The Effect Of Brush-Induced Injury On Molecular Expression Profile Of Rabbit Airway Epithelium During Regeneration And Repair
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Date
2014-09
Authors
Latahir, Ahmad Zaeri
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Abstract
Tracheal epithelium is equipped with well-balanced defensive features to keep the body away from harmful particles. Upon an injury, the cellular repairing process undergone with the involvement of cell migration and spreading, proliferation, and redifferentiation. Molecular aspect also play important role in the repairing process. The genes involved in the process can be grouped based on their roles, which include migration, proliferation, inflammation, stem cell marker and cell-specific gene marker. Understanding the interaction between cellular and molecular components during repair will give a better view on how cells behave and function. Thus, in this study, cellular and molecular responses following tracheal brushing-induced injury using rabbit model were investigated. Thirty six New Zealand white rabbits were divided into uninjured and injured (0.5, 1, 6, 12 hours and 1, 2, 3, 4, 7, 14, and 21 days) groups. After brushing, the animals were maintained before being sacrificed according to their respective time point. The trachea was retrieved and preserved before subjected to haematoxylin and eosin (H&E) staining, Alcian blue-periodic acid Schiff (AB-PAS) staining, and real time PCR. The technique used in this study was proved to cause removal of the epithelium. The extent of damage reduced as the time point increased and completely closed on day 21. The number of goblet cells was observed increased and reached the peak level after 3 days and subsided on later time point. Molecular analysis revealed that the genes were expressed differentially
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in various time points. Genes related to cellular proliferative and migration activity expressed in the early time and reduced on the other end. We have successfully developed a blinded brushing technique to cause injury on the tracheal airway epithelium. This technique is practical, inexpensive, and can reduce the risk of infection. Reduction of the extent of damage over the time point indicated that regeneration process was undertaken. By cellular response, the number of goblet cells increased on the early time point, giving an instant response by providing a mucus protection barrier. This finding was in accordance to the proliferative and migration activity related gene expression. The activity was crucial in the early time point to encourage the remaining cells to repopulate the damaged area. We also found that the basal cells were predominantly involved during the early stage of repair, while ciliated cells rose on the later stage. Interestingly, the Clara cells did not participate in repairing the injured tracheal epithelium. Our technique provides approach in achieving sustainability goal with its suitability use in simple laboratory setting. The number of goblet cells increase on the early time point provides importance defensive mechanism against harm particles. Molecular together with various functions in regulating the protective mechanism and while on the later stage contribute to formation of regenerated epithelium.
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Keywords
The Effect Of Brush-Induced Injury , Of Rabbit Airway Epithelium