Abstract
Objective To explore the mechanism of macrophage activation in murine model of Hirschsprung-associated enterocolitis. Methods According to drawing lot method, specific-pathogen-free level laboratory 2-week-old and 3-week-old EdnrB -/- mice were selected as experimental group (n=3), wild-type mice of comparable age as control group (n=3) and 2-week-old EdnrB -/- mice treated with clodronate liposomes (CLOD) as treatment group (n=3). All intestinal samples were prepared into expansion and narrow sections according to their morphological changes. The expressions of CD68 and inducible nitric oxide synthase (iNOS) were detected by immunofluorescent staining for assessing the macrophage and the activation level of intestinal macrophage. And reverse transcription-polymerase chain reaction (RT-qPCR) was employed for detecting the expressions of iNOS and TNF-α at the mRNA level. Results As compared with 2-week-old wild-type murine intestinal expansion colonic sections, the positive cells of CD68 in 2-week-old EdnrB -/- mice slightly increased [(1.31±0.81)% vs.(5.71±1.96)%, P<0.05]. However, for expansion colonic sections of 3-week-old EdnrB -/- mice, the positive cells of CD68 [(12.81±2.28)% vs. (2.25±1.17)%, P<0.05] and Inos [(7.81±2.35)% vs. (1.22±0.66)%, P<0.05] significantly increased. The expansion colonic sections of CLOD-treated 3-week-old EdnrB -/- mice group, the positive cells of CD68 and iNOS were (2.52±0.49)% and (1.29±0.38)%. As compared with expansion colonic sections of 3-week-old EdnrB -/-, there were significant differences. In 2-week-old EdnrB -/- mice group, RT-qPCR showed that iNOS, TNF-α and IL-1β had no differences as compared with control group at the level of mRNA. Yet in 3-week-old EdnrB -/- mice, these cytokines significantly increased. In CLOD-treated 3-week-old EdnrB -/- mice group, inflammatory injury markedly declined and the expressions of cytokines decreased obviously as compared with 3-week-old EdnrB -/- mice group. Conclusions Intestinal macrophage activation and secretion of proinflammatory cytokines may be associated with the occurrence and development of Hirschsprung-associated enterocolitis. Key words: Hirschsprung disease; Enterocolitis; Macrophage activation
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