Yin Yinyu, Li Yiping, Pan Jian, Tang Ruze, Zhu Jie, Qin Zhenfang, Xu Xiaobing, Wang Jian
Department of General Surgery, Xuzhou Children's Hospital, Xuzhou, Jiangsu 221006, P.R. China.
Department of General Surgery, Children's Hospital of Soochow University, Suzhou, Jiangsu 215025, P.R. China.
Exp Ther Med. 2017 Sep;14(3):2221-2227. doi: 10.3892/etm.2017.4783. Epub 2017 Jul 12.
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% O and 95% N 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.
本研究测定了新生期诱导坏死性小肠结肠炎(NEC)小鼠回肠中MYPT1、CPI-17和MLC20表达水平的变化,为包括NEC期间平滑肌变化的发病机制模型提供依据。一组7日龄BALB/c小鼠经配方奶喂养(40 μl/g,每天5次),并进行缺氧处理(5% O₂和95% N₂,持续10分钟,每天2次),持续4天以诱导NEC并建立小鼠模型。另一组7日龄BALB/c小鼠作为对照组,在整个处理期间由母鼠哺育。模型建立后,处死两组小鼠,收集末端回肠组织并进行蛋白质免疫印迹分析和免疫组织化学检测。结果显示,NEC组小鼠回肠中MYPT1和pMYPT1的表达水平低于对照组(P<0.01)。与对照组相比,NEC组中CPI17和pCPI17的水平更高。NEC组中MLC20的表达水平低于对照组(P<0.01),但NEC组中pMLC20的水平更高(P<0.05)。免疫组织化学结果显示,NEC组中MYPT1、CPI-17和MLC20的染色强度低于对照组,且NEC组阳性细胞比例也较低(P<0.01)。综上所述,我们的结果表明,NEC的发生伴随着MYPT1和pCPI-17蛋白水平的变化,这可能调节回肠平滑肌收缩。