Biochanin Research Articles

Phytoestrogen isoflavone intervention to engage the neuroprotective effect of glutamate oxaloacetate transaminase against stroke.

In the pathophysiologic setting of cerebral ischemia, excitotoxic levels of glutamate contribute to neuronal cell death. Our previous work demonstrated the ability of glutamate oxaloacetate transaminase (GOT) to metabolize neurotoxic glutamate in the stroke-affected brain. Here, we seek to identify small-molecule inducers of GOT expression to mitigate ischemic stroke injury. From a panel of phytoestrogen isoflavones, biochanin A (BCA) was identified as the most potent inducer of GOT gene expression in neural cells. BCA significantly increased GOT mRNA and protein expression at 24 h and protected against glutamate-induced cell death. Of note, this protection was lost when GOT was knocked down. To validate outcomes in vivo, C57BL/6 mice were intraperitoneally injected with BCA (5 and 10 mg/kg) for 4 wk and subjected to ischemic stroke. BCA levels were significantly increased in plasma and brain of mice. Immunohistochemistry demonstrated increased GOT protein expression in the brain. BCA attenuated stroke lesion volume as measured by 9.4T MRI and improved sensorimotor function-this protection was lost with GOT knockdown. BCA increased luciferase activity in cells that were transfected with the pERRE3tk-LUC plasmid, which demonstrated transactivation of GOT. This increase was lost when estrogen-related receptor response element sites were mutated. Taken together, BCA represents a natural phytoestrogen that mitigates stroke-induced injury by inducing GOT expression.-Khanna, S., Stewart, R., Gnyawali, S., Harris, H., Balch, M., Spieldenner, J., Sen, C. K., Rink, C. Phytoestrogen isoflavone intervention to engage the neuroprotective effect of glutamate oxaloacetate transaminase against stroke.

Antioxidant Effects of Biochanin A in Streptozotocin Induced Diabetic Rats

ABSTRACT Bioflavonoid-containing diets have been reported to be beneficial in diabetes. In the current study, the effect of Biochanin A (BCA) on blood glucose, antioxidant enzyme activities and oxidative stress markers in diabetic rats were investigated. 30 male Wistar rats were divided into five groups. Two of them were selected as control; group1: control (receiving 0.5%DMSO), and group2: Control+BCA (receiving 10 mg/kg.bw BCA). Diabetes was induced in other rats with injection of (55 mg/kg.bw) streptozotocin; group3: diabetic control (receiving 0.5%DMSO), groups 4 and 5 were treated with 10 and 15 mg/kg.bw BCA respectively. After 6 weeks the following results were obtained. Fasting blood glucose (FBG), Triglyceride (TG), total cholesterol (TC), low density lipoprotein cholesterol (LDL-C), very low density lipoprotein cholesterol (VLDL-C) and malondialdehyde (MDA) levels significantly increased and body weight, high density lipoprotein cholesterol (HDL-C), superoxide dismutase (SOD) and catalase (CAT) activity and total antioxidant status (TAS) significantly decreased in diabetic rats as compared to control rats. Oral administration of BCA in 10 and 15 mg/kg.bw, FBG, TG, TC, LDL-C, VLDL-C were decreased significantly in all treated rats. MDA was decreased in all treated rats but it was significant just in 15 mg/kg.bw BCA. HDL, CAT, SOD, and TAS were significantly increased in treated group with 15 mg/kg.bw. The obtained results indicated hypoglycemic and hypolipidemic effect of BCA. Also BCA reduced oxidative stress in diabetic rats.

Potential Lipid-Lowering Mechanisms of Biochanin A.

Extensive studies have demonstrated that biochanin A (BCA) has a significant hypolipidemic effect. However, its mechanism of action is not clear. In this context, the effect of BCA on a high-fat diet (HFD)-induced hyperlipidemia in mice was determined. The results showed that treatment with a medium dose of biochanin A (BM) significantly decreased low-density lipoprotein cholesterol (LDL-C) 85% (from 1.196 ± 0.183 to 0.181 ± 0.0778 mM) and total cholesterol (TC) 39% (from 5.983 ± 0.128 to 3.649 ± 0.374 mM) levels, increased lipoprotein lipase (LPL) 96% (from 1.421 ± 0.0982 to 2.784 ± 0.177 U/mg protein) and hepatic triglyceride lipase (HTGL) 78% (from 1.614 ± 0.0848 to 2.870 ± 0.0977 U/mg protein) activities, significantly improved fecal lipid levels, and lowered the epididymal fat index in hyperlipidemic mice compared with the HFD control mice (p < 0.05). In vitro, the high antioxidant capacity of BCA was determined by the FRAP assay, ABTS•+ scavenging method, and an ROS assay. In RAW 264.7 macrophages, a dose of 10 μM BCA significantly increased the cholesterol efflux by 18.7% compared with the control cells. Moreover, molecular docking of BCA on cholesterol ester transfer protein (CETP) (Asn24 and Thr27 at the N-terminal; Ala274 and Phe270 at the C-terminal) gave new insights into the role of BCA in preventing cholesterol ester transport.

Investigating Interaction Between Biochanin A and Human Serum Albumin by Multi-spectroscopic and Molecular Simulation Methods

Biochanin A (BCA), the most abundant isoflavone in chickpeas, presents a wide range of biological activities, such as hypolipidaemic, anti-oxidative, anti-proliferative, and estrogen-like effects. We investigated the interaction between BCA and human serum albumin (HSA) via several techniques. UV–Vis absorption spectroscopy verified the conformational variation of HSA after BCA addition, and fluorescence spectroscopy revealed the relevant binding parameters. Circular dichroism spectroscopy was used to estimate the secondary structural changes of HSA with and without BCA. Molecular docking and dynamics simulations were then applied to study the characteristics of HSA with BCA. Energy decomposition analysis was used to prove that Trp214 in subdomain IIA of HSA is the most likely binding site of BCA. Van der Waals forces and hydrophobic interactions may play important roles during the binding process. All of our results showed that BCA presents significant binding affinity to HSA, thus confirming that the role of HSA has as an efficient transporter of biomolecules.

Effect of biochanin A on corn grain (Zea mays) fermentation by bovine rumen amylolytic bacteria.

The objective was to determine the effect of biochanin A (BCA), an isoflavone produced by red clover (Trifolium pratense L.), on corn fermentation by rumen micro-organisms. When bovine rumen bacterial cell suspensions (n=3) were incubated (24h, 39°C) with ground corn, amylolytic bacteria including group D Gram-positive cocci (GPC; Streptococcus bovis; enterococci) proliferated, cellulolytic bacteria were inhibited, lactate accumulated and pH declined. Addition of BCA (30μgml-1 ) inhibited lactate production, and pH decline. BCA had no effect on total amylolytics, but increased lactobacilli and decreased GPC. The initial rate and total starch disappearance was decreased by BCA addition. BCA with added Strep. bovis HC5 supernatant (containing bacteriocins) inhibited the amylolytic bacteria tested (Strep. bovis JB1; Strep. bovis HC5; Lactobacillus reuteri, Selenemonas ruminatium) to a greater extent than either addition alone. BCA increased cellulolytics and dry matter digestibility of hay with corn starch. These results indicate that BCA mitigates changes associated with corn fermentation by bovine rumen bacteria exvivo. BCA could serve as an effective mitigation strategy for rumen acidosis. Future research is needed to evaluate the effect of BCA on mitigating rumen acidosis invivo.

Enhancing the oral bioavailability of biochanin A by encapsulation in mixed micelles containing Pluronic F127 and Plasdone S630.

Biochanin A (BCA), a natural dietary isoflavone, has been reported to show anticancer activities. However, its low biological availability and poor aqueous solubility limit its usefulness as a chemotherapeutic agent. We developed BCA-loaded micelles with Pluronic F127 and Plasdone S630 (BCA-FS). The optimized, spherical-shaped BCA-FS was obtained at a ratio of 1:1 (F127:S630). The particle size was 25.17±1.2 nm, and the zeta potential was −10.9±0.24 mV. BCA solubility in water increased to 5.0 mg/mL after encapsulation, and the drug-loading efficiency was 5.88%±0.76%. In vitro release experiments showed a delayed release of BCA from the mixed micelles. Furthermore, the BCA absorption permeability across a Caco-2 cell monolayer from the apical side to the basolateral side increased by 54% in BCA-FS. A pharmacokinetics evaluation showed a 2.16-fold increase in the relative oral bioavailability of BCA-FS compared with raw BCA, indicating that the mixed micelles may promote absorption in the gastrointestinal tract. A gastrointestinal safety assay was used to assess the reliability and safety of BCA-FS. On the basis of these findings, we conclude that this simple nanomicelle system could be leveraged to deliver BCA and other hydrophobic drugs.

The Benzopyrone Biochanin-A as a reversible, competitive, and selective monoamine oxidase B inhibitor

BackgroundMonoamine oxidase-B (MAO-B) inhibitors are widely used in the treatment of Parkinson’s disease. They increase vital monoamine neurotransmitters in the brain. However, there is a need for safer natural reversible MAO inhibitors with MAO-B selectivity. Our previous studies showed that Psoralea corylifolia seeds (PCS) extract contains compounds that inhibit monoamine oxidase-B.MethodsIn this study, six of PCS constituents sharing a benzopyrone structure were investigated. The compounds Biochanin-A (BIO-A), isopsoralen, 6-prenylnaringenin, neobavaisoflavone, psoralen, and psoralidin, were tested for their ability to inhibit recombinant human MAO-A and B (hMAO-A and hMAO-B) isozymes. The ability of these compounds to inhibit MAO-A and MAO-B were compared to that of PCS ethanolic extract (PCSEE) using spectrophotometric assays and confirmed by luminescence assays. The highly potent and selective MAO-B inhibitor, BIO-A, was further investigated for both isozymes reversibility and enzyme kinetics. Molecular docking studies were used to predict the bioactive conformation and molecular interactions of BIO-A with both isozymes.ResultsThe data obtained indicate that benzopyrones inhibited hMAO-A and hMAO-B with different degrees as confirmed with the luminescence assay. BIO-A inhibited hMAO-B with high potency and selectivity in the present study (IC50 = 0.003 μg/mL) and showing 38-fold more selectivity than PCSEE (hMAO-B IC50 = 3.03 μg/mL, 17-fold selectivity) without affecting hydrogen peroxide. Furthermore, BIO-A reversibly and competitively inhibited both hMAOs with significantly lower inhibitory constant (Ki) in hMAO-B (3.8 nM) than hMAO-A (99.3 nM). Our docking studies indicated that the H-bonds and hydrophobic interactions at the human MAO-A and MAO-B active sites contributed to the reversibility and selectivity of BIO-A.ConclusionsThe data obtained indicate that BIO-A is a potent, reversible and selective MAO-B inhibitor and may be recommended for further investigation in its possible use in the therapeutic management of Parkinson’s and Alzheimer’s diseases.

Biochanin A (an Isoflavone Produced by Red Clover) Promotes Weight Gain of Steers Grazed in Mixed Grass Pastures and Fed Dried‐Distillers’ Grains

Biochanin A (BCA) is an isoflavone produced by red clover (Trifloium pratense L.) that can inhibit hyper‐ammonia‐producing bacteria (HAB) to reduce deamination in the rumen and increase the feed amino acids available for gastric digestion. An ex vivo experiment was conducted to evaluate the effect of dried distiller's grains (DDG) and BCA (30 μg mL−1) on HAB in bovine rumen fluid. Following a 24‐h incubation, HAB and ammonia concentrations in the media increased with addition of DDG. However, DDG fermentations with BCA had 100‐fold fewer HAB and 37% less ammonia than DDG‐only fermentations. A grazing experiment was conducted with crossbred steers grazing a mixture of predominately endophyte‐free tall fescue [Lolium arundinaceum (Schreb.) Darbysh], Kentucky bluegrass (Poa pratense L.), and orchardgrass (Dactylis glomerata L.) in the early and late growing season. Average daily gains (ADG) were compared among treatments of pasture‐only control, daily feeding of 1.4 kg DDG steer−1, and daily feeding of 1.4 kg DDG plus BCA (6.3 g steer−1, representing ∼30% red clover diet). Averaged over the early and late growing seasons, feeding DDG (0.83 ± 0.05 kg d−1) increased ADG by 15% over the pasture‐only treatment (0.72 ± 0.03 kg d−1), but addition of BCA to the DDG (0.93 ± 0.04 kg d−1) provided an additive increase in ADG of 29% over the control treatment. Results of the ex vivo and grazing experiments provide evidence that BCA inhibits HAB in the rumen and that the reduced deamination in the rumen can enhance weight gain performance of steers.

Biochanin A improves hepatic steatosis and insulin resistance by regulating the hepatic lipid and glucose metabolic pathways in diet-induced obese mice.

Natural compounds that regulate peroxisome proliferator-activated receptor alpha (PPARα) have been reported to have beneficial effects in obesity-mediated metabolic disorders. In this study, we demonstrated that biochanin A (BA), an agonist of PPAR-α, improved hepatic steatosis and insulin resistance by regulating hepatic lipid and glucose metabolism. C57BL/6 mice were fed a normal chow diet, a high-fat diet (HFD), and an HFD supplemented with 0.05% BA for 12 weeks. Histological and biochemical examinations indicated that BA prevented obesity-induced hepatic steatosis and insulin resistance in HFD-fed mice. BA stimulated the transcriptional activation of PPAR-α in vitro and increased the expression of PPAR-α and its regulatory proteins in the liver. CE-TOF/MS analyses indicated that BA administration promoted the recovery of metabolites involved in phosphatidylcholine synthesis, lipogenesis, and beta-oxidation in the livers of obese mice. BA also suppressed the levels of gluconeogenesis-related metabolites and the expression of the associated enzymes, glucose 6-phosphatase and pyruvate kinase. Taken together, these results showed that BA ameliorated metabolic disorders such as hepatic steatosis and insulin resistance by modulating lipid and glucose metabolism in diet-induced obesity. Thus, BA may be a potential therapeutic agent for the prevention of obesity-mediated hepatic steatosis and insulin resistance.

Evaluations of the Inhibitory effects of Psoralea Corylifolia Benzopyrone Derivatives in Human Monoamine Oxidase A and B

The classical inhibitors of monoamine oxidases (MAOs) are effectively used in the therapeutic management of neurological disorders with rare, but serious, side‐effects. In our investigation for safe and natural MAO‐B inhibitors, Psoralea corylifolia Linn seeds (PCS) ethanolic extract (PCSEE) and seven of its benzopyrone constituents were investigated for their recombinant human (h) MAO‐A and B inhibition and selectivity. The extensively extracted PCSEE selectively inhibited MAO‐B (IC50 of 3.03 μg/ml) more than A. PCS benzopyrones showed weak to potent hMAO‐A and B hydrogen peroxide (H2O2) byproduct inhibition. MAO‐A inhibition potencies of PCSEE common compounds were in biochanin‐A (BIO‐A) &gt; isopsoralen &gt; genistein &gt; 6‐prenylnaringenin &gt; neobavaisoflavone &gt; psoralen. Meanwhile, MAO‐B inhibition potencies were in BIO‐A &gt; genistein &gt; 6‐prenylnaringenin &gt; isopsoralen &gt; neobavaisoflavone &gt; psoralen. Psoralidin was inactive in both isozymes. Compared to PCSEE and its tested benzopyrones, BIO‐A and genistein, showed the most potent hMAO‐A and B inhibition, equal or more potent than PCSEE. Furthermore, the most potent MAOI, BIO‐A, reversibly inhibited both isozymes. BIO‐A reversibility was supported by docking and kinetics studies. BIO‐A was predicted to form reversible interactions with FAD of MAO‐A, and with threonine 201 and p phenylalanine 168 in MAO‐B active site. Michaelis‐Menten kinetics studies also showed a competitive mode of BIO‐A inhibition on both isozymes. The obtained data indicate that PCSEE benzopyrones may contribute to its MAOs inhibitory activities and could be utilized as a natural neuroprotective source in neurological disorders.Support or Funding InformationSupported by NIH grants from NIMHD G12 MD007582 and P20 MD006738

Biochanin A Ameliorates Arsenic-Induced Hepato- and Hematotoxicity in Rats.

Biochanin A (BCA) is a natural organic compound of the phytoestrogenic isoflavone class that has antioxidant and metal chelator properties in the presence of transition metal ions, however, its efficacy in animal models is still obscure. Therefore, the objective of this study was to investigate the protective effects of BCA against arsenic-induced hepatic injury and hematotoxicity in rats. The results suggest that arsenic intoxicated rats showed significantly higher levels of plasma hepatic markers than normal control rats. Furthermore, an increase in lipid peroxidation with depletion of reduced glutathione (GSH) and activities of superoxide dismutase (SOD) and catalase (CAT) occurred in the livers of rats exposed to arsenic. Administration of BCA (20 mg/kg·bw/day) and selenium (3 mg/kg·bw/day) resulted in a significant reversal of hepatic and oxidative stress markers in arsenic-intoxicated rats. A low dose of BCA (10 mg/kg·bw/day) did not show any preventive effect, while a high dose of BCA (40 mg/kg·bw/day) partially prevented all hepatotoxicity events. These biochemical perturbations were supported by histopathological observations of the liver. Our results suggest that administration of BCA (20 mg/kg·bw/day) attenuated the arsenic hepatotoxicity, a property that could contribute to the therapeutic approaches for chronic liver diseases.

In vivo prediction of CYP-mediated metabolic interaction potential of formononetin and biochanin A using in vitro human and rat CYP450 inhibition data.

Formononetin (FMN) and Biochanin A (BCA) are the principal isoflavones present in commercially available extracts of red clover that are widely been consumed for various health benefits. We investigated the in vitro effects of FMN and BCA on catalytic activity of human/rat cytochrome P450 enzymes to assess the drug interaction potential of red clover. IC50 and Ki values of FMN and BCA for CYPs were determined in human/rat liver microsomes. FMN and BCA showed concentration-dependent inhibition of CYP1A2 activity with IC50 values of 13.42 and 24.98μM in human liver microsomes and 38.57 and 11.86μM in rat liver microsomes, respectively. The mode of inhibition of human CYP1A2 by FMN was found to be competitive with apparent Ki value of 10.13±1.96μM. FMN also inhibited human CYP2D6. BCA exerted moderately inhibitory effects on human CYP2C9. The predicted in vivo inhibition for CYP1A2 was insignificant (R value <1.1) at hepatic level while at intestinal level, it was significant (R value >11). The inhibitory effects on other CYPs were found to be minimal. Red clover may be considered safe to be consumed along with co-prescribed medications; however, precaution must be taken while co-administering it with CYP1A2 substrates.

Isolation of biochanin A, an isoflavone, and its selective sensing of copper(II) ion

Biochanin A (BnA) a natural o-methylated isoflavone, is isolated from the leaves of Eupatorium adenaphorum and is effectively utilized as a sensor probe for copper ion with good selectivity and sensitivity. The effect with the presence of other metal ions is also investigated and BnA is found to be selective towards Cu2+ ion over other metal ions such as Zn2+, Sn2+, Ni2+, Ag+, Bi2+, Pb2+, Co3+, In3+, Mn2+, Fe2+, Li+, Hg2+ and Ca2+ at pH 7.1 with a detection limit upto 1.0×10−6molL−1. The sensor probe BnA exhibits selective fluorescence quenching via an intramolecular charge transfer (ICT) upon addition of Cu2+. The proposed mechanism is strongly supported by TD-DFT calculations and the 1:1 complex (BnA–Cu2+) formation is confirmed by ESI-MS and Job's plot.

Therapeutic efficacy of biochanin A against arsenic-induced renal and cardiac damage in rats

The present study was investigated the effects of biochanin A (BCA) on arsenic toxicity in rats. For this purpose, rats were orally treated with arsenic in the form of sodium meta-arsenite alone (10mg/kg body weight (bw)/day) and co-administered selenium (10mg/kgbw/day) and BCA at different doses (10, 20 and 40mg/kgbw/day) for 6 weeks. Arsenic altered the oxidative stress indices in both renal and cardiac tissues. There was an increase in plasma renal markers, triglyceride, lipoproteins with no alterations in cholesterol levels were noted in arsenic-intoxicated rats. Non-significant changes of phospholipids and free fatty acids levels in the tissues of arsenic-exposed rats. The biochemical disturbances were well correlated with the histological findings in the kidney, but not in the heart. The administration of BCA and selenium significantly reversed the alterations in the above-mentioned parameters in arsenic-intoxicated rats. Our findings revealed the beneficial effects of BCA against arsenic toxicity.

Surface molecularly imprinted silica for selective solid-phase extraction of biochanin A, daidzein and genistein from urine samples

Selective molecularly imprinted silica polymer (SiO2MIP) for extraction of biochanin A, daidzein and genistein was synthesized using the surface molecular imprinting technique with the silica gel as a support. Biochanin A (BCA) was used as a template, 3-aminopropyltriethoxysilane (APTES) as a functional monomer, and tetraethoxysilicane (TEOS) as a cross-linker. Non-imprinted polymer with the sol–gel process (SiO2NIP) was also prepared for comparison. The synthesized polymers were characterized by Fourier transform infrared spectrometry (FTIR), scanning electron microscopy (SEM) and a standard Brunauer-Emett-Teller (BET) and Barret–Joyner–Halenda (BJH) analysis. The obtained results indicated the structural differences between imprinted and non-imprinted polymers. Finally, the SiO2MIP and SiO2NIP were adopted as the adsorbents of solid phase extraction for isolation and preconcentration of biochanin A and its structural analogues–daidzein and genistein from aqueous and urine samples. The performance analysis revealed that SiO2MIP displayed better affinity to the three investigated isoflavones compared with SiO2NIP. The recoveries of spiked samples for studied analytes ranged from 65.7% to 102.6% for molecularly imprinted silica polymer and 8.9–16.0% for non-imprinted sorbents.

Isoflavones enhance interleukin-17 gene expression via retinoic acid receptor-related orphan receptors α and γ

The retinoic acid receptor-related orphan receptors α and γ (RORα and RORγ), are key regulators of helper T (Th)17 cell differentiation, which is involved in the innate immune system and autoimmune disorders. In this study, we investigated the effects of isoflavones on RORα/γ activity and the gene expression of interleukin (IL)-17, which mediates the function of Th17 cells. In doxycycline-inducible CHO stable cell lines, we found that four isoflavones, biochanin A (BA), genistein, formononetin, and daidzein, enhanced RORα- or RORγ-mediated transcriptional activity in a dose-dependent manner. In an activation assay of the Il17a promoter using Jurkat cells, these compounds enhanced the RORα- or RORγ-mediated activation of the Il17a promoter at concentrations of 1 × 10(-6)M to 1 × 10(-5)M. In mammalian two-hybrid assays, the four isoflavones enhanced the interaction between the RORα- or RORγ-ligand binding domain and the co-activator LXXLL peptide in a dose-dependent manner. In addition, these isoflavones potently enhanced Il17a mRNA expression in mouse T lymphoma EL4 cells treated with phorbol myristate acetate and ionomycin, but showed slight enhancement of Il17a gene expression in RORα/γ-knockdown EL4 cells. Immunoprecipitation and immunoblotting assays also revealed that BA enhanced the interaction between RORγt and SRC-1, which is a co-activator for nuclear receptors. Taken together, these results suggest that the isoflavones have the ability to enhance IL-17 gene expression by stabilizing the interactions between RORα/γ and co-activators. This also provides the first evidence that dietary chemicals can enhance IL-17 gene expression in immune cells.

PEGylated nanostructured lipid carriers (PEG–NLC) as a novel drug delivery system for biochanin A

With the aim to develop a lipid nanoparticle for biochanin A (BCA) by emulsion-evaporation and low temperature-solidification technique. The results revealed that BCA–PEG–NLC not only have small mean particle (148.5 ± 2.88 nm) with narrow polydispersity index (PI) (0.153 ± 0.01), encapsulation capacity (99.62 ± 0.06%), payload (9.06 ± 0.01%), zeta potential (−19.83 ± 1.19 mV), but also slower release rate compared with BCA suspension over 48 h by the dialysis method (n=3). The crystallinity of lipid matrix within BCA–PEG–NLC was evaluated by differential scanning calorimetry (DSC) which verified the BCA successfully into the nanoparticles. Particularly, in pharmacokinetic, the BCA–PEG–NLC of Cmax values and AUC (area under curve) was higher than BCA suspension (approximately 15.8 and 2.9 times, respectively), meanwhile, the mean residence time (MRT) was significantly longer. Furthermore, in vitro cytotoxicity BCA–PEG–NLC showed higher cytotoxicity against MCF-7 cell line compared with BCA suspension. This study suggested that PEG–NLC is a novel anti-cancer nanoparticle, which could provide attractive treatment for a wide variety of tumors and improved the oral bioavailability of poorly water-soluble drug.

The impact of dietary isoflavonoids on malignant brain tumors.

Poor prognosis and limited therapeutic options render malignant brain tumors one of the most devastating diseases in clinical medicine. Current treatment strategies attempt to expand the therapeutic repertoire through the use of multimodal treatment regimens. It is here that dietary fibers have been recently recognized as a supportive natural therapy in augmenting the body's response to tumor growth. Here, we investigated the impact of isoflavonoids on primary brain tumor cells. First, we treated glioma cell lines and primary astrocytes with various isoflavonoids and phytoestrogens. Cell viability in a dose-dependent manner was measured for biochanin A (BCA), genistein (GST), and secoisolariciresinol diglucoside (SDG). Dose–response action for the different isoflavonoids showed that BCA is highly effective on glioma cells and nontoxic for normal differentiated brain tissues. We further investigated BCA in ex vivo and in vivo experimentations. Organotypic brain slice cultures were performed and treated with BCA. For in vivo experiments, BCA was intraperitoneal injected in tumor-implanted Fisher rats. Tumor size and edema were measured and quantified by magnetic resonance imaging (MRI) scans. In vascular organotypic glioma brain slice cultures (VOGIM) we found that BCA operates antiangiogenic and neuroprotective. In vivo MRI scans demonstrated that administered BCA as a monotherapy was effective in reducing significantly tumor-induced brain edema and showed a trend for prolonged survival. Our results revealed that dietary isoflavonoids, in particular BCA, execute toxicity toward glioma cells, antiangiogenic, and coevally neuroprotective properties, and therefore augment the range of state-of-the-art multimodal treatment approach.

Biochanin-A ameliorates behavioural and neurochemical derangements in cognitive-deficit mice for the betterment of Alzheimer’s disease

Biochanin-A (BCA), a potent phytoconstituent, has been previously used as an antitumour, a dopaminergic neuron protective agent, an antioxidant, an anticholinergic and on other pharmacological activities including neuroprotection. The present study was aimed to evaluate the behavioural and neurochemical evidence of BCA in cognitive-deficit mice in scopolamine challenged and natural aged-induced amnesia models in young and aged mice, respectively. BCA has exhibited decrease in the transfer latency and increase in step through latency significantly (p < 0.001) in scopolamine-treated and natural aged mice of exteroceptive behavioural models such as elevated plus maze and passive shock avoidance paradigm, respectively. A decrease in acetylcholinesterase activity of whole brain was seen in scopolamine and aged mice with standard piracetam (Pira; p < 0.001) and BCA in dose-dependent manner. The antioxidant property of BCA was proven by increase in GSH (p < 0.01) and decrease in thiobarbituric acid reactive substances level significantly in a scopolamine-challenged and aged mice. The scopolamine-treated mice exhibited significant (p < 0.01) increase in the content of noradrenalin and dopamine, which is a sign of dementia, and these excess increased neurotransmitters were reversed by BCA 40 mg kg(-1) (p < 0.05), BCA 20 mg kg(-1) (p > 0.05), BCA 10 mg kg(-1) (p < 0.05) and standard Pira (p < 0.05) when compared with scopolamine group. Furthermore, in histopathology of hippocampus, the Pira and BCA-treated mice were protected from the formation of pyknotic neurons, increases in the viable cells count and decreases in the number of degenerative cells compared with the scopolamine group. Hence, BCA could be potential enough for the betterment of Alzheimer's disease.

Biochanin A Protects against Acute Carbon Tetrachloride-Induced Hepatotoxicity in Rats

Biochanin A (BCA) is an isoflavone found in red clover possessing multiple pharmacological activities including antimicrobial, antioxidant, and anticancer ones. The present study aimed to assess its hepatoprotective potential at different doses in a carbon tetrachloride (CCl4)-induced hepatotoxicity model in rats. The effects on hepatic injury were explored by measuring serum levels of alanine aminotransferase, aspartate aminotransferase, and alkaline phosphatase. Furthermore, the serum levels of glucose, urea, creatinine, total bilirubin, total proteins, triglycerides, and total cholesterol were determined. The metabolic capacity of the liver was assessed by measuring changes in cytochrome P450 2E1 activity. The underlying mechanisms were substantiated by measuring oxidative stress markers as catalase, superoxide dismutase, glutathione peroxidase, glutathione transferase, glutathione reductase, reduced glutathione, total antioxidant capacity, and lipid peroxidation, as well as inflammation markers such as nitric oxide, inducible nitric oxide synthase, cyclooxygenase2, tumor necrosis factor-α, and leukocyte-common antigen. The results were confirmed by histopathological examination, and the median lethal dose was determined to confirm the safety of the drug. BCA successively protected against CCl4-induced damage, normalizing many parameters to that of the control group. The study indicates that BCA possesses multimechanistic hepatoprotective activity that can be attributed to its antioxidant, anti-inflammatory, and immunomodulatory actions.