Objective To explore the effects of microRNA-223-3p(MiR-223-3p) on the inflammatory response and cell damage in the intestinal tissue of neonatal rats with necrotizing enterocolitis(NEC) and its possible mechanism. Methods The NEC model of neonatal rats was constructed and randomly divided into a model group, a negative control group, and a MiR-223-3p overexpression group. The normal rats of same number were taken as the normal group. Hematoxylin-eosin(HE) staining and TUNEL staining were used to detect intestinal histopathological changes and cell apoptosis; ELISA method was used to detect the levels of interleukin-6(IL-6), interleukin-1β(IL-1β) and interleukin-18(IL-18) in serum and intestinal tissues; SYBR Green Ⅰ real-time fluorescent quantitative PCR method was used to detect the expression of MiR-223-3p in intestinal tissues; Western Blot was used to detect the expression of NLRP3 and caspase-1 proteins in intestinal tissue. Dual luciferase reporter gene experiment was used to verify the target relationship between MiR-223-3p and NLRP3. Results Compared with the normal group, the levels of IL-6, IL-1β, IL-18 in the serum and intestinal tissues of rats in the model group and the negative control group were all significantly increased(F=215.525, 276.499, 354.826, 204.410, 261.474, 280.667, P<0.05), the expression of NLRP3 and caspase-1 proteins in the intestinal tissues and the apoptosis rate were all significantly increased(F=181.745, 137.553, P<0.05), the expression of MiR-223-3p in the intestinal tissue was significantly reduced(F=170.180, P<0.05), and there was obvious pathological damage in the intestinal tissue. Compared with the negative control group, the MiR-223-3p overexpression group was able to up-regulate the expression of MiR-223-3p in intestinal tissues, reduce the levels of IL-6, IL-1β, IL-18 and cell apoptosis in rat serum and intestinal tissues, inhibit the expression of NLRP3 and caspase-1 proteins(P<0.05), and the pathological damage of intestinal tissue was significantly improved. The results of the dual luciferase reporter gene experimental analysis showed that MiR-223-3p and NLRP3 was able to targetingly be combined. Conclusion MiR-223-3p reduces inflammation and cell damage in the intestinal tissue of NEC neonatal rats by targeting NLRP3.
Key words
MiR-223-3p /
NLRP3 /
necrotizing enterocolitis /
neonatal rat /
inflammatory response /
apoptosis
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