Study on the expression of NLRP3 and related inflammatory factors in necrotizing enterocolitis in newborn rats

ZHAO Ming; QIN Miao; SUN Meng-ya; JIANG Hong; LI Xiang-hong; JIANG Jian; LIU Yan

doi:10.11852/zgetbjzz2020-1858

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Chinese Journal of Child Health Care ›› 2021, Vol. 29 ›› Issue (8) : 852-856. DOI: 10.11852/zgetbjzz2020-1858

Basic Expeximental Artictes

Study on the expression of NLRP3 and related inflammatory factors in necrotizing enterocolitis in newborn rats

ZHAO Ming, QIN Miao, SUN Meng-ya, JIANG Hong, LI Xiang-hong, JIANG Jian, LIU Yan

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Abstract

Objective To investigate the expression level of nucleotide-binding oligomerization domain like receptor protein 3(NLRP3) and related inflammatory factors in newborn rats with necrotizing enterocolitis. Methods Twenty-four 3-day-old Sprague-Dawley (SD) rats were selected and randomly divided into control group (n=12) and experimental group (n=12). The rats were decapitated on the seventh day of life, their intestinal tissues of the caecal part were retrieved for related indexes. HE staining was used to observe the changes of ileocecal intestinal histopathology and double-blind method was used to score the pathology. Apoptosis in ileocecal tissue was observed by TUNEL staining. The ultrastructure of cells was observed by electron microscopy. IL-18, IL-1β and IL-6 levels were detected by ELISA. The expression levels of Caspase-1, Bcl-2 and Bim were detected by Western-blot, and NLRP3 mRNA level was detected by RT-PCR. Results 1) HE staining showed that the intestinal wall villi of rats in the experimental group were seriously damaged, most of the villi were broken in the middle and roots, and some of the crypts could not be recognized with visible wall necrosis. The intestinal wall villi damage in the control group was less than that in the experimental group. 2) TUNEL staining showed that the number of apoptosis increased in the ileocecal tissue of the experimental group and decreased in the control group (t＝30.54,P<0.05). 3) Under electron microscope, mitochondria of the experimental group were swollen, the ridge structure was disordered, and some brittle fractures were observed. Mitochondrial injury in control group was significantly reduced compared with that in experimental group. 4) The expression levels of IL-18, IL-1β and IL-6 in the ileocecal tissues of rats in the experimental group was significantly increased compared with those in the control group (t=23.38, 29.78, 26.90, P<0.05). 5) The expression levels of Caspase-1 and Bim in the experimental group were significantly higher than those in the control group (t=15.12, 5.82, P<0.05), while Bcl-2 expression was significantly lower in the experimental group(t＝6.03, P<0.05). 6) The mRNA expression level of NLRP3 in the experimental group was significantly higher than that in the control group (t＝9.21, P<0.05). Conclusions NLRP3 plays a crucial role in the development of NEC in newborn rats. Blocking the activation of NLRP3 is expected to be an effective way to prevent neonatal NEC.

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nucleotide-binding oligomerization domain like receptor protein 3 / inflammatory factor / neonatal rats / necrotizing enterocolitis

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ZHAO Ming, QIN Miao, SUN Meng-ya, JIANG Hong, LI Xiang-hong, JIANG Jian, LIU Yan. Study on the expression of NLRP3 and related inflammatory factors in necrotizing enterocolitis in newborn rats[J]. Chinese Journal of Child Health Care. 2021, 29(8): 852-856 https://doi.org/10.11852/zgetbjzz2020-1858

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