脑啡肽酶作为诊断心力衰竭生物学标记的研究

Study on neprilysin as a biological marker for diagnosis of heart failure

目的研究脑啡肽稳定性是否适合作为心力衰竭生物学标志,以及研究其在血清中含量与活性的关系。方法抽取心力衰竭患者空腹血液2ml,于非抗凝管中。采集后的血液4℃保存,并在4h内3 000rpm离心10分钟,缓慢吸出上层血清,转入EP管中;将30个心力衰竭患者的血清放入一个50ml离心管中,涡旋混匀后分装,每管200μl,将获得的多管混合血清分组,分别放入-80℃、-20℃和4℃中保存,以-80℃中的样本作为对照组,测定4℃保存8h、12h、16h和24h的样本中NEP含量。同样,以-80℃中的样本作为对照组,测定-20℃保存7d、14d、21d和28d的样本中NEP含量;分别用NEP含量检测试剂盒和NEP酶活性检测试剂盒测试40例患者血清中的NEP。通过相关性分析比对,血清中NEP含量及活性关系。结果分离后的血清在4℃中保存24h之内,-20℃保存的血清28d之内其含量下降并不显著(P>0.05);血清中NEP含量与酶活性成正相关,相关系数R2为0.7017,二者相关性较强。结论 NEP在人血样本中较稳定,可以满足临床检测对样本储存及保留复检的要求。结合之前的研究结果,NEP有望成为心力衰竭诊断的生物学标记。

Objective To study whether the stability of neprilysin is suitable as a biological marker of heart failure,and to study its relationship between serum content and activity.Methods 2 ml of fasting blood from patients with heart failure was taken and placed in a non-anticoagulated tube.The collected blood is stored at 4℃,centrifuge at 3000 rpm for 10 minutes,slowly aspirate the upper serum and transfer to the EP tube;place the serum of 30 heart failure patients in a 50 ml centrifuge tube.Separate serum,200μl per tube,store in-80℃,-20℃ and 4℃,respectively.The sample in-80℃ was used as a control group,and the NEP content in the sample was measured at 4℃for 8 hours,12 hours,16 hours,and 24 hours.Similarly,the sample in-80℃ was used as a control group,and the NEP content in the samples stored at-20℃for 7 days,14 D,21 Dand 28 Dwas determined;The NEP in the serum of 40 patients was tested by NEP content detection kit and NEP enzyme activity detection kit,respectively.The relationship between NEP content and activity in serum was compared by correlation analysis.Results The serum after isolation is stable,stored at 4℃for 24 hours,-20℃for 28 days.The serum NEP content was positively correlated with the enzyme activity,and the correlation coefficient R2 was 0.7017.The correlation between the two was strong.Conclusion NEP is more stable in human blood samples,which can meet the requirements of clinical testing for sample storage and retention re-examination.Combined with previous research results,NEP is expected to become a biological marker for the diagnosis of heart failure.