龈沟液弹性蛋白酶洗提和保存方法的初步研究

A preliminary study on the elution and storage of elastase in gingival crevicular fluid

目的 :观察不同保存温度、时间和不同缓冲液以及不同洗提方法对龈沟液弹性蛋白酶 (GCF-EA)活性的影响。方法 :采用底物反应法分别于取样即刻和 1、2、3、4周时检测 - 2 0℃和 - 70℃保存的PBS组和Tris-HCl组以震荡法和震荡 +离心法洗提的GCF样本中EA的活性。结果 :不同洗提方法间洗提效果的差异具有显著性 (0 .0 1<P <0 .0 2 ) ,“震荡 +离心法”优于单纯“震荡法” ;而洗提时采用PBS或Tris -HCl作为缓冲液 ,对洗提效果无明显影响 (P >0 .5 )。在相同时间内 ,PBS组或Tris -HCl组 - 2 0℃保存的GCF样本EA活性与- 70℃者间的差异无显著性 (P >0 .0 5 ) ,但 - 2 0℃保存者有略高于 - 70℃者的倾向 ,且 - 2 0℃比 - 70℃条件下保持EA活性的时间略长。结论 :将滤纸条上的GCF样本进行洗提时 ,采用“震荡 +离心法”优于“震荡法”。GCF样本的保存过程中 ,缓冲液的种类以及保存温度可影响GCF -EA的活性。以PBS作为缓冲液、- 2 0℃保存可能更利于其中EA活性的稳定。

AIM:To observe the effects of different storage time and temperature,different buffer and elution on elastase(EA) activity in gingival crevicular fluid(GCF).METHODS:EA activities in GCF treated with different methods were measured with a chromogenic low molecular weight substrate.RESULTS:EA activities of GCF samples eluted by 'shaking for 1 hour and centrifuging for 10 minutes' were significantly higher than that by 'shaking for 1 hour'only while no differences were found between the effects of eluting with different buffers.In same storage time the EA activity showed a better stability when GCF samples were kept at -20℃ than -70℃.CONCLUSION:The eluting effect of shaking and centrifuging is superior to shaking only when GCF sample absorbed on paper strip is eluted.EA activity of GCF sample can be influenced by the category of buffer and the storage temperature.The elastase activity showed a better stability when samples were kept in PBS at -20℃. [