Investigation of Extracellular Matrix Protein Expression Dynamics Using Murine Models of Systemic Inflammation
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Extracellular matrix (ECM) proteins form the structural support for migration of leukocytes and provide multiple signals to assist in their functions during inflammatory conditions. Presence of pro-inflammatory mediators in the tissues results in the remodelling of matrices which could modify the functions of extravasated leukocytes. Previous reports have shown changes in the expression of ECM proteins during local inflammatory responses. In this study, we have investigated the time- and tissue-specific expression profile of key ECM proteins in systemic inflammation using lipopolysaccharide (LPS)-induced endotoxemia and cecal ligation and puncture (CLP) mouse models. The results show that compared to naïve tissues, within 12 h following CLP surgery, a 20–30-fold increase was observed in the expression of collagen-IV (Col-IV) transcripts in the mesentery tissues with a 2.4-fold increase in the protein by 24 h. However, Western blot band intensities indicated that vimentin and fibrinogen were remarkably expressed in more quantity compared to Col-IV. Secondly, in CLP group of mice, fibrinogen showed 6–40-fold increase in mRNA level in various tissues with about 2-fold increase in the protein level compared to respective naïve tissues. Similar studies in the LPS-injected mice showed up to 2–3 fold increase in the expression of Col-IV, fibrinogen and vimentin at protein level in the lungs. In such animals, although similar pattern was observed for fibrinogen in kidney and liver tissues, the mesentery showed prominent changes in Col-IV and vimentin mRNA compared to CLP. Further, bioinformatics analysis showed multiple pathways which could be associated with vimentin, Col-IV and fibrinogen under inflammatory conditions both in human and mouse. The current study will help in better understanding of possible signalling from ECM proteins in inflammatory microenvironment and may contribute in development of cell adhesion-based therapeutics.
KEY WORDSextra cellular matrix inflammation sepsis
PC, SD, CB and PD performed the in vivo experiments. CB and SD performed qRT-PCR and Western blot experiments and analysed the data. PD and PC assisted in RNA isolation from animal tissues and performing experiments. PK performed the bioinformatics studies and analysed the data. PPS conceived and directed the study. CB, SD and PPS wrote the manuscript.
The Department of Biotechnology, Govt. of India (102/IFD/SAN/1671/2014-2015) to P.P.S., University Grants Commission, Govt. of India fellowship to CB (Sr. No-2061330682) and SPD (Sr. No-2061430670), Indian Council of Medical Research, Govt. of India fellowship to PD, and, Ministry of Human Resource Development, Govt. of India fellowship to PC (MHR 02-23-200-429) provided funding.
Compliance with Ethical Standards
Conflict of Interest
The authors declare that they have no conflicts of interest.
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