Baskaran Asweni, Chua Kek Heng, Sabaratnam Vikineswary, Ravishankar Ram Mani, Kuppusamy Umah Rani
Institute of Biological Sciences, Faculty of Science, University of Malaya, 50603, Kuala Lumpur, Malaysia.
Mushroom Research Centre, Faculty of Science, University of Malaya, 50603, Kuala Lumpur, Malaysia.
BMC Complement Altern Med. 2017 Jan 13;17(1):40. doi: 10.1186/s12906-016-1546-6.
Pleurotus giganteus (Berk. Karunarathna and K.D. Hyde), has been used as a culinary mushroom and is known to have medicinal properties but its potential as an anti-inflammatory agent to mitigate inflammation triggered diseases is untapped. In this study, the molecular mechanism underlying the protective effect of ethanol extract of P. giganteus (EPG) against lipopolysaccharide (LPS) and combination of LPS and hydrogen peroxide (HO)-induced inflammation on RAW 264.7 macrophages was investigated.
The effect of EPG on nitric oxide (NO) production as an indicator of inflammation in RAW 264.7 macrophages was estimated based on Griess reaction that measures nitrite level. The expressions of inducible nitric oxide synthase (iNOS), cyclooxygenase-2 (COX-2), NF-kB activating protein (NKAP), signal transducer and activator of transcription 3 protein (STAT 3) and glutathione peroxidase (GPx) genes were assessed using real time reverse transcription polymerase chain reaction (RT-PCR) approach.
EPG (10 μg/ml) showed the highest reduction in the LPS-induced NO production in RAW 264.7 macrophages and significantly suppressed (p < 0.05) the expression iNOS, STAT 3 and COX-2. There was a significant increase (p < 0.05) in combination of LPS and HO- induced iNOS production when compared to the LPS-induced iNOS production in RAW 264.7 macrophages and this concurred with the NO production which was attenuated by EPG at 10 μg/ml. A significant (p < 0.05) down regulation was observed in the combination of LPS and HO-induced iNOS and GPx expression by EPG.
Our data suggest that the anti-inflammatory activity of EPG is mediated via the suppression of the STAT 3 and COX-2 pathways and can serve as potential endogenous antioxidant stimulant.
巨大侧耳(Berk. Karunarathna和K.D. Hyde),已被用作食用蘑菇,并且已知具有药用特性,但其作为减轻炎症引发疾病的抗炎剂的潜力尚未被开发。在本研究中,研究了巨大侧耳乙醇提取物(EPG)对脂多糖(LPS)以及LPS与过氧化氢(HO)联合诱导RAW 264.7巨噬细胞炎症的保护作用的分子机制。
基于测量亚硝酸盐水平的格里斯反应,评估EPG对RAW 264.7巨噬细胞中作为炎症指标的一氧化氮(NO)产生的影响。使用实时逆转录聚合酶链反应(RT-PCR)方法评估诱导型一氧化氮合酶(iNOS)、环氧化酶-2(COX-2)、NF-κB激活蛋白(NKAP)、信号转导和转录激活因子3蛋白(STAT 3)以及谷胱甘肽过氧化物酶(GPx)基因的表达。
EPG(10μg/ml)在RAW 264.7巨噬细胞中对LPS诱导的NO产生显示出最大程度的降低,并显著抑制(p<0.05)iNOS、STAT 3和COX-2的表达。与RAW 264.7巨噬细胞中LPS诱导的iNOS产生相比,LPS与HO联合诱导的iNOS产生显著增加(p<0.05),这与10μg/ml的EPG减弱的NO产生一致。EPG对LPS与HO联合诱导的iNOS和GPx表达有显著(p<0.05)下调作用。
我们的数据表明,EPG的抗炎活性是通过抑制STAT 3和COX-2途径介导的,并且可以作为潜在的内源性抗氧化剂刺激物。