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目的:建立测定大鼠心肌组织中三磷酸腺苷、二磷酸腺苷、一磷酸腺苷(ATP、ADP、AMP)含量的高效液相色谱法,并观察慢性压力负荷性心衰大鼠心肌组织中腺苷酸含量的变化。方法:SD大鼠随机分为心衰模型组和假手术对照组,按腹主动脉缩窄法建立慢性压力负荷性心衰大鼠模型。色谱柱为VP-ODSC18柱(250mm×4.6mm,5μm);流动相为50mmol.L-1磷酸盐缓冲液(pH6.5);流速为1.0mL·min-1;柱温25℃;检测波长259nm;进样量20μL。结果:ATP、ADP、AMP的标准液与峰面积均呈良好的线性关系(r=0.9997～0.9999),平均回收率分别为96.30%,97.64%和103.28%。日内和日间精密度分别在1.15%和4.13%以下。心衰大鼠与假手术对照大鼠相比,ATP和ADP的含量明显下降,而AMP明显升高。结论:本法操作简便,准确快速,可以用于测定大鼠心肌组织中腺苷酸的含量。
OBJECTIVE: To establish a high performance liquid chromatographic method for the determination of adenosine triphosphate, adenosine diphosphate and adenosine monophosphate (ATP, ADP, AMP) in rat myocardium and to observe the effect of adenosine in myocardium of rats with chronic pressure overload heart failure Changes in acid content. Methods: SD rats were randomly divided into heart failure model group and sham operation control group. The rat model of chronic stress-induced heart failure was established according to abdominal aorta constriction. The column was VP-ODSC18 (250 mm × 4.6 mm, 5 μm). The mobile phase was 50 mmol·L-1 phosphate buffer (pH 6.5), the flow rate was 1.0 mL · min-1, the column temperature was 25 ℃, 259nm; Injection volume 20μL. Results: The calibration curves of ATP, ADP and AMP showed a good linearity (r = 0.9997 ~ 0.9999) with the average recoveries of 96.30%, 97.64% and 103.28%, respectively. Intra- and inter-day precision was 1.15% and 4.13% respectively. Compared with the sham operation control rats, the content of ATP and ADP in heart failure rats decreased significantly, but AMP increased obviously. Conclusion: This method is simple, accurate and rapid, and can be used to determine the content of adenylate in myocardial tissue of rats.