白介素-1β在羊膜腔内注射脂多糖导致的新生鼠肺泡化阻滞中的作用
Effects of interleukin-1β on intra-amniotic lipopolysaccharide-induced alveolar arrest in neonatal rats

目的 ·探讨白介素-1β（IL-1β）在羊膜腔内注射脂多糖（LPS）导致的新生鼠肺泡化阻滞中的作用。方法 ·通过SD孕鼠羊膜腔内注射LPS建立新生鼠支气管肺发育不良（BPD）模型。孕鼠（孕19 d）随机分为生理盐水（Saline）组、LPS组和LPS+anti-IL1β组。随机取出生1、3、7 d新生鼠肺组织，经苏木精-伊红（H-E）染色后观察病理学改变，并通过real-time PCR和ELISA检测肺组织中IL-1β mRNA及蛋白表达水平。培养大鼠骨髓源性原代巨噬细胞，给予LPS干预，经全转录组测序检测与IL-1β相关的多个基因的表达变化。结果 ·与Saline组相比，LPS组肺泡和次级突起数量减少，平均内衬间隔增大；肺组织IL-1β mRNA表达及蛋白表 达量均明显增加。应用anti-IL-1β可改善LPS引起的新生鼠肺组织病理变化。LPS可导致Gbp5、Ccl3、Nod2、Ccr5、Mefv、Casp4、Ifnar1等基因表达明显上调，而Lgals9、Gstp1基因表达明显下调，其中Gbp5、Ccl3、Nod2、Ccr5、Casp4、Ifnar1、Lgals9可正向调节IL-1β表达。结论 ·在BPD模型中LPS通过上调巨噬细胞中IL-1β的表达，诱导新生鼠肺泡化阻滞。全转录组测序揭示LPS可以通过多种路径调控巨噬细胞中IL-1β的表达。
： Objective · To investigate the effects of interleukin-1β (IL-1β) on neonatal rat alveolar arrest induced by intra-amniotic injection of lipopolysaccharide (LPS). Methods · A neonatal SD rat model of bronchopulmonary dysplasia (BPD) was constructed by intra-amniotic injection of LPS in pregnant rats. The pregnant rats (E19) were randomly assigned to Saline group, LPS group and LPS+anti-IL-1β group. The lungs of the neonatal rats were randomly collected 1, 3 and 7 days after birth. Pathological changes in the lungs were observed by hematoxylin-eosin (H-E) staining, and expression of IL-1β mRNA and protein was detected by real-time PCR and ELISA, respectively. Rat bone marrow derived primary macrophage was cultured in vitro, and given LPS intervention, then genes related with IL-1β were detected through whole transcriptome sequencing. Results · Compared with the Saline group, the alveolar counts and secondary septa counts significantly decreased, and mean liner intercept significantly increased in LPS group. Moreover, the expression of IL-1β mRNA and protein in lungs significantly increased in LPS group. The LPS-induced pathological changes of lung tissues in neonatal rats were improved by anti-IL-1β. LPS could up-regulate the expression of genes including Gbp5, Ccl3, Nod2, Ccr5, Mefv, Casp4 and Ifnar1, but down-regulate Lgals9 and Gstp1. Among these genes Gbp5, Ccl3, Nod2, Ccr5, Casp4, Ifnar1 and Lgals9 could positively regulate IL-1β production. Conclusion · LPS can induce alveolar arrest through up-regulating the expression of IL-1β in macrophages in neonatal rat BPD model. Whole transcriptome sequencing reveals that LPS can regulate the expression of IL-1β in macrophages through several paths