摘要
目的:挖掘哮喘的潜在ceRNA网络并探索平喘颗粒抑制哮喘气道血管新生的机制。方法:利用GSE117038数据集,分析文件中差异表达的lncRNA和mRNA,通过比对、预测构建lncRNA-miRNA-mRNA关联,构建ceRNA调控网络。通过屋尘螨(HDM)诱导构建哮喘小鼠模型,并给予平喘颗粒灌胃治疗。分别采用HE染色、Masson染色和免疫组化法检测肺组织病理改变和VEGFA蛋白的表达,采用qRT-PCR法检测肺组织中ceRNA调控网络中关键基因的表达。体外实验观察平喘颗粒含药血清对脐静脉内皮细胞形态、增殖、迁移和ceRNA调控网络中关键基因表达的影响。结果:差异分析共得到816个差异lncRNA,其中下调86个,上调730个,3 171个差异mRNA,其中下调354个,上调2 817个。通过预测lncRNA结合的miRNA共206个,miRNA靶向调控的mRNA共1 269,取交集后得到300个差异mRNA,并构建ceRNA网络。体内实验表明,与模型组比较,平喘颗粒能够显著抑制哮喘气道炎症、胶原沉积和VEGFA蛋白的表达,同时降低肺组织中lncRNA MALAT1和VEGFA mRNA的表达(P<0.05)。体外实验表明,过表达lncRNA MALAT1后HUVEC细胞增殖和迁移数量显著增加(P<0.05)。与MALAT1过表达组比较,平喘颗粒组能够显著抑制HUVEC细胞增殖和迁移,抑制HUVEC细胞lncRNA MALAT1和VEGFA mRNA的表达,增加miR-140-5p的表达(P<0.05)。结论:平喘颗粒能够下调肺组织中lncRNA MALAT1的表达,通过内源性竞争机制进而增加miR-140-5p与VEGFA的结合,从而促进其降解,达到抑制血管新生的作用。
Objective:To explore the potential ceRNA network of asthma and explore the mechanism of Pingchuan Granules inhibiting airway angiogenesis.Methods:Differentially expressed lncRNA and mRNA were analyzed by using the microchip data of GSE117038 dataset,and the lncRNA-miRNA-mRNA association was constructed by comparison and prediction,and ceRNA network was finally constructed.Asthmatic mice were induced by house dust mite and treated with Pingchuan Granules.HE,Masson and immunohistochemistry staining were used to detect the pathological changes and the expression of VEGFA protein in lung tissue.qRT-PCR was used to detect the expression of key genes in ceRNA network.The effects of Pingchuan Granules containing serum on the morphology,proliferation,migration of HUVEC and the expression of key genes in ceRNA regulatory network were observed in vitro.Results:A total of 816 differentially expressed lncRNA were obtained,including 86 down regulated,730 up-regulated and 3171 differentially expressed mRNA,of which 354 were down-regulated and 2817 were up-regulated.It was predicted that there were 206 miRNAs that lncRNA could bind to,and 1269 miRNAs were targeted by miRNA.After the intersection,300 differentially expressed mRNA were obtained and the ceRNA network was constructed.In vivo experiments showed that Pingchuan Granules could significantly inhibit airway inflammation,collagen deposition and VEGFA protein expression,and reduce the expression of lncRNA MALAT1 and VEGFA mRNA in lung tissue,with significant difference when compared with the model group(P<0.05).The proliferation and migration of HUVEC cells were significantly increased after overexpression of lncRNA MALAT1(P<0.05).Compared with the overexpression group,Pingchuan Granules could significantly inhibit the proliferation and migration of HUVEC cells,inhibit the expression of lncRNA MALAT1 and VEGFA mRNA,and increase the expression of miR-140-5 p(P<0.05).Conclusion:Pingchuan Granules can down regulate the expression of lncRNA MALAT1 in lung tissue and increase the binding of miR-140-5 p with VEGFA through endogenous competition mechanism,thus promoting its degradation and inhibiting angiogenesis.
作者
袁星星
王雪慧
王瑶
李竹英
YUAN Xing-xing;WANG Xue-hui;WANG Yao;LI Zhu-ying(Heilongjiang University of Chinese Medicine,Harbin 150040,China;Nangang Branch of Heilongjiang Academy of Traditional Chinese Medicine,Harbin 150006,China;First Affiliated Hospital of Heilongjiang University of Chinese Medicine,Harbin 150040,China)
出处
《中华中医药杂志》
CAS
CSCD
北大核心
2021年第4期2006-2012,共7页
China Journal of Traditional Chinese Medicine and Pharmacy
基金
国家自然科学基金面上项目(No.82074365)
黑龙江省自然科学基金面上项目(No.H2017068)
哈尔滨市应用技术研究与开发项目(No.2017RAXXJ053)。
