[關鍵詞]
[摘要]
目的 研究黃河三角洲貝殼堤島葉底珠內生真菌代謝產物醋酸乙酯粗提物的體外抗炎鎮(zhèn)痛和抗氧化活性,篩選鑒定具有活性的內生真菌。方法 以黃河三角洲貝殼堤島葉底珠為實驗材料分離內生真菌,通過1,1-二苯基-2-三硝基苯肼(DPPH)法、二甲苯致小鼠耳廓腫脹法以及醋酸扭體法測定其代謝產物醋酸乙酯粗提物的抗氧化活性、抗炎作用以及鎮(zhèn)痛作用。通過雷氏鹽剩余比色法、Folin-Ciocalteu法、亞硝酸鈉-硝酸鋁比色法檢測活性菌株的總生物堿、總酚、總黃酮的含量,SPSS 16.0軟件分析3種生物活性與3種成分含有量的相關性?;诤颂求wRNA基因(即rDNA)內轉錄間隔區(qū)(Internal Transcribed Spacer,ITS)的多態(tài)性的序列分析,通過分子生物學方法對篩選出的活性菌株進行鑒定。結果 共分離得到內生真菌59株,其中8株表現(xiàn)出不同的抗氧化活性,10株對小鼠耳腫脹有不同程度的抑制作用,8株對醋酸誘導的疼痛有不同程度的抑制作用;總酚含量和抗氧化作用、總生物堿含量和抗炎活性間具有顯著相關性;初步鑒定生物活性較高的8株內生真菌菌株為Chaetomium globosum、Chaetomium sp.、Fusarium sp.、Fusarium solani、Cladosporium ramotenellum、Leptosphaeria sp.、Phoma multirostrata和Phoma sp.。結論 黃河三角洲貝殼堤島葉底珠內生真菌具有良好的體外抗炎鎮(zhèn)痛和抗氧化活性,總酚和生物堿含量是影響生物活性的主要因素。
[Key word]
[Abstract]
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 anti-inflammatory 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.
[中圖分類號]
R965.1
[基金項目]
國家級大學生創(chuàng)新創(chuàng)業(yè)訓練計劃項目(201910449012、201710449009);山東省自然科學基金項目(ZR2016CL01、ZR2016BL16);濱州學院博士基金(2016Y02、2016Y17)