Isobavachin, a main bioavailable compound in Psoralea corylifolia, alleviates lipopolysaccharide-induced inflammatory responses in macrophages and zebrafish by suppressing the MAPK and NF-κB signaling pathways.

ETHNOPHARMACOLOGICAL RELEVANCE: Psoralea corylifolia L. (PC) is widely used in traditional medicines to treat inflammatory and infectious diseases. Isobavachin (IBC) is a bioavailable prenylated flavonoid derived from PC that has various biological properties. However, little information is available on its anti-inflammatory effects and mechanisms of action. AIM OF THE STUDY: In this study, we aimed to determine the anti-inflammatory effects of IBC in vitro and in vivo by conducting a mechanistic study using murine macrophages. MATERIALS AND METHODS: We evaluated the modulatory effects of IBC on the production of pro-inflammatory cytokines and mediators in murine macrophages. In addition, we examined whether IBC inhibits lipopolysaccharide (LPS)-induced inflammatory responses in a zebrafish model. Alterations in inflammatory response-associated genes and proteins were determined using quantitative reverse transcriptional polymerase chain reaction (RT-qPCR) and Western blotting analysis. RESULTS: IBC markedly reduced the overproduction of inflammatory mediators, pro-inflammatory cytokines, inducible nitric oxide synthase (iNOS), cyclooxygenase-2 (COX-2), phosphorylation of mitogen-activated protein kinase (MAPK) and nuclear translocation of nuclear factor-kappa B (NF-κB) in macrophages induced by lipopolysaccharides (LPS). In addition, excessive NO, ROS, and neutrophil level induced by LPS, were suppressed by IBC treatment in a zebrafish inflammation model. CONCLUSIONS: Collectively, bioavailable IBC inhibited on the inflammatory responses by LPS via MAPK and NF-κB signaling pathways in vitro and in vivo, suggesting that it may be a potential modulatory agent against inflammatory disorders.

Network pharmacology-based identification of bioavailable anti-inflammatory agents from Psoralea corylifolia L. in an experimental colitis model.

ETHNOPHARMACOLOGICAL RELEVANCE: In traditional oriental medicine, the dried seeds of Psoralea corylifolia L. (PC) have been used to treat various diseases, including gastrointestinal, urinary, orthopedic, diarrheal, ulcer, and inflammatory disorders. AIM OF THE STUDY: Although its various biological properties are well-known, there is no information on the therapeutic effects and bioavailable components of PC against inflammatory bowel disease. Therefore, we focused on the relationship between hydroethanolic extract of PC (EPC) that ameliorates colitis in mice and bioactive constituents of EPC that suppress pro-inflammatory cytokines in macrophages. MATERIALS AND METHODS: We investigated the therapeutic effects of EPC in a dextran sulfate sodium-induced colitis mouse model and identified the orally absorbed components of EPC using UPLC-MS/MS analysis. In addition, we evaluated and validated the mechanism of action of the bioavailable constituents of EPC using network pharmacology analysis. The effects on nitric oxide (NO) and inflammatory cytokines were measured by Griess reagent and enzyme linked immunosorbent assay in lipopolysaccharide (LPS)-induced macrophages. RESULTS: In experimental colitis, EPC improved body weight loss, colon length shortening, and disease activity index. Moreover, EPC reduced the serum levels of pro-inflammatory cytokines and histopathological damage to the colon. Network pharmacological analysis identified 13 phytochemicals that were bioavailable following oral administration of EPC, as well as their potential anti-inflammatory effects. 11 identified EPC constituents markedly reduced the overproduction of NO, tumor necrosis factor-α, and/or interleukin-6 in macrophages induced by LPS. The LPS-induced expression of the nuclear factor kappa-light-chain-enhancer of activated B cells reporter gene was reduced by the 4 EPC constituents. CONCLUSIONS: The results indicate that the protective activity of EPC against colitis is a result of the additive effects of each constituent on the expression of inflammatory cytokines. Therefore, it suggests that 11 bioavailable phytochemicals of EPC could aid in the management of intestinal inflammation, and also provides useful insights into the clinical application of PC for the treatment of inflammatory bowel diseases.

Individual and combined effects of chromium and ultraviolet-B radiation on defense system, ultrastructural changes, and production of secondary metabolite psoralen in a medicinal plant Psoralea corylifolia L.

The present study focuses on the effects of individual and combined stress of chromium (Cr) and ultraviolet-B (UV-B) radiation on Psoralea corylifolia L. The experiment comprised four sets: (i) control, (ii) eUV-B (elevated UV-B i.e., ambient + 7.2 kJ m-2 day-1 UV-B), (iii) Cr (chromium; 30 mg kg-1 soil), and (iv) Cr + eUV-B (chromium and elevated UV-B; Cr 30 mg kg-1 and ambient + 7.2 kJ m-2 day-1 UV-B). The eUV-B and Cr individually and in combination showed the variable responses on ultrastructure, physiology and biomass however, the impact was more prominent under individual Cr treatment followed by Cr + eUV-B and eUV-B. Higher bioconcentration factor and the lowered translocation factor consequently led to a higher reduction in the below ground biomass and the lesser reduction in above ground biomass under Cr + eUV-B treatment as compared to individual Cr treatment. In addition, higher induction in the enzymatic (glutathione reductase, ascorbate peroxidase, superoxide dismutase, and glutathione-S-transferase) and non-enzymatic antioxidants (glutathione reduced) were found to be responsible for efficient scavenging of hydrogen peroxide and superoxide radical leading to lowered MDA content under combined treatment as compared to Cr treatment. Deposition of Cr as electron dense granules in the cytoplasm, vacuoles, and cell wall under Cr and Cr + eUV-B is contemplated as one of the cellular mechanisms of P. corylifolia against the toxicity of Cr. Psoralen increased under all treatments with a maximum increase under Cr + eUV-B treatment. Taken together our results accentuated that P. corylifolia can be grown in an area contaminated with Cr and has a higher influx of UV-B for the attainment of psoralen considering its pharmaceutical perspectives.

Psoralea corylifolia L. seed extract attenuates dexamethasone-induced muscle atrophy in mice by inhibition of oxidative stress and inflammation.

ETHNOPHARMACOLOGICAL RELEVANCE: The seeds of Psoralea corylifolia (PCS), also called "Boh-Gol-Zhee" in Korean, have been used in traditional medicine. PCS is effective for the treatment of vitiligo, cancer, inflammatory diseases, neurodegenerative diseases, kidney diseases, and musculoskeletal diseases. AIM OF THE STUDY: In this study, we validated the beneficial effects of PCS extract on dexamethasone (DEX)-induced muscle atrophy in mice. MATERIALS AND METHODS: DEX (20 mg/kg/day, 10 days) was intraperitoneally injected into C57BL/6 male mice to induce muscular atrophy. Oral administration of PCS extract (200 or 500 mg/kg/day) was started 2 days before DEX injection and continued for 12 days. RESULTS: PCS extract inhibited DEX-induced decrease in body and muscle weight, grip strength, and cross-sectional area of the tibialis anterior. PCS extract significantly increased the mRNA and protein expression levels of myosin heavy chain 1, 2A, and 2X in DEX-administered mice. DEX administration significantly increased the levels of muscle atrophy factors atrogin-1, muscle RING-finger protein-1, and myostatin, which were inhibited by the PCS extract. Additionally, PCS extract increased the expression of muscle regeneration factors, such as myoblast determination protein 1, myogenin, and embryonic myosin heavy chain, and muscle synthesis markers, such as protein kinase B and mammalian target of rapamycin signaling molecules. PCS extract also significantly decreased the DEX-induced production of 4-hydroxynonenal, an oxidative stress marker. Furthermore, PCS extract recovered superoxide dismutase 2, glutathione peroxidase, and catalase activities, which were significantly reduced by DEX administration. Moreover, DEX-induced activation of nuclear factor kappa-light-chain-enhancer of activated B cells and expression of cytokines, such as tumor necrosis factor α and monocyte chemoattractant protein-1, significantly decreased after PCS extract administration. CONCLUSIONS: Here, we demonstrated that PCS extract administration protected against DEX-induced muscle atrophy. This beneficial effect was mediated by suppressing the expression of muscle degradation factors and increasing the expression of muscle regeneration and synthesis factors. This effect was probably due to the inhibition of oxidative stress and inflammation. These results highlight the potential of PCS extract as a protective and therapeutic agent against muscle dysfunction and atrophy.

Design and Engineering of "Green" Nanoemulsions for Enhanced Topical Delivery of Bakuchiol Achieved in a Sustainable Manner: A Novel Eco-Friendly Approach to Bioretinol.

In the present work, we establish novel "environmentally-friendly" oil-in-water nanoemulsions to enhance the transdermal delivery of bakuchiol, the so-called "bioretinol" obtained from powdered Psoralea corylifolia seeds via a sustainable process, i.e., using a supercritical fluid extraction approach with pure carbon dioxide (SC-CO2). According to Green Chemistry principles, five novel formulations were stabilized by "green" hybrid ionic surfactants such as coco-betaine-surfactin molecules obtained from coconut and fermented rapeseed meal. Preliminary optimization studies involving three dispersion stability tests, i.e., centrifugation, heating, and cooling cycles, indicated the most promising candidates for further physicochemical analysis. Finally, nanoemulsion colloidal characterization provided by scattering (dynamic and electrophoretic light scattering as well as backscattering), microscopic (transmission electron and confocal laser scanning microscopy), and spectroscopic (UV-Vis spectroscopy) methods revealed the most stable nanocarrier for transdermal biological investigation. In vitro, topical experiments provided on human skin cell line HaCaT keratinocytes and normal dermal NHDF fibroblasts indicated high cell viability upon treatment of the tested formulation with a final 0.02-0.2 mg/mL bakuchiol concentration. This excellent biocompatibility was confirmed by ex vivo and in vivo tests on animal and human skin tissue. The improved permeability and antiaging potential of the bakuchiol-encapsulated rich extract were observed, indicating that the obtained ecological nanoemulsions are competitive with commercial retinol formulations.

Correlation analysis between extracts and endogenous metabolites to characterise the influence of salt-processing on compatibility mechanism of 'Psoraleae Fructus & Foeniculi Fructus'.

ETHNOPHARMACOLOGICAL RELEVANCE: 'Salt-processed Psoraleae Fructus & salt-processed Foeniculi Fructus' (sPF&sFF) is a common Chinese medicinal combination for treating diarrhoea. However, it is not clear how sPF and sFF work together, and why salt-processing is necessary. AIM OF THE STUDY: To investigate the compatibility mechanism of sPF&sFF and the influence of salt-processing on it. MATERIALS AND METHODS: Firstly, the metabolomics approach was appliedto screen the differential components between four (s)PF&(s)FF extracts, i.e., sPF&sFF, sPF&FF, PF&sFF, and PF&FF extracts. Then, an in vivo metabolomics study was carried out to filter critical metabolites reflecting the curative effects of (s)PF&(s)FF, and construct a metabolic network. Finally, a correlation analysis between chemical components in extracts and critical metabolites in vivo was performed to find out the synergistic and/or antagonistic effects between herbs as well as the influence of salt-processing. RESULTS: Salt-processing had a direct influence on the contents of chemical components in sPF and sFF extracts, and there existed positive/negative correlations between the content change of chemical components and the effects of critical metabolites. Therefore, salt-processing indirectly affected on these correlations and was (i) conducive to the positive effects of sPF and sFF on bile acids, making sFF play a synergistic role, thereby, sPF&sFF could perform better than sPF and other three combinations and effectively relieve the symptoms of fatty diarrhoea, osmotic diuresis, malnutrition, and weight loss; (ii) conducive to the positive effects of sPF on triacylglycerol, 12(S)-hydroxyeicosatetraenoic acid, cholesterol, and arachidonic acid, and adverse to that of sFF, making sFF play an antagonistic role, thereby, sPF&sFF could prevent a series of side effects caused by over-regulation and suitably relieve the symptoms of osmotic diuresis, polyuria, malnutrition, and weight loss; and (iii) adverse to the positive effects of sPF and sFF on thromboxane A2, sphinganine and sphingosine, making sFF play a synergistic role, thereby, sPF&sFF could prevent a series of side effects and moderately relieve the symptoms of metabolic diarrhoea and polyuria. CONCLUSIONS: Salt-processing indirectly affected on the correlations between chemical components in extracts and critical metabolites in vivo, and exhibited both conducive and adverse effects on the efficacy, making sPF and sFF cooperate with each other to moderately repair the metabolic disorders. Thereby, sPF&sFF could suitably relieve the diarrhoea and polyuria symptoms in the model and exert the most appropriate efficacy. Moreover, this novel strategy provided a feasible approach for further studying the compatibility mechanism of herbs.

The ethanol extraction of prepared Psoralea corylifolia induces apoptosis and autophagy and alteres genes expression assayed by cDNA microarray in human prostate cancer PC-3 cells.

Prostate cancer is the most common male reproductive system cancer. The prevalence of prostate cancer in Europe and the United States is higher than that in the Asian region. However, the treatment of prostate cancer remains unsatisfactory. Psoralea corylifolia has been used to cure this disease as Chinese medicine in the Asian region. In this study, we analyzed the components of ethanol extraction of unprepared and prepared P. corylifolia by HPLC. Psoralen and isopsoralen content from the prepared P. corylifolia is twofold higher than that from unprepared, so we use the prepared extraction in this study. However, the effects of the ethanol extraction of P. corylifolia (PCE) on PC-3 human prostate cancer cells remain unclear. PC-3 cells were treated with PCE for different time periods and cells were examined for cell morphological change and total viable cells by using contrast phase microscopy and flow cytometer, respectively. Results indicated that PCE induced cell morphological changes and cytotoxic effect in PC-3 cells in dose-dependent manners. PCE induced chromatin condensation of PC-3 cells dose-dependently. PCE also induced apoptosis and autophagy in PC-3 by western blotting and acridine orange (AO) staining, respectively. Furthermore, a complementary DNA microarray analysis demonstrated that PCE treatment led to 944 genes upregulation and 872 genes downregulation. For example, the DNA damage-associated gene DNA-damage-inducible transcript 3 (DDIT 3) had a 62.1-fold upregulation and CDK1 2.68-fold downregulation. The differential genes were classified according to the Gene Ontology. Furthermore, GeneGo software was used for the key genes involved and their possible interaction pathways. Those genes were affected by P. corylifolia, which provided information for the understanding of the antiprostate cancer mechanism at the genetic level and provide additional targets for the treatments of human prostate cancer.

Arbuscular mycorrhizal fungi alter the content and composition of secondary metabolites in Bituminaria bituminosa L.

Secondary metabolites may be affected by arbuscular mycorrhizal fungi (AMF), which are beneficial symbionts associated with the roots of most plant species. Bituminaria bituminosa (L.) C.H.Stirt is known as a source of several phytochemicals and therefore used in folk medicine as a vulnerary, cicatrising, disinfectant agent. Characteristic metabolites found in B. bituminosa are furanocoumarins and pterocarpans, which are used in cosmetics and as chemotherapeutic agents. Here we address the question whether AMF inoculation might affect positively the synthesis of these phytochemicals. B. bituminosa plants were inoculated with different AMF and several metabolites were assessed during full vegetative stage and flowering phase. Pigments (chlorophylls and carotenoids), polyphenols and flavonoids were spectrophotometrically determined; specific isoflavones (genistein), furanocoumarins (psoralene and angelicin), pterocarpans (bitucarpin A and erybraedin C) and plicatin B were assessed with HPLC; leaf volatile organic compounds were analysed using SPME and identified by GC-MS. During the vegetative stage, the inoculated plants had a high amount of furanocoumarins (angelicin and psoralen) and pterocarpans (erybraedin C and bitucarpin A). The analysis of volatile organic compounds of inoculated plants showed different chemical composition compared with non-mycorrhizal plants. Given the important potential role played by furanocoumarins and pterocarpans in the pharmaceutical industry, AMF inoculation of B. bituminosa plants may represent a suitable biotechnological tool to obtain higher amounts of such metabolites for pharmaceutical and medicinal purposes.

Enhanced daidzin production from jasmonic and acetyl salicylic acid elicited hairy root cultures of Psoralea corylifolia L. (Fabaceae).

Daidzin (7-O-glucoside of daidzein) has several pharmacological benefits in herbal remedy, as antioxidant and shown antidipsotropic activity. Hairy root culture of Psoralea corylifolia L. was developed for biomass and enhanced daidzin production using signalling compounds such as jasmonic acid (JA) and acetyl salicylic acid (ASA). Best response of 2.8-fold daidzin (5.09% DW) with 1 µM JA treatment after second week and 7.3-fold (3.43% DW) with 10 µM JA elicitation after 10th week was obtained from hairy roots compared to untreated control. ASA at 10 µM promoted 1.7-fold increase in daidzin (1.49% DW) content after seventh week compared to control (0.83% DW). Addition of 25 µM ASA resulted in 1.44% DW daidzin (1.5-fold increase) with 0.91% DW in control after fifth week and 1.44% DW daidzin (2.3-fold increase) after eighth week when compared to untreated control (0.62% DW). Reduced biomass with increased daidzin content was facilitated by elicited hairy root cultures.

Separation and peroxisome proliferator-activated receptor-γ agonist activity evaluation of synthetic racemic bavachinin enantiomers.

Bavachinin, isolated from Psoralea corylifolia seeds, has been reported to demonstrate peroxisome proliferator-activated receptor-γ (PPAR-γ) agonist activity. However, isolated bavachinin is actually a mixture of S and R configurations, with an enantiomeric excess value of approximately 24.3%. For further study on the structure-activity relationships of bavachinin, investigating the PPAR-γ agonist activity of the two enantiomers is crucial. Considering the limited availability, racemic bavachinin was prepared in this study using chemical synthesis. The enantiomers of racemic bavachinin were then separated using supercritical fluid chromatography. This concise strategy yielded (S)- and (R)-bavachinin in optical purity as high as ⩾97.5%. The PPAR-γ agonist activity of the two enantiomers was evaluated using a time-resolved fluorescence resonance energy transfer-based competitive binding assay method; IC50 values of (S)- and (R)-bavachinin were 616.7 and 471.2 nM, respectively. The interaction between the compounds and PPAR-γ was further explored using a molecular docking method. This study suggests that (S)- and (R)-bavachinin demonstrate similar PPAR-γ agonist activities.

Psoralea corylifolia L. seed extract ameliorates streptozotocin-induced diabetes in mice by inhibition of oxidative stress.

Pancreatic beta-cell death is known to be the cause of deficient insulin production in diabetes mellitus. Oxidative stress is one of the major causes of beta-cell death. In this study, we investigated the effects of Psoralea corylifolia L. seed (PCS) extract on beta-cell death. Oral administration of PCS extract resulted in a significant improvement of hyperglycemia in streptozotocin-induced diabetic mice. PCS extract treatment improved glucose tolerance and increased serum insulin levels. To study the mechanisms involved, we investigated the effects of PCS extract on H2O2-induced apoptosis in INS-1 cells. Treatment with PCS extract inhibited cell death. PCS extract treatment decreased reactive oxygen species level and activated antioxidative enzymes. Among the major components of PCS extract, psoralen and isopsoralen (coumarins), but not bakuchiol, showed preventive effects against H2O2-induced beta-cell death. These findings indicate that PCS extract may be a potential pharmacological agent to protect against pancreatic beta-cell damage caused by oxidative stress associated with diabetes.

Psoralidin, a coumestan analogue, as a novel potent estrogen receptor signaling molecule isolated from Psoralea corylifolia.

A novel biological activity of psoralidin as an agonist for both estrogen receptor (ER)α and ERß agonist has been demonstrated in our study. Psoralidin has been characterized as a full ER agonist, which activates the classical ER-signaling pathway in both ER-positive human breast and endometrial cell lines as well as non-human cultured cells transiently expressing either ERα or ERß. The estrogenic activity was determined using the relative expression levels of either reporter or the endogenous genes dependent on the agonist-bound ER to the estrogen response element (ERE). Psoralidin at 10 µM was able to induce the maximum reporter gene expression corresponding to that of E2-treated cells and such activation of the ERE-reporter gene by psoralidin was completely abolished by the cotreatment of a pure ER antagonist, implying that the biological activities of psoralidin are mediated by ER. Psoralidin was also able to induce the endogenous estrogen-responsive gene, pS2, in human breast cancer cells MCF-7. It was observed that activation of the classical ER-signaling pathway by psoralidin is mediated via induction of ER conformation by psoralidin and direct binding of the psoralidin-ER complex to the EREs present in the promoter region of estrogen-responsive genes, as shown by chromatin immunoprecipitation assay results. Finally, molecular docking of psoralidin to the ligand binding pocket of the ERα showed that psoralidin is able to mimic the binding interactions of E2, and thus, it could act as an ER agonist in the cellular environment.

Synthesis of cyclobakuchiols A, B, and C by using conformation-controlled stereoselective reactions.

Cyclohexanone with the pMeOC6H4 and CH2=C(Me) substituents at the C3 and C4-positions was prepared from (+)-ß-pinene and converted to the allylic picolinate by a Masamune-Wittig reaction followed by reduction and esterification. Allylic substitution of this picolinate with Me2CuMgBr·MgBr2 in the presence of ZnI2 proceeded with γ regio- and stereoselectively to afford the quaternary carbon center on the cyclohexane ring with the CH2=CH and Me groups in axial and equatorial positions, respectively. This product was converted to cyclobakuchiol A by demethylation and to cyclobakuchiol C by epoxidation of the CH2=C(Me) group. For the synthesis of cyclobakuchiol B, the enantiomer of the above cyclohexanone derived from (-)-ß-pinene was converted to the cyclohexane-carboxylate, and the derived enolate was subjected to the reaction with CH2=CHSOPh followed by sulfoxide elimination to afford the intermediate with the quaternary carbon center with MeOC(=O) and CH2=CH groups in axial and equatorial positions. The MeOC(=O) group was transformed to the Me group to complete the synthesis of cyclobakuchiol B.

Elemental distribution in tissue components of N2-fixing nodules of Psoralea pinnata plants growing naturally in wetland and upland conditions in the Cape Fynbos of South Africa.

There is little information on in situ distribution of nutrient elements in N2-fixing nodules. The aim of this study was to quantify elemental distribution in tissue components of N2-fixing nodules harvested from Psoralea pinnata plants grown naturally in wetland and upland conditions in the Cape Fynbos. The data obtained from particle-induced X-ray emission revealed the occurrence of 20 elements (Si, P, S, Cl, K, Ca, Ti, Mn, Fe, Ni, Cu, Zn, As, Br, Rb, Sr, Y, Zr, Mo and Ba) in nodule components. Although, in upland plants, the concentrations of S, Fe, Si, Mn and Cu showed a steady increase from the middle cortex to the medulla region of P. pinnata nodules, in wetland plants, only S, Fe and Mn showed an increase in concentration from the middle cortex to the bacteria-infected medulla of P. pinnata nodules. By contrast, the concentrations of Cl, K, Ca, Zn and Sr decreased from middle cortex to nodule medulla. The alkaline earth, alkali and transition elements Rb, Sr, Y and Zr, never before reported in N2-fixing nodules, were found to occur in root nodules of P. pinnata plants grown in both wetland and upland conditions.

Anti-oxidant modulation in response to gamma radiation induced oxidative stress in developing seedlings of Psoralea corylifolia L.

The seeds of Psoralea corylifolia L., an important medicinal herb in Indian and Chinese Pharmacopeia were exposed to gamma rays (2.5, 5, 10, 15 and 20 kGy) from Co(60) source at dose rate of 1.65 kGy h(-1). Enzymatic and non-enzymatic anti-oxidant responses were verified according to the developmental stages and gamma dose applied. Plants grown from seeds exposed to higher gamma doses exhibit higher activity of the antioxidants such as [Ascorbate peroxidase (APX, 1.11.1.1), superoxide dismutase (SOD, 1.15.1.1), glutathione reductase (GR, 1.6.4.2) and MDA content till flowering and declined thereafter. In contrast, CAT (1.11.1.6) activity declined in dose and age dependent manner. The correlation of gamma dose applied and oxidative stress was inferred from the increased enzymes activities and depression in total glutathione pool in seedlings developed from irradiated seeds. Nevertheless, the maintenance of high anti-oxidant capacity, psoralen accumulation seems to be an important strategy during acclimation of P. corylifolia to gamma radiation stress. Pronounced accumulation of psoralen following 15 and 20 kGy at post-flowering stage where oxidative stress is triggered modulates lipid peroxidation and proline accumulation. Further, in psoralen producing plants an increase in psoralen content can be used as a biomarker which specifies plant is under stress.

Enhancement in furanocoumarin content and phenylalanine ammonia lyase activity in developing seedlings of Psoralea corylifolia L. in response to gamma irradiation of seeds.

Gamma irradiation of seeds is known to be an important factor in stimulating biochemical and physiological processes. The aim of the present study was to investigate phenylpropanoids and associated enzymes responsible for the production of active metabolites. Furanocoumarin content was estimated in seeds of Psoralea corylifolia L. during two successive generations (G(1) and G(2)) where as phenylalanine ammonia lyase (PAL) activity was measured in leaves at different developmental stages of P. corylifolia L. raised from seeds irradiated with variable doses of gamma rays. Maximum accumulation of psoralen and isopsoralen was observed at 15 and 20 kGy doses during G(1) and G(2) generations, respectively. Psoralen proved to be the dominating metabolite in terms of its concentration, while isopsoralen was accumulated at relatively lower concentrations in successive generations. PAL activity was induced maximally following 15 and 20 kGy in G(1) plants and was preceded by psoralen and isopsoralen accumulation which peaked at the same dose rates in both generations. These effects were transmitted and prevalent in the next generation, that is, G(2) (indirectly irradiated). These long-term changes in plant metabolomics demonstrate genomic instability induced by gamma irradiation. However, no detrimental effects were seen at any irradiation dose in seeds. Furanocoumarin concentrations were also enhanced at 15 and 20 kGy. The present study further points out the persistence of changes in the biosynthesis of coumarin derivatives in the next generation. However, accumulation of these metabolites does not lead to any lethal effects.

Impact of nutrient components on production of the phytoestrogens daidzein and genistein by hairy roots of Psoralea corylifolia.

Transformed hairy roots of Psoralea corylifolia were established by infection with Agrobacterium rhizogenes LBA 9402. The aim of this work was to elucidate the effects of media constituents on production of the phytoestrogenic isoflavones daidzein and genistein. A. rhizogenes strain LBA 9402 harboring Ri plasmid was used to transform stem segments of in vitro seedlings. The resultant hairy roots were confirmed by polymerase chain reaction (PCR) and exhibited Ri T-DNA. Transformed hairy root clones were cultured in Murashige and Skoog's (MS) medium altered with different concentrations of NH(4) (+) and NO(3) (-) and their growth and production of isoflavones were assessed. Biomass and productivity increased when MS medium was supplemented with NH(4) (+) and NO(3) (-) at a ratio of 20:10. Increased yield of daidzein was obtained when sucrose level in the culture medium increased, whereas decreased level of sucrose favored genistein production. The hairy roots produced the highest levels of daidzein (2.06% dry wt.) and genistein (0.37% dry wt.) in the presence of low concentrations of PO(4) (3-). Hairy roots secreted trace amounts of daidzein and genistein into the culture medium. The present results demonstrated that the productivity of daidzein was 2.2-fold more than that of untransformed roots.

Psoralen production in hairy roots and adventitious roots cultures of Psoralea coryfolia.

Psoralea corylifolia is an endangered plant producing various compounds of medical importance. Adventitious roots and hairy roots were induced in cultures prepared from hypocotyl explants. Psoralen content was evaluated in both root types grown either in suspension cultures or on agar solidified medium. Psoralen content was approximately 3 mg g(-1) DW in suspension grown hairy roots being higher than in solid grown hairy roots and in solid and suspension-grown adventitious roots.

Studied enhancement strategies for phytoestrogens production in shake flasks by suspension culture of Psoralea corylifolia.

This study proposed secondary metabolites incremental yield due to manipulation of nutrient components into the culture medium. To validate this, the effects of nutrients such as carbon, phosphate and nitrogen on growth and production of phytoestrogens daidzein and genistein by suspension cultures of Psoralea corylifolia was investigated for the first time. The maximum production of daidzein and genistein was achieved when sucrose and maltose used as a sole source of carbon. Suspension cell cultures enriched with sucrose (3%) stimulated accumulation of isoflavones daidzein (1.76% dry wt) and genistein (0.25% dry wt) compared to glucose, fructose and maltose. Sucrose feeding strategy significantly stimulated biomass growth and isoflavones (2.79% dry wt of daidzein and 0.32% dry wt of genistein) production rate. Reduced concentrations of phosphate (0.625 mM) promoted daidzein (1.89% dry wt) and genistein (0.26% dry wt) production by suspension cell cultures, whereas high amount (5mM) in medium was inhibited isoflavones production. It was observed that medium fortified with NH(4)(+) and NO(3)(-) alone inhibited production of isoflavones. The maximum production obtained of daidzein (2.20% dry wt) and genistein (0.29% dry wt) when medium comprised with NH(4)(+)/NO(3)(-) at ratio 20:40 mM as a nitrogen source. Similar nutrient components ratio when altered NH(4)(+)/NO(3)(-); 40:20mM) resulted in approximately 3-fold decrease in production. HPLC analysis revealed that suspension cells cultures leached out trace amount of daidzein and genistein into the culture medium.

Psoralea corylifolia L. inhibits mitochondrial complex I and proteasome activities in SH-SY5Y cells.

The growing interest in alternative medicines, including traditional medicinal plants, has caused some health concerns due to poor awareness in the general population of the possible side effects from inappropriate practices. Psoralea corylifolia L. has been used in Chinese and Indian traditional medicine for the treatment of various inflammatory diseases of the skin and to improve vitality. Our data show that the extract obtained from the seeds of Psoralea corylifolia L. decreased mitochondrial complex I and proteasome activities; and oxidative stress might be an early event.