Apoptotski kapacitet u kunića sa eksperimentalnim emfizemom pluća

Abstract

In order to better understand pathogenesis of pulmonary emphysema, the model of experimentally induced pulmonary emphysema in Chinchilla rabbits was used for the estimation of apoptotic clearance of pulmonary tissue. Bronchoalveolar lavage was performed in three groups of animals: experimental group-E on hypercholesterolemic diet (4% edible oil solution of crystalline cholesterol), control group-C1 on standard diet for that animal species and animals on oily diet-C2. Apoptotic detection in cytocentrifuge preparations of lung washings was evaluated by in situ TUNEL. The property of alveolar macrophages to engulf apoptotic cells was estimated by light microscopy including 300 features (related subsequent steps: adsorption, internalization and intracellular processing of free apoptotic bodies) and was evaluated by scoring and indexing method. Internalization of apoptotic bodies by alveolar macrophages, as well as free apoptotic bodies were decreased in E compared to both C1 and C2 group (p<0.01 and p<0.05 respectively). Intracellular processing of apoptotic bodies by alveolar macrophages is significantly decreased in C2 in comparison with E (p<0.05) and C1 group (p<0.01). Apoptotic capacity of pulmonary tissue is significantly decreased in C2 in comparison with C1 group (p<0.01). The results implicate that immuno-metabolic competence of pulmonary tissue might be essentially associated with tissue remodeling in pulmonary emphysema.

U cilju boljeg razumevanja patogeneze plućnog emfizema, u radu je korišć en eksperimentalni model emfizema pluća na činčila kunićima za procenu apoptotskog kapaciteta plućnoga tkiva. Bronholaveolarna lavaža je urađena na tri grupe životinja: eksperimentalnoj grupi-E na hiperholesterolskoj dijeti (4% uljani rastvor kristalnog holesterola), kontrolnoj grupi-C1 na standardnoj dijeti za tu životinjsku vrstu i grupi životinja na uljanoj dijeti-C2. Određivanje apoptotskih parametara cito-centrifužnih preparata bronhoalveolarnog lavata vršeno je posle bojenja preparata TUNEL in situ citohemijskim metodom. Sposobnost alveolarnih makrofaga da odstrane apoptotske ćelije fagocitozom procenjivana je svetlosnom mikroskopijom na 300 prikaza po preparatu (prikazi uključuju: adsorpciju, internalizaciju i intracelularno procesiranje apoptotskih tela) i evaluirana metodom indeksiranja i skora. Internalizacija apoptotskih tela alveolarnim makrofazima, kao i relativni procenat slobodnih apoptotskih tela, bili su signifikantno smanjeni u E grupi poredeći sa C1 (p<0.01) i C2 grupom (p<0.05). Intracelularno procesiranje apoptotskih tela alveolarnim makrofazima bilo je signifikantno smanjeno u C2 u odnosu na E (p<0.05) i C1 grupu (p<0.01). Apoptotski kapacitet tkiva pluća bio je signifikantno smanjen u C2 u poređenju sa C1 grupom (p<0.01). Ovi rezultati ukazuju da imuno-metabolička kompetentnost plućnog tkiva može biti suštinski povezana sa remodelovanjem tkiva pluća u eksperimentalnom emfizemu pluća.