However, since its standardization by Rittirsch in 2009, it has been possible to reproduce CLP exactly. Moreover, animals subjected to CLP simulate a true infectious septic state, and the decreased cardiovascular function and cardiac output induced by LPS are avoided. In the CLP procedure used in the present study, the large ligation and needle size were necessary selleckbio for high grade sepsis induced DIC, and ensured 85% mortality at 72 h post procedure. Development of DIC in the experimental mice was confirmed by the decreased platelet count and fibrinogen concentration, the prolongation of PT and aPTT, the elevation of D dimer and the autopsy evidence of throm bosis in end organs. The evolution of multiple organ dysfunction was corroborated by the increased ALT and creatinine.
Six hours post CLP was identified as the early stage of sepsis induced Inhibitors,Modulators,Libraries DIC in the model, because all significant changes in clinical markers had occurred by this time. CD63 and CD62P are expressed on the platelet surface and secreted into plasma when platelets become activated. In resting Inhibitors,Modulators,Libraries platelets CD63 is stored in lysosomes and CD62P in a granule. Therefore, the percentage platelet surface expression of Inhibitors,Modulators,Libraries CD62P and CD63 indicating platelet activation were significantly higher 2 h post CLP. Simultaneously, the platelet secreted factors TSP, PF4, TIMP 1 and TCK 1 changed significantly during the first 6 h post CLP and were therefore regarded as meaningful biomarkers Inhibitors,Modulators,Libraries of the early stage of sepsis induced DIC. G CSF and CXCR4 were excluded because their levels in the experimental mice began to differ significantly after 6 h post CLP.
TSP was first isolated from platelets that had been stimulated with thrombin, and was designated as a matricellular glycoprotein with multiple functions in many biological processes including Inhibitors,Modulators,Libraries angiogenesis, apoptosis and immune regulation. PF4, also known as chemokine C X C motif ligand 4, is released from activated plate lets during platelet aggregation and promotes blood coagulation. TIMP 1, as natural inhibitor of matrix metalloproteinases has been shown to be involved in degradation of the extracellular matrix and to have an effect on the fibrinolytic system. TCK 1 belongs to the family of CXC chemokines and has been shown to be a chemoattractant for pro B cells engineered to express the CXCR2 receptor.
In the early stage of the mouse CLP model, TSP, PF4, TIMP 1and TCK 1 levels selleck compound exhibited differing trends on protein microarray analysis. PF4 levels in the CLP group peaked 2 h post CLP and decreased rapidly to the level of the sham group, which would made it difficult to capture abnor mal PF4 levels in clinical practice. TSP levels in the CLP group increased and peaked sharply at 1 h post CLP, then decreased but remained significantly higher than in the sham group.