Down-regulation of key regulatory factors in sphingosine-1-phosphate (S1P) pathway in human lung fibroblasts transfected with selected microRNAs.

IF 1.5 Q4 BIOCHEMISTRY & MOLECULAR BIOLOGY Molecular Biology Research Communications Pub Date : 2024-01-01 DOI:10.22099/mbrc.2024.49810.1951
Abdolamir Allameh, Mostafa Atashbasteh, Esmaeil Mortaz, Bahareh Naeeni, Majid Jafari-Khorchani
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Abstract

Sphingosine 1 phosphate (S1P) is involved in the pathogenesis of asthma by stimulation of the alpha-smooth muscle actin (SMA) expression and remodeling of fibroblasts. This study was designed to determine the effects of selected micro RNAs in regulation of S1P and related metabolic pathways in a human lung fibroblast cell line. The fibroblast cell line (CIRC-HLF, C580) was cultured and transfected with individual viral vectors carrying miR124, mi125b, mi133b or mi130a. After 48 hours, expression level of miRNAs and their target genes, sphingosine kinase 1(SPHK1), sphingosine 1-phosphate lyase 1 (SGPL1), sphingosine 1- phosphate receptor 1 (S1PR1) and sphingosine 1- phosphate receptor 2 (S1PR2) were determined. Expression of miRNA and mRNA determined by reverse transcriptionquantitative polymerase chain reaction (qPCR) showed that the expression level of the miRNAs was significantly higher in human lung fibroblasts following transfection compared to controls (vector backbone without miRNA). The expressions of miRNAs-targeted genes were significantly downregulated in transfected fibroblasts compared to control cells (p<0.05). Data show that miR 124, miR 125b, miR 133b and miR130a by targeting regulatory genes in S1P-pathway can down-regulate key factors such as SPHK1, SGPL1, S1PR1 and S1PR2 genes in lung fibroblasts. The results showed that S1P pathway and key factors are suppressed in lung fibroblasts expressing miR124, miR125b, miR130a or miR133b. It appears that suppression of any of the intermediate factors in S1P by miRNA can affect the regulation of the entire S1P pathway.

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转染特定 microRNAs 的人肺成纤维细胞中鞘磷脂-1-磷酸(S1P)通路关键调控因子的下调。
磷酸鞘氨醇 1(S1P)通过刺激α-平滑肌肌动蛋白(SMA)的表达和成纤维细胞的重塑参与了哮喘的发病机制。本研究旨在确定所选微 RNA 在调节 S1P 及相关代谢途径方面对人肺成纤维细胞系的影响。培养成纤维细胞系(CIRC-HLF,C580)并用携带 miR124、mi125b、mi133b 或 mi130a 的病毒载体转染。48 小时后,测定 miRNA 及其靶基因鞘磷脂激酶 1(SPHK1)、1-磷酸鞘磷脂裂解酶 1(SGPL1)、1-磷酸鞘磷脂受体 1(S1PR1)和 1-磷酸鞘磷脂受体 2(S1PR2)的表达水平。反转录定量聚合酶链反应(qPCR)测定的 miRNA 和 mRNA 的表达表明,与对照组(不含 miRNA 的载体骨架)相比,转染后 miRNA 在人肺成纤维细胞中的表达水平显著升高。与对照组(不含 miRNA 的载体骨架)相比,转染成纤维细胞中 miRNAs 靶向基因的表达明显下调(p
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来源期刊
Molecular Biology Research Communications
Molecular Biology Research Communications BIOCHEMISTRY & MOLECULAR BIOLOGY-
CiteScore
3.00
自引率
0.00%
发文量
12
期刊介绍: “Molecular Biology Research Communications” (MBRC) is an international journal of Molecular Biology. It is published quarterly by Shiraz University (Iran). The MBRC is a fully peer-reviewed journal. The journal welcomes submission of Original articles, Short communications, Invited review articles, and Letters to the Editor which meets the general criteria of significance and scientific excellence in all fields of “Molecular Biology”.
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