Free radical scavenging and anti-isolated human LDL oxidation activities of Butea superba Roxb. extract.

BACKGROUND: Butea superba Roxb. (B. superba), is an herbal plant traditionally used for rejuvenation. Additionally, there have been reports on its antioxidant properties. Low-density lipoproteins (LDL) oxidation is the leading cause of cardiovascular diseases (CVDs). Natural products with antioxidant properties have the potential to inhibit LDL oxidation. However, no work has been done about the anti-isolated human LDL oxidation of B. superba extract (BSE). This study aimed to investigate the antioxidant potential of BSE and its ability to prevent isolated human (LDL) oxidation induced by free radical agents. METHODS: The antioxidant properties were investigated by antioxidant assays, including 2,2-diphenyl-1-picrylhydrazyl (DPPH), 2,2-azinobis-(3-ethylbenzothiazoline)-6-sulfonic acid (ABTS), ferric reducing ability power (FRAP), nitric oxide (NO) and peroxynitrite scavenging assay. More so, anti-isolated human LDL oxidation activities were evaluated by 2,2'-azobis (2-amidinopropane) dihydrochloride (AAPH) and 3-morpholinosydnonimine hydrochloride (SIN-1) induced LDL oxidation assay. RESULTS: BSE exhibited a significant (p < 0.05) antioxidant activity in all the test systems, demonstrating its potential as a potent free radical scavenger. It displayed scavenging effects on DPPH (p < 0.05; IC50 = 487.67 ± 21.94 µg/ml), ABTS (p < 0.05; IC50 = 30.83 ± 1.29 µg/ml). Furthermore, it generated significantly (p < 0.05) increased antioxidant capacity in a dose-dependent manner in FRAP assay and exhibited significantly (p < 0.01) higher percent NO scavenging activity than gallic acid. Besides, BSE at 62.5 µg/ml exhibited a considerable percent peroxynitrite scavenging of 71.40 ± 6.59% after a 2 h period. Moreover, BSE demonstrated anti-isolated human LDL oxidation activity induced by AAPH and SIN-1 (p < 0.05) and revealed scavenging activity similar to ascorbic acid (p > 0.05). Identifying the main constituents of BSE revealed the presence of genistein, daidzein, and biochanin A through Liquid Chromatography-Mass Spectrometer/Mass Spectrometer (LC-MS/MS) analysis. CONCLUSION: This is the first report that the presence of isoflavones in BSE could play an important role in its antioxidation and isolated human LDL oxidation scavenging properties. These findings suggest the potential for developing antioxidant herbal supplements. However, further studies must be investigated, including efficacious and safe human dosages.

Butein-Enriched Fractions of Butea monosperma (Lam.) Taub. Flower Decrease Weight Gains and Increase Energy Expenditure in High-Fat Diet-Induced Obese Mice.

Butea monosperma (Lam.) Taub. has been applied to treat inflammatory, metabolic, and infectious diseases. However, the antiobesity effects of B. monosperma (Lam.) Taub. flower (BMF) and the underlying mechanisms have not been determined. In this study, we analyzed the various extraction procedures, investigated the antiobesity effects, and identified the main chemical constituents of BMF. The BMF was subjected to acid hydrolysis in 5% H2SO4 in methanol at 50°C for 48 h and partitioned with ethyl acetate. The acid-hydrolyzed BMF ethyl acetate extracts (BMFE) strongly induced the expression of uncoupling protein 1 (Ucp1) and other thermogenic genes in C3H10T1/2 adipocytes. Daily oral administration of 70 mg/kg BMFE (BMFE70) to mice with diet-induced obesity resulted in less body weight gain, increased glucose tolerance, higher rectal temperature, and increased oxygen consumption. Qualitative and quantitative analyses along with treatments in Akt1 knockout mouse embryonic fibroblasts indicate that butein is a major active ingredient of BMFE, which stimulates Ucp1 gene expression. These data show the effects of butein-containing B. monosperma flower extract on thermogenesis and energy expenditure, further suggesting the potential role of BMFE as a functional ingredient in obesity and related metabolic diseases.

Dual Function Antimicrobial Loaded Lectin Carrier: A Strategy to Overcome Biomolecular Interference without Detectable Resistance.

The disposition of a drug in a biological system may be altered by complex biological fluids; especially, protein binding to drugs influences their activity. Herein, we demonstrated a convenient method involving the noncovalent formulation of butea monosperma seed lectin (BMSL) with an antimicrobial lipid, cationic N-acylethanolamine (cNAE) to mitigate the serum protein interference. Fluorescence spectroscopy and molecular docking study revealed that cNAEs readily formed noncovalent complexes with serum protein, bovine serum albumin. The resulting complexes interfered with the antimicrobial activity of cNAEs. Strikingly, the noncovalent conjugates developed with BMSL and cNAEs (BcNAE) overcame the interference from serum protein and displayed remarkable antimicrobial activity against uropathogenic Escherichia coli (UPEC). Strikingly, the minimum inhibitory concentration (MIC) of the lectin conjugates (7.81 µM) was 4-fold lower than the MIC of pure cNAE. Mechanistic studies showed that BcNAE depolarized the bacterial membrane and affected the integrity to exert the antimicrobial activity. The membrane directed activities of BcNAE on UPEC efficiently eliminated the development of resistance even after 25 passages. The hemocompatibility results and the biosafety assessed in a zebrafish model suggested that BcNAE was nontoxic with good selectivity to bacteria. While testing the therapeutic efficacy against UPEC infected zebrafish, we found that 1× MIC cNAE is ineffective due to interference from biological fluids, which is in agreement with in vitro studies. Remarkably, the infected fish treated with 1× MIC BcNAE conjugates were rescued from infection and restored to the normal life in less than 9 h. Bacterial colony count assay revealed that BcNAE was more efficient in overcoming the biological fluid interference and eliminated the bacterial burden in infected zebrafish. Histopathology analysis supported that BcNAE treatment restored the pathological changes induced by UPEC and, thus, increased survival. The high antimicrobial intensity with limited chance for resistance development and potential to overcome biomolecular interference with a lack of toxicity enhance the merits of exploring lectin conjugates against infectious pathogens.

A Novel Serum Glycobiomarker for Diagnosis and Prognosis of Cholangiocarcinoma Detected by Butea monosperma Agglutinin.

Plant lectins are widely used in medical glycosciences and glycotechnology. Many lectin-based techniques have been applied for the detection of disease-associated glycans and glycoconjugates. In this study, Butea monosperma agglutinin (BMA), a lectin purified from seeds of the medicinal plant Butea monosperma, was used for the detection of cholangiocarcinoma (CCA)-associated glycans. Expression of BMA-binding N-acetyl galactosamine/galactose (GalNAc/Gal)-associated glycan (BMAG) in CCA tissues was determined using BMA lectin histochemistry; the results showed that BMAG was undetectable in normal bile ducts and drastically increased in preneoplastic bile ducts and CCA. The study in hamsters showed that an increase of BMAG was associated with carcinogenesis of CCA. Using an in-house double BMA sandwich enzyme-linked lectin assay, BMAG was highly detected in the sera of CCA patients. The level of serum BMAG in CCA patients (N = 83) was significantly higher than non-CCA controls (N = 287) and it was applicable for diagnosis of CCA with 55.4% sensitivity, 81.9% specificity, and 76.0% accuracy. A high level of serum BMAG (≥82.5 AU/mL) was associated with unfavorable survival of CCA patients; this information suggested the potential of serum BMAG as a poor prognostic indicator of CCA. In summary, BMAG was aberrantly expressed in preneoplastic bile ducts and CCA, it was also highly detected in patient serum which potentially used as a marker for diagnosis and prognostic prediction of CCA.

Antitumor potential of the protein phosphatase inhibitor, cantharidin, and selected derivatives.

Cantharidin is a potent natural protein phosphatase monoterpene anhydride inhibitor secreted by several species of blister beetle, with its demethylated anhydride analogue, (S)-palasonin, occurring as a constituent of the higher plant Butea frondosa. Cantharidin shows both potent protein phosphatase inhibitory and cancer cell cytotoxic activities, but possible preclinical development of this anhydride has been limited thus far by its toxicity. Thus, several synthetic derivatives of cantharidin have been prepared, of which some compounds exhibit improved antitumor potential and may have use as lead compounds. In the present review, the potential antitumor activity, structure-activity relationships, and development of cantharidin-based anticancer drug conjugates are summarized, with protein phosphatase-related and other types of mechanisms of action discussed. Protein phosphatases play a key role in the tumor microenvironment, and thus described herein is also the potential for developing new tumor microenvironment-targeted cancer chemotherapeutic agents, based on cantharidin and its naturally occurring analogues and synthetic derivatives.

UHPLC-QTOF-MS/MS based phytochemical characterization and anti-hyperglycemic prospective of hydro-ethanolic leaf extract of Butea monosperma.

Butea monosperma is one of the extensively used plants in traditional system of medicines for many therapeutic purposes. In this study, the antioxidant activity, α-glucosidase and α-amylase inhibition properties of freeze drying assisted ultrasonicated leaf extracts (hydro-ethanolic) of B. monosperma have been investigated. The findings revealed that 60% ethanolic fraction exhibited high phenolic contents, total flavonoid contents, highest antioxidant activity, and promising α-glucosidase and α-amylase inhibitions. The UHPLC-QTOF-MS/MS analysis indicated the presence of notable metabolites of significant medicinal potential including apigenin, apigenin C-hexoside C-pentoside, apigenin C-hexoside C-hexoside, apigenin-6,8-di-C-pentoside and genistin etc., in B. monosperma leave extract. Docking studies were carried out to determine the possible role of each phytochemical present in leaf extract. Binding affinity data and interaction pattern of all the possible phytochemicals in leaf extract of B. monosperma revealed that they can inhibit α-amylase and α-glucosidase synergistically to prevent hyperglycemia.

Butea monosperma (Lam.) Taub. Bark fractions protect against free radicals and induce apoptosis in MCF-7 breast cancer cells via cell-cycle arrest and ROS-mediated pathway.

Butea monosperma (Lam.) Taub. is an ethnomedicinal tree of remedial value in the treatment of diabetes, bone fractures, and liver and neurological disorders. However, the information available on DNA-protective and anti-proliferative potential of bark of this tree is scarce. In the present study, the extract/fractions obtained from bark of B. monosperma were evaluated for antioxidant, DNA-protective, and anti-proliferative activities, along with their phytochemical profiling for identifying major constituents present in them. Different extract/fractions, namely, Bmth (methanol), Bhex (hexane), Bchl (chloroform), and Beac (ethyl acetate), were prepared and evaluated for antioxidant activity using in vitro assays. Extract/fractions were also evaluated for anti-proliferative and apoptotic activity in human breast carcinoma cell line MCF-7, using in vitro assays, namely, MTT, clonogenic, and neutral comet assay, along with confocal microscopy and flow cytometry. Among all extract/fractions, a pronounced antioxidant activity was exhibited by Bchl and Beac fractions, in DPPH· (EC50 213.2 and 161.5 µg/mL, respectively), ABTS+· (EC50 139.3 and 44.1 µg/mL, respectively), and reducing power assay (EC50 86.7 and 84.5 µg/mL, respectively). Both fractions protected plasmid DNA against hydroxyl radical induced damage in plasmid nicking assay. Bchl and Beac were also observed to inhibit the growth of MCF-7 cells (GI50 203.7 and 246.5 µg/mL, respectively). Both fractions induced apoptosis in MCF-7 cells, by arresting the cell cycle in G1 and sub-G1 phase, respectively, enhancing ROS levels, decreasing mitochondrial membrane potential, and inducing double-strand DNA breaks. HPLC analysis revealed high kaempferol content in Bchl, and catechin, epicatechin, and gallic acid in Beac.

Revealing the Significance of the Glycan Binding Property of Butea monosperma Seed Lectin for Enhancing the Antibiofilm Activity of Silver Nanoparticles against Uropathogenic Escherichia coli.

The incompetence of conventional antibiotics against bacteria residing in biofilms demands newer therapeutic intervention. In this study, we demonstrated that the interaction between silver nanoparticles (AgNPs) and Butea monosperma seed lectin (BMSL) forms efficient surface-functionalized AgNPs with excellent antibiofilm competency against uropathogenic Escherichia coli (UPEC). The minimum biofilm inhibitory concentration (MBIC) of AgNPs and the BMSL-AgNP conjugate (BAgNP) against UPEC was 75 and 9.37 µM, respectively. The eight-fold reduction in the MBIC of AgNPs was attributed to lectin functionalization. The chemical modification of serine amino acids affects the hemagglutination activity of BMSL but not its interaction with the AgNPs. At the same time, AgNPs surface-functionalized with modified BMSL display poor antibiofilm activity. Molecular docking studies revealed that BMSL binds to galactose with a free energy of -5.72 kcal/mol, whereas the serine residue-modified BMSL showed the lowest free energy values, suggesting incompetence for binding galactose. These results showcase that the sugar binding site of BMSL aids in the adhesion of AgNPs to the biofilm matrix and disturbs the formation of the biofilm, which was confirmed by light microscopy using crystal violet staining. At 37.5 µM, BAgNPs also have the capability to eradicate preformed biofilm. As a proof of concept, UPEC biofilm prevention and eradication were demonstrated on a urinary catheter. A scanning electron microscopy study showed that BAgNPs prevent bacterial colonization and thereby curtail biofilm growth. In addition to antibiofilm activity, BAgNPs exert antibacterial activity at 18.75 µM, which is four-fold lower than the MIC of AgNPs. A mechanistic study revealed that BAgNPs affect the integrity of the bacterial outer membrane and generate an imbalance in the antioxidant defense, which induces cell death. The results highlight that lectin functionalization can be extended to other nanoparticles and different antibiotics to enhance their efficacy against drug-resistant bacteria.

Biological activities of three medicinal plants from district Mirpur, AJK, Pakistan.

Present research work was aimed to investigate the biological activities i.e. antibacterial, antifungal, antioxidant, cytotoxic and antitumor activities of crude methanolic extract of Anagallis arvensis L., Butea monosperma (Lam.) Kuntze and Coronopus didymus (L.) Pers. against Gram positive strains (Bacillus subtilis, Staphylococcus aureus) and gram negative strains (Vibrio cholera, Enterobacter aerogenes, Klebsiella pneumonia, Agrobacterium tumefaciens, Escherichia coli) were screened. Best activity was observed against K. pneumonia and S. aureus by A. arvensis compared with other strains. Butea monosperma exhibited considerable activity against S. aureus, V. cholera, E. aerogenes and K. pneumonia compared with other strains. Methanolic extract of A. arvensis L. inhibited fungal growth against A. niger up to 30.2%. B. monosperma inhibited the growth of A. niger up to 43.5% and against A. fumigatus 27.3%. C. didymus inhibited the A. fumigates up to 27.3% and against A. niger, it inhibited 48%. Brine shrimps lethality bioassay was used to evaluate the cytotoxic activity and LD50 value was calculated by using probit analysis. Potato disc bioassay was designed to screen antitumor activity and data was analyzed by one way ANOVA.

Protective effects of Butea frondosa leaves against stress induced immune impairment in sprague dawley rats.

Stress is thought to impair immune function through emotional or behavioral manifestations thus the present study was done to assessed the effect of ethanolic extract of Butea frondosa (BF) leaves on behaviour, immunomodulatory activity and brain acetyl cholinesterase activity in normal and stress induced male rats. Neuroprotective effects of BF, doses (100,200,400mg/kg p.o) were measured by assessing the changes in the behaviour and the immunity of the rats. In stress control, the results indicated that the retention transfer latency, time spent in a closed arm, agglutination, total leukocytes counts (TLC), total paw edema ,size of spleen , decreased significantly (p<0.01) while glucose level, size of the kidney and the liver, AChE activity increased significantly (p<0.01) in comparison with normal control. In BF (200mg/kg) treated rats, the results indicated that the time spent in a closed arm (p<0.01), agglutination (p<0.01), TLC (p<0.01), total paw edema (p<0.05), size of spleen(p<0.01), increased significantly while glucose level (p<0.01), size of the kidney and the liver (p<0.01), AChE activity (p<0.01) decreased significantly in comparison with stress control. This study therefore concluded that the ethanolic extract of BF (200mg/kg) showed a protective effect against the stress induced impaired immune system and the psychological disorders.

Development of Ultra Performance Liquid Chromatography Tandem Mass Spectrometry Method for Simultaneous Identification and Quantitation of Potential Osteogenic Phytochemicals in Butea monosperma.

An ultra performance liquid chromatography coupled with hybrid triple-quadrupole linear ion trap tandem mass spectrometry (UPLC-ESI-QqQLIT-MS-MS) method in multiple reaction monitoring mode was developed for identification and simultaneous determination of potential osteogenic compounds in ethanol extracts of different plant parts of Butea monosperma collected from different geographical regions. The chromatographic separation was carried out on an Acquity UPLC CSH C18 column (1.7 µm, 2.1 × 100 mm) with 0.1% (v/v) formic acid in water and methanol as mobile phase under gradient conditions in 8 min. The developed method was validated according to the guidelines of international conference on harmonization. The correlation coefficients of all the calibration curves were ≥0.9995 and recoveries ranged from 95.2 to 105.8% (RSD ≤ 1.95%). Relative standard deviations of intra-day, inter-day precisions and stability were ≤1.74, 1.84 and 2.8%, respectively. The quantitative results showed remarkable differences in the content of all potential osteogenic compounds in different parts of the plant as well as samples from different geographical regions. Quantitative variations studied from principal component analysis indicated tentative markers for B. monosperma cultivars which can discriminate sample of different geographical regions.

Breast cancer amelioration by Butea monosperma in-vitro and in-vivo.

ETHNOPHARMACOLOGICAL RELEVANCE: Butea monosperma belonging to family Fabaceae is used in the Indian traditional medicine (Ayurveda) for various ailments including abdominal tumors and possess anti-estrogenic activity. AIM OF THE STUDY: The present study is aimed at investigating the chemo-preventive potential of Butea monosperma in breast cancer and elucidating it's mechanism of action by assessing its effect on key processes like apoptosis, angiogenesis and metastasis. METHODS: Cytotoxic potential of methanol extract of Butea monosperma flower (MEBM) was tested in MCF-7 (estrogen receptor positive), MDA-MB-231 (triple negative) and MDA-MB-453 (HER2 positive) human breast cancer cells by MTT assay. Chemo-preventive potential was evaluated in-vivo in Methylnitrosourea (MNU) induced mammary cancer in nulliparous Sprague-Dawley rats. The mechanism for anticancer potential was screened by in-vitro studies involving Annexin V- FITC assay (apoptosis), Chick Chorioallantoic Membrane assay (angiogenesis) and Migration assay (metastasis). Statistical analysis was done by one way and two way ANOVA (for Growth Rate and feed consumption efficiency) followed by post hoc Bonferroni's test with P value < 0.05. RESULTS: It is observed that the exposure of MEBM, at various concentrations and time intervals to different cell lines, resulted in decreased cell proliferation. The IC50 value of MCF-7 cells was found significantly less than that of MDA-MB-231 and MDA-MB-453 cells, which indicated that the extract of said medicinal plant were more potent inhibitors of estrogen positive breast cancer cells than other types of breast cancer cells in vitro. Corroborative evidences were acquired in MNU actuated mammary carcinogenesis where MEBM constricted tumor parameters, decreased expression of estrogen and progesterone, nucleic acid content and increased latency period. MEBM also induced apoptosis, inhibited angiogenesis and metastasis in-vitro. CONCLUSION: Selective cytotoxic activity in MCF-7 estrogen positive breast cancer cells and inhibition of growth of mammary carcinoma in-vivo by methanol extract of Butea monosperma flowers (MEBM) suggests chemo-prevention through modulation of estrogen and progesterone receptor, apoptotic, anti-angiogenesis and anti-metastatic activity.

Effect of sodium salt of Butrin, a novel compound isolated from Butea monosperma flowers on suppressing the expression of SIRT1 and Aurora B kinase-mediated apoptosis in colorectal cancer cells.

The infrequent manifestation of SIRT1 and Aurora B kinase has shown to play a pivotal role in colorectal cancer (CRC) progression by regulating Wnt signaling pathway. The present study investigates the signaling events that regulate the modulation of SIRT1 and Aurora B kinase expression and it's mediated cell proliferation in SW480 human primary adenocarcinoma CRC cells using Butea monosperma floral compounds (BMFC). In this, cell viability, mitochondrial mediated apoptosis, cell cycle arrest and inhibition of Wnt pathway were examined. Interestingly, the active novel compound, sodium salt of butrin, from BMFC significantly enhances the apoptosis activity, where SIRT1 and Aurora B kinase were ectopically overexpressed in CRC cells. Moreover, mRNA and protein expressions analysis indicates that the expression of GSK-3ß, ß-catenin, cyclin D1, pAKT, TGF-3ß, SIRT1 and Aurora B kinase were down regulated in BMFC treated cells. These findings provide valuable information that the active BMFC having great impact on SIRT1 and Aurora B kinase mediated Wnt signaling down regulation in SW480 CRC cells.

Superbanone, A New 2-Aryl-3-benzofuranone and Other Bioactive Constituents from the Tube Roots of Butea superba.

Phytochemical investigation from the tube roots of Butea superba, led to the isolation and identification of a new 2-aryl-3-benzofuranone named superbanone (1), one benzoin, 2-hydroxy-1-(2-hydroxy-4-methoxyphenyl)-2-(4-methoxyphenyl)ethanone (2), eight pterocarpans (3 - 10), and eleven isoflavonoids (11 - 21). Compound 2 was identified for the first time as a natural product. The structure of the isolated compounds was elucidated using spectroscopic methods, mainly 1D- and 2D-NMR. The isolated compounds and their derivatives were evaluated for α-glucosidase inhibitory and antimalarial activities. Compounds 3, 7, 8, and 11 showed promising α-glucosidase inhibitory activity (IC50  = 13.71 ± 0.54, 23.54 ± 0.75, 28.83 ± 1.02, and 12.35 ± 0.36 µm, respectively). Compounds 3 and 11 were twofold less active than the standard drug acarbose (IC50  = 6.54 ± 0.04 µm). None of the tested compounds was found to be active against Plasmodium falciparum strain 94. On the basis of biological activity results, structure-activity relationships are discussed.

Inhibitory Activities of Butanol Fraction from Butea monosperma (Lam.) Taub. Bark Against Free Radicals, Genotoxins and Cancer Cells.

The present study was undertaken to investigate antioxidant, antigenotoxic, and antiproliferative activity of butanol fraction (Bmbu) from bark of medicinal plant Butea monosperma. Antioxidant potency of Bmbu was examined by various in vitro assays. It was also investigated for antigenotoxic activity using Escherichia coli. PQ37 employing SOS chromotest. Further, cytotoxic and apoptosis inducing activity of Bmbu was evaluated in MCF-7 breast cancer cells. Bmbu showed potent free radical scavenging ability in ABTS assay (IC50 56.70 µg/ml) and anti-lipid peroxidation ability (IC50 40.39 µg/ml). 4NQO and H2 O2 induced genotoxicity was suppressed by Bmbu in SOS chromotest by 74.26% and 82.02% respectively. It also inhibited the growth of MCF-7 cells with GI50 value of 158.71 µg/ml. Induction of apoptosis in MCF-7 cells by Bmbu treatment was deciphered using confocal microscopy, flow cytometry, and neutral comet assay. Bmbu treatment increased cell population in sub-G1 phase (69.6%) indicating apoptotic cells. Further, Bmbu treatment resulted in increased reactive oxygen species generation and decreased mitochondrial membrane potential indicating involvement of mitochondrial dependent pathway of apoptosis. HPLC profiling showed the presence of polyphenols such as ellagic acid, catechin, quercetin, and gallic acid as its major constituents. Consequently, it is suggested that the phytoconstituents from this plant may be further exploited for development of novel drug formulation with possible therapeutic implication.

Hepatoprotective activity of Butea monosperma bark against thioacetamide-induced liver injury in rats.

For thousands of years, the plant-based natural products have been a source of curative agents for various ailments. Butea monosperma (Fabaceae) has an important place in Indian traditional system of medicine for curing number of disorders. The present study deals with evaluation of hepatoprotective properties of ethyl acetate fraction (Beac) from B. monosperma bark in rat model. In preliminary antioxidant studies, Beac demonstrated pronounced superoxide scavenging (IC50 88.85µg/ml) and anti-lipid peroxidation (IC50 131.66µg/ml) potential. In animal studies, Beac showed protective effect against thioacetamide-induced pathophysiology in liver of male Wistar rats. The levels of different parameters related to hepatic functions were altered by thioacetamide treatment (300mg/g bw) in rats. The pre-treatment of rats with Beac (50, 100 and 200mg/kg bw) was able to normalize the biochemical markers viz. serum bilirubin, SGOT, SGPT, albumin and ALP along with liver antioxidative molecules viz. SOD, CAT, GSH and GR. Results of histopathological and colorimetric studies revealed that Beac treatment also restored the markers of fibrosis i.e. collagen and hydroxyproline towards normal level. Beac considerably inhibited thioacetamide-induced expression of p-PI3K, p-Akt and p-mTOR in hepatocytes as revealed from immunohistochemical studies. This finding is the first evidence of inhibitory action of B. monosperma bark on these pro-carcinogenic proteins. HRMS analysis revealed the presence of quercetin, buteaspermin B and ononin in Beac fraction of Butea monosperma. From the results, it can be concluded that B. monosperma bark is a rich source of phytochemicals with in vitro and in vivo protective activities which deserves further mechanistic studies for its use as a hepatoprotective agent in the prevention of hepatic inflammation and its related malignancies.

Toxicology of the aqueous extract from the flowers of Butea monosperma Lam. and it's metabolomics in yeast cells.

Due to lack of scientific evidence for the safety of Butea monosperma (Fabaceae), our study aimed to carry out its toxicological profile and to identify its metabolic pattern in yeast cell. The effect of aqueous extract of B. monosperma flower on glucose uptake in yeast cell was evaluated through optimizing pH, temperature, incubation time, substrate concentration and kinetic parameters. Further, the metabolic pattern of extract as such and in yeast cell were analyzed by gas chromatography-mass spectrometry. Mice were administered aqueous extract up to 6000 and 4000 mg/kg for acute oral and intraperitoneal toxicity, respectively, while up to 4500 mg/kg for sub-acute oral toxicity (30 days). Elongation in the lag and log phase was observed in yeast cells supplemented with extract as compared to control. A maximum of 184.9% glucose uptake was observed whereas kinetic parameters (Km and Vmax) were 1.38 and 41.91 mol/s, respectively. Out of 75 metabolites found in the extract, 14 and 18 metabolites were utilized by yeast cell after 15 and 30 min of incubation, respectively. The LD50 of extract administered through intraperitoneal route was estimated to be 3500 mg/kg. The extract did not elicit any significant difference (P ≥ 0.05) in weight gain, food consumption, water intake, hematological, biochemical parameters and histological changes as compared to the normal control. Results ascertained the safety of B. monosperma flower extract which can be explored as potential candidates for the development of anti-diabetic phytopharmaceuticals.

The synergy effect of daidzein and genistein isolated from Butea superba Roxb. on the reproductive system of male mice.

Butea superba Roxb. (BS) has been used in Thai men as an aphrodisiac, and prevent erectile dysfunction. Nevertheless, the active ingredients, dosages, have not been cleared. Hence, this study was to investigate the effect of compounds from the BS on the reproductive parameters of male mice. The results revealed that BS was extracted to afford biochanin A and genistein, which were first reported on BS, and daidzein. The mice were treated by daidzein and genistein alone and in combination. The results showed that the sperm number and motility, cholesterol and testosterone level of all isoflavones-treated groups were significantly higher than controls (p < 0.01). Obviously, daidzein plus genistein exhibited a synergistic effect, which is also the first report, and resulted in significantly displayed higher levels of these parameters compared to others. So, the synergistic activity of these isoflavones may be useful in improving libido, erectile capacity and assist infertility of poor spermatozoa in men.

An isoflavone cladrin prevents high-fat diet-induced bone loss and inhibits the expression of adipogenic gene regulators in 3T3-L1 adipocyte.

OBJECTIVE: This study evaluates the effect of isoflavone cladrin on high-fat diet (HFD)-induced bone loss and adipogenesis. METHODS: Thirty-two 4-week-old male C57BL/6J mice were divided into four groups: a standard diet group, a HFD group and HFD group with cladrin (5 and 10 mg/kg per day orally) for 12 weeks. The effect of cladrin on bone micro-architecture, bone marrow cell lineages and hyperlipidaemia were assessed. For assessing anti-adipogenic activity of cladrin, 3T3-L1 cells were used. KEY FINDINGS: Cladrin attenuated HFD-induced hyperlipidaemia and bone loss by preserving bone micro-architecture and strength. Effect of cladrin was found at the level of bone marrow progenitor cells. Gene expression profile of cladrin-treated mice bone showed upregulation of osteoblast and downregulation of adipogenic transcription factors and increased OPG/RANKL ratio. Cladrin inhibited cellular lipid accumulation through downregulation of transcription factors such as PPAR-γ and C/EBP-α and modulated the expression of major adipokines involved behind obesity stimulation without eliciting cell cytotoxicity in 3T3-L1 adipocytes. CONCLUSION: We conclude that cladrin may improve obesity-induced bone loss and hyperlipidaemia in mice fed HFD and adipogenesis in 3T3-L1 cells by modifying adipokines and could offer clinical benefits as a supplement to treat obesity-induced disorders.

A profile of protein-protein interaction: Crystal structure of a lectin-lectin complex.

Proteins may utilize complex networks of interactions to create/proceed signaling pathways of highly adaptive responses such as programmed cell death. Direct binary interactions study of proteins may help propose models for protein-protein interaction. Towards this goal we applied a combination of thermodynamic kinetics and crystal structure analyses to elucidate the complexity and diversity in such interactions. By determining the heat change on the association of two galactose-specific legume lectins from Butea monosperma (BML) and Spatholobus parviflorus (SPL) belonging to Fabaceae family helped to compute the binding equilibrium. It was extended further by X-ray structural analysis of BML-SPL binary complex. In order to chart the proteins interacting mainly through their interfaces, identification of the nature of forces which stabilized the association of the lectin-lectin complex was examined. Comprehensive analysis of the BMLSPL complex by isothermal titration calorimetry and X-ray crystal structure threw new light on the lectin-lectin interactions suggesting of their use in diverse areas of glycobiology.