竹叶椒抗炎内生真菌的分离

doi:10.5246/jcps.2021.07.045

中国药学(英文版) ›› 2021, Vol. 30 ›› Issue (7): 570-577.DOI: 10.5246/jcps.2021.07.045

竹叶椒抗炎内生真菌的分离

赵爱红¹^,^#, 王天霞¹^,^#, 刘晓艳², 赵伟红¹, 马建苹¹, 杨秀伟², 郭涛³^,^*(), 张友波²^,^*()

1. 兰州理工大学甘肃省高校中藏药筛选评价及深加工省级重点实验室, 甘肃兰州 730050
2. 北京大学医学部药学院天然药物学系, 天然药物及仿生药物国家重点实验室, 北京 100191
3. 河南中医药大学药学院, 河南郑州 450046

收稿日期:2020-12-15 修回日期:2021-02-10 接受日期:2021-02-16 出版日期:2021-07-27 发布日期:2021-07-27
通讯作者: 郭涛, 张友波
作者简介:
+ Tel.: +86-10-82801569, E-mail: ybzhang@bjmu.edu.cn
+ E-mail: gt010010@163.com
基金资助:
The Key Laboratory of Herbal-Tebitan Drug Screening and Deep Processing of Gansu Province (Grant No. 20180807) and Undergraduate innovation and entrepreneurship training program of Lanzhou University of Technology (Grant No. DC2020249).

Isolation of endophytic fungi with anti-inflammatory effect in vitro from Zanthoxylum armatum

Aihong Zhao¹^,^#, Tianxia Wang¹^,^#, Xiaoyan Liu², Weihong Zhao¹, Jianping Ma¹, Xiuwei Yang², Tao Guo³^,^*(), Youbo Zhang²^,^*()

1 Key Laboratory of Herbal-Tebitan Drug Screening and Deep Processing of Gansu Province, Lanzhou University of Technology, Lanzhou 730050, China
2 State Key Laboratory of Natural and Biomimetic Drugs and Department of Natural Medicines, School of Pharmaceutical Sciences, Peking University, Beijing 100191, China
3 School of Pharmacy, Henan University of Chinese Medicine, Zhengzhou 450046, China

Received:2020-12-15 Revised:2021-02-10 Accepted:2021-02-16 Online:2021-07-27 Published:2021-07-27
Contact: Tao Guo, Youbo Zhang
About author:
# These authors contributed equally to this work.

摘要/Abstract

摘要：

内生真菌富含具有多种生物活性的次生代谢产物。本研究从竹叶椒中分离鉴定了8种内生真菌, 并通过体外实验对其抗菌活性进行了筛选。结果发现4种内生真菌提取物(Fusarium oxysporum Schlecht. (Z-1)、Alternaria arborescens E.G Simmons (Z-3)、Mucor fragilis Bainier (Z-4)和Aspergillus terreus Thom (Z-6))对LPS诱导的RAW264.7细胞一氧化氮过量生成具有显著抑制作用。其中, 真菌 Fusarium oxysporum Schlecht提取物具有最好的抗炎活性, 其IC₅₀值为174.0 μg/mL。真菌 Fusarium oxysporum Schlecht的抗炎活性与其抑制多种细胞炎症因子的释放有关。实时荧光定量PCR结果显示, 真菌Fusarium oxysporum Schlecht对LPS诱导的RAW264.7细胞中Il-1β、Il-6、iNOS等炎症因子的mRNA表达具有显著抑制作用。总之, 竹叶椒中多种内生真菌具有抗炎活性, 其中Fusarium oxysporum Schlecht有望发展成为新的抗炎活性药物。

关键词: 内生真菌, 竹叶椒, 抗炎, 实时荧光定量聚合酶链反应

Abstract:

Endophytic fungus is a rich source of bioactive metabolites with multiple activities. In the present study, eight endophytic fungal strains (Z-1 to Z-8) from Zanthoxylum armatum DC. were isolated and identified, and the anti-inflammatory effect of their extracts was evaluated in vitro. The results showed that four endophytic fungal extracts (Fusarium oxysporum Schlecht. (Z-1), Alternaria arborescens E.G Simmons (Z-3), Mucor fragilis Bainier (Z-4) and Aspergillus terreus Thom (Z-6)) displayed good activities in inhibition of excessive production of nitric oxide in lipopolysaccharide-activated mouse macrophage RAW264.7 cells. Among them, the extracts of Fusarium oxysporum Schlecht showed the maximum inhibition activity with a half maximal inhibitory concentration of 174.0 μg/mL. The underlying mechanism of the anti-inflammatory effect of Fusarium oxysporum Schlecht extract was associated with its inhibition on the release of several inflammatory factors. The mRNA expression of interleukin 1β, interleukin 6, inducible nitric oxide synthase in lipopolysaccharide-activated mouse macrophage RAW264.7 cells were all inhibited by Z-1 extract detected by real-time polymerase chain reaction. In summary, several endophytic fungal from Zanthoxylum armatum have good anti-inflammatory activity and Fusarium oxysporum Schlecht possesses a promising potential to be developed to a novel anti-inflammatory agent.

Key words: Endophytic fungi, Zanthoxylum armatum, Anti-inflammation, Real-time polymerase chain reaction

Supporting:

赵爱红, 王天霞, 刘晓艳, 赵伟红, 马建苹, 杨秀伟, 郭涛, 张友波. 竹叶椒抗炎内生真菌的分离[J]. 中国药学(英文版), 2021, 30(7): 570-577.

Aihong Zhao, Tianxia Wang, Xiaoyan Liu, Weihong Zhao, Jianping Ma, Xiuwei Yang, Tao Guo, Youbo Zhang. Isolation of endophytic fungi with anti-inflammatory effect in vitro from Zanthoxylum armatum[J]. Journal of Chinese Pharmaceutical Sciences, 2021, 30(7): 570-577.

图/表 5

Table 1. Sequences of the primers.

Figure 1. Basis of identification of endophytic fungi from Zanthoxylum armatum with ITS sequence.

Table 2. IC50 of the endophytic fungi on the production of NO in LPS-stimulated RAW264.7 cells.

Figure 2. IC50 of the endophytic fungi from Zanthoxylum armatum on NO excessive production in RAW264.7 cells. (A) Z-1; (B) Z-3; (C) Z-4; (D) Z-6.

Figure 3. Regulation of Z-1 on the mRNA expression of inflammatory cytokines in RAW264.7 cells. (A) mRNA expression of Il-1β; (B) mRNA expression of Il-6; (C) mRNA expression of Cox-2; (D) mRNA expression of iNos. *P < 0.05, **P < 0.01, ***P < 0.001. Significance was determined by using one-way ANOVA. Data are presented as the mean ± SEM, N = 3 per group.

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竹叶椒抗炎内生真菌的分离

Isolation of endophytic fungi with anti-inflammatory effect in vitro from Zanthoxylum armatum

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