Sharifi Laleh, Aghamohammadi Asghar, Aletaha Somaye, Bigdeli Razieh, Asgary Vahid, Bokaie Saied, Asgardoon Mohammad Hossein, Azizi Gholamreza, Mirshafiey Abbas
Uro-Oncology Research Center, Tehran University of Medical Sciences, Tehran, Iran.
Research Center for Immunodeficiencies, Pediatrics Center of Excellence, Children's Medical Center, Tehran University of Medical Sciences, Tehran, Iran.
Endocr Metab Immune Disord Drug Targets. 2019;19(2):144-149. doi: 10.2174/1871530319666181126153752.
Inhibition of Toll-like receptors (TLRs) signaling plays a crucial role in suppressing the inflammation and available data presenting G2013 as an immunomodulatory agent, therefore, we designed this study to answer whether G2013 can affect the signaling pathway of TLR2 and TLR4.
Cytotoxicity study of G2013 was performed by MTT assay. HEK293 TLR2 and HEK293 TLR4 cell lines were cultured and treated with low dose (5µg/ml) and high dose (25µg/ml) of G2013 for 24 hours. Gene expressions of MyD88, Tollip, and NF-κB were defined by quantitative real-time PCR.
The cytotoxicity assay showed that the concentrations lesser than 125μg/ml of G3012 had no apparent cytotoxicity, however, the concentrations of 5µg/ml and 25µg/ml could suppress the mRNA expression of MyD88, Tollip and NF-κB in HEK293 TLR2 and HEK293 TLR4 cell lines.
in our study, we verified the linkage between the immunosuppressive property of G2013 and TLR2, TLR4 signaling cascade; but so far, the specific target of G2013 and its molecular mechanism has not been detected yet. We recommend further studies on other Patten Recognition Receptors (PRRs)and other mechanisms of inflammation like oxidative stress to be conducted in the future.
Toll样受体（TLRs）信号通路的抑制在抑制炎症中起着关键作用，并且有数据表明G2013是一种免疫调节剂，因此，我们设计了本研究以回答G2013是否能影响TLR2和TLR4的信号通路。
通过MTT法进行G2013的细胞毒性研究。培养HEK293 TLR2和HEK293 TLR4细胞系，并用低剂量（5μg/ml）和高剂量（25μg/ml）的G2013处理24小时。通过定量实时PCR确定MyD88、Tollip和NF-κB的基因表达。
细胞毒性试验表明，浓度低于125μg/ml的G3012没有明显的细胞毒性，然而，5μg/ml和25μg/ml的浓度可以抑制HEK293 TLR2和HEK293 TLR4细胞系中MyD88、Tollip和NF-κB的mRNA表达。
在我们的研究中，我们验证了G2013的免疫抑制特性与TLR2、TLR4信号级联之间的联系；但到目前为止，尚未检测到G2013的具体靶点及其分子机制。我们建议未来对其他模式识别受体（PRRs）和其他炎症机制如氧化应激进行进一步研究。
