Diffusa Extracts Research Articles

Biochemical and molecular docking-based strategies of Acalypha indica and Boerhavia diffusa extract by targeting bacterial strains and cancer proteins

Antibiotic-resistant microbes have emerged around the world, presenting a risk to health. Plant-derived drugs have become a potential source for the production of antibiotic-resistant drugs and cancer therapies. In this study, we investigated the antibacterial, cytotoxic and antioxidant properties of Acalypha indica and Boerhavia diffusa, and conducted in silico molecular docking experiments against EGFR and VEGFR-2 proteins. The metabolic extract of A. indica inhibited Streptococcus iniae and Staphylococcus sciuri with inhibition zones of 21.66 ± 0.57 mm and 20.33 ± 0.57 mm, respectively. The B. diffusa leaf extract produced inhibition zones of 20.3333 ± 0.5773 mm and 20.33 ± 0.57 mm against Streptococcus iniae and Edwardsiella anguillarum, respectively. A. indica and B. diffusa extracts had toxicities of 162.01 μg/ml and 175.6 μg/ml, respectively. Moreover, B. diffusa (IC50 =154.42 µg/ml) leaf extract exhibited moderately higher antioxidant activity compared with the A. indica (IC50 = 218.97 µg/ml) leaf extract. Multiple interactions were observed at Leu694, Met769 and Leu820 sites for EGFR and at Asp1046 and Cys1045 sites for VEGFR during the molecular docking study. CID-235030, CID-70825 and CID-156619353 had binding energies of −7.6 kJ/mol, −7.5 kJ/mol and −7.6 kJ/mol, respectively, with EGFR protein. VEGFR-2 protein had docking energies of −7.5 kJ/mol, −7.6 kJ/mol and −7.3 kJ/mol, respectively, for CID-6420353, CID-156619353 and CID-70825 compounds. The MD simulation trajectories revealed the hit compound; CID-235030 and EGFR complex, CID-6420353 and VEGFR-2 exhibit stable profile in the root mean square deviation (RMSD), radius of gyration (Rg), solvent accessible surface area (SASA), hydrogen bond and root mean square fluctuation (RMSF) and the binding free energy by MM-PBSA method. This study indicates that methanol extracts of A. indica and B. diffusa may play a crucial role in developing antibiotic-resistant and cancer drugs. Communicated by Ramaswamy H. Sarma

Biochemical and molecular anticancer approaches for Boerhaavia diffusa root extracts in oral cancer.

Boerhaavia diffusa is a medicinal herb with anti-inflammatory, antiproliferative, anticancer, and immunomodulatory properties, found across India. The present study is designed to investigate the therapeutic potential for B. diffusa root extracts in oral cancer cell line. The aqueous and methanolic extracts of B. diffusa were prepared using Soxhlet apparatus. In order to determine the phytochemical constituents of B. diffusa, the extracts were subjected to gas chromatography-mass spectrometry analysis. The antioxidant potential of B. diffusa extracts was assessed by 2,2-Diphenyl-picrylhydrazyl, ferric ion-reducing antioxidant power, catalase and peroxidase assays. The effective concentration of B. diffusa root on cell viability was analyzed by [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide] assay. The ability of B. diffusa root extracts to modify the cell-cycle phases was performed by FACS analysis. The apoptotic inducing potential of B. diffusa in oral cancer cells was confirmed by acridine orange-ethidium bromide and 4',6-diamidino-2-phenylindole staining. The protein profile of apoptotic processes was validated by the Western blot analysis; docking studies were also performed. We observed that antioxidant activity was higher in B. diffusa methanolic extract compared with aqueous extract. The results showed that the methanolic and aqueous extracts of B. diffusa exhibited significant cytotoxic effect with IC50 value of 36 μg/ml and 30 μg/ml, respectively. The apoptotic DNA fragmentation and the apoptotic inducing potential in KB oral cancer cell line were higher for the methanolic extract compared with the aqueous extract. These results were also confirmed by in-silico analysis. The results indicate that extracts obtained from the roots of B. diffusa inhibit the progression of oral cancer. These compounds of pharmacological importance can be either used alone or in combination with other drugs to treat oral cancer.

Novel Antiviral Efficacy of Hedyotis diffusa and Artemisia capillaris Extracts against Dengue Virus, Japanese Encephalitis Virus, and Zika Virus Infection and Immunoregulatory Cytokine Signatures

Currently, there are no specific therapeutics for flavivirus infections, including dengue virus (DENV) and Zika virus (ZIKV). In this study, we evaluated extracts from the plants Hedyotis diffusa (HD) and Artemisia capillaris (AC) to determine the antiviral activity against DENV, ZIKV, and Japanese encephalitis virus (JEV). HD and AC demonstrated inhibitory activity against JEV, ZIKV, and DENV replication and reduced viral RNA levels in a dose–responsive manner, with non-cytotoxic concentration ranging from 0.1 to 10 mg/mL. HD and AC had low cytotoxicity to Vero cells, with CC50 values of 33.7 ± 1.6 and 30.3 ± 1.7 mg/mL (mean ± SD), respectively. The anti-flavivirus activity of HD and AC was also consistent in human cell lines, including human glioblastoma (T98G), human chronic myeloid leukemia (K562), and human embryonic kidney (HEK-293T) cells. Viral-infected, HD-treated cells demonstrated downregulation of cytokines including CCR1, CCL26, CCL15, CCL5, IL21, and IL17C. In contrast, CCR1, CCL26, and AIMP1 were elevated following AC treatment in viral-infected cells. Overall, HD and AC plant extracts demonstrated flavivirus replication inhibitory activity, and together with immunoregulatory cytokine signatures, these results suggest that HD and AC possess bioactive compounds that may further be refined as promising candidates for clinical applications.

Biological Synthesis of Monodisperse AuNPs@Damiana with Enhanced Antiseptic Activity Against Gram-Negative Bacteria

Turnera diffusa commonly called Damiana is a wild shrub traditionally used as aphrodisiac, diuretic, and for cancer and diabetes treatments. Its extracts are rich in flavonoid, phenolic, and terpenoid compounds that have been reported as potential reductors for metal ions. In this sense, this study reports a novel and facile method for green gold nanoparticle (AuNPs) synthesis using T. diffusa extracts. The water-soluble organic compounds were obtained by hydrothermal route and the AuNPs@Damiana samples were fully characterized. The results indicated high efficiency of T. diffusa extracts to reduce and stabilize the AuNPs which showed spherical shape, 24 nm diameter, negative surface charge, and high colloidal stability at biological pH and alkaline media. Additionally, its antimicrobial activity was evaluated against Salmonella Enteritidis, Escherichia coli ATCC 8739, and Listeria monocytogenes, showing an enhanced microbial inhibition on Gram-negative bacteria.

Effect of a multi-ingredient based food supplement on sexual function in women with low sexual desire

BackgroundStudies have demonstrated that women with low desire and low excitement have negative feelings regarding their physical and emotional satisfaction, as well as their happiness. In this study, we evaluate the efficacy of Libicare® - a multi-ingredient food supplement - to improve sexual function in postmenopausal women.MethodsThis was an exploratory, prospective, non-controlled, observational study. Postmenopausal women aged 45–65 with a risk of sexual dysfunction (Female Sexual Function Index (FSFI) < 25.83) were included during routine clinical visits and treated with 2 tablets of Libicare® daily for 2 months. Libicare® is an oral food supplement containing Trigonella foenum graecum, Turnera diffusa, Tribulus terrestris, and Ginkgo biloba dry extracts. Primary endpoint: change vs. baseline in FSFI score. Secondary endpoints: 1) changes in testosterone and serum steroid levels of free testosterone and sex hormone-binding globulin (SHBG) levels and 2) tolerability.ResultsA total of 29 patients (mean age: 54.69 years) were included. FSFI mean (SD) score showed a significant increase: 20.15 (4.48) vs 25.03 (6.94), baseline vs final; p = 0.0011, paired t-test. Most patients (86.2%) increased their FSFI score. All FSFI domains, except dyspareunia, showed significant increases. The highest increase was observed in the desire domain (p = 0.0004). Testosterone and SHBG levels were assessed in 21 patients. A significant increase in testosterone level was observed: 0.41 (0.26) vs. 0.50 (0.34) pg/mL, baseline vs. final; p = 0.038, Wilcoxon test. 52.4% of patients increased their testosterone levels. Finally, a significant decrease was observed in SHBG level: 85 (32.9) vs. 73 (26.8) nmol/L, baseline vs. final; p = 0.0001; paired t-test. 95.2% of patients decreased their SHBG levels.ConclusionIn this pilot study, a significant improvement in sexual function and related hormone levels was observed with Libicare®. Further studies must be conducted to confirm these exciting results.Trial registrationCurrent Controlled Trial ISRCTN12928573. Date of registration: 28/March/2019. Retrospectively registered.

Cytotoxic Properties of Damiana (Turnera diffusa) Extracts and Constituents and A Validated Quantitative UHPLC-DAD Assay.

In our continuing search for new cytotoxic agents, we assayed extracts, fractions, and pure compounds from damiana (Turnera diffusa) against multiple myeloma (NCI-H929, U266, and MM1S) cell lines. After a first liquid-liquid solvent extraction, the ethyl acetate layer of an acetone (70%) crude extract was identified as the most active fraction. Further separation of the active fraction led to the isolation of naringenin (1), three apigenin coumaroyl glucosides 2–4, and five flavone aglycones 5–9. Naringenin (1) and apigenin 7-O-(4″-O-p-E-coumaroyl)-glucoside (4) showed significant cytotoxic effects against the tested myeloma cell lines. Additionally, we established a validated ultra-high performance liquid chromatography diode array detector (UHPLC-DAD) method for the quantification of the isolated components in the herb and in traditional preparations of T. diffusa.

Selective 1D-TOCSY and chemometrics to evaluate authenticity of Turnera diffusa and related botanical extracts

Nuclear magnetic resonance (NMR) spectroscopy has emerged as a powerful tool for the development of new analytical methods in metabolomics and metabolic profiling of a plethora of biological samples, including natural products. NMR-based metabolomics enables characterisation of metabolites and biomarkers in plant extracts. Selective 1D- Total Correlation Spectroscopy TOCSY (1D-TOCSY) has been used in metabolomic research as a direct method for spectroscopic separation and identification of chemical compounds in complex mixtures. In this work, a method for the specific identification of the biomarker compound hepatodamianol in Turnera diffusa plants and related botanical extracts, based on NMR and multivariate analysis, was developed. For this purpose, methanolic extracts from T. diffusa samples were prepared and analysed using 1H-NMR. Hepatodamianol selective 1D-TOCSY spectra were recorded to obtain the spectroscopic fingerprint of carbohydrate moieties, and then compared against authenticated T. diffusa extracts and commercial samples. 1H-NMR data were collected to generate a PCA statistical model to explore the chemical variability between samples of T. diffusa extracts. The selective 1D-TOCSY experiment was successfully applied for unambiguous identification of hepatodamianol in T. diffusa extracts and commercial botanical remedies to verify authenticity of botanical material. This combined methodology based on NMR metabolomics is a useful analytical tool to establish if T. diffuse is present in herbal preparations claiming to be prepared from this plant. Prospectively, this method could be extended to analyse traditional remedies and herbal medicines, as a quality control proof to assess the presence of bioactive compounds.

168 Effect of a Trigonella foenum graecum-based food supplement on sexual function in women with low sexual desire. Pilot study

To evaluate the efficacy of Libicare® - a Trigonella foenum graecum-based food supplement - to improve sexual function in peri and posmenopausal women. An exploratory, prospective, non-controlled, observational study. Women aged 45 -65 years with low sexual desire (FSFI < 25.83) were included during routine clinical visits and treated with 2 tablets of Libicare® daily for 2 months. Libicare® is an oral food supplement containing Trigonella foenum graecum, Turnera Diffusa, Tribulus Terrestris and Ginkgo Biloba dry extracts. Primary endpoint: change vs baseline in FSFI score. Secondary endpoints: 1) changes in testosterone and SHBG levels and 2) tolerability. A total of 29 patients (mean age: 53.9 years) were included. FSFI mean (SD) score showed a significant increase: 20.15 (4.48) vs 25,03 (6.94), baseline vs final; p=0.0002, t-test. Most of patients (89.7%) increased FSFI score. Testosterone and SHBG levels were assessed in 21 patients. A significant increase in testosterone level was observed: 0,41 (0.26) vs 0,50 (0.34) pg/ml, baseline vs final; p=0.038, Wilcoxon test. 52.4% of patients increased their testosterone levels. Finally, a significant decrease was observed in SHBG level: 85 (32.9) vs 73 (26.8) nmol/l, baseline vs final; p=0.0001; paired t-test. 95.2% of patients decrease their SHBG levels.

Bush mint (Hyptis suaveolens) and spreading hogweed (Boerhavia diffusa) medicinal plant extracts differentially affect activities of CYP1A2, CYP2D6 and CYP3A4 enzymes

Ethno-pharmacological relevanceHyptis suaveolens (L) Poit and Boerhavia diffusa Linn are medicinal herbal plants commonly found in the tropics and sub-tropics. They are used to treat various conditions among them boils, dyslipidaemia, eczema, malaria, jaundice and gonorrhoea. Thus, the herbal medicinal extracts are now found as part of some commercial herbal formulations. There has not been adequate characterization of these medicinal herbs on their effects on drug metabolising enzymes. Aim of the studyTo investigate the effects of extracts of Hyptis suaveolens (HS) and Boerhavia diffusa (BD) on activity of drug metabolising enzymes, CYP1A2, CYP2D6 and CYP3A4, as well predict their potential for herb-drug interaction. A secondary aim was to identify constituent compounds such as polyphenolics, in the crude extract preparations of Hyptis suaveolens and Boerhavia diffusa and measure them for activity. Materials and methodsCYP450 inhibition assays using recombinant CYP450 (rCYP) and fluorescence screening employing individual isozymes (CYP1A2, CYP2D6 and CYP3A4) were used to determine reversible- and time-dependent inhibition (TDI) profiles of extracts of Hyptis suaveolens and Boerhavia diffusa. Inhibition kinetic parameters, Ki and Kinact were also estimated. UPLC-MS employing a Synapt G2 (ESI negative) coupled to a PDA detector was used to identify polyphenolic compounds in crude extracts of Hyptis suaveolens and Boerhavia diffusa. ResultsThe inhibitory potency of Hyptis suaveolens and Boerhavia diffusa extracts varied among the different enzymes, with CYP1A2 (3.68 ± 0.10µg/mL) being the least inhibited by HS compared to CYP2D6 (1.39 ± 0.01µg/mL) and CYP3A4 (2.36 ± 0.57µg/mL). BD was most potent on CYP3A4 (7.36 ± 0.94µg/mL) compared to both CYP2D6 (17.79 ± 1.02µg/mL) and CYP1A2 (9.48 ± 0.78µg/mL). Extracts of Hyptis suaveolens and Boerhavia diffusa exhibited TDIs on all CYPs. The most prominent phenolic candidates identified in both medicinal herbs using UPLC-MS analysis included caffeic acid, rutin, quercetin, citric acid, ferulic acid and gluconic acid. These phenolic compounds are thought to potentially give HS and BD their therapeutic effects and inhibitory characteristics affecting CYP450 activities. In vivo predictions showed the potential for HS and BD extracts to cause significant interactions if co-administered with other medications. ConclusionsThe study reveals that crude aqueous extracts of HS and BD potentially inhibit drug metabolising isozymes CYP1A2, CYP2D6 and CYP3A4 in a reversible and time-dependent manner. Thus care should be taken when these extracts are co-administered with drugs that are substrates of CYP1A2, CYP2D6 and CYP3A4.

Aqueous Oldenlandia diffusa extracts inhibits colorectal cancer cells via activating AMP-activated protein kinase signalings.

Here we evaluated the anti-cancer activity of aqueous Oldenlandia diffusa (OD) extracts (ODE) in colorectal cancer (CRC) cells. We showed that ODE exerted potent anti-proliferative, cytotoxic and pro-apoptotic activities against a panel of established CRC lines (HCT-116, DLD-1, HT-29 and Lovo) and primary (patient-derived) human CRC cells. ODE activated AMP-activated protein kinase (AMPK) signaling, which led to subsequent mTORC1 inhibition and Bcl-2/HIF-1α downregulation in CRC cells. In ODE-treated CRC cells, AMPKα1 formed a complex with p53. This might be important for p53 activation and subsequent cancer cell apoptosis. Inhibition of AMPK signaling, though dominant negative (dn) mutation or shRNA/siRNA knockdown of AMPKα1 attenuated ODE-exerted CRC cytotoxicity. In vivo, i.p. administration of ODE inhibited HCT-116 xenograft tumor growth in SCID mice. In addition, AMPK activation, mTORC1 inhibition and p53 activation were observed in ODE-treated HCT-116 xenograft tumors. These results suggest that ODE inhibits CRC cells in vitro and in vivo, possibly via activation of AMPK-dependent signalings.

Phytochemical analysis of hydroethanolic extract of Turnera diffusa Willd and evaluation of its effects on astrocyte cell death.

ABSTRACTObjective To evaluate the phytochemical composition of hydroethanolic extracts from powdered aerial parts of Turnera diffusa Willd (Turneraceae; T. diffusa), as well as its toxicity in astrocytes.Methods Chemical analyses of hydroethanolic extract from powdered aerial parts of T. diffusa were carried out using HPLC-DAD-ESI-MS/MS. In vitro assays using astrocytes culture were performed to evaluate cell death.Results Flavone-C, O-diglycosides, such as, luteolin-8-C-[6-deoxy-2-O-rhamnosyl]-xylo-hexos-3-uloside, apigenin-8-C-[6-deoxy-2-O-rhamnosyl]-xylo-hexos-3-uloside and apigenin-7-O-6”-p-coumaroylglucoside were the main compounds found in this hydroethanolic extract. Concentration time-effect demonstrated the toxicity of this extract at a concentration of 1,000µg/mL in astrocyte culture, after 6 and 24 hours of incubation.Conclusion In phytochemical analyses, important antioxidants (mainly flavonoids) were observed. T. diffusa extracts presented cytotoxic effect in high concentrations, leading to increased cell death in astrocyte culture.

Oxygen radical absorption capacity of Lippia origanoides (thymol and phellandrene chemotypes) and Turnera diffusa essential oils and extracts

Damiana (Turnera diffusa) and Mountain oregano (Lippia origanoides) are shrubs that grow wild in semi-arid terrains of Central and northern South America. Three L. origanoides chemotypes have been found, characterized by their different essential oil main constituents (thymol, carvacrol and phellandrene). GC-MS analysis of essential oils (EO) and extracts (obtained with supercitical CO2) from stems and leaves of T. diffusa and L. origanoides (thymol and phellandrene chemotypes) showed that the secondary metabolites common to these aromatic plants were: α-pinene (0.5 – 1.8%), limonene (0.2 – 37.8%), linalool (0.2 – 1.1%), trans-β-caryophyllene (1.0 – 26.4%), α-humulene (4 – 2.4%), caryophyllene oxide (3 – 7.2%), and germacrene D (0.05 – 37.68%). Dehydrofuquinone (52.3%) and drima-7,9-diene (4.4%) were the main components of damiana oil. The oxygen radical absorption capacity (ORAC) of a L. origanoides (phellandrene chemotype) extract obtained with supercritical CO2 showed a remarkably high value (4130 ± 29 micromol Trolox/g substance), attributed to its high pinocembrin content. Lower ORAC values were obtained for L. origanoides (thymol chemotype) EO (3710 ± 10 micromol Trolox/g sample), followed by L. origanoides (phellandrene chemotype) EO (1260 ± 29 micromol Trolox/g sample) and damiana EO (813 ± 9 micromol Trolox/g sample). EO were obtained by microwave radiation-assisted hydrodistillation. Extracts were obtained with CO2 at 400 bar, 50 g/min and 10% ethanol in a Thar SFE-2000 – 2-FMC50 system; extract fractions were collected at 40 and 80 bar. The separation and identification of secondary metabolites was performed on an Agilent Technologies GC 6890 with 5973 mass selective detector (EI, 70 eV). Chromatographic columns DB-5 and DB-WAX (60 m × 0.25 mm, ID × 0.25 mm) were used. The ORAC assay was performed in a Turner Biosystems multiplate reader. Trolox® was used as a reference standard for these measurements.

Effective 2D-RPLC/RPLC enrichment and separation of micro-components from Hedyotis diffusa Willd. and characterization by using ultra-performance liquid chromatography/quadrupole time-of-flight mass spectrometry

An effective method aiming at enrichment and analysis of micro-components in traditional Chinese medicine (TCM) was developed. One fraction (fraction E) from the extract of Hedyotis diffusa Willd. was selected as test sample, which was isolated by using the XAD-4 macroporous resin. To study the micro-components, a two-dimensional reverse-phase liquid chromatography (2D-RPLC/RPLC) method was developed, comprising Click OEG and C18 stationary phases as the first and second dimensions, respectively. Of the eight sub-fractions isolated from the first dimension, three sub-fractions (fractions II–IV) containing micro-components were further separated with the second dimension. The 2D-RPLC/RPLC system was proved to possess high orthogonality. Furthermore, the micro-components were characterized by using ultra-performance liquid chromatography–diode array detector/quadrupole time-of-flight mass spectrometry (UPLC–DAD/Q-TOF MS) with electrospray ionization (ESI) source. With the optimized separation and characterization method, a large number (>400) of micro-components were enriched and detected from the extracts of H. diffusa Willd., the majority of which has not been isolated from the herb before. Among these isolated micro-components, 38 compounds involving 24 phenylpropanoids, 7 flavonoids and 7 iridoid glucosides (IGs), were identified or tentatively identified from the H. diffusa extracts on the basis of spectral data of the authentic standards and the fragmentation characteristics information available in literatures. The proposed method made it possible to effectively screen and analyze the micro-components in TCMs or other complex natural medicines.

Quantitative Analysis of Boeravinones in the Roots of Boerhaavia Diffusa by UPLC/PDA

Boerhaavia diffusa is a perennial herb belonging to Nyctaginaceae. Various classes of chemical constituents such as phenolics (boeravinones), terpenoids and organic acids have been reported in B. diffusa roots. As boeravinones have been proposed as putative active constituents for the anti-cancer, spasmolytic and anti-inflammatory activities exhibited by B. diffusa extracts, it is worthwhile developing and validating an ultra-performance liquid chromatography (UPLC) method for analysis of boeravinones in B. diffusa roots. To develop and validate a simple, accurate, robust and rapid UPLC analytical method for quality control of B. diffusa roots. Samples for analysis were prepared by refluxing powdered root material with methanol for 2 h. The extracts were concentrated, dried and stored at -20°C until their use. A UPLC with photodiode array (PDA) method was developed and validated for the quantification of boeravinones in the roots of B. diffusa. The separation of boeravinones was achieved using a BEH Shield C18 -column (2.1 × 100 mm, 1.7 µm) with gradient elution of methanol and water (0.1% acetic acid), at a flow rate of 0.4 mL/min and detection was carried out at λmax 273 nm. The UPLC method developed showed good linearity (r(2) ≥ 0.9999), accuracy and precision. The UPLC method developed provided a selective, sensitive and rapid analytical method for the quantification of boeravinones in B. diffusa roots. All the validation parameters were found to be within the permissible limits as per International Conference on Harmonisation guidelines.

Phytotoxicity of organic extracts of Turnera ulmifolia L. and Turnera diffusa Willd. ex Schult. in cucumber seeds

We evaluated the phytotoxic effects that the hexane, ethyl acetate and methanol extracts from leaves and branches of the species Turnera ulmifolia L. and Turnera diffusa Willd. ex Schult. (Turneraceae), at concentrations of 1.25, 2.50, 3.75 and 5.00 mg ml-1, have on seed germination and seedling development in cucumber (Cucumis sativus L.). None of the extracts tested prevented germination, although the ethyl acetate extracts of T. diffusa, at 3.75 and 5.00 mg ml-1, reduced the mean germination speed and time to germination. Hexane extracts of both species reduced the main root length, number of secondary roots and hypocotyl length. In the ethyl acetate and methanol extracts, the number of secondary roots and hypocotyl length varied by species and concentration. Ethyl acetate extracts of T. ulmifolia at < 2.50 mg ml-1 stimulated growth of the main root and hypocotyl, as did T. ulmifolia methanol extracts at 1.25 and 3.75 mg ml-1; all other extract/concentration combinations had an inhibitory effect on those parameters. The hexane and ethyl acetate extracts of T. diffusa inhibited the formation of secondary roots and of the main root, although significant inhibition of hypocotyl growth was observed only at > 2.50 mg ml-1. Comparatively, T. diffusa extracts inhibited development to a greater degree, thus presenting greater allelopathic potential, than did T. ulmifolia extracts.

Effects of Panax ginseng, Turnera diffusa and Heteropterys tomentosa extracts on hippocampal apoptosis of aged rats

ABSTRACTObjective:To verify if the medicinal plants Panax ginseng C.A. Mey, Turnera diffusa Willd. ex Schult., and Heteropterys tomentosa O. Mach., which are amply used by the population as tonics and cognition enhancers, could have a protective effect on cell death by apoptosis, since this could be one of the mechanisms of action of these substances.Methods:Aged male Wistar rats (n=24) were divided into four groups. Over 30 days, three groups received treatments with hydroalcoholic extracts of the plants, and one group received saline solution. A fifth group with young adult male Wistar rats (n=4) received saline solution during the same period. Using the TUNEL technique, the percentage of apoptosis in the hippocampus of these animals was evaluated.Results:No differences were observed between the percentage of apoptotic cells in the hippocampus of aged animals and of young control animals. The percentage of apoptosis in the hippocampus of aged animals treated chronically with the extracts from the three plants also did not differ from the percentage of apoptosis in the hippocampus of the control group of aged animals.Conclusion:Treatment with the hydroalcoholic extracts of Panax ginseng, Turnera diffusa, and Heteropterys tomentosa did not influence the apoptosis of the hippocampal cells of aged rats.

Biological Screening of Triherbal Formulation on Chemically Induced Hepatocellular Carcinoma

The hepatoprotective activity of a triherbal formulation (LAB) comprising of 50 % ethanolic extract of Lawsonia innermis, Aeagel marmelos, Boerrhavia diffusa was studied using Swiss albino rats. The animals received a single intraperitoneal injection of N-nitrosodiethylamine 200mg/kg body wt followed by subcutaneous injection of CCl4 in a dose of 3 ml/kg body wt. The administration of Lawsonia innermis, Aeagel marmelos, Boerrhavia diffusa extracts and cisplatin decreased the liver weight and average liver weight, which shows the rehabilitating capability of extracts in respect with anticancer potency in comparison with the very much effective in preventing NDEA-induced multistage hepatocarcinogenesis possibly through antioxidant and antigenotoxic nature, which was confirmed by various liver injury and biochemical tumor markers enzymes and molecular events. LAB dose dependently and significantly the increase in serum hepatic enzyme levels after NDEA&amp; CCl4 treatment compared to the toxin control group. On administration of LAB, it dose dependently resulted in an elevation in the levels of serum CAT, GPX, GSH, GST, SOD and total protein and decrease in lipid peroxidation compared to the toxin control group. It also increased the attenuated the total protein concentrations compared to the toxin control group. The results of this study confirmed the antioxidant and hepatoprotective activity of the triherbal formulation against carbon tetrachloride&amp; N-nitrosodiethylamine induced hepatotoxicity in Swiss albino rats. © 2011 IGJPS. All rights reserved.

Correlation between Chromatographic Fingerprint and Antioxidant Activity ofTurnera diffusa(Damiana)

In the present contribution, the partial least squares (PLS) method was used to establish a correlation between the antioxidant activity (obtained by DPPH assay) and chromatographic profiles of TURNERA DIFFUSA extracts. Chromatograms were obtained using HPLC-DAD. A model was constructed using 40 samples with 2550 X variables corresponding to the responses obtained at different times; the Y variables consisted of experimental values of antioxidant activity of each extract (measured as EC₅₀). Prior to this analysis, alignment of chromatograms was performed based on consideration of seven high-intensity signals present in all samples. The PLS1 model was validated by cross-validations; its capacity was evaluated using correlation parameters R², root mean square error of calibration (RMSEC), and root mean square error of prediction (RMSEP). The best results were achieved with zero order chromatograms using five-point smoothing (R² = 0.96, RMSEC = 3.31, and RMSEP = 7.86). Under these conditions, the optimal number of components was five. The model was applied to the prediction of antioxidant activity of commercial products; no significant differences were found between the experimental and predicted antioxidant activities for 83 % of them.

Quantitative Determination of β-Arbutin and Seven Flavonoids from Turnera diffusa (Damiana) Extracts and Dietary Supplements Claiming to Contain Damiana by Using HPLC-UV Method

Turnera diffusa Willd (Turneraceae), common name damiana, is an aromatic shrub with small yellow flowers. The leaves and sometimes the stems of damiana have medicinal uses. Evaluation of herbal dietary supplements marketed on the internet for recreational use shows that 10% of the most common products were claiming to contain damiana in the product ingredients [1,2]. An HPLC/UV method permitting the simultaneous determination of 8 compounds isolated from T. diffusa has been developed. A separation was achieved within 45 minutes by using the C-18 material column. The mobile phase was comprised of acetonitrile/methanol (90:10, v/v) containing 0.1% acetic acid and 50 mM ammonium acetate (pH = 4.2) at a flow rate of 1 mL/min and the column temperature was maintained at 30°C. The method was validated for linearity, repeatability, limits of detection (LOD) and limits of quantification (LOQ). The developed method was applied for the quantitative determination of eight compounds [β-Arbutin (1), apigenin 7-O-β-D-glucoside (2), tricin 7-O-β-D-glucoside (3), turneradiffusin (4), diffusavone (5), turneradin (6), apigenin 7-O-β-D-(5-O-β-coumaroyl)-glucoside (7) and echinaticin (8)] for two different species of Turnera and dietary supplements. The eight compounds in two different species of Turnera and dietary supplements were further confirmed by LC-ESI/MS. Acknowledgements: This research is funded in part by “Science Based Authentication of Dietary Supplements” Funded by the Food and Drug Administration grant number 2 U01 FD 002071-07. The authors would like to thank Annette Ford, University of Mississippi for extraction of samples.

식중독유발 세균의 증식에 미치는 백화사설초 추출물의 영향

This study was performed to investigate the antimicrobial effect of the Hedyotis diffusa extracts against food-borne pathogens. First, the Hedyotis diffusa was extracted with methanol at room temperature and the fractionation of the methanol extracts was carried out by using petroleum ether, chloroform, and ethyl acetate, and methanol respectively. The antimicrobial activity of the Hedyotis diffusa extracts was determined by using a paper disc method against food-borne pathogens and food spoilage bacteria. The methanol extracts of Hedyotis diffusa showed the highest antimicrobial activity against Staphylococcus aureus and Shigella flexneri. Synergistic effect in inhibition was observed when Hedyotis diffusa extract was mixed Sophora subprostrata extract as compared to each extracts alone. Finally, the growth inhibition curves were determined by using ethyl acetate extracts of Hedyotis diffusa against Staphylococcus aureus and Salmonella Typhimurium. The methanol extract of Hedyotis diffusa had strong antimicrobial activity against Staphylococcus aureus at the concentration of 4,000 ppm. At this concentration, the growth of Staphylococcus aureus was retarded more than 36 hours and up to 24 hours for Salmonella Typhimurium. In conclusion, the methanol extracts of Hedyotis diffusa inhibit efficiently Staphylococcus aureus and Salmonella Typhimurium.