Backgrounds Necrotizing enterocolitis (NEC) was one of the main causes of morbidity and mortality in neonates. Our objective was to detect the mechanism of miR‐124 in small bowel tissues of NEC. Methods Hematoxylin and eosin (H&E) staining was used to detect the repair of the damaged tissues in rat NEC model. Terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) staining was used to evaluate the cell apoptosis level in intestinal tissue. Reverse transcription polymerase chain reaction (RT‐PCR) was used to detect the messenger RNA (mRNA) expression level of miR‐124, Rho‐associated coiled‐coil‐containing protein kinase 1 (ROCK1), myosin phosphatase target subunit 1 (MYPT1), and Toll‐like receptor 9 (TLR9) in NEC tissues and IEC‐6 cells. Luciferase reporter assay was used to verify whether ROCK1 is a direct target of miR‐124. Results miR‐124 was overexpressed in the NEC tissues, while ROCK1 and MYPT1 was downregulated in the NEC tissues. Inhibition of miR‐124, suppressed the intestinal cell apoptosis and promoted the expression of ROCK1 and MYPT1. What is more, overexpression of miR‐124 could inhibit the expression of ROCK1, TLR9, and MYPT1. Luciferase assay confirmed that miR‐124 can regulate the transcriptional activity of ROCK1 through binding its 3′‐UTR region. Conclusion miR‐124 was a promoter of NEC, which promotes the intestine cell apoptosis and inflammatory cell infiltration through the inhibition of TLR9 expression by targeting ROCK1.
Background: To investigate the expressions of fibrinogen (Fib) and Interleukin-12 (IL-12) in serum of neonatal necrotizing enterocolitis (NEC), and to analyze the correlation between the two and their relationship with clinicopathological features. Methods: Forty two children with NEC treated in Xuzhou Children’s Hospital, Xuzhou Medical University Xuzhou, China from 2016-2019 were selected as an observation group and 40 children who underwent physical examination at the same period as a control group. The expression levels of Fib and IL-12 in the serum of two groups were detected by ELISA. The correlation between Fib and IL-12 in the observation group and the correlation among the expressions of Fib, IL-12, the clinicopathological features and common examination indexes of the children with NEC were investigated by Pearson correlation analysis. Results: The levels of Fib and IL-12 in the serum of the children in observation group were significantly higher than those in the control group were (P<0.05). There was a significant positive correlation between the levels of Fib and IL-12 in the serum of the children in observation group (P<0.05). The expression levels of Fib, IL12 were not significantly correlated with sex and age of NEC children, but correlated with vomiting, diarrhea, bloody stool and bradycardia in NEC children (P<0.05). Fib and IL-12 were positively correlated with erythrocyte level (P<0.05) and negatively correlated with platelet level. Conclusion: The expressions of Fib and IL-12 in the serum of NEC children can objectively predict the severity of NEC.
The present study determined the changes in the expression levels of MYPT1, CPI-17 and MLC20 in the ileum of mice with neonatal induced necrotizing enterocolitis (NEC) to provide a basis for a pathogenesis model that includes smooth muscle changes during NEC. A group of 7-day-old BALB/c mice were fed with formula (40 µl/g, 5 times/day) and given hypoxia treatments (5% O2 and 95% N2 for 10 min, twice daily) for 4 days to induce NEC and establish a mouse model. A control group of 7-day-old BALB/c mice were left with their mother for the duration of the treatment. After establishing the model, the two groups of mice were sacrificed, and the terminal ileum tissue was collected and subjected to western blot analysis and immunohistochemistry. The results showed the expression levels of MYPT1 and pMYPT1 in the ileum of the mice in the NEC group were lower than those in the control group (P<0.01). The levels of CPI17 and pCPI17 were higher in the NEC group compared with those in the control group. The expression level of MLC20 in NEC group was lower than that in the control group (P<0.01), but the level of pMLC20 in the NEC group was higher (P<0.05). The results of immunohistochemistry showed that the staining intensities of MYPT1, CPI-17 and MLC20 in the NEC group were lighter than those in the control group, and the proportion of positive cells was also lower in the NEC group (P<0.01). Taken together our results suggest that establishment of NEC is accompanied by changes in the protein levels of MYPT1 and pCPI-17, which can regulate smooth muscle contraction in the ileum.
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