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The enhancement of Arg1 and activated ERβ expression in microglia HMC3 by induction of 96% ethanol extract of Marsilea crenata Presl. leaves

Burhan Ma’arif, Mangestuti Agil and Hening Laswati

Abstract

Background

Phytoestrogens have a high potential to overcome the neuroinflammation caused by estrogen deficiency. Marsilea crenata Presl. is a plant known to contain phytoestrogens. This research aimed to report the activity of a 96% ethanol extract of M. crenata leaves in inducing activation of microglia HMC3 cell to M2 polarity, which has anti-inflammatory characteristics.

Methods

The study was done by culturing microglia HMC3 cell in 24-well microplate and inducing it with IFN-γ for 24 h to activate the cell to M1 polarity, which has proinflammatory characteristics. The 96% ethanol extract was added with various doses of 62.5, 125, and 250 ppm. Genistein, 50 μM, was used as a positive control. The analysis of the immunofluorescence of Arginase-1 (Arg1) and ERβ as markers was done using a convocal laser scanning microscope.

Results

The result of Arg1 shows a significant difference in Arg1 expression in the microglia HMC3 cell line between the negative control and all treatment groups at p < 0.05, with the best result at 250 ppm, whereas for ERβ, the results show, at doses of 125 and 250 ppm, that the 96% ethanol extract of M. crenata leaves decrease the activated ERβ expression at p < 0.05, with the best result at 250 ppm. The Arg1 and activated ERβ expression have a weak negative relationship with the Pearson correlation test.

Conclusions

The 96% ethanol extract of M. crenata leaves has an antineuroinflammation activity through the induction of Arg1 and activated ERβ expression in microglia HMC3 cell, with the best dose at 250 ppm.

Acknowledgments

This research is supported by the Research and Community Service Funds, Ministry of Research, Technology and Higher Education Republic of Indonesia 2019.

  1. Author contributions: All authors have accepted responsibility for the entire content of this manuscript and approved its submission.

  2. Research funding: None declared.

  3. Competing interests: The authors declare there is no conflict of interest in this research.

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Received: 2019-09-27
Accepted: 2019-10-21
Published Online: 2020-01-25

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