奇楠沉香精油挥发性成分分析及其体外抗炎活性研究

Analysis of Volatile Components of Essential Oil of Qi-Nan Agarwood and Its Anti-inflammatory Activity in vitro

本研究基于挥发性成分分析结合细胞炎症模型探究奇楠沉香精油(Qi-Nanessentialoil,QEO)对RAW264.7细胞抗炎活性的影响。采用GC-MS方法,分析QEO挥发性成分;通过DPPH自由基清除率和总抗氧化能力测定,检测QEO的抗氧化活性;利用生物信息学预测QEO与炎症相关的靶点和信号通路;应用荧光探针法检测一氧化氮(NO)和活性氧(ROS)水平;应用ELISA法检测TNF-α、IL-6和IL-1β的含量;应用Western blot检测P-P65、P65、IκB-α和P-IκB-α的表达水平。结果表明:QEO主要挥发性成分为2-(2-苯乙基)-色酮(50.08%)、石竹素(8.45%)、绿叶烷(6.46%)、榄香醇(3.88%)、2-烯丙基-6-甲基苯酚(2.78%)。QEO在浓度为1 mg/mL时对DPPH自由基的清除率为51.98%,其总抗氧化能力在1~10mg/mL浓度范围内随着浓度增大而增大。QEO与炎症的交集靶点有533个,KEGG富集分析显示主要涉及MAPK、NF-κB和PI3K-Akt等信号通路。QEO显著降低NO、ROS、TNF-α、IL-6和IL-1β的产生和表达水平(P<0.001);同时,QEO显著抑制P-P65和P-IκB-α蛋白表达(P<0.001)。以上研究结果表明,QEO可通过减少ROS的产生,抑制NF-κB信号通路的激活,从而显著减少细胞炎症因子的释放,以达到抗炎作用,具有开发成炎症抑制剂的潜力。

The aim of the study was to explore the anti-inflammatory activity of Qi-Nan essential oil(QEO)based on component analysis combined with the cellular inflammation model.The volatile components of QEO were analyzed by GC-MS.The antioxidant activity of QEO was detected by the measurement of DPPH radical scavenging and total antioxidant capacity.Bioinformatics was used to predict the targets and signaling pathways of QEO related to inflammation.NO and ROS levels were detected by fluorescent probe method.ELISA was used to detect the levels of inflammatory factors TNF-α,IL-6 and IL-1β.Western blot was used to detect the expressions of P-P65,P65,IκB-αand P-IκB-α.The main components of Qinan essential oil were 2-phenethyl-4H-chromen-4-one(50.08%),(-)-caryophyllene oxide(8.45%),patchulane(6.46%),elemol(3.88%),2-methyl-6-(2-propenyl)-phenol(2.78%).The DPPH radicals scavenging of QEO was 51.98%at a concentration of 1 mg/mL and the total antioxidant capacity increased with the increase of QEO in the concentration range of 1-10 mg/mL.There were 533 intersecting targets of QEO and inflammation,and KEGG enrichment analysis showed that the main signaling pathways involved were MAPK,NF-κB and PI3K-Akt pathways.QEO significantly reduced the production and expression levels of NO,ROS,TNF-α,IL-6 and IL-1β(P<0.001).Meanwhile,the results of western blot showed that QEO also inhibited the expression of P-P65 and P-IκB-αproteins(P<0.001).Therefore,QEO can achieve anti-inflammatory activity through reducing the production of ROS,thereby inhibiting the activation of NF-κB signaling pathway and further drastically decrease the release of pro-inflammatory cytokines.The results demonstrate that QEO has the potential to be developed as an inflammation inhibitor.