胞嘧啶核苷二磷酸胆碱对新生大鼠坏死性小肠结肠炎的治疗作用

Cytidine diphosphate choline for treatment of necrotizing enterocolitis in neonatal rats

目的探讨胞嘧啶核苷二磷酸胆碱(CDP-胆碱)对新生大鼠坏死性小肠结肠炎(NEC)的治疗作用。方法将40只清洁级SD新生大鼠按随机数字表法分4组,每组10只:空白对照组(母鼠喂养且无任何干预措施)、NEC对照组(配方奶喂养,低温+缺氧刺激法建模,腹腔注射0.9%氯化钠注射液)、CDP-胆碱治疗组(配方奶喂养,低温+缺氧刺激法建模,腹腔注射CDP-胆碱1次/d)、CDP-胆碱对照组(母鼠喂养,腹腔注射CDP-胆碱1次/d)。实验第7天处死大鼠并取肠段进行HE染色及Shiou评分,同时采用免疫荧光法检测自噬基因Beclin-1及微管相关蛋白LC3Ⅱ荧光强度,ELISA法检测TNF-α、IL-1β、IL-6等炎症因子水平。结果NEC对照组大鼠肠绒毛普遍水肿,黏膜及黏膜下层可见以中性粒细胞为主的大量炎症细胞,毛细血管有扩张,血管外可见红细胞,部分肠上皮细胞消失;Shiou评分及肠道组织TNF-α、IL-1β、IL-6水平均明显高于其他3组(均P<0.05);肠道组织Beclin-1、LC3Ⅱ荧光强度明显增强。空白对照组、CDP-胆碱治疗组和CDP-胆碱对照组大鼠肠黏膜上皮连续、完整,细胞与腺体排列规则,未见炎症细胞浸润;Shiou评分及肠道组织TNF-α、IL-1β、IL-6水平两两比较,差异均无统计学意义(均P>0.05);肠道组织Beclin-1、LC3Ⅱ荧光强度差异不明显。结论CDP-胆碱对新生大鼠NEC有明显治疗作用。

Objective To explore the efficacy of cytosine diphosphate choline(CDP-choline)for treatment of necrotizing enterocolitis(NEC)in neonatal rats.Methods Forty SD newborn rats were divided into 4 groups randomly with 10 rats in each group:rats in blank control group received breast feeding had no any intervention;rats in NEC model group were fed with formula milk and treated with hypothemia,hypoxia and intraperitoneal injection of 0.9%sodium chloride to induce NEC;rats in CDP-choline treatment group were fed with formula and given intraperitoneal injection of CDP-choline qd after the NEC modeling,and rats in CDP-choline control group had breast feeding and had no any intervention after NEC modeling.The rats were sacrificed on d7 and the intestinal samples were taken for HE staining and evaluation with Shiou score.The immunofluorescence method was used to detect the fluorescence intensity of autophagy gene Beclin-1 and microtubule-associated protein LC3Ⅱ,and ELISA method was used to detect TNF-α,IL-1β,and IL-6.Results The intestinal villi of rats in the NEC control group were generally edematous,and a large number of inflammatory cells,mainly neutrophils,were seen in the mucosa and submucosa,capillaries were dilated,red blood cells were seen outside the vessels,and some intestinal epithelial cells disappeared.Shiou score and the intestinal tissue levels of TNF-α,IL-1β,and IL-6 were significantly higher than those of the other three groups(all P<0.05),and the fluorescence intensity of Beclin-1 and LC3Ⅱin intestinal tissue was significantly increased.The intestinal mucosal epitheliumof rats in the blank control group,CDP-choline treatment group and CDP-choline control group was continuous and complete,the cells and glands were arranged regularly,and there was no inflammatory cell infiltration.There were no significant differences in Shiou score and intestinal tissue TNF-α,IL-1βand IL-6 levels(all P>0.05),as well as in the fluorescence intensity of Beclin-1 and LC3Ⅱamong these three groups.Conclusion CDP-choline exerts protective and therapeutic effects on neonatal rats with NEC.