Box-Behnken Design-Based Optimization of Phytochemical Extraction from Diplazium esculentum (Retz.) Sw. Associated with Its Antioxidant and Anti-Alzheimer's Properties.

A previous study reported that the ethanolic extract of the edible fern, Diplazium esculentum (Retz.) Sw. (DE), obtained from a non-optimized extraction condition exhibited anti-Alzheimer's disease (AD) properties through the inhibition of a rate-limiting enzyme in amyloid peptide formation, ß-secretase-1 (BACE-1). Nevertheless, a non-optimized or suboptimal extraction may lead to several issues, such as a reduction in extraction efficiency and increased time and plant materials. In this study, extraction of the DE was optimized to obtain appropriate BACE-1 inhibition using a Box-Behnken design (BBD) and response surface methodology (RSM). Data revealed that the optimal extraction condition was 70% (v/v) aqueous ethanol, 50 min extraction time, 30 °C extraction temperature, and 1:30 g/mL solid/liquid ratio, giving BACE-1 inhibition at 56.33%. In addition, the extract also exhibited significant antioxidant activities compared to the non-optimized extraction. Metabolomic phytochemical profiles and targeted phytochemical analyses showed that kaempferol, quercetin, and their derivatives as well as rosmarinic acid were abundant in the extract. The optimized DE extract also acted synergistically with donepezil, an AD drug suppressing BACE-1 activities. Data received from Drosophila-expressing human amyloid precursor proteins (APPs) and BACE-1, representing the amyloid hypothesis, showed that the optimized DE extract penetrated the fly brains, suppressed BACE-1 activities, and improved locomotor functions. The extract quenched the expression of glutathione S transferase D1 (GSTD1), inositol-requiring enzyme (IRE-1), and molecular chaperone-binding immunoglobulin (Bip), while donepezil suppressed these genes and other genes involved in antioxidant and endoplasmic reticulum (ER) stress response, including superoxide dismutase type 1 (SOD1), activating transcription factor 6 (ATF-6), and protein kinase R-like endoplasmic reticulum kinase (PERK). To sum up, the optimized extraction condition reduced extraction time while resulting in higher phytochemicals, antioxidants, and BACE-1 inhibitors.

Enhancing Therapeutic Efficacy of Donepezil, an Alzheimer's Disease Drug, by Diplazium esculentum (Retz.) Sw. and Its Phytochemicals.

Alzheimer's disease (AD) is the most common type of dementia and a significant concern to global public health due to the prevalence of aging populations. Donepezil is one of only a few medications approved for use as an anti-AD agent but all have adverse side effects. Reducing the dosage of AD drugs with plant extracts (phytotherapy) while maintaining efficacy is one strategy to minimize adverse side effects. We previously reported the anti-AD properties of an edible fern, Diplazium esculentum (Retz.) Sw. (DE), which inhibited key enzymes involved in AD pathogenesis including acetylcholinesterase (AChE), butyrylcholinesterase (BChE), and ß-secretase 1 (BACE-1). This study aimed to determine whether DE exhibited a synergistic effect with donepezil. The enzyme inhibitory assay showed that DE extract and its bioactive compounds, kaempferol, and quercetin, slightly impeded AChE inhibition with donepezil, while DE extract and quercetin showed synergistic or additive effects with donepezil against BChE and BACE-1, respectively. DE extract combined with donepezil also improved eye phenotypes in a Drosophila model of AD by preventing ommatidia atrophia and bristle breakages. Furthermore, the DE extract exhibited no genotoxic activities, as determined by the Ames test. Our data revealed that DE extract showed promise when combined with donepezil during AD treatment by targeting BChE and BACE-1.

Targeting Melanoma with a phytochemical pool: Tailing Makisterone C.

BACKGROUND AND OBJECTIVE: According to World Health Organization, melanoma claims the lives of about 48000 people worldwide each year. The purpose of this study was to identify potential phytochemical pool from Diplazium esculentum against proteins that contribute to melanoma development. METHODS: The research was carried to locate potentially bioactive molecules and conduct a theoretical analysis of active ingredients from DE to impact melanoma. Network pharmacology, pharmacokinetics, protein network interaction, gene enrichment, survival, and infiltration analysis were conducted. Furthermore, molecular docking and molecular dynamics simulation was carried out for makisterone C-MAPK1, MAPK3, and AKT1 complexes. RESULTS: The potential phytochemical pool were identified (stigmast-5-en-3-ol, esculentic acid, rutin, and makisterone C) and based on network pharmacology and molecular docking studies, makisterone-C was proposed to be the most promising ingredient. Furthermore, the investigation revealed 14 genes as critical "hubs" involved in combating melanoma that are manipulated by the above-mentioned 4 active ingredients and modulate multiple signaling in melanoma development. CONCLUSION: This study insights into the potential anti-melanoma effects of phytochemical pool from Diplazium esculentum using network pharmacology analysis, molecular docking, and simulation tailing makisterone C as a lead moiety and suggests the need for makisterone C further evaluation in intervening melanoma progression.

Phytochemical profiling and biological activities of Diplazium esculentum (Retz.) Sw.: an edible vegetable fern.

OBJECTIVES: Diplazium esculentum (Retz.) Sw. is an edible vegetable fern of the Himalayan region with high nutritional and therapeutic value owing to its richness in various secondary metabolites and both macro and micronutrients. CONTENT: This updated review discusses the general traditional use, ethnopharmacology, phytochemistry, nutritional value, pharmacology, and toxicity concerns of D. esculentum. SUMMARY: The plant parts, viz. rhizomes, shoots, fronds and leaves, have immense ethnomedicinal importance, being traditionally used to cure several health disorders. Among other pharmacological effects, this botanical reveals excellent anti-inflammatory, analgesic, antifungal, antibacterial, antioxidant, anti-leishmanial, antioxidant, anaphylactic, antipyretic, anthelmintic and hepatoprotective activities, directly attributed to the presence of many secondary metabolites. From a pharmacological point of view, the excellent antioxidant potential of D. esculentum suggests its promising use for nutraceutical or functional food formulation purposes. OUTLOOK: Considering the evidences on popular ethnomedicinal uses of D. esculentum as an edible vegetable, its immense bio-potential, and multiple pharmacological roles, there is a huge need to evaluate its therapeutic applications in light of standard clinical trials.

Potential of the Ethanol Extract of Aerial Parts of Diplazium esculentum (Retz) SW. AS Larvicide against Anopheles gambiae Giles and Culex quinquefasciatus Say

Plant products have been tested as insecticides against mosquitoes as they are promising candidates to replace conventional insecticides. This study was carried out to evaluate the larvicidalpotential of ethanol extract of the aerial parts of Diplazium esculentumagainst Anopheles gambiaeand Culex quinquefasciatus. Ethanol extract of the aerial parts of D. esculentumwas screened for its phytochemical constituents and used for larvicidal assay. A stock solution of the extract (5g in Original Research Article Umohata et al.; IJTDH, 41(3): 40-47, 2020; Article no.IJTDH.5566841100ml of water) was prepared. From the stock solution, 0.45, 0.60, 0.75, 0.90 and 1.05%w/v concentrations of the extract were obtained for the study. Each concentration of the extract had 3 replicates. The control was also replicated. Twenty (20) third instar larvae each of Anopheles gambiae andCulex quinquefasciatuswere separately exposed to each extract concentration for a duration of 48 hours. Larval nutrient was added to each experimental set up. Observations were made after 24 and 48 hours exposure period.Phytochemical screening revealed the presence of some plant metabolites. Mortality of larvae exposed to the extract increased with increased concentration and exposure time. This study revealed a differential susceptibility of larvae of the two mosquito species to the extract as evident by the 24h LC50values obtained which were 0.355 and 2.468%w/v for An. gambiaeand Cx.quinquefasciatus respectively. Exposure of An. gambiaelarvae to the extract resulted in 100% mortality even with the least concentration of 0.45%w/v after 48 hours exposure period while the highest concentration of extract (1.05%w/v) resulted in 53.33% mortality of Cx.quinquefasciatuslarvae, after an exposure period of 48 hours. Results obtained from this study suggest that the aerial parts of D. esculentumif further explored would be useful in the control of An. gambiae andCx.quinquefasciatus.

Steroids from Diplazium esculentum: Antiplasmodial activity and molecular docking studies to investigate their binding modes

@#Diplazium esculentum is an edible fern commonly consumed by the local community in Malaysia either as food or medicine. Isolation work on the ethyl acetate extract of the stem of D. esculentum resulted in the purification of two steroids, subsequently identified as stigmasterol (compound 1) and ergosterol5,8-endoperoxide (compound 2). Upon further testing, compound 2 displayed strong inhibitory activity against the Plasmodium falciparum 3D7 (chloroquine-sensitive) strain, with an IC50 of 4.27±1.15 µM, while compound 1 was inactive. In silico data revealed that compound 2 showed good binding affinity to P. falciparum-Sarco endoplasmic reticulum calcium-dependent ATPase (PfATP6); however, compound 1 did not show an antiplasmodial effect due to the lack of a peroxide moiety in the chemical structure. Our data suggested that the antiplasmodial activity of compound 2 from D. esculentum might be due to the inhibition of PfATP6, which resulted in both in vitro and in silico inhibitory properties.

In vitro propagation of a rare medicinal fern of Western Ghats – Diplazium esculentum (Reytz.).

Present study aimed for in vitro culture of circinate part of young leaves of D. esculentum which is amongst the leafy vegetables consumed as vegetable by Paniya and Chetti tribes of Western Ghats. The circinate part of young leaves (crosiers), excised before the beginning of foliar expansion, was inoculated on half strength Murashige and Skoog (MS) medium supplemented with auxins indole-3-butyric acid (IBA) or α-napthalene acetic acid (NAA) or 2,4-Dichlorophenoxyacetic acid (2,4-D) and cytokinin 6- benzylaminopurine (BA) in a range 0.5 to 2.5 mg L-1. Combinations of different concentrations of 2,4 D + BA, IBA + BA as well as of NAA+ BA were also tested in half strength MS medium with 3% sucrose and with pH 5.8. The best morphogenic response was obtained with half strength MS medium supplemented with 2,4-D 0.5 mg L-1 and BA 2.5 mg L-1, 3% sucrose, at pH 5.8. For rooting of the microshoots, half strength MS medium supplemented with 2,4-D ( 2 and 1 mg L-1 ) exhibited best results. Present study reports the successful in vitro culturing of D. esculentum.