Methodological issues affecting the determination of plasma matrix metalloproteinase (MMP)-2 and MMP-9 activities

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• 作者：Souza-Tarla ; Caroline D. ; Uzuelli ; Juliana A. ; Machado ; Alcyone A. ; Gerlach ; Raquel F. ; Tanus-Santos ; Jose E.
• 刊名：Clinical Biochemistry
• 出版年：2005
• 出版时间：May, 2005
• 年：2005
• 卷：38
• 期：5
• 页码：410-414
• 全文大小：241 K

文摘

Objectives:

To examine whether a linear relationship exists between the volume of plasma loaded in each gel lane and the gelatinolytic activities of plasma matrix metalloproteinase (pMMP)-2 and pMMP-9, and the influences of plasma storage temperature, fasting, and a variable number of repeated freeze–thaw cycles.

Design and methods:

Plasma from venous blood collected into EDTA tubes was stored for 1 month at −20°C or at −70°C until assayed for gelatin zymography of pMMP-2 and pMMP-9. The effects of food intake, repeated freeze–thaw cycles, and the gelatinolytic activity produced by serial dilutions of plasma were examined.

Results:

Storage temperature, fasting, and up to five repeated freeze–thaw cycles had no effect on the activities of pMMPs (all P < 0.05). A linear relationship was found with plasma volumes below 0.75 μL/lane for MMP-2, and from 0.75 μL/lane to 2 μL/lane for MMP-9.

Conclusions:

Different volumes of plasma should be studied when pMMP-2 and pMMP-9 activities are studied. Storage temperature, repeated freeze–thaw cycles, and food intake do not affect the activities of pMMPs.