C18 （4.6 mm×250 mm, 5μm）column was used, the mobile phase was 100 mmol.L^-1KH2PO4-K2 HPO4 buffer solution （pH=6.0）, the flow rate was 1.0 ml. min^-1, the column temperature was 35~C, the detection wavelength was 254 nm. Results： The linear range was 2.5-320 μg,ml^-1 （r=0.999 9）for ATP, ADP, AMP.The average recoveries （n=6） were ATP 97.5%, ADP 105%, AMP 99.5% respectively ;the relative standard deviations （RSDs） of within-batches were 0.9%, 1.0%, 0.8% ;the RSDs of between-batches were 3.1%, 3.3%, 3.2%. Conclusion： This analytical method is simple,rapid and accurate. Dioscin can significantly improve the myocardium energy metabolism in ischemia-reperfusion rat.
出处
《天津医科大学学报》
2008年第4期409-411,429,共4页
Journal of Tianjin Medical University
基金
国家天然药物工程技术中心资助(200703)
