Antifungal Activity of Six Medicinal Plants of Pakistan Against Selected Fungi

Antifungal properties of Cucumis sativus, Portulaca oleracea, Malus baccata, Saxifraga flagillaris, Geranium wallichianum. and Monotheca buxifolia were evaluated against Alternaria, Acremonium, Verticillium, Pythium, and Trichoderma, using agar well-diffusion method. The crude ethanolic extract of G. wallichianum showed the highest antifungal activity followed by P. oleracae and S. flagillaris. The dichloromethane fraction of G. wallichianum showed the highest antifungal activity against Acremonium (28 mm), Alternaria (20.50 mm) and Trichoderma (20 mm). The n-hexane fraction of C. sativus showed the maximum antifungal activity (20 mm) against Pythium. The present findings demonstrated that six selected plants contain precious natural products for treating infectious diseases and can be used to isolate chemical compounds for future drug sources. Bangladesh J. Bot. 50(2): 441-443, 2021 (June)

Metabolic fingerprinting, antioxidant characterization, and enzyme-inhibitory response of Monotheca buxifolia (Falc.) A. DC. extracts

Background Ethnobotanical and plant-based products allow for the isolation of active constituents against a number of maladies. Monotheca buxifolia is used by local communities due to its digestive and laxative properties, as well as its ability to cure liver, kidney, and urinary diseases. There is a need to explore the biological activities and chemical constituents of this medicinal plant. Methods In this work, the biochemical potential of M. buxifolia (Falc.) A. DC was explored and linked with its biological activities. Methanol and chloroform extracts from leaves and stems were investigated for total phenolic and flavonoid contents. Ultrahigh-performance liquid chromatography coupled with mass spectrometry (UHPLC–MS) was used to determine secondary-metabolite composition, while high-performance liquid chromatography coupled with photodiode array detection (HPLC–PDA) was used for polyphenolic quantification. In addition, we carried out in vitro assays to determine antioxidant potential and the enzyme-inhibitory response of M. buxifolia extracts. Results Phenolics (91 mg gallic-acid equivalent (GAE)/g) and flavonoids (48.86 mg quercetin equivalent (QE)/g) exhibited their highest concentration in the methanol extract of stems and the chloroform extract of leaves, respectively. UHPLC–MS analysis identified a number of important phytochemicals, belonging to the flavonoid, phenolic, alkaloid, and terpenoid classes of secondary metabolites. The methanol extract of leaves contained a diosgenin derivative and polygalacin D, while kaempferol and robinin were most abundant in the chloroform extract. The methanol extract of stems contained a greater peak area for diosgenin and kaempferol, whereas this was true for lucidumol A and 3-O-cis-coumaroyl maslinic acid in the chloroform extract. Rutin, epicatechin, and catechin were the main phenolics identified by HPLC–PDA analysis. The methanol extract of stems exhibited significant 2,2-diphenyl-1-picrylhydrazyl (DPPH) and 2,2′-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid (ABTS) radical-scavenging activities (145.18 and 279.04 mmol Trolox equivalent (TE)/g, respectively). The maximum cupric reducing antioxidant capacity (CUPRAC) (361.4 mg TE/g), ferric-reducing antioxidant power (FRAP) (247.19 mg TE/g), and total antioxidant potential (2.75 mmol TE/g) were depicted by the methanol extract of stems. The methanol extract of leaves exhibited stronger inhibition against acetylcholinesterase (AChE) and glucosidase, while the chloroform extract of stems was most active against butyrylcholinesterase (BChE) (4.27 mg galantamine equivalent (GALAE)/g). Similarly, the highest tyrosinase (140 mg kojic-acid equivalent (KAE)/g) and amylase (0.67 mmol acarbose equivalent (ACAE)/g) inhibition was observed for the methanol extract of stems. Conclusions UHPLC–MS analysis and HPLC–PDA quantification identified a number of bioactive secondary metabolites of M. buxifolia, which may be responsible for its antioxidant potential and enzyme-inhibitory response. M. buxifolia can be further explored for the isolation of its active components to be used as a drug.

Genetic diversity in the natural population of threatened fruit- bearing plant species Monotheca buxifolia (Falc.) A.D.

Monotheca buxifolia is an ethnomedicinally and economically important threatened fruit bearing plant species in Malakand Division Pakistan. The genetic diversity among the 92 various genotypes of Monotheca buxifolia was carried out using sodium dodecyl sulfate poly acrylamide gel electrophoresis (SDS-PAGE) method. A considerable amount of inter districts genetic diversity (66.70%) was observed among the genotypes of M. buxifolia. Protein profiling was conducted on 12% gel electrophoresis. A total of 6 protein bands were observed in M. buxifolia genotypes. SDS-PAGE practice is a convenient scheme for the examination of both genetic diversity and relationship. Particularly, L-4 and L-5 were monomorphic in the inter districts Monotheca buxifolia genotypes and was recognized as species specific. The remaining other loci were polymorphic. In this investigation, the high inter and intra- districts specific diversity was observed demonstrating SDS-PAGE is an authoritative procedure for categorizing the genetically diverse germplasms in M. buxifolia. The findings from this study could be useful in the identification and selection of suitable M. buxifolia genotypes for future conservation programmes. Today, there is still a need to examine the genetic diversity and protect genetic resources, in particular wild species, for possible benefits in plant conservation programmes. To the best of our knowledge, this is the first ever report that addresses genetic variability in M. buxifolia.

Bioactive Compounds from Monotheca Buxifolia Inhibit the Growth of Hepatocellular Carcinoma Cells

The natural products and conventional chemotherapeutic drugs are believed to increase the cure rates of anti-cancer treatment while reducing their toxicity. The current study investigates the cytotoxic and apoptogenic effects of bioactive compounds from Monotheca buxifolia on Hep G2 cell lines. The effect on the viability of Hep G2 cells was evaluated by MTT assay; Morphological changes were studied, the apoptotic activity was demonstrated through Annexin-V-FITC/ PI, a molecular dynamics simulation study was conducted to explore the binding pattern of the compounds in the active site of the PPRAδ protein. The isolated compounds lauric acid, oleanolic acid, and bis(2-ethylhexyl) phthalate inhibited the growth of hepatocellular cancer cells, as determined by MTT assay and annexin V-FITC/PI. The IC50 value for lauric acid was 56.4584 ± 1.20 µg/ml, that for oleanolic acid was 31.9421 ± 1.03 µg/ml, and that for bis(2-ethylhexyl) phthalate was 83.8019 ± 2.18 µg/ml. After 24 h of treatment, 29.5% of Hep G2 cells treated with lauric acid, 52.1% of those treated with oleanolic acid, and 22.4% of those treated with bis(2-ethylhexyl) phthalate were apoptotic. Morphological assay and Hoechst staining microscopy revealed the morphological alterations of cell membrane accompanied by nuclear condensation after treatment. The high fluctuation indicates the high potency and adopting various interactions, and vice versa, the oleanolic acid showed highly residues fluctuation, which remains stable in the active site of PPARδ protein and involved in various interactions while remaining locally fluctuated in the binding site the other two compounds. In conclusion, a significant apoptogenic effect was exhibited by lauric acid, oleanolic acid, and bis(2-ethylhexyl) phthalate against HepG2 cells in inducing apoptosis. Our findings indicate that these bioactive compounds hold promise as potential therapeutic for hepatocellular carcinoma.