miR-29b通过抑制卵泡抑素样蛋白1减轻新生儿呼吸窘迫综合征小鼠肺损伤  

miR-29b alleviates lung injury in neonatal respiratory distress syndrome mouse by inhibiting follistatin-like protein 1

作　　者：罗玲英[1] 汤淑斌[1] 范小刚[1] Luo Lingying;Tang Shubin;Fan Xiaogang(Department of Neonatology,Baoji maternal and Child Health Hospital,Baoji 721000,Shaanxi)

机构地区：[1]宝鸡市妇幼保健院新生儿科,宝鸡721000

出　　处：《中国组织化学与细胞化学杂志》2024年第5期420-428,共9页Chinese Journal of Histochemistry and Cytochemistry

基　　金：陕西省重点研发计划项目(2021SF-211)。

摘　　要：目的分析miR-29b在新生儿呼吸窘迫综合征(neonatal respiratory distress syndrome,NRDS)小鼠模型中的表达,并探讨其在NRDS过程中的调节机制。方法建立气管内滴注LPS(5 mg/kg)诱导的NRDS小鼠模型,以及LPS(100 ng/mL)处理的人肺微血管内皮细胞(human pulmonary microvascular endothelial cell,HPMEC)模型。通过实时定量聚合酶链反应(qRT-PCR)检测miR-29b和卵泡抑素样蛋白1(follistatin-like protein 1,FSTL-1)的表达水平;采用Western blot、流式细胞术和酶联免疫吸附分析(ELISA)评估细胞凋亡及炎症损伤情况;利用荧光素酶活性分析验证miR-29b与FSTL-1的相互作用;通过组织病理学和TUNEL分析评估小鼠肺组织损伤,免疫组化检测肺组织中FSTL-1的表达。结果在体内外NRDS模型中均观察到miR-29b表达下调。上调miR-29b基因可显著抑制LPS处理的人肺微血管内皮细胞的凋亡及炎症反应,而FSTL-1的加入则减弱miR-29b对细胞凋亡和炎症反应的抑制作用。荧光素酶活性分析进一步证实miR-29b可与FSTL-1结合。此外,miR-29b基因上调可改善NRDS小鼠模型的肺组织病理学结构,减少细胞凋亡并减轻炎症损伤。结论miR-29b通过调节FSTL-1的表达,减轻LPS诱导的NRDS小鼠肺损伤,从而在NRDS的发生和进展中发挥重要的保护作用。Objective To analyze the expression of miR-29b in a neonatal respiratory distress syndrome(NRDS)mouse model and explore its regulatory mechanism in the process of NRDS.Methods The NRDS mouse model was established by intratracheal instillation of LPS(5 mg/kg),and a human pulmonary microvascular endothelial cell(HPMEC)model was induced with LPS(100 ng/mL).The expression levels of miR-29b and follistatin-like protein 1(FSTL-1)were measured using quantitative real-time PCR(qRT-PCR).Western blot,flow cytometry,and enzyme-linked immunosorbent assay(ELISA)were used to assess cell apoptosis and inflammatory damage.Luciferase activity assays were conducted to verify the interaction between miR-29b and FSTL-1.Histopathological analysis and TUNEL assays were used to evaluate lung tissue damage in the animal model,and immunohistochemistry was performed to detect FSTL-1 expression in lung tissue.Results Downregulation of miR-29b was observed in both in vitro and in vivo models of NRDS.Upregulation of miR-29b significantly inhibited apoptosis and inflammation in LPS-treated HPMEC,and the addition of FSTL-1 weakened its inhibitory effects.Luciferase activity assays confirmed the interaction between miR-29b and FSTL-1.Moreover,upregulation of miR-29b improved the histopathological structure of lung tissue,reduced apoptosis,and alleviated inflammation in the NRDS mouse model.Conclusion Upregulation of miR-29b mitigates LPS-induced lung injury in NRDS mice by regulating FSTL-1,playing a protective role in the development and progression of NRDS.

关 键 词：miR-29b 卵泡抑素样蛋白1 呼吸窘迫综合征 新生小鼠 肺损伤 炎症 

分 类 号：R722[医药卫生—儿科]