ABSTRACT
Long noncoding RNAs (lncRNAs) are expressed in different species and different tissues, and perform different functions, but little is known about their involvement in the synthesis or secretion of follicle-stimulating hormone (FSH). In general, we have revealed lncRNA‒microRNA (miRNA)‒messenger RNA (mRNA) interactions that may play important roles in rat primary pituitary cells. In this study, a new lncRNA was identified for the first time. First, we analyzed the gene expression of lncRNA-m18as1 in different tissues and different stages by reverse transcription-quantitative polymerase chain reaction (RT-qPCR) and observed the localization of lncRNA-m18as1 with fluorescence in situ hybridization, which indicated that this lncRNA was distributed mainly in the cytoplasm. Next, we used RT-qPCR and enzyme-linked immunosorbent assay (ELISA) to analyze the regulation of FSH synthesis and secretion after overexpression or knockdown of lncRNA-m18as1 and found that lncRNA-m18as1 was positively correlated with FSH synthesis and secretion. In addition, mothers against decapentaplegic homolog 2 (Smad2) was highly expressed in our sequencing results. We also screened miR-18a-5p from our sequencing results as a miRNA that may bind to lncRNA-m18as1 and Smad2. We used RNA immunoprecipitation-qPCR (RIP-qPCR) and/or dual luciferase assays to confirm that lncRNA-m18as1 interacted with miR-18a-5p and miR-18a-5p interacted with Smad2. Fluorescence in situ hybridization (FISH) showed that lncRNA-m18as1 and miR-18a-5p were localized mainly in the cytoplasm. Finally, we determined the relationship among lncRNA-m18as1, miR-18a-5p, and the Smad2/3 pathway. Overall, we found that lncRNA-m18as1 acts as a molecular sponge of miR-18a-5p to regulate the synthesis and secretion of FSH through the Smad2/3 pathway.
Subject(s)
Animals , Rats , Cell Line, Tumor , Cell Proliferation , Follicle Stimulating Hormone/metabolism , Gene Expression Regulation, Neoplastic , In Situ Hybridization, Fluorescence , MicroRNAs/metabolism , RNA, Long Noncoding/metabolismABSTRACT
Objective To study the characteristics of EEG after tramsient ischemic attack and to offer reference for screening procedure of aircrew and astronaut selection. Method The dominant frequency uncertainty of alpha band EEG in 12 pilots(males; age 30±5) with transient ischemic episodes in middle cerebral artery(MCA) territories and in 20 normal healthy pilots was analyzed with frequency-fluctuation analysis. Result The dominant probability of the main frequency coinciding with sites affected by transient ischaemic attack(TIA) in patient pilots was higher than that in healthy pilots (P<0.01),and the dominant probability ratio logarithmic index I≥0 in all patient pilots with normal EEG, but I<0 in all healthy pilots. It was also found that not only I≥0, but the second component shifted to lower frequency(8 Hz) in patients with slight focal EEG alterations,i.e. slowing of frequency. The relative entropy values (percentage) were decreased significantly in pilots with TIA as compared with healthy pilots (P＜0.05). Conclusion The dominant frequency uncertainty analysis of alpha band showed clear superiority of computerized evaluation over routine visual assessment for the diagnosis of minor cerebral ischemia. It offers not only a possibility of studying pathophysiological functional parameter, but also the reference for screening procedure in aircrew and astronaut selection.