上海交通大学学报（医学版）

上海交通大学学报（医学版） ›› 2019, Vol. 39 ›› Issue (7): 714-.doi: 10.3969/j.issn.1674-8115.2019.07.005

• 论著·基础研究 • 上一篇    下一篇

鲨肝醇对支气管肺发育不良新生大鼠肺泡化阻滞的改善作用

谢小华 1，陈泽 1，刘成博 1，杨奕辉 2，张拥军 1   

  1. 1.上海交通大学医学院附属新华医院新生儿科，上海 200092；2. 上海市嘉定区中心医院儿科，上海 201880
  • 出版日期:2019-07-28 发布日期:2019-08-26
  • 通讯作者: 张拥军，电子信箱：zhangyongjun@sjtu.edu.com。
  • 作者简介:谢小华（1991—），女，硕士生；电子信箱： Xiexh7166@163.com。
  • 基金资助:
    国家自然科学基金面上项目（81470201）

Improvement of batyl alcohol on alveolarization arrest in newborn rats with bronchopulmonary dysplasia

XIE Xiao-hua1, CHEN Ze1, LIU Cheng-bo1, YANG Yi-hui2, ZHANG Yong-jun1   

  1. 1. Department of Neonatology, Xinhua Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai 200092, China; 2. Department of Pediatrics, Jiading District Central Hospital, Shanghai 201880, China
  • Online:2019-07-28 Published:2019-08-26
  • Supported by:
    National Natural Science Foundation of China, 81470201

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摘要： 目的 ·探讨鲨肝醇（ batyl alcohol，BTA）对脂多糖（ lipopolysaccharide，LPS）诱导的新生大鼠支气管肺发育不良（bronchopulmonary dysplasia，BPD）的病理学改变的改善作用及其作用机制。方法 ·将孕 16.5 d的 SD大鼠随机分为 Saline组、LPS组、LPS+BTA组，羊膜腔注射 LPS建立新生鼠 BPD模型； LPS+BTA组在注射 LPS同时及出生后 1～ 7 d每日注射 BTA干预，其他 2组注射等体积生理盐水。取出生第 1、3、7日新生鼠肺组织，通过苏木精 -伊红（ H-E）染色和间苯二酚碱性品红染色观察肺泡化阻滞的改变；利用荧光定量 PCR和 ELISA法分别检测新生鼠肺组织中炎症因子白细胞介素 1β（IL-1β）mRNA水平及蛋白表达量。体外培养小鼠巨噬细胞系 RAW264.7，检测 LPS诱导及 BTA干预后 IL-1β mRNA水平及蛋白表达量。分离 SD大鼠骨髓巨噬细胞，给予 LPS刺激和 BTA干预，利用全转录组测序技术筛选 BTA抑制炎症的可能靶点。结果 · H-E染色结果显示，出生后第 1、3、7日 LPS+BTA组较 LPS组肺泡数量增多、次级间隔数量增多、平均内衬间隔减小（均 P<0.05）；且经 BTA干预后肺组织 IL-1β mRNA及蛋白表达量均较 LPS组下调（均 P<0.05）。体外实验结果显示， LPS刺激后巨噬细胞 IL-1β mRNA和蛋白均较 Saline组显著增加（均 P0.000），加入 BTA后两者均较 LPS组减少（均 P<0.05）。全转录组测序结果显示， BTA抑制血小板应答蛋白 1（thrombospondin， Thbs1）、髓样细胞触发受体 1（triggering receptor expressed on myeloid cells 1，Trem1）、表面抗原分化簇 274（cluster of differentiation 274，Cd274）等促炎症因子基因的表达，促进补体 C1q C链（complement C1q C chain，C1qc）、RT1-Da（RT1 class Ⅱ，locus Da）、 RT1-Db1（RT1 class Ⅱ，locus Db1）等抗炎因子基因的表达。结论 · BTA可下调巨噬细胞中 IL-1β的表达，抑制炎症反应，进而改善 LPS诱导的 BPD新生鼠肺组织病理学改变。

关键词: 支气管肺发育不良, 炎症, 鲨肝醇, 白细胞介素 1&, beta, 肺泡化阻滞

Abstract:

Objective · To investigate whether batyl alcohol (BTA) can improve the pathology of bronchopulmonary dysplasia (BPD) in newborn rats inducedlipopolysaccharide (LPS) and the mechanism. Methods · Pregnant SD rats (16.5 d) were randomly assigned into Saline group, LPS group, and LPS+BTA group. Amniocentesis injection of LPS was performed to establish neonatal bronchopulmonary dysplasia (BPD) rat model. In LPS+BTA group, LPS and BTA were injected at the same time. After birth, LPS+BTA group was injected with BTA continuously everyday for 7 days. The other two groups were injected with normal saline of equal volume. Lung tissues of neonatal rats on the first, third and seventh day after birth were stainedhematoxylineosin (H-E) and resorcin-fuchsin respectively, to observe alveolarization arrest. The mRNA and protein levels of interleukin 1β (IL-1β) in newborn rats lungs were detectedreal-time PCR and ELISA. In vitro, momacrophages RAW264.7 were cultured to detect IL-1β mRNA levels and protein levels after treatment with LPS and BTA. SD rat bone marrow macrophages were isolated and treated with LPS and BTA. RNA-sequence was taken to screen for possible targets of BTA inhibition of inflammation. Results · The results of H-E staining showed that LPS+BTA group had a milder pathology of BPD, with more secondary septa counts, more alveolar counts, and smaller mean linear intercept (all P<0.05); after BTA intervention the levels of IL1β mRNA and protein in lung tissues of neonatal rats were significantly lower than those in LPS group (both P<0.05). In vitro, IL-1β mRNA and protein increased after LPS stimulation (both P0.000), but decreased in the LPS+BTA group (both P<0.05). RNA-sequence results showed that BTA inhibited the s of some inflammatory factors, such as thrombospondin1 (Thbs1), triggering receptor expressed on myeloid cells 1 (Trem1), and cluster of differentiation 274 (Cd274), and promoted the s of some anti-inflammatory factors, such as complement C1q C chain (C1qc), RT1 class Ⅱ, locus Da (RT1-Da), and RT1 class Ⅱ, locus Db1 (RT1-Db1). Conclusion · BTA can improve lung pathology of neonatal rats with BPDdownregulating the of IL-1β and reducing inflammatory response.

Key words: bronchopulmonary dysplasia (BPD), inflammation, batyl alcohol (BTA), interleukin-1&beta, (IL-1&beta;), alveolarization arrest

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