目的 探究大黄素对新生坏死性小肠结肠炎模型大鼠细胞焦亡及胞内模式识别受体3（NLRP3）-白细胞介素1β（IL-1β）通路的影响。方法 将72只新出生1 d的SD大鼠，随机分为对照组、模型组及大黄素高、中、低剂量（20、10、5 mg/kg）组，每组15只（对照组12只）。除对照组外，其余大鼠均采用配方奶粉人工喂养＋缺氧＋冷刺激方法制备新生坏死性小肠结肠炎模型。每日观察大鼠一般状态，定时测量和记录大鼠体质量；苏木精-伊红（HE）染色观察肠组织病理学变化；酶联免疫吸附法（ELISA）检测肠组织匀浆中IL-1β、IL-18水平；实时荧光定量PCR检测NLRP3、IL-1β、IL-18 mRNA表达水平；Western blotting检测肠组织Caspase-1、P10、P20蛋白表达水平。结果 与对照组相比，模型组大鼠出现腹胀、喂养困难、活动减少、黑便、血便现象，肠组织损伤严重；病理损伤评分，IL-1β和IL-18水平，IL-1β、IL-18、NLRP3 mRNA表达水平，P10、P20蛋白表达水平均显著增加（P<0.05），大鼠体质量、Caspase-1蛋白表达明显降低（P<0.05）；与模型组相比，大黄素高、中、低剂量组大鼠腹胀、黑便、血便情况明显减轻，无喂养困难的现象，肠组织损伤评分降低，IL-1β、IL-18水平和mRNA表达水平明显降低，NLRP3 mRNA表达水平明显降低，P10、P20蛋白表达水平明显降低（P<0.05），大鼠体质量、Caspase-1蛋白表达明显升高（P<0.05）。结论 细胞焦亡可能参与了新生儿坏死性小肠结肠炎的发病过程，大黄素可能通过抑制NLRP3-IL-1β信号通路，减少新生坏死性小肠结肠炎模型大鼠肠细胞焦亡。

Objective To investigate the effects of emodin on pyroptosis and NOD-like receptor family, pyrin domain containing 3 (NLRP3)-interleukin-1β (IL-1β) pathway in neonatal necrotizing enterocolitis (NEC) rats. Methods A total of 72 one-day-old SD rats were randomly divided into control group, NEC model group, high, medium and low dose emodin (20, 10, 5 mg/kg) groups, including 12 rats in control group and 15 rats in each group. Except for the control group, the rest of the rats were fed with formula milk powder + hypoxia + cold stimulation method to prepare NEC model. The general state of rats were observed every day, the body weight of rats were measured and recorded regularly. Hematoxylin eosin (HE) staining was used to observe the pathological changes of intestinal tissue; the contents of IL-1β and IL-18 in intestinal homogenate were detected by enzyme-linked immunosorbent assay (ELISA). The mRNA levels of NLRP3, IL-1β, and IL-18 were detected by real-time fluorescence quantitative PCR; and the protein levels of Caspase-1, P10 and P20 were detected by Western blotting. Results Compared with those in the control group, the rats in model group showed abdominal distension, feeding difficulty, reduced activity, black stool and bloody stool, and serious intestinal tissue damage, the pathological injury score, the content and mRNA level of IL-1β and IL-18, the expression of NLRP3 mRNA, P10, and P20 protein were significantly higher (P<0.05), the body weight and Caspase-1 protein expression were significantly lower (P<0.05). Compared with those in model group, the abdominal distension, black stool and bloody stool of rats in high, medium and low dose emodin groups were significantly reduced, there was no feeding difficulty. The tissue injury score, the content and mRNA level of IL-1β and IL-18, and the expression of NLRP3 mRNA, P10 and P20 protein were significantly lower (P<0.05), and the body weight and Caspase-1 protein expression were significantly higher (P<0.05). Conclusion Pyroptosis may be involved in the pathogenesis of NEC. Emodin may reduce the pyroptosis of Intestinal cells in NEC model rats by inhibiting NLRP3-IL-1β signaling pathway.

R285.5

国家自然科学青年科学基金资助项目（81500430）