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

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 ERß 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 ERß, the results show, at doses of 125 and 250 ppm, that the 96% ethanol extract of M. crenata leaves decrease the activated ERß expression at p < 0.05, with the best result at 250 ppm. The Arg1 and activated ERß 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 ERß expression in microglia HMC3 cell, with the best dose at 250 ppm.

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

Background 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. Methods 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. Results 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. Conclusions 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.

1-triacontanol cerotate isolated from Marsilea quadrifolia Linn. safeguards hippocampal CA3 neurons and augments special memory deficit in chronic epileptic rats / Cerotato de 1-triacontanol aislado de Marsilea quadrifolia Linn. protege las neuronas CA3 del hipocampo y mejora el déficit de memoria especial en ratas epilépticas crónicas

SUMMARY: Currently many people with epilepsy do not have seizure control even with the best available medications. Moreover various antiepileptics have adverse cognitive impact with other side effect. Thus, need for new antiepileptic drugs still remains challenge. However, many of the natural components have antiepileptic action and this fact remains scientifically unexplored. This study was designed to check the behavioral and neuro-pathological outcome of 1-Triacontanol cerotate (1TAC), isolated from Marsilea quadrifolia Linn. (MQ) on chronic Pentylenetetrazol (PTZ) kindling model of epilepsy in rats. Two-month-old adult male Wistar rats (n=60) were randomly divided into six groups; Group I (Cage Control), II (Vehicle Control), III (Positive Control), IV (Standard drug treated), V (1TAC: 40 mg/kg) & VI (1TAC: 80 mg/kg). To induce kindling a 35 mg/kg dose of PTZ was injected i.p. in every 48 hrs for 30 days in Group III to VI. Spatial memory performance was tested using Morris water maze, following which brains were further processed for histopathological investigations. Interestingly, 1TAC was able to minimize the loss of pyramidal cells in hippocampal CA3 region. These cellular changes were behaviorally responded as improved special learning and memory, a better spatial navigation and object place configuration. The current study strongly implicates that 1TAC from MQ has potent neuroprotective role and augments special memory deficit in chronic epileptic rats. The isolated component which attenuates spatial memory performance could be beneficial outcome to retain cognitive blunting in chronic epilepsy.

RESUMEN: Actualmente, muchas personas con epilepsia no cuentan con un control adecuado de las convulsiones, incluso con los mejores medicamentos disponibles. Además, varios antiepilépticos tienen un impacto cognitivo adverso además de efectos secundarios. Por lo tanto, la necesidad de nuevos fármacos antiepilépticos sigue siendo un desafío. Sin embargo, muchos de los componentes naturales tienen acción antiepiléptica y este hecho permanece científicamente inexplorado. Este estudio se diseñó para verificar el resultado conductual y neuro-patológico del cerotato de 1-triacontanol (1TAC), aislado de Marsilea quadrifolia Linn. (MQ) en el modelo de epilepsia en ratas del pentilenetetrazol (PTZ) crónico (PTZ). Ratas Wistar adultas de dos meses de edad (n = 60) se dividieron aleatoriamente en seis grupos; Grupo I (Control de jaula), II (Control de vehículo), III (Control positivo), IV (Medicamento estándar de tratamiento), V (1TAC: 40 mg / kg) y VI (1TAC: 80 mg / kg). Para inducir la inflamación se inyectó una dosis de 35 mg / kg de PTZ i.p. en cada 48 horas durante 30 días en los grupos III a VI. El rendimiento de la memoria espacial se probó utilizando el laberinto de agua de Morris, después de lo cual se procesaron los cerebros para investigaciones histopatológicas. Curiosamente, 1TAC pudo minimizar la pérdida de células piramidales en la región CA3 del hipocampo. Estos cambios celulares respondieron de manera conductual como una mejora del aprendizaje especial y la memoria, una mejor navegación espacial y la configuración del lugar del objeto. El estudio actual implica fuertemente que 1TAC de MQ tiene un potente papel neuroprotector y mejora el déficit de memoria especial en ratas epilépticas crónicas. El componente aislado que atenúa el rendimiento de la memoria espacial podría ser un resultado beneficioso para retener la reducción cognitiva en la epilepsia crónica.

Biogenic synthesis of Marsilea quadrifolia gold nanoparticles: a study of improved glucose utilization efficiency on 3T3-L1 adipocytes.

This study aims mainly to provide an insight and understanding of the effect of glucose utilization efficiency of biogenic gold nanoparticles (GNPs) synthesized through the mediation of Marsilea quadrifolia (M. quadrifolia) methanol extract on 3T3-L1 adipocytes. The biosynthesized GNPs were characterized by UV visible spectrophotometry and FTIR. Simultaneously, the nature, stability, and morphological characteristics were analyzed by XRD, TG-DTA, SEM-EDS, HRTEM, and SAED. The results of characterization studies were used to assess the properties of GNPs. The in vitro cytotoxicity screening indicates that 100 µM of biogenic GNPs were displayed 71.23 ± 1.56% of cellular viability in 3T3-L1 adipocyte cells. Subsequently, increased glucose utilization of biosynthesized GNPs based on a dose-dependent manner on 3T3-L1 has also been demonstrated. The effect of GNPs (30 µg) on glucose uptake was higher than that of insulin and metformin. Moreover, the observed results clearly highlight that the biogenic GNPs have higher efficiency of glucose utilization and cellular viability in 3T3-L1 adipocytes with lower toxicity.

Isolation and identification of polyphenols from Marsilea quadrifolia with antioxidant properties in vitro and in vivo.

Marsilea quadrifolia is an edible aquatic medicinal plant used as a traditional health food in Asia. Four new polyphenols including kaempferol 3-O-(2″-O-E-caffeoyl)-ß-d-glucopyranoside (1), kaempferol 3-O-(3″-O-E-caffeoyl)-α-l-arabinopyranoside (3), 4-methy-3'-hydroxypsilotinin (4) and (±)-(E)-4b-methoxy-3b,5b-dihydroxyscirpusin A (18) together with 14 known ones (2, 5-17) were isolated from the ethanol extract of M. quadrifolia. Structures of the new compounds were elucidated by extensive spectroscopic analyses. In DPPH and oxygen radical absorbance capacity antioxidant assays, some compounds showed stronger antioxidant activities and quercetin (9) was the most potent antioxidant in both assays. In a restraint-induced oxidative stress model in mice, quercetin significantly attenuated the increase in plasma ALT and AST levels as well as liver MDA content of restrained mice. Liver SOD activity was also significantly increased by quercetin, indicating a significant in vivo antioxidant activity. As a rich source of polyphenols with strong antioxidant activities, M. quadrifolia may be developed to a product for relieving oxidative stress.

1-Triacontanol cerotate; isolated from Marsilea quadrifolia Linn. ameliorates reactive oxidative damage in the frontal cortex and hippocampus of chronic epileptic rats.

ETHNOPHARMACOLOGICAL RELEVANCE: Marsilea quadrifolia Linn. (MQ) has been used for insomnia and epileptic disorders in traditional Indian medicine. The present study is to isolate the active component responsible for antiepileptic property of MQ by evaluating its ability to minimize the reactive oxidative damage in brain due to chronic epilepsy in rat. MATERIALS AND METHODS: 1-Triacontanol cerotate (1TAC) was isolated after chromatography on a silica gel from dried petroleum ether fraction of methanolic extract of MQ. Acute oral toxicity studies of 1TAC were carried out and efficacy of 1TAC on malondialdehyde (MDA) and reduced glutathione (GSH) production in different brain areas of chronic pentylenetetrazole (PTZ) induced epileptic rats were evaluated. RESULTS: Our results showed that PTZ-kindled chronic epileptic rats had an increase MDA and decreased GSH concentration in the frontal cortex as well as hippocampus, compared to the normal control. MDA and GSH concentrations in those brain areas were normalized after treatment with sodium valproate (SV) in 200 mg kg(-1)bw; as well as 1TAC in 40 and 80 mg kg(-1)bw doses. CONCLUSION: Production of reactive oxygen species (ROS) is known to worsen epileptogenesis. The isolated component 1TAC which reduced the reactive oxidative damage in hippocampus and frontal cortex of PTZ kindled rats could be responsible for antiepileptic property of MQ. Its action is found to be dose dependent, with 80 mg kg(-1)bw showing even better efficacy than 200 mg kg(-1)bw of SV.

Evaluation of anti-epileptic property of Marsilea quadrifolia Linn. in maximal electroshock and pentylenetetrazole-induced rat models of epilepsy.

OBJECTIVE: To study the anti-epileptic effects of methanolic extract of Marsilea quadrifolia Linn. (MQ) in maximal electroshock (MES) and pentylenetetrazole (PTZ) induced rat models of epilepsy. METHOD: A total of 84 adult male Wistar rats were used. An acute oral toxicity study was conducted on 36 rats and the remaining were used for other experiments. Each model had 24 rats which were allotted into four groups (n = 6). Group I (Control) received 1% carboxymethyl cellulose solution, Group II (Positive control) received phenytoin 300 mg kg(-1) b.w. in the MES model; sodium valproate 200 mg kg(-1) b.w. in the PTZ model, Group III (MQ) received 400 mg kg(-1) b.w. MQ extract and Group IV (MQ) received 600 mg kg(-1) b.w. MQ extract. Hind limb extension (HLE) time and recovery time were noted in the MES model. Latency for myoclonic jerk, seizures and EEG was recorded in the PTZ model. RESULTS: When compared to control, the phenytoin received group did not show HLE. In MQ pre-treated groups only 50% of rats showed HLE. Sodium valproate and various doses of MQ significantly increased the latency for onset of clonus and seizures. PTZ-induced EEG alterations were significantly attenuated by MQ administration and this was comparable to that of the sodium valproate effect. CONCLUSION: Marsilea quadrifolia Linn. showed significant anti-epileptic efficacy against various epilepsy models.

Effect of standardized extract of Marsilea minuta on learning and memory performance in rat amnesic models.

CONTEXT: Marsilea minuta Linn (Marsileaceae) is a common Indian hydrophytic plant. Traditionally, the plant has been used as a sedative for the treatment of insomnia and other mental disorders. Background information of this plant has encouraged us to investigate its antiamnesic activity in rat. OBJECTIVE: Standardized ethanol extract of M. minuta was investigated for their putative role in learning and memory performance in normal and amnesic rats. MATERIALS AND METHODS: Ethanol extract of M. minuta (EMM) was standardized for marsiline using HPLC. The effect of standardized extract of M. minuta (1.15% w/w marsiline) was tested in amnesic rat using elevated plus maze (EPM) and passive avoidance (PA) test. Amnesia was induced after scopolamine (1 mg/kg, s.c.) and electroconvulsive shock (150 mA, 0.2 s) treatment. Behavioral studies were further substantiated with acetylcholinesterase (AChE) activity and radioligand muscarinic receptor binding studies in rat brain regions. RESULTS: Oral administration of EMM at 200 and 400 mg/kg/day for 3 days significantly reversed the amnesia whereas, no per se effect was observed. In comparison to control, AChE activity in frontal cortex and hippocampus was found to be significantly (P < 0.05) inhibited by EMM. EMM at doses 200 and 400 mg/kg has significantly (P < 0.05) increased (+34 % and +40 % change in affinity, respectively) the binding of 3H-QNB in frontal cortex indicating the up regulation of the muscarinic receptors. DISCUSSION AND CONCLUSION: These findings suggest that standardized extract of M. minuta have excellent antiamnesic activity, probably mediating through central cholinergic system.

Anticonvulsant effect of Marsilea quadrifolia Linn. on pentylenetetrazole induced seizure: a behavioral and EEG study in rats.

ETHNOPHARMACOLOGICAL RELEVANCE: Marsilea quadrifolia Linn (MQ) extract has been used traditionally as sedative and antiepileptic drug in India. AIM OF THIS STUDY: To investigate the anticonvulsive potential of MQ extracts by using behavior and electroencephalographic (EEG) analysis on pentylenetetrazole (PTZ) induced seizure model in rats. MATERIALS AND METHODS: For anticonvulsant effect, 60minutes after administration of MQ, behavior and EEG were analyzed during PTZ (60mg/kg) induced seizures. Changes of EEG power, latency of onset of seizure, seizure severity score, and duration of epileptic seizure were determined. RESULTS: Both the water and ethanol extract of MQ increased the latency of seizure but also decreased duration of epileptic seizure and seizure severity score. This reduction of seizure severity was also observed in EEG recording and EEG power analysis. The effectiveness of MQ ethanol extract is better than MQ water extract. CONCLUSION: Both water and ethanol extract of MQ were effective in reducing the severity of behavioral and EEG seizures induced by PTZ in rats. This study justifies the traditional use of this plant in epilepsy.

Cholinesterase inhibition activity of Marsilea quadrifolia Linn. an edible leafy vegetable from West Bengal, India.

Maesilea quadrifolia Linn. (Marsileaceae) is a leafy vegetable well known in India. The current study aims to explore the phytochemical profile of M. quadrifolia and investigate its anti-cholinesterase potential. The methanol extract of the plant was subjected to qualitative and quantitative phytochemical screening (total alkaloidal content, saponin content and phenol content) and its anti-cholinesterase potential was tested by TLC bioautography and other screening methods using acytylcholinesterase (AChE) and butyrylcholinesterase (BChE). The study revealed that the extract contains various classes of phytoconstituents including steroids, saponins, alkaloids and other polyphenols. Total alkaloid, phenolic and saponin contents were found to be 19.3 mg g⁻¹ and 158.5 ± 1.02 mg g⁻¹ as gallic acid equivalents and 2.63 mg g⁻¹ of the extract, respectively. The TLC bioautography method exhibited the inhibition of both enzymes. In a microtiter plate assay, the IC50 values of the extract for AChE and BChE were found to be 51.89 ± 0.24 µg mL⁻¹ and 109.43 ± 2.82 µg mL⁻¹, respectively. These findings suggest that M. quadrifolia is a potential lead as an AChE and BChE inhibitor, which may be useful in the management of Alzheimer's disease.

Anti-aggressive activity of a standardized extract of Marsilea minuta Linn. in rodent models of aggression.

The present study was undertaken to evaluate in vivo anti-aggressive potential of a standardized extract of Marsilea minuta Linn. (Marsileaceae). The standardized extract of Marsilea minuta was evaluated for its potential effects against defensive and offensive aggressive behavior models of rodents. Marsilea minuta extract was orally administered at three dose levels (100, 200, and 400 mg/kg BW) once daily for 14 consecutive days as a suspension in polyethylene glycol (PEG), diazepam (2.5 mg/kg, p.o.) was used as a standard anti-aggressive agent. Control group animals were given an equal volume of vehicle (10%, v/v, PEG suspension). Anti-aggressive activity was evaluated using the following validated models of aggression, viz.: foot shock-induced aggression, isolation-induced aggression and resident-intruder aggression, in rodents. As a result, Marsilea minuta extract showed dose dependant anti-aggressive activity in the aforementioned, validated models of aggression. This suggests that the extract from Marsilea minuta has a promising anti-aggressive activity qualitatively comparable to that of diazepam.

Abscisic acid regulation of heterophylly in Marsilea quadrifolia L.: effects of R-(-) and S-(+) isomers.

The plant hormone abscisic acid (ABA) induces a developmental switch in the aquatic fern Marsilea quadrifolia, causing the formation of aerial type characteristics, including the elongation of petioles and roots, a change in leaf morphology, the expansion of leaf surface area, and the shortening of the internodes. A number of ABA-responsive heterophylly (ABRH) genes are induced early during the transition. Using optically pure isomers of ABA, it was found that both the natural S-(+)-ABA and the unnatural R-(-)-ABA are capable of inducing a heterophyllous switch and regulating ABRH gene expression. When dose responses are compared, the unnatural ABA gives stronger morphogenic effects than the natural ABA at the same concentration, it is effective at lower concentrations, and its optimal concentration is also lower compared with the natural ABA. Deuterium-labelled ABA enantiomers were used to trace the fate of applied ABA and to distinguish the applied compound and its metabolites from the endogenous counterparts. In tissues, the supplied (+)-ABA was metabolized principally to dihydrophaseic acid, while the supplied (-)-ABA was converted at a slower rate to 7'-hydroxy abscisic acid. Treatment with either enantiomer resulted in increased biosynthesis of ABA, as reflected in the accumulation of endogenous dihydrophaseic acid. Taken together, these results suggest two distinct mechanisms of action for (-)-ABA: either (-)-ABA is intrinsically active, or its activity is due to the stimulation of ABA biosynthesis.