Hwang Dukhyun, Son Byeng-Wha, Shin Pyung-Gyun, Choi Jae-Sue, Seo Yong-Bae, Kim Gun-Do
Department of Microbiology, College of Natural Sciences, Pukyong National University, Busan, South Korea.
Department of Chemistry, College of Natural Sciences, Pukyong National University, Busan, South Korea.
J Pharm Pharmacol. 2015 Sep;67(9):1297-305. doi: 10.1111/jphp.12421. Epub 2015 Apr 23.
The purpose of this study is to investigate anti-inflammatory effects of toluhydroquinone in lipopolysaccharide (LPS)-stimulated RAW264.7 cells.
Toluhydroquinone was purified from a fungal strain, Aspergillus sp. We investigated that levels of nitric oxide (NO) using Griess reagent, production of prostaglandin E2 (PGE2 ) and pro-inflammatory cytokines using ELISA assay. We conducted Western blot analysis to investigate regulatory effects of toluhydroquinone on expression of inducible nitric oxide synthase (iNOS), cyclooxyganse-2 (COX-2), nuclear factor-κB (NF-κB), Akt and mitogen-activated protein kinases (MAPKs) in LPS-stimulated RAW264.7 cells. The translocation of NF-κB was detected by immunofluorescence staining.
Toluhydroquinone inhibited production of NO and PGE2 via suppressing protein expression of iNOS and COX-2, respectively. Secretion and expression of inflammatory cytokines were down-regulated by toluhydroquinone as well. Toluhydroquinone reduced phosphorylation of Akt, NF-κB and MAPKs. Moreover, toluhydroquinone inhibited translocation of NF-κB from the cytosol into the nucleus.
We revealed that inhibitory effects of toluhydroquinone on expression of inflammatory mediators are induced through inactivation of Akt, NF-κB and MAPKs. Thus, our results suggest that toluhydroquinone may be used for a potential anti-inflammatory reagent.
本研究旨在探讨甲苯氢醌对脂多糖(LPS)刺激的RAW264.7细胞的抗炎作用。
从真菌菌株曲霉属中纯化出甲苯氢醌。我们使用格里斯试剂研究一氧化氮(NO)水平,使用酶联免疫吸附测定法研究前列腺素E2(PGE2)和促炎细胞因子的产生。我们进行蛋白质印迹分析,以研究甲苯氢醌对LPS刺激的RAW264.7细胞中诱导型一氧化氮合酶(iNOS)、环氧化酶-2(COX-2)、核因子-κB(NF-κB)、Akt和丝裂原活化蛋白激酶(MAPKs)表达的调节作用。通过免疫荧光染色检测NF-κB的易位。
甲苯氢醌分别通过抑制iNOS和COX-2的蛋白表达来抑制NO和PGE2的产生。甲苯氢醌还下调了炎性细胞因子的分泌和表达。甲苯氢醌降低了Akt、NF-κB和MAPKs的磷酸化。此外,甲苯氢醌抑制了NF-κB从细胞质向细胞核的易位。
我们发现甲苯氢醌对炎症介质表达的抑制作用是通过Akt、NF-κB和MAPKs的失活诱导的。因此,我们的结果表明甲苯氢醌可能用作潜在的抗炎试剂。