Department of Pharmacy, Jashore University of Science and Technology, Jessore, 7408, Jashore, Bangladesh; School of Biomedical Science, Charles Sturt University, Booroma St, Wagga Wagga, NSW, Australia.
Department of Pharmacy, Faculty of Basic Medicine and Health Sciences, University of Science and Technology Chittagong, Foy's Lake, Chittagong, Bangladesh.
J Ethnopharmacol. 2021 Jun 12;273:113975. doi: 10.1016/j.jep.2021.113975. Epub 2021 Feb 27.
Functional disability associated with rheumatoid arthritis (RA), a chronic inflammatory autoimmune disease is a challenging concern in healthcare systems. Along with environmental factors and epigenetic disorders, multiple pathways are reported as prominent mechanism for the progression of RA symptoms including; pain, swelling and stiffness of joints. Elaeocarpus floribundus Blume has been used as a folklore medicine for RA from ancient times. This plant harbours a suite of endophytic fungi that produce a range of metabolites of potential interest. Thus, for the establishment of a scientific basis for this folklore use, it is essential to find out the involvement, if any, of the endophytic fungi living in this plant and the metabolites they elaborate, for the management of RA.
This study was designed to isolate, identify and evaluate the in vitro anti-inflammatory and in vivo antinociceptive and antiarthritic activities of the compounds produced by the endophytic fungi living in different parts of Elaeocarpus floribundus Blume.
Endophytic fungi from different parts of the plant were isolated and cultured for the production of secondary metabolites. Chromatographically fractionated fungal extracts were assessed for anti-inflammatory and antinociceptive activities. For the evaluation of anti-inflammatory activity, in vitro cyclooxygenase (COX1/COX2) and 5-lipoxygenase (5-LOX) inhibitory assays were performed. For the evaluation of in vivo antinociceptive activity, hot plate acetic acid induced writhing, and formalin induced paw licking methods were adopted, whereas complete Freund's adjuvant (CFA) induced poly-arthritic method was adopted for the evaluation of antiarthritic activity. The most effective fraction was analyzed by liquid chromatography-mass spectroscopy (LC-MS) in search of the bioactive extracellular metabolites.
Five endophytic fungi viz. Aspergillus fumigatus, Aspergillus niger, Rhizoctonia oryzae, Rhizopus oryzae, and Syncephalastrum racemosum were isolated. COX1/COX2 and 5-LOX inhibitory assays state that the Aspergillus niger fraction possesses the greatest activity against these enzymes of inflammatory process. In vivo antinociceptive showed significant (***P<0.001) reduction of pain in a dose dependent manner. As well, significant (***P<0.001) reduction of paw volume was observed in CFA induce poly-arthritic test. LC/MS analysis of the Aspergillus niger fraction revealed the presence of bioactive compounds including tensyuic acid, hexylitaconic acid, chlorogenic acid, nigragillin, TMC-256C1, asnipyrone B, asperenone, fumaric acid and fusarubin, all having reported pharmacological activities.
The present study demonstrates that secondary metabolites produced by endophytic fungi living in various parts of Elaeocarpus floribundus Blume had potential to relief pain and inflammation. The endophytes were found to contain multiple biomolecules effective in rheumatoid arthritis. These findings provide a rationale for the folklore use of the plant in the management of rheumatoid arthritis.
类风湿关节炎（RA）是一种慢性炎症性自身免疫性疾病，与功能障碍相关，这是医疗保健系统面临的一个挑战。除了环境因素和表观遗传紊乱外，多种途径被报道为 RA 症状进展的主要机制，包括：疼痛、关节肿胀和僵硬。从古代开始，海红豆（Elaeocarpus floribundus Blume）就被用作治疗 RA 的民间药物。这种植物蕴藏着一套内生真菌，它们产生一系列具有潜在价值的代谢物。因此，为了为这种民间用途建立科学基础，有必要了解生活在这种植物中的内生真菌及其产生的代谢物，如果有的话，是否参与了 RA 的管理。
本研究旨在分离、鉴定和评估生活在海红豆不同部位的内生真菌产生的化合物的体外抗炎和体内镇痛及抗关节炎活性。
从植物的不同部位分离内生真菌并进行培养以生产次生代谢物。对色谱分离的真菌提取物进行抗炎和镇痛活性评估。为了评估抗炎活性，进行了体外环氧化酶（COX1/COX2）和 5-脂氧合酶（5-LOX）抑制试验。为了评估体内镇痛活性，采用了热板乙酸诱导扭体和甲醛诱导舔爪方法，而采用完全弗氏佐剂（CFA）诱导多关节炎方法评估抗关节炎活性。采用液相色谱-质谱联用（LC-MS）分析最有效的馏分，寻找具有生物活性的细胞外代谢物。
共分离出 5 株内生真菌，分别为烟曲霉（Aspergillus fumigatus）、黑曲霉（Aspergillus niger）、稻瘟病菌（Rhizoctonia oryzae）、根霉（Rhizopus oryzae）和密粘褶菌（Syncephalastrum racemosum）。COX1/COX2 和 5-LOX 抑制试验表明，黑曲霉（Aspergillus niger）部分对这些炎症过程中的酶具有最大的活性。体内镇痛试验表明，疼痛呈剂量依赖性显著（***P<0.001）减轻。在 CFA 诱导的多关节炎试验中，也观察到爪体积的显著（***P<0.001）减少。黑曲霉（Aspergillus niger）部分的 LC/MS 分析显示存在生物活性化合物，包括 Tensyuic 酸、己基衣康酸、绿原酸、Nigragillin、TMC-256C1、Asnipyrone B、Asperenone、富马酸和 Fusarubin，它们都具有报道的药理活性。
本研究表明，生活在海红豆不同部位的内生真菌产生的次生代谢物具有缓解疼痛和炎症的潜力。内生真菌中含有多种对类风湿关节炎有效的生物分子。这些发现为该植物在类风湿关节炎治疗中的民间用途提供了依据。
