Department of Cellular and Molecular Biology, Kish International Campus, University of Tehran, Kish, Iran; Department of Immunology, School of Public Health, Tehran University of Medical Sciences, Tehran, Iran.
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.
Int Immunopharmacol. 2018 Feb;55:323-329. doi: 10.1016/j.intimp.2018.01.003. Epub 2018 Jan 5.
This research aimed to study the anti-inflammatory and immunomodulatory effects of guluronic acid (G2013) on gene expression of TLR4, MyD88, SHIP1, SOCS1, NF-κB, and assessment of the level of IL-1β as a pro-inflammatory cytokine in HEK-Blue hTLR4 cell line.
The cytotoxicity of G2013 was assessed by the MTT assay. The mRNA expression levels of the mentioned genes were measured by qRT-PCR. IL-1β concentration in culture media was determined using ELISA method.
MTT assay demonstrated that G2013 (before the concentration of 125μg/ml) had no cytotoxic effect on HEK-Blue hTLR4 cells. Our results indicated that the low and high doses of this drug could significantly reduce the gene expression of TLR4 and MyD88, as compared to the control group (p<0.05). Moreover, it was found that the low dose of this drug could significantly increase the gene expression of SHIP1 and SOCS1, as compared to the control group (p<0.05). Furthermore, the study findings revealed that the level of NF-κB gene expression significantly reduced, in both doses of G2013 compared to the control group (p<0.05, p<0.01, respectively). Our data showed that the level of IL-1β in culture media decreased by both doses of this drug in comparison to control group (p<0.05).
This study indicates that G2013 is able to induce SHIP1, SOCS1 and reduce TLR4, MyD88, NF-κB at the level of gene expression and decrease IL-1β as a pro-inflammatory cytokine which might be recommended for reduction of inflammatory reactions.
本研究旨在研究古洛糖醛酸（G2013）对 TLR4、MyD88、SHIP1、SOCS1、NF-κB 基因表达的抗炎和免疫调节作用，并评估作为促炎细胞因子的 IL-1β 的水平在 HEK-Blue hTLR4 细胞系中的变化。
通过 MTT 测定法评估 G2013 的细胞毒性。通过 qRT-PCR 测定所述基因的 mRNA 表达水平。使用 ELISA 法测定培养基中 IL-1β 的浓度。
MTT 测定法表明，G2013（浓度在 125μg/ml 之前）对 HEK-Blue hTLR4 细胞没有细胞毒性作用。我们的结果表明，与对照组相比，该药物的低剂量和高剂量均可显著降低 TLR4 和 MyD88 的基因表达（p<0.05）。此外，还发现与对照组相比，该药物的低剂量可显著增加 SHIP1 和 SOCS1 的基因表达（p<0.05）。此外，研究结果表明，与对照组相比，G2013 的两种剂量均可显著降低 NF-κB 基因表达水平（p<0.05，p<0.01）。我们的数据显示，与对照组相比，该药物的两种剂量均可降低培养基中 IL-1β 的水平（p<0.05）。
本研究表明，G2013 能够诱导 SHIP1 和 SOCS1，并降低 TLR4、MyD88 和 NF-κB 的基因表达水平，同时降低作为促炎细胞因子的 IL-1β，这可能有助于减轻炎症反应。
