| Objective:Neonatal necrotizing enterocolitis (NEC) is a hot spot in the newborns in intensive.There are many basic and clinical problem has not been solved, this study through two experiments, on the one hand, through the different frequency and intensity of hypoxia reoxygenation and cold stimulation, to seek for this batch of rat model can be made, on the other hand, on the basis of the one experiment, research on the NEC mignonette element with the newborn rat model of intestinal injury prevention and control mechanism, seek an effective way for further clinical discussion.Methods:First xperiment for building mode selection, the newborn rats were randomly divided into 4 groups.group A as the normal control group, giving normal rat breast feeding, group B given lower frequency and intensity of hypoxia reoxygenation and cold stimulation, group C for medium frequency and intensity of hypoxia reoxygenation and cold stimulation, group D given A higher frequency and intensity of hypoxia reoxygenation and cold stimulation. After four days to die, we observated the newborn rat’s general situation in the process of experiment, weight changes before and after, and im histopathological grading, to compare the differences between groups in terms, to choose suitable bulding mode for the second experiment. We used the appropriate building way in Experiment 2.The newborn rats were randomly divided into 3 groups, group A as the control group, in addition to breastfeeding after building NEC, every day to give sterilized saline lavage. group B of mignonette element processing, in addition to breastfeeding after building NEC, mignonette element filled the stomach every day; For mignonette intervention group, group C in the building at the same time, every day to be inserted mignonette.Then,we observed the general situation of rats, weight changes before and after, intestinal tissue samples of changes to the naked eye.The intestinal tissue was detected the expression of TLR4 and NF-kB p65 protein by immunohistochemical method, and by ELIS A method determined the content of TNF-aand IL-10.Results:In the first experiment, the fourth day, group D appeared one death, intestinal tissue changes to the naked eye, B. C, D groupas compared with group A all had different degrees of change,group C,D obviously. Pathology score A= B< C< D. Building success, each group (alpha = 0.017) compared with group A, B of power and no statistical differences with group A (P> 0.017), the success rate of building C, D group were with group A statistically significant (P< 0.017). Group C can be well balanced with NEC of the subsequent experiments in the clinical, pathological features and degree of tolerance, so group C building was selected scheme of the second experiment. In experiment 2, intestinal tissue specimens to the naked eye observation saw groups of rats pneumatosis visible lumenof different degree,and in group A part of the bowel had distortion. Immunohistochemical determination of TLR4 expression in intestinal tissue, mean of A> B> C, three groups were statistically significant difference (P< 0.05); In the expressions of the NF-kB P65, A> B> C, group A between B and C of difference had statistical significance (P< 0.05); there was no statistically significant difference between Band C groups (P> 0.05).Determination of content of TNF-a, A>B、 C, group A between and B,C groups of difference had statistical significance (P< 0.05),there was no statistically significant difference betweenBand C groups (P> 0.05).Content of IL-10, mean of C>B＞A, had statistically significant difference between the 3 groups (P< 0.05).Conclusions: Moderate with the newborn rat NEC model successfully established, NEC creates opportunities for further study. In the study of mignonette element, group A (NECmodel control group) the expression level of TLR4, NF - kB and TNF-ahad a significant rise. All involved in the newborn rat inflammatory reaction mechanism of NEC, and plays an important role in the NEC insufflate the pathological damage. Also found that when TLR4 expression increases, NF - kB p65 activity increases, The expression of serum TNF -αalso proportional relationship with both, prompt the TLR4/ NF - kB pathways play an important role in immune regulation in the pathogenesis of NEC. Give mignonette after treatment or intervention, blind to the naked eye color better, TLR4, NF-kB and the expression of TNF-αconcentration have dropped significantly, and mignonette early intervention,TLR4mRNA express more apparently.Prompt mignonette can inhibit the expression of TLR4, cut the NF-kB protein expression, thus reducing the release of TNF-aproinflammatory factor. The expression of IL-10 in B and C group was significantly higher in group A, show mignonette but also by raising inflammation inhibiting factor expression of IL-10, avoid bowel mucosa further damage. The study from the level of cell and molecular biology discuss mignonette element on newborn rat model of NEC intestinal injury control mechanism, so as to provide the oretical basis for further clinical prevention and treatment. |
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