A theoretical timeline for myocardial infarction: immunohistochemical evaluation and western blot quantification for Interleukin-15 and Monocyte chemotactic protein-1 as very early markers

Journal of Translational Medicine

Vittorio Fineschi²  Irene Riezzo¹  Margherita Neri¹  Marco Di Paolo³  Emanuela Turillazzi¹ 

[1] Department of Forensic Pathology, University of Foggia, Ospedale Colonnello D’Avanzo, Viale degli Aviatori, n. 1, 71100 Foggia, Italy;Department of Forensic Pathology, University “La Sapienza” of Rome, Viale Regina Elena 336, 00161 Roma, Italy;Department of Forensic Pathology, University of Pisa, via Roma 55, 56100 Pisa, Italy

关键词: Western blotting;    Timing;    Myocardial infarction;    Immunohistochemistry;    Cytokines;   

Others  :  1148783
DOI  :  10.1186/1479-5876-12-188

 received in 2014-03-31, accepted in 2014-06-11,  发布年份 2014

【 摘 要 】

Background

Experimental and human studies have demonstrated that innate immune mechanisms and consequent inflammatory reaction play a critical role in cardiac response to ischemic injury. Thus, the detection of immuno-inflammatory and cellular phenomena accompanying cardiac alterations during the early inflammatory phase of myocardial infarction (MI) may be an excellent diagnostic tool. Current knowledge of the chronology of the responses of myocardial tissue following the occurrence of ischemic insult, as well as the existence of numerous studies aiming to identify reliable markers in dating MI, induced us to investigate the myocardial specimens of MI fatal cases in order to better define the age of MI.

Methods

We performed an immunohistochemical study and a Western blot analysis to evaluate detectable morphological changes in myocardial specimens of fatal MI cases and to quantify the effects of cardiac expression of inflammatory mediators (CD15, IL-1β, IL-6, TNF-α, IL-15, IL-8, MCP-1, ICAM-1, CD18, tryptase) and structural and functional cardiac proteins.

Results

We observed a biphasic course of MCP-1: it was strongly expressed in the very early phase (0-4 hrs), to diminish in the early period (after 6-8 hrs). Again, our choice of IL-15 is explained by the synergism with neutrophilic granulocytes (CD15) and our study shows the potential for striking cytokine synergy in promoting fast, local neutrophil response in damaged tissues. A progressively stronger immunoreaction for the CD15 antibody was visible in the areas where the margination of circulating inflammatory cells was detectable, up to very strong expression in the oldest ones (>12 hours). Further, the induction of CD15, IL-15, MCP-1 expression levels was quantified by Western blot analysis. The results were as follows: IL-15/β-actin 0.80, CD15/β-actin 0.30, and MCP-1/β-actin 0.60, matching perfectly with the results of immunohistochemistry. Control hearts from traumatic death cases did not show any immunoreactivity to the pro-inflammatory markers, neither were there any reactions in Western blot analysis.

Conclusions

Essential markers (i.e. IL-15, MCP-1) are suitable indicators of myocardial response to ischemic insult involving very early phase reaction (inflammatory response and cytokine release). In the very near future, proteomics may help clinicians and pathologists to better understand mechanisms relating to cardiac repair and remodeling and provide targets for future therapies.

【 授权许可】

   
2014 Turillazzi et al.; licensee BioMed Central Ltd.

【 图 表 】

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