Mortazavi-Jahromi Seyed Shahabeddin, Jamshidi Mehdi Malek, Farazmand Ali, Aghazadeh Zahra, Yousefi Mehdi, Mirshafiey Abbas
Department of Cellular and Molecular Biology, Kish International Campus, University of Tehran, Kish, Iran; School of Biology, University College of Science, University of Tehran, Tehran, Iran.
Department of Immunology, School of Public Health, Tehran University of Medical Sciences, Tehran, Iran.
Pharmacol Rep. 2017 Jun;69(3):479-484. doi: 10.1016/j.pharep.2017.01.021. Epub 2017 Jan 24.
Impaired expression and function of microRNAs (miRNAs) are involved in the pathogenesis of many autoimmune and inflammatory diseases. Moreover, there is a close relationship between TLRs and miRNAs and impairment in regulating their expression which can play a vital role in the immunopathogenesis of many inflammatory reactions. This research aimed to study the pharmaceutical effects of M2000 (β-d-mannuronic acid) on the expression of miR-146a and its two target molecules (IRAK1 and TRAF6), and the transcription factor NF-κB in the HEK-Blue hTLR2 cell line.
The cytotoxicity of M2000 was assessed by the MTT assay, and the qRT-PCR technique was employed in the presence and absence of M2000 treatment to measure gene-expression levels of miR-146a, IRAK1, TRAF6, and NF-κB.
MTT assay indicated that M2000 (before the concentration of 500μg/ml) had no cytotoxic effect on HEK-Blue hTLR2 cells. Our results showed that M2000 at low and high doses (5 and 25μg/well) could significantly reduce gene expression levels of miR-146a (p<0.01). Furthermore, it was found that this medication at two different doses could considerably decrease IRAK1 and TRAF6 gene expression (p<0.001). Moreover, this study revealed that expression level of NF-κB also significantly declined at these two doses (p<0.01).
This study for the first time shows that M2000 as a novel NSAID with immunosuppressive properties is able to modify TLR signaling through suppressing the adaptor molecules IRAK1 and TRAF6, the transcription factor NF-κB and miR-146a as a new therapeutic approach.
微小RNA（miRNA）的表达和功能受损与许多自身免疫性和炎性疾病的发病机制有关。此外，Toll样受体（TLR）与miRNA之间存在密切关系，且其表达调控受损在许多炎症反应的免疫发病机制中起关键作用。本研究旨在探讨M2000（β - D - 甘露糖醛酸）对人胚肾 - 蓝色hTLR2细胞系中miR - 146a及其两个靶分子（白细胞介素 - 1受体相关激酶1（IRAK1）和肿瘤坏死因子受体相关因子6（TRAF6））以及转录因子核因子κB（NF - κB）表达的药物作用。
采用MTT法评估M2000的细胞毒性，并在有或无M2000处理的情况下，运用qRT - PCR技术检测miR - 146a、IRAK1、TRAF6和NF - κB的基因表达水平。
MTT法表明，M2000（浓度在500μg/ml之前）对人胚肾 - 蓝色hTLR2细胞无细胞毒性作用。我们的结果显示，低剂量和高剂量（5和25μg/孔）的M2000均可显著降低miR - 146a的基因表达水平（p<0.01）。此外，发现该药物在两种不同剂量下均可显著降低IRAK1和TRAF6的基因表达（p<0.001）。而且，本研究表明这两种剂量下NF - κB的表达水平也显著下降（p<0.01）。
本研究首次表明，M2000作为一种具有免疫抑制特性的新型非甾体抗炎药，能够通过抑制衔接分子IRAK1和TRAF6、转录因子NF - κB以及miR - 146a来调节TLR信号传导，这是一种新的治疗方法。
