| Abstract | CILJ ISTRAŽIVANJA: Procijeniti mogući proupalni učinak hiperbaričnog tretmana u štakora određujući izražaj biljega CD11b i CD15s na leukocitima. Opisati promjene u izražaju biljega CD77 na štakorskim endotelnim (CD34+) stanicama bubrega, pluća i srca nakon izlaganja hiperbaričnom tlaku.
MATERIJALI I METODE: Eksperimentalno istraživanje provedeno je na Katedri za medicinsku kemiju i biokemiju Medicinskog fakulteta u Splitu. Mužjaci štakorskog soja Sprague-Dawley, starosti 4 tjedna bili su podijeljeni u 2 skupine: ispitivanu skupinu koja je podvrgnuta hiperbaričnom tretmanu (n=9) i netretiranu kontrolnu skupinu (n=5). Štakori su podvrgnuti hiperbaričnom tlaku smjese zraka koji odgovara dubini zarona od 65 m, u trajanju od 30 min. Tijekom tri dana, nakon izlaganja hiperbariji, štakorima je uzet uzorak krvi za određivanje leukocitnih biljega CD15s i CD11b protočnom citometrijom. Trećeg dana životinje su žrtvovane produljenom anestezijom dietileterom te su im nakon toga izvađeni bubrezi, srce i pluća. Pripremljene stanice bubrega, pluća i srca obilježene su protutijelima za CD34 i CD77 te su analizirane protočnom citometrijom. Za statističku obradu korišten je neparametrijski Mann Whitney U test.
REZULTATI: Postotci CD15s+CD11b- leukocita i ukupnih CD15s leukocita bili su značajno povišeni u skupini tretiranih štakora naspram kontrolne skupine. Postotak CD77+CD34- stanica bubrega štakora u ispitivanoj (HB) skupini bio je značajno manji nego u kontrolnoj skupini, kao i udio ukupnih CD77+ stanica. Uočeno je značajno smanjenje % CD77+CD34+ bubrežnih stanica u HB skupini u odnosu na kontrolnu skupinu. Ukupni udio CD34+ bubrežnih stanica bio je značajno povišen u odnosu na kontrolu. Postotak CD77+CD34-, % ukupnih CD77+, % CD77+CD34+, kao i % ukupnih CD34+ stanica pluća štakora u HB skupini bio je značajno viši nego u kontrolnoj skupini. Postotak ukupnih CD77+, % ukupnih CD34+, % CD77+CD34+, kao i % CD77-CD34+ stanica srca štakora u HB skupini bio je značajno viši nego u kontrolnoj skupini.
ZAKLJUČAK: Rezultati povećanog postotka leukocita koji imaju izražen CD15s, ligand za endotelni selektin, nakon hiberbaričnog tretmana, ukazuju na njegovu ulogu u prevenciji oštećenja endotela. Rezultati ovog istraživanja ukazuju na moguću ulogu CD77 u mehanizmu odgovornim za razvoj hiperbarične diureze. Hiperbarični tretman značajno je povećao postotak plućnih i srčanih endotelnih stanica što može biti dio mehanizma popravka hiperoksijom ozlijeđenog endotela. |
| Abstract (english) | AIM OF RESEARCH: To estimate the potential pro-inflammatory effect of hyperbaric treatment in rats by determination of CD11b and CD15s on leukocytes. To describe the changes in the expression of CD77 in rat's kidney, pulmonary and cardiac endotelial (CD34+) cells.
MATHERIALS AND METHODS: Experimental research was conducted at Department for Medical Chemistry and Biochemistry, University of Split School of Medicine. Male Sprague-Dawley rats, 4 weeks old, were divided into 2 groups: the examination group, that undergone hyperbaric treatment (n=9) and untreated control group (n=5). The rats were exposed to hyperbaric pressure of air composition that equals the depth of immersion of 65 m, in duration of 30 min. During three days, after the treatment, a blood sample was collected from the rats for determination of leukocyte CD11b and CD15s markers. By the end of the third day from the beginning of hyperbaric treatment rats were sacrified in prolonged anesthesia with diethylether and their kidneys, lungs and heart were extracted afterwards. Furthermore, prepared isolated renal, pulmonary and cardiac cells were incubated with antibodies for CD34 and CD77, and were analysed by flow cytometry. For statistical analysis non-parametric Mann Whitney U test was used.
RESULTS: The percentage of CD15s+CD11b- leukocytes and total CD15s leukocytes were significantly increased in examination group compared to control group. The percentage of CD77+CD34+ kidney cells in examination group was significantly lower compared to control group, as well as the percentage of total CD77+ cells. A significant decrease in % CD77+CD34+ kidney cells was observed in examination group compared to control group. Total share of CD34+ kidney cells was significantly higher compared to control group. The percentage of CD77+CD34-, % of total CD77+, % CD77+CD34+, as well as the % of totalCD34+ pulmonary cells in examination group was significantly higher compared to control group. The percentage of total CD77+, % of total CD34+, % of CD77+CD34+, as well as % of CD77-CD34+ cardiac cells in examination group was significantly higher compared to control group.
CONCLUSION: The results of increased percentage of leukocytes that have CD15s, ligand for endothelial selectin, after hyperbaric treatment, suggest its role in the prevention of endothelial damage. The results of this experiment indicate a possible role of CD77 in the mechanism responsible for hyperbaric diuresis. Hyperbaric treatment significantly increased the percentage of pulmonary and cardiac endothelial cells which may be a part of hyperoxia-induced endothelial damage repair mechanism. |
