Skip to content
Licensed Unlicensed Requires Authentication Published by De Gruyter January 22, 2020

Antineuroinflammation activity of n-butanol fraction of Marsilea crenata Presl. in microglia HMC3 cell line

  • Burhan Ma’arif , Denis Mery Mirza , Mu’akibatul Hasanah , Hening Laswati and Mangestuti Agil EMAIL logo



Neuroinflammation is one of the main causes of neurodegenerative events. Phytoestrogen is a group compounds that have an estrogen-like structure or function. Phytoestrogen has a high potential to overcome neuroinflammation caused by estrogen deficiency in postmenopausal women. Marsilea crenata Presl. is a plant known to contain phytoestrogens. This research aimed to analyze the activity of an n-butanol fraction of M. crenata leaves in inhibiting the classical pathway activation of microglia HMC3 cell line to M1 polarity, which has proinflammatory characteristics.


Microglia HMC3 cell line was cultured in Eagle’s minimum essential medium and induced with IFN-γ for 24 h to activate the cell to M1 polarity in 24-well microplates. The n-butanol fraction was added with various doses of 62.5, 125, and 250 ppm and genistein 50 μM as a positive control. The expression of major histocompatibility complex II (MHC II) as a marker was tested using a confocal laser scanning microscope.


The result of MHC II measurement shows a significant difference in the MHC II expression in the microglia HMC3 cell line between the negative control and all treatment groups at p<0.05, indicating a non-monotonic dose-response profile.


The best dosage to inhibit MHC II expression was 250 ppm with the value of 200.983 AU. It is then concluded that n-butanol fraction of M. crenata leaves has antineuroinflammation activity due to its phytoestrogens.

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

  1. Research funding: None declared.

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

  3. Competing interests: Authors state no conflict of interest.


[1] Arcuri C, Mecca C, Bianchi R, Giambanco I, Donato R. The pathophysiological role of microglia in dynamic surveillance, phagocytosis and structural remodeling of the developing CNS. Front Mol Neurosci 2017;10:191.10.3389/fnmol.2017.00191Search in Google Scholar

[2] Matt SM, Johnson RW. Neuro-immune dysfunction during brain aging: new insights in microglial cell regulation. Curr Opin Pharmacol 2015;26:96–101.10.1016/j.coph.2015.10.009Search in Google Scholar

[3] Whitney NP, Eidem TM, Peng H, Huang Y, Zheng JC. Inflammation mediates varying effects in neurogenesis: relevance to the pathogenesis of brain injury and neurodegenerative disorders. J Neurochem 2009;108:1343–59.10.1111/j.1471-4159.2009.05886.xSearch in Google Scholar

[4] Lee WL, Tsui KH, Seow KM, Cheng MH, Su WH, Chen CP, et al. Hormone therapy for postmenopausal women – an unanswered issue. Gynecol Minim Invasive Ther 2013;2:13–7.10.1016/j.gmit.2012.12.003Search in Google Scholar

[5] Beral V. Breast cancer and hormone-replacement therapy in the Million Women Study. Lancet 2003;362:419–27.10.1016/S0140-6736(03)14596-5Search in Google Scholar

[6] Agarwal S, Alzahrani FA, Ahmed A. Hormone replacement therapy: would it be possible to replicate a functional ovary. Int J Mol Sci 2018;19:1–16.10.3390/ijms19103160Search in Google Scholar PubMed PubMed Central

[7] Vinogradova Y, Coupland C, Hippisley-Cox J. Use of hormone replacement therapy and risk of venous thromboembolism: nested case-control studies using the QResearch and CPRD database. Br Med J 2019;364:1–14.10.1136/bmj.k4810Search in Google Scholar PubMed PubMed Central

[8] Sirotkin AV, Harrath AH. Phytoestrogen and their effects. Eur J Pharmacol 2014;741:230–6.10.1016/j.ejphar.2014.07.057Search in Google Scholar PubMed

[9] 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

[10] Laswati H. Green clover potentiates delaying the increment of imbalance bone remodeling process in postmenopausal women. Fol Med Indones 2011;47:112–7.Search in Google Scholar

[11] 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 App Pharm Sci 2018;8:55–9.Search in Google Scholar

[12] Ma’arif B, Mirza DM, Suryadinata A, Muchlisin MA, Agil M. Metabolite profiling of 96% ethanol extract from Marsilea crenata Presl. leaves using UPLC-QToF-MS/MS and anti-neuroinflammatory predicition activity with molecular docking. J Trop Pharm Chem 2019;4:6.10.25026/jtpc.v4i6.213Search in Google Scholar

[13] 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

[14] Tiyaningsih DA. Studi Makroskopis, Mikroskopis dan Skrining Fitokimia Marsilea crenata Presl. Skripsi. Surabaya: Universitas Airlangga, 2007.Search in Google Scholar

[15] Yacoeb AM, Nurjanah, Arifin M, Sulistiono W, Kristiono SS. Deskripsi histologis dan perubahan komposisi kimia daun dan tangkai semanggi (Marsilea crenata Presl., Marsileaceae) akibat perebusan. J Pengolah Has Perikan Indones 2010;12:81–95.Search in Google Scholar

[16] Nurjanah, Azka A, Abdullah A. Aktivitas Antioksidan dan Komponen Bioaktif Semanggi Air (Marsilea crenata). Jurnal Inovasi dan Kewirausahaan 2012;1:152–8.Search in Google Scholar

[17] Ganai AA, Farooqi H. Bioactivity of genistein: a review of in vitro and in vivo studies. Biomed Pharmacother 2015;76:30–8.10.1016/j.biopha.2015.10.026Search in Google Scholar PubMed

[18] Jyote, Agrawal SS, Saxsena S, Sharma A. Phytoestrogen “genistein”: its extraction and isolation from soy bean seeds. Int J Pharmacogn Phytochem Res 2014;7:1112–6.Search in Google Scholar

[19] 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;68:253–62.10.6061/clinics/2013(02)OA21Search in Google Scholar

[20] Cherry J, Olschowka J, O’banion K. Neuroinflammation and M2 microglia: the good, the bad, and the inflamed. J Neuroinflamm 2014;11:98.10.1186/1742-2094-11-98Search in Google Scholar PubMed PubMed Central

[21] Yu J, Bi X, Yu B, Chen D. Review isoflavones: anti-inflammatory benefit and possible caveats. Nutrients 2016;8:1–16.10.3390/nu8060361Search in Google Scholar PubMed PubMed Central

[22] Taylor CR, Rudbeck L. Immunohistochemical staining methods, handbook 6th ed., Dako Denmark: IHC, 2013.Search in Google Scholar

[23] 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 2010;16:515–30.10.1017/S1431927610000371Search in Google Scholar PubMed

[24] 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;157:188–211.10.1016/j.pneurobio.2015.12.008Search in Google Scholar PubMed PubMed Central

[25] Villa A, Vegeto E, Poletti A, Maggi A. Estrogens, neuroinflammation and neurodegeneration. Endocr Rev 2016;37:372–402.10.1210/er.2016-1007Search in Google Scholar PubMed PubMed Central

[26] Tang Y, Le W. Differential roles of M1 and M2 microglia in neurodegenerative diseases. New York: Springer Science and Business Media, 2015.10.1007/s12035-014-9070-5Search in Google Scholar PubMed

[27] Shih R, Wang C-Y, Yang C-M. NF-kB signaling pathways in neurological inflammation: a mini review. Front Mol Neurosci 2015;8:77.10.3389/fnmol.2015.00077Search in Google Scholar PubMed PubMed Central

[28] Au A, Feher A, McPhee L, Jessa A, Oh S, Einstein G. Estrogens, inflammation and cognition. Front Neuroendocrinol 2016;40:87–100.10.1016/j.yfrne.2016.01.002Search in Google Scholar PubMed

[29] 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;33:378–455.10.1210/er.2011-1050Search in Google Scholar PubMed PubMed Central

[30] Cui J, Shen Y, Li R. Estrogen synthesis and signaling pathways during aging: from periphery to brain. Trends Mol Med 2013;19:197–209.10.1016/j.molmed.2012.12.007Search in Google Scholar PubMed PubMed Central

[31] Kalaitzidis D, Gilmore TD. Review: transcription factor cross-talk: the estrogen receptor and NF-kB. Trends Endocrin Metab 2005;16:46–52.10.1016/j.tem.2005.01.004Search in Google Scholar PubMed

Received: 2019-09-06
Accepted: 2019-10-06
Published Online: 2020-01-22

© 2019 Walter de Gruyter GmbH, Berlin/Boston

Downloaded on 27.2.2024 from
Scroll to top button