胶原酶与不同药物相配伍对其酶活性影响的研究

Research on activity effect of collagenase's compatibility with different drugs

目的:观察注射用胶原酶与不同药物相配伍后酶活性变化的情况,探讨胶原酶与不同药物配伍对酶活性的影响。方法:将注射用胶原酶制剂与12种药物分别配伍后,将样品管、底物对照管和酶对照管各加入无水乙醇0.5 ml终止酶反应,冷却过滤。取滤液稀释后置于TU-1800 pc紫外可见分光光度计,将样品管的吸收度减去底物对照管和酶对照管的吸收度,再从标准曲线上查得亮氨酸的μg数,以胶原酶在37℃、pH7.5条件下作用于胶原蛋白,每小时从胶原蛋白中释放的多肽相当于茚三酮显色1μmol亮氨酸量为一个活性单位来计算注射用胶原酶制剂的活性。结果:12种药物对注射用胶原酶的活性均有不同程度的影响,轻者导致胶原酶的活性下降,重者导致胶原酶的活性丧失。结论:胶原酶的化学本质是蛋白质,任何能改变蛋白质性质的条件,都会使酶的活性下降或失活。忽视胶原酶的化学本质而随意配伍的不规范做法,有悖于酶促反应动力学的基本原理,在临床工作中不应提倡。

Objective： To observe the change of the activity of collagenase＇ s compatibility with different drugs. Methods： The collagenase was mixed with twelve kinds of drugs. Adding anhydrous alcohol 0. 5 ml to the sample tube, the subtrate control tube and the enzyme control tube respectively to stop enzyme reaction, cooling and filtering. Putting filtered solution which was diluted into TU-1800PC type ultraviolet photometer,subtracting the absorption ratio of the subtrate control tube and the absorption ratio of the enzyme control tube from that of the example tube, checking leucine＇ s μg value from the standard curve. On the condition of 37 ℃ pHT. 5, collagenase effected on collagen. Released peptide from collagen per hour was equivalent to 1 μmol leucine quantity from ninhydrin color reaction as one unit to calculate collagenase solution preparation＇ s activity. Resuits： Twelve kinds of drugs had different degree effect on the collagenase＇ s activity. It could cause collagenase＇ s activity to decline in slight condition and loss in severe condition. Conclusion：Collagenase is protein in chemical nature. Any conditions which change protein＇ s character can lead to enzyme＇ s activity declined or lost. This kind of un-standard way which ignore enzyme＇ s chemical nature and compatibility will violate enzymatic reaction dynamic＇ s basic theory, shouldn＇ t be advocated in clinical practice.