叶底珠内生真菌提取物的抗炎镇痛和抗氧化活性研究

Study on anti-inflammatory,analgesic and antioxidative activities of extracts from Securinega suffruticosa

目的研究黄河三角洲贝壳堤岛叶底珠内生真菌代谢产物醋酸乙酯粗提物的体外抗炎镇痛和抗氧化活性,筛选鉴定具有活性的内生真菌。方法以黄河三角洲贝壳堤岛叶底珠为实验材料分离内生真菌,通过1,1-二苯基-2-三硝基苯肼(DPPH)法、二甲苯致小鼠耳廓肿胀法以及醋酸扭体法测定其代谢产物醋酸乙酯粗提物的抗氧化活性、抗炎作用以及镇痛作用。通过雷氏盐剩余比色法、Folin-Ciocalteu法、亚硝酸钠-硝酸铝比色法检测活性菌株的总生物碱、总酚、总黄酮的含量,SPSS 16.0软件分析3种生物活性与3种成分含有量的相关性。基于核糖体RNA基因(即rDNA)内转录间隔区(Internal Transcribed Spacer,ITS)的多态性的序列分析,通过分子生物学方法对筛选出的活性菌株进行鉴定。结果共分离得到内生真菌59株,其中8株表现出不同的抗氧化活性,10株对小鼠耳肿胀有不同程度的抑制作用,8株对醋酸诱导的疼痛有不同程度的抑制作用;总酚含量和抗氧化作用、总生物碱含量和抗炎活性间具有显著相关性;初步鉴定生物活性较高的8株内生真菌菌株为Chaetomium globosum、Chaetomium sp.、Fusarium sp.、Fusarium solani、Cladosporium ramotenellum、Leptosphaeria sp.、Phoma multirostrata和Phoma sp.。结论黄河三角洲贝壳堤岛叶底珠内生真菌具有良好的体外抗炎镇痛和抗氧化活性,总酚和生物碱含量是影响生物活性的主要因素。

Objective In order to study the in vitro anti-inflammatory,analgesic and antioxidant activity of crude extracts of endophytic fungi metabolites of the Securinega suffruticosa(Pall.)Rehd from seashell islands of Yellow River Delta,and to screen and identify bioactive endophytic fungi.Methods The endophytic fungi were isolated by using the S.suffruticosa from seashell islands of Yellow River Delta.The antioxidant activity,anti-inflammatory activity and analgesic activity of the crude metabolite extracts were determined by DPPH,xylene-induced mouse ear swelling method and acetic acid writhing test.The active components of the active strain were detected.SPSS16.0 software was used to analyze the correlation between the three biological activities and the content of the three components.The selected active strains were identified by molecular biology methods.Results A total of 59 endophytic fungi strains were isolated.Among them,eight strains showed different antioxidant activities,ten strains showed different inhibitory effects on ear swelling of mice and eight strains showed different inhibitory effects on pain induced by acetic acid.There was significant correlation between total phenol content and antioxidant activity,total alkaloid content and antiinflammatory activity.Eight endophytic fungal strains with high bioactivity were identified as Chaetomium globosum,Chaetomium sp.,Fusarium sp.,Fusarium solani,Cladosporium ramotenellum,Leptosphaeria sp.,Phoma multirostrata,and Phoma sp..Conclusion The endophytic fungi of the S.suffruticosa from seashell islands of Yellow River Delta have good anti-inflammatory,analgesic and antioxidant activities in vitro.The total phenol content and alkaloid content are the main factors that affect the biological activity.