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.
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.
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.
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.
This research is supported by the Research and Community Service Funds, Ministry of Research, Technology and Higher Education Republic of Indonesia 2019.
Author contributions: All authors have accepted responsibility for the entire content of this manuscript and approved its submission.
Research funding: None declared.
Competing interests: The authors declare there is no conflict of interest in this research.
 Nurjanah AA, Abdullah A. Aktivitas antioksidan dan komponen bioaktif semanggi air (Marsilea crenata). Jurnal Inovasi dan Kewirausahaan 2012;1:152–8.Search in Google Scholar
 Ma’arif B, Agil M, Laswati H. Phytochemical assessment on n-hexane extract and fractions of Marsilea crenata Presl. leaves through GC-MS. Trad Med J 2016;21:77–85.Search in Google Scholar
 Laswati H. Green clover potentiates delaying the increment of imbalance bone remodeling process in postmenopausal women. Folia Medica Indonesiana 2011;47:112–7.Search in Google Scholar
 Agil M, Ma’arif B, Aemi NY. Antiosteoporotic activity of n-hexane fraction from Marsilea crenata Presl. leaves in increasing trabecular vertebrae bone density of female mice. Jurnal Tumbuhan Obat Indonesia 2018;11:1–7.Search in Google Scholar
 Ma’arif B, Agil M, Laswati H. Alkaline phosphatase activity of Marsilea crenata Presl. extract and fractions as marker of MC3T3-E1 osteoblast cell differentiation. J Appl Pharm Sci 2018;8:55–9.Search in Google Scholar
 Ososki AL, Kennelly EJ. Phytoestrogens: a review of the present state of research. Phytother Res 2003;17:845–69.Search in Google Scholar
 Yang TS, Wang SY, Yang YC, Su CH, Lee FK, Chen SC, et al. Effects of standardized phytoestrogen on Taiwanese menopausal women. Taiwan J Obstet Gynecol 2012;51:229–35.Search in Google Scholar
 Roy S, Awasthi H. Herbal Medicines as neuroprotective agent: a mechanistic approach. Int J Pharm Pharm Sci 2017;9:1–7.Search in Google Scholar
 Cui J, Shen Y, Li R. A review: estrogen synthesis and signaling pathways during aging: from periphery to brain. Trends Mol Med 2013;19:197–209.Search in Google Scholar
 Au A, Feher A, McPhee L, Jessa A, Oh S, Einstein G. Estrogens, inflammation and cognition. Front Neuroendocrinol 2016;40:87–100.Search in Google Scholar
 Engler-Chiurazzi EB, Brown CM, Povroznik JM, Simpkins JW. Estrogens as neuroprotectants: estrogenic actions in the context of cognitive aging and brain injury. Prog Neurobiol 2017;157:188–211.Search in Google Scholar
 Pfeffer LM. The role of nuclear factor Kb in the interferon response. J Interferon Cytokine Res 2011;31:553–9.Search in Google Scholar
 Vagaska B, New SE, Alvarez-Gonzalez C, D’Acquisto FS, Gomez G, Bulstrode NW, et al. MHC-class-II are expressed in a subpopulation of human neural stem cells in vitro in an IFNγ-independent fashion and during development. Sci Rep 2016;6:242–51.Search in Google Scholar
 Chen L, Lei L, Ding P, Tang Q, Wu Y. Osteogenic effect of Drynariae rhizoma extracts and Naringin on MC3T3-E1 cells and an induced rat alveolar bone resorption model. Arch Oral Biol 2011;56:1655–62.Search in Google Scholar
 Yu J, Bi X, Yu B, Chen D. Review isoflavones: anti-inflammatory benefit and possible caveats. Nutrients 2016;8:1–16.Search in Google Scholar
 Tafzi F, Andarwulan N, Giriwonob PE, Dewid FN. Uji Efikasi Ekstrak Metanol Daun Torbangun (Plectranthus amboinicus) pada Sel Epitel Kelenjar Susu Manusia MCF-12A. Jurnal Ilmu Kefarmasian Indonesia 2017;15:17–24.Search in Google Scholar
 Taylor CR, Rudbeck L. Immunohistochemical staining methods, Handbook 6th ed. Dako Denmark: IHC, 2013.Search in Google Scholar
 Stanciu SG, Hristu R, Boriga R, Stanciu GA. On the suitability of SIFT technique to deal with image modifications specific to confocal scanning laser microscopy. Microsc Microanal J 2010;6:55–530.Search in Google Scholar
 Cherry J, Olschowka J, O’banion K. Neuroinflammation and M2 microglia: the good, the bad, and the inflamed. J Neuroinflamm 2014;11:98.Search in Google Scholar
 Cioffi M, Esposito K, Vietri MT, Gazzerro P, D’Auria A, Ardovino I, et al. Cytokine pattern in postmenopause. Maturitas 2002;41:187–92.Search in Google Scholar
 Abu-Taha M, Rius C, Hermenegildo C, Noguera I, Cerda-Nicolas JM, Issekutz AC, et al. Menopause and ovariectomy cause a low grade of systemic inflammation that may be prevented by chronic treatment with low doses of estrogen or losartan. J Immunol 2009;183:1393–402.Search in Google Scholar
 Mizuno T. Neuron-microglia interaction in neuroinflammation. Clin Exp Neuroimmunol 2015;6:225–31.Search in Google Scholar
 Shih R, Wang C-Y, Yang C-M. NF-kB signaling pathways in neurological inflammation: a mini review. Front Mol Neurosci 2015;8:77.Search in Google Scholar
 Straub RH. The complex role of estrogens in inflammation. Endocr Rev 2007;28:521–74.Search in Google Scholar
 Ganai AA, Farooqi H. Bioactivity of genistein: a review of in vitro and in vivo studies. Biomed Pharmacother 2015;76:30–8.Search in Google Scholar
 Jyote, Agrawal SS, Saxsena S, Sharma A. Phytoestrogen “Genistein”: it’s extraction and isolation from soy bean seeds. Int J Pharmacogn Phytochem Res 2014;7:1121–6.Search in Google Scholar
 Rosmani S, Omar SZ, Ali , Kan NL, Musameh NI, Das S, Kasim NM. Effect of phytoestrogen genistein on the development of the reproductive system of Sprague Dawley rats. Clinics 2012;68:253–62.Search in Google Scholar
 Yang Z, Ming XF. Functions of arginase isoforms in macrophage inflammatory responses: impact on cardiovascular diseases and metabolic disorders. Front Immunol 2014;5:533.Search in Google Scholar
 Tada H, Amino N. Anti-receptor antibody and autoimmune disease. Rinsho Byori 1998;46:311–6.Search in Google Scholar
 Vandenberg LN, Colborn T, Hayes TB, Heindel JJ, Jacobs Jr. DR, Lee DH, et al. Hormones and endocrine-disrupting chemicals: low-dose effects and nonmonotonic dose responses. Endocr Rev 2012;33:378–455.Search in Google Scholar
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