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

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-γfor 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.