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Antioxidative properties of 4-methylumbelliferone are related to antibacterial activity in the silkworm (Bombyx mori) digestive tract

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Abstract

Umbelliferones have gained significant attention due to their tumor-inhibitory effects in vitro. This study was undertaken to examine the impact of umbelliferones in an invertebrate model organism, Bombyx mori, to assess the underlying antimicrobial activities via antioxidation in vivo. Oral administration of 4 mM 4-methylumbelliferone (4-MU), a model umbelliferone drug, in B. Mori larvae caused a rapid increase in reactive oxygen species, such as hydrogen peroxide (H2O2) and antimicrobial activity in the digestive tract. In addition, a significant increase in total antioxidant capacity as well as superoxide anion radical-inhibiting activity and reduced glutathione were detected. The antioxidant defense system was activated following induction of H2O2, resulting in a significant rise in catalase (50–66 %) and glutathione peroxidase (175 %) activities, which were helpful in defending digestive tract cells against oxidative injury. These results help in understanding the anticancer mechanism of 4-MU based on its antioxidation in organisms.

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Abbreviations

4-MU:

4-Methylumbelliferone

ROS:

Reactive oxygen species

T-AOC:

Total antioxidant capacity

CAT:

Catalase

SOD:

Superoxide dismutase

GR:

Glutathione reductase

GPx:

Glutathione peroxidase

GCL:

γ-Glutamate–cysteine ligase

GSH:

Reduced glutathione

GSSG:

Oxidized glutathione

NOX-1:

NADH oxidase

DHE:

Dihydroethidium

DCFH-DA:

2′,7′-Dichlorofluorescin diacetate

MAPK:

Mitogen-activated protein kinases

ERK:

Extracellular signal-related kinase

MEK:

MAPK–ERK kinase

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Acknowledgments

We wish to acknowledge Associate Professor Linchuan Zhao for his help and advice during our research. The present work was supported by the National High-Tech R&D Program of China (863 Program) (Grant No. 2011AA100306), National Natural Science Foundation of China (Grant No. 31172264), Provincial Key Technology R&D Program of Jiangsu (Project No. BE2011327-1), and Priority Academic Program Development of Jiangsu Higher Education Institutions.

Conflict of interest

The authors declare no conflict of interest.

Author information

Authors and Affiliations

  1. Department of Applied Biology, School of Biology and Basic Medical Sciences, Medical College, Soochow University, Suzhou, 215123, China

    Yan Fang, Hua Wang, Wenjuan Zhu, Lu Wang, Hengjiang Liu, Xu Xu, Weimin Yin, Yanghu Sima & Shiqing Xu

  2. National Engineering Laboratory for Modern Silk, Institute of Agricultural Biotechnology and Ecology, Soochow University, Suzhou, 215123, China

    Yan Fang, Hua Wang, Wenjuan Zhu, Lu Wang, Hengjiang Liu, Xu Xu, Weimin Yin, Yanghu Sima & Shiqing Xu

Authors
  1. Yan Fang
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  2. Hua Wang
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  3. Wenjuan Zhu
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  4. Lu Wang
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  5. Hengjiang Liu
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  6. Xu Xu
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  7. Weimin Yin
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  8. Yanghu Sima
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  9. Shiqing Xu
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Corresponding authors

Correspondence to Yanghu Sima or Shiqing Xu.

Additional information

Communicated by I. D. Hume.

Y. Fang and H. Wang are co-first authors, who contributed equally to this work.

Electronic supplementary material

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360_2014_840_MOESM1_ESM.tif

FigureS1. Diagram of the study timeline.(A)The process of preparing the tissues. The digestive tract in the picture is a whole gut. We isolated the extro-partment of the digestive tract for experiments.(B)Negative: Negative control of no injection. Buffer: Dorsal injection with 50μlof 10 % DMSO. 4-MU: Oral injection with 50μlof 4 mM 4-MU dissolved in 10 %DMSO. (C) High-performance liquid chromatography measurements of the digestive tract 8 min after an oral injection of 4-MU. (a) Direct loading of 4-MU. (b) Buffer, (c) 4-MU and (d) Mobile phase proportion in gradient elution. The column was Sepax HP-C18(4.6 × 150 mm, 5 μm) at room temperature. Sample size was 20 μl. Eluent flow was 1.0 ml/min. Determination was A320 nm. (TIFF 10191 kb)

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Fang, Y., Wang, H., Zhu, W. et al. Antioxidative properties of 4-methylumbelliferone are related to antibacterial activity in the silkworm (Bombyx mori) digestive tract. J Comp Physiol B 184, 699–708 (2014). https://doi.org/10.1007/s00360-014-0840-1

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  • DOI: https://doi.org/10.1007/s00360-014-0840-1

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