Detection of in vivo matrix metalloproteinase activity using microdialysis sampling and liquid chromatography/mass spectrometry.
Detection of in vivo matrix metalloproteinase activity using microdialysis sampling and liquid chromatography/mass spectrometry.
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DOI：
10.1021/ac901703g
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发表时间：
2009-12-15
影响因子：
7.4
通讯作者：
Stenken, Julie A.
中科院分区：
文献类型：
--
作者：
Wang, Ying;Zagorevski, Dmitri V.;Lennartz, Michelle R.;Loegering, Daniel J.;Stenken, Julie A.
Matrix metalloproteinases (MMPs) are a family of endoproteases that break down extracellular matrix and whose upregulation contributes to several diseases. An LC/MS/MS method was developed to quantify MMP-1 and -9 substrates and their N-terminal peptide products in samples obtained from implanted microdialysis sampling probes. In vitro studies with purified human MMP-1 and MMP-9 were used to optimize the assay and determine the effectiveness of the local delivery of a broad spectrum MMP inhibitor, GM 6001. Localized delivery of GM 6001 at 10 μM was sufficient to completely inhibit product formation in vitro. In vivo studies in male Sprague-Dawley rats were performed with microdialysis probes implanted into the subcutaneous tissue. Directly after microdialysis probe implantation, infusions of the MMP-1 and MMP-9 substrates (50 μM each) resulted in recovered product concentrations of approximately 2 μM. During a 50 μM GM 6001 co-infusion with the substrates, a 30% and 25% reduction in product formation for the MMP-1 and MMP-9 substrates was obtained, respectively. Blank dialysates were negative for enzymatic activity that could cleave the MMP substrates. This method allowed for the activity of different MMPs surrounding the microdialysis probe to be observed during in vivo sampling.
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影响因子：
12.3
作者：
Somerville RP;Oblander SA;Apte SS
通讯作者：
Apte SS
DOI：
10.1152/ajpheart.2001.281.3.h987
发表时间：
2001-09-01
影响因子：
4.8
作者：
Etoh, T;Joffs, C;Spinale, FG
通讯作者：
Spinale, FG
影响因子：
2.9
作者：
Gogly, B;Groult, N;Pellat, B
通讯作者：
Pellat, B
影响因子：
7.4
作者：
Li, Qiang;Zubieta, Jon-Kar;Kennedy, Robert T.
通讯作者：
Kennedy, Robert T.
影响因子：
2.9
作者：
KLEINER, DE;STETLERSTEVENSON, WG
通讯作者：
STETLERSTEVENSON, WG