张喜;段效军;李林瑞;陈艳萍;

• 1:湖南省儿童医院呼吸内科

摘要(Abstract)：

目的探究lncRNA NEAT1通过负调控miR-29a促进低氧低糖诱导的肺纤维化进程的机制。方法收集健康者和缺血再灌注引起急性肺损伤患者的血清样本,通过qRT-PCR检测血清样本中NEAT1与miR-29a的表达。构建肺纤维化细胞模型和动物模型,进行离体和在体的RNA干扰,通过显微镜观察细胞的形态学改变,Hoechst染色检测细胞凋亡,HE染色和Masson染色评估肺组织病理改变,肺组织羟脯氨酸(HYP)含量判定肺组织纤维化程度,Western blot分析细胞中纤维化标志蛋白col1、col3a1、α-SMA、TGF-β的表达,qRT-PCR检测NEAT1、miR-29a的表达;再通过RIP和双荧光素酶报告基因验证NEAT1与miR-29a之间的相互作用及靶向关系。结果在缺血再灌注引起急性肺损伤患者的血清样本中,NEAT1表达上调,miR-29a表达下调。在细胞模型和动物模型中,同样发现NEAT1上调和miR-29a下调。在低氧低糖诱导的肺纤维化细胞模型中,发现敲除NEAT1可以抑制因低氧低糖造成的肺泡上皮细胞形态由鹅卵石型向长梭型变化,同时抑制肺泡上皮细胞的凋亡,降低纤维化标志蛋白col1、col3a1、α-SMA、TGF-β的表达;RIP实验揭示了NEAT1与miR-29a直接结合;双荧光素酶报告基因检测发现,NEAT1靶向调控miR-29a;抑制miR-29a可以逆转敲除NEAT1对低氧低糖诱导肺泡上皮细胞凋亡和降低纤维化标志蛋白col1、col3a1、α-SMA、TGF-β表达的抑制作用,进而促进肺纤维化进程。结论 lncRNA NEAT1通过靶向抑制miR-29a促进低氧诱导的肺纤维化进程,为低氧诱导肺纤维化的机制和新药的研发提供新思路。

关键词(KeyWords)： lncRNA NEAT1;miR-29a;缺氧;肺纤维化;凋亡

Abstract:

Keywords:

基金项目(Foundation): 湖南省卫生健康委2020年度科研立项课题(20200641)

作者(Author): 张喜;段效军;李林瑞;陈艳萍;

Email:

DOI: 10.14053/j.cnki.ppcr.202103004

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