PRKCQ-AS1通过Akt信号促进气道平滑肌细胞增殖的研究  

PRKCQ-AS1 promotes airway smooth muscle cell proliferation through Akt signaling

作　　者：吴露依 胡方启 WU Lu-yi;HU Fang-qi(Anqing Medical College Clinical Research Center,Anqing Municipal Hospital,Anqing Anhui 246000,China)

机构地区：[1]安庆医药高等专科学校临床科研中心,安庆市立医院,安徽安庆246000

出　　处：《中国药理学通报》2025年第1期88-94,共7页Chinese Pharmacological Bulletin

基　　金：安徽省高校自然科学研究项目(No KJ2021A1291)。

摘　　要：目的探讨PRKCQ-AS1在气道平滑肌细胞(airway smooth muscle cells,ASMCs)的作用。方法首先通过转录组测序技术(RNA-seq)检测哮喘儿童肺泡灌洗液中的差异表达基因(differentially expressed genes,DEGs)。随后,分别构建PRKCQ-AS1慢病毒敲低和过表达载体并转染ASMCs。qRT-PCR方法检测了ASMCs中PRKCQ-AS1的表达;用CCK-8和流式细胞术分别检测ASMCs的增殖和凋亡;酶联免疫吸附测定(enzyme-linked immunosorbent assay,ELISA)检测ASMCs培养上清液中TNF-α和IL-1β的含量;lncRNA芯片检测PRKCQ-AS1敲低后的DEGs并进行KEGG通路富集;Western blot检测p-Akt和Akt的水平。结果RNA-seq结果显示,PRKCQ-AS1是上调最明显的DEGs之一。PRKCQ-AS1过表达促进了ASMCs的增殖和迁移并抑制了其凋亡,同时,PRKCQ-AS1过表达使上清液中炎性因子TNF-α和IL-1β的水平升高,而PRKCQ-AS1敲低则呈现相反的结果。对PRKCQ-AS1敲低后得到的606个DEGs进行KEGG信号通路富集分析,结果显示PI3K-Akt信号被明显富集。Western blot结果表明,PRKCQ-AS1敲低抑制了Akt的活化;相反,PRKCQ-AS1过表达则促进了Akt的水平。结论PRKCQ-AS1可能通过增强Akt信号促进ASMCs炎症反应,进而加剧了ASMCs的增殖和迁移,最终导致哮喘的发展。Aim To explore the role of PRKCQ-AS1 in airway smooth muscle cells(ASMCs).Methods Differentially expressed genes(DEGs)in alveolar lavage fluid of children with asthma were first detected by transcriptome sequencing technology(RNA-seq).The expression of PRKCQ-AS1 in ASMCs was detected by qRT-PCR;the proliferation and apoptosis of PRKCQ-AS1 were detected by CCK-8 and flow cytometry,respectively;the levels of TNF-αand IL-1βin the culture supernatant of ASMCs were detected by enzyme-linked immunosorbent assay(ELISA);and lncRNA microarray detected DEGs after PRKCQ-AS1 knockdown and KEGG pathway enrichment;Western blot to detect the levels of p-Akt and Akt.Results RNA-seq results showed that PRKCQ-AS1 was one of the most significantly up-regulated DEGs.PRKCQ-AS1 overexpression promoted proliferation and migration and inhibited apoptosis of ASMCs.At the same time,PRKCQ-AS1 overexpression increased the levels of the inflammatory factors TNF-αand IL-1βin the supernatant,whereas PRKCQ-AS1 knockdown showed the opposite results.KEGG signaling pathway enrichment analysis of 606 DEGs obtained after PRKCQ-AS1 knockdown showed that PI3K-Akt signaling was significantly enriched.Western blot results indicated that PRKCQ-AS1 knockdown inhibited Akt activation;on the contrary,PRKCQ-AS1 overexpression promoted Akt levels.Conclusions PRKCQ-AS1 promoted the inflammatory response of ASMCs by enhancing Akt signaling probably,which in turn exacerbated the proliferation and migration of ASMCs,ultimately leading to the development of asthma.

关 键 词：PRKCQ-AS1 ASMCs 哮喘 Akt信号 细胞实验 炎性细胞因子 

分 类 号：R562.25[医药卫生—呼吸系统]