Alba Root Research Articles

Root-soil contact dynamics of Vicia faba in sand

AimsRoot shrinkage in drying soil has been shown repeatedly. The aim of this study was to investigate the dynamics of root-soil contact and its relationship with plant water status during soil drying.MethodsThe development of root-soil contact of Vicia faba L. during a drying period was studied. Plants (N = 4) were grown in cylinders filled with a sandy soil. Samples were repeatedly scanned with an X-ray CT scanner to visualize root-soil contact. Soil matric potential, transpiration rate, and stomatal conductance were measured daily.ResultsRoot-soil contact was lower in taproots than in lateral roots at any time. Transpiration rate and stomatal conductance decreased before roots started to shrink. Root-soil contact decreased significantly over the course of the drying period, starting at soil matric potentials below −20 kPa. Root shrinkage did not differ significantly between taproots and laterals.ConclusionsThis study confirms previous findings with Lupinus albus roots in that roots shrink after transpiration rate decreases. The dynamics of root shrinkage are governed by soil water availability and transpirational demand.

Optimization of Imidazolium-Based Ionic Liquid-Microwave Assisted Extraction for Oxyresveratrol Extraction from Morus alba Roots

Objective: The application of ionic liquids as solvent in the microwave assisted extraction (MAE) method has been developed for extraction of secondary metabolites from plants. However, oxyresveratrol extraction from Morus alba roots using IL-MAE method has never been reported. Purpose of this research was to get the optimum condition of imidazoliumbased ionic liquid-microwave assisted extraction (IL-MAE) for oxyresveratrol extraction from M. alba roots. Methods: M. alba roots were extracted with 3 kinds of ionic liquids (1-butyl-3-methylimidazolium bromide ([Bmim]Br), 1-hexyl-3-methylimidazolium bromide ([Hmim]Br)and 1-butyl-3-methylimidazolium sulfate ([Bmim]2[SO4]), respectively by microwave with 3 different microwave power (P10%, P30% and P50%) for 10 min. Then, fractionated with ethyl acetate and salted out by additionof 4 different salts (Na2CO3, NaHCO3, K2HPO4, and NaCl). Oxyresveratrol concentration of the extracts was analyzed with TLC-densitometry and HPLC. Results: Based on the results of this study, the most optimum condition of IL-MAE for oxyresveratrol from M. alba roots was using [Bmim]Br concentration 1.5mol/L with microwave power 10% for 10 min. The salt 0.01 mol/L NaHCO3 was optimum for the back extraction of oxyresveratrol from [Bmim]Br solution. Under optimized conditions, oxyresveratrol concentration in M. alba roots extract using IL-MAE method was 735.25 ppm. Conclusion: This IL-MAE method was suitable to be applied as an alternative technique to extract oxyresveratrol from the plants. Key words: Ionic liquid, Microwave-assisted extraction, IL-MAE, Oxyresveratrol, Morus alba, Roots extract.

Protein Tyrosine Phosphatase 1B Inhibition and Glucose Uptake Potentials of Mulberrofuran G, Albanol B, and Kuwanon G from Root Bark of Morus alba L. in Insulin-Resistant HepG2 Cells: An In Vitro and In Silico Study.

Type II diabetes mellitus (T2DM) is the most common form of diabetes and has become a major health problem across the world. The root bark of Morus alba L. is widely used in Traditional Chinese Medicine for treatment and management of diabetes. The aim of the present study was to evaluate the enzyme inhibitory potentials of three principle components, mulberrofuran G (1), albanol B (2), and kuwanon G (3) in M. alba root bark against diabetes, establish their enzyme kinetics, carry out a molecular docking simulation, and demonstrate the glucose uptake activity in insulin-resistant HepG2 cells. Compounds 1–3 showed potent mixed-type enzyme inhibition against protein tyrosine phosphatase 1B (PTP1B) and α-glucosidase. In particular, molecular docking simulations of 1–3 demonstrated negative binding energies in both enzymes. Moreover, 1–3 were non-toxic up to 5 µM concentration in HepG2 cells and enhanced glucose uptake significantly and decreased PTP1B expression in a dose-dependent manner in insulin-resistant HepG2 cells. Our overall results depict 1–3 from M. alba root bark as dual inhibitors of PTP1B and α-glucosidase enzymes, as well as insulin sensitizers. These active constituents in M. alba may potentially be utilized as an effective treatment for T2DM.

Anti-inflammatory activities and potential mechanisms of phenolic acids isolated from Salvia miltiorrhiza f. alba roots in THP-1 macrophages

Ethnopharmacological relevanceThe roots of Salvia miltiorrhiza f. alba (Lamiaceae) (RSMA) are used as the Danshen, a traditional Chinese medicine, to treat the vascular diseases at local clinics, especially for the remedy of thromboangiitis obliterans (TAO) more than 100 years. Phenolic acids are one of the major effective constituents of RSMA, and some studies have linked phenolic acids with anti-inflammatory functions. Aim of the studyThe purpose of this research was to isolate phenolic acids from RSMA and investigate their anti-inflammatory effects and potential mechanisms. Materials and methodsNine already known compounds were obtained from RSMA. Their structures were elucidated through the spectroscopic analysis and comparing the reported data. The anti-inflammatory effects and potential mechanisms were investigated in LPS-stimulated THP-1 cells, using salvianolic acid B (SalB) as the positive control. The enzyme-linked immunosorbent assays (ELISA) were used to determine the secretory protein levels of interleukin-1β (IL-1β), interleukin-6 (IL-6) and tumor necrosis factor-α (TNF-α). And quantitative real-time polymerase chain reaction (qRT-PCR) was used to analyze the mRNA levels of these inflammatory cytokines. The expression of TLR4, p65, p-p65, IκBα, and p-IκBα were measured using western blot. ResultsAll these compounds, except for rosmarinic acid (5) and isosalvianolic acid (6) for IL-6 protein levels, rosmarinic acid-o-β-D-glucopyranoside (3) for IL-6 mRNA, and rosmarinic acid-o-β-D-glucopyranoside (3), rosmarinic acid (5) and isosalvianolic acid (6) for TNF-α mRNA levels, remarkably inhibited the production of TNF-α, IL-1β, and IL-6 at the concentration of 5 and 25 μM in the mRNA and protein levels. Lithospermic acid (7) showed the strongest inhibitory effect among them and was similar to that of SalB. In particular, lithospermic acid (7) and SalB markedly downregulated the expressions of TLR4, p-p65, and p-IκBα induced by LPS in THP-1 macrophages. ConclusionsAll the phenolic acids displayed anti-inflammatory properties and the potential mechanisms involved the TLR4/NF-κB signal pathway. Results of this study indicate that phenolic acids may be effective constituents of RSMA to treat vascular diseases associated with inflammation.

Quadruple high-resolution α-glucosidase/α-amylase/PTP1B/radical scavenging profiling combined with high-performance liquid chromatography–high-resolution mass spectrometry–solid-phase extraction–nuclear magnetic resonance spectroscopy for identification of antidiabetic constituents in crude root bark of Morus alba L.

In this paper, quadruple high-resolution α-glucosidase/α-amylase/PTP1B/radical scavenging profiling combined with HPLC-HRMS-SPE-NMR were used for studying the polypharmacological properties of crude root bark extract of Morus alba L. This species is used as an anti-diabetic principle in many traditional treatment systems around the world, and the crude ethyl acetate extract of M. alba root bark was found to inhibit α-glucosidase, α-amylase and protein-tyrosine phosphatase 1B (PTP1B) with IC50 values of 1.70 ± 0.72, 5.16 ± 0.69, and 5.07 ± 0.68 μg/mL as well as showing radical scavenging activity equaling a TEAC value of (3.82 ± 0.14) × 104 mM per gram extract. Subsequent investigation of the crude extract using quadruple high-resolution α-glucosidase/α-amylase/PTP1B/radical scavenging profiling provided a quadruple biochromatogram that allowed direct correlation of the HPLC peaks with one or more of the tested bioactivities. This was used to target subsequent HPLC-HRMS-SPE-NMR analysis towards peaks representing bioactive analytes, and led to identification of a new Diels-Alder adduct named Moracenin E as well as a series of Diels-Alder adducts and isoprenylated flavonoids as potent α-glucosidase and α-amylase inhibitors with IC50 values in the range of 0.60–27.15 μM and 1.22–69.38 μM, respectively. In addition, these compounds and two 2-arylbenzofurans were found to be potent PTP1B inhibitors with IC50 values ranging from 4.04 to 21.67 μM. The high-resolution radical scavenging profile also revealed that almost all of the compounds possess radical scavenging activity. In conclusion the quadruple high-resolution profiling method presented here allowed a detailed profiling of individual constituents in crude root bark extract of M. alba, and the method provides a general tool for detailed mapping of bioactive constituents in polypharmacological herbal remedies.

Protective effect of Paeoniae radix alba root extract on immune alterations in mice with atopic dermatitis

ABSTRACTAtopic dermatitis is a progressive inflammatory disease characterized by type 2 helper T cell (TH2) reactivity. The aim of this study was to examine the therapeutic effects of Paeoniae radix alba root extract using a murine model of atopic dermatitis. Atopic dermatitis was induced in a murine model characterized by immune alterations skewed toward TH2 reactivity and pathophysiological dermal alterations which resemble human atopic dermatitis. The root extract at 1% or 6% was applied to the mouse dorsal skin for 3 weeks following induction of atopic dermatitis. Splenocytes were stimulated with immobilized anti-CD3 for 48 h to measure cytokine production. Levels of serum IgE, IgG1, and IgG2a were quantitated. Epidermal thickness and numbers of skin mast cells were determined. Mice in which atopic dermatitis was induced displayed increased numbers of skin mast cells, increased frequency of scratching, elevated serum IgE levels, increased ratios of IgG1 to IgG2a, and ratios of IL-4 to IFN-γ. The frequency of scratching was significantly decreased following application of 1% or 6% extract for 1 week. The root extract also reversed TH2 skewing, as serum IgE levels, ratio of serum IgG1 to IgG2a, and ratio of IL-4 to IFN-γ production by in vitro-stimulated splenocytes were all suppressed following application of 1% or 6% extract for 3 weeks. Taken together Paeoniae radix alba root extract is suggested to reverse the immunological alterations and skin manifestations symptoms found in atopic dermatitis.

Effect of AmLexin (Acacia catechu & Morus alba) on Redox Balance and Subjective Pain in Healthy Runners

IntroductionIntense exercise is known to increase both oxidative stress and post‐exercise pain. Muscle soreness and soft tissue pain is a frequent complaint among recreational exercisers and a significant obstacle to continued participation in regular physical activity.ObjectiveOur study examined the effects of an herbal dietary supplement (AmLexin; combination of Acacia catechu heartwood extract and Morus alba root bark extract) versus a look‐alike placebo on redox balance (plasma oxidative stress and oxidative capacity) and subjective pain levels following intense exercise.Subjects & MethodsTwenty‐four (N=24) healthy men and women were recruited to participate in a progressive 8‐week running program followed by a 13.1‐mile timed run (half‐marathon). Subjects were randomly divided in double‐blind fashion to receive either the supplement at 200 mg b.i.d. (N=12) or placebo (N=12) for 9 weeks (8 weeks pre‐race during training and 1‐week post‐race). We measured range of motion (ROM) and pain/stiffness (WOMAC & VAS) weekly during 8‐weeks of training and daily for 6‐days post‐race, and plasma redox status on days 1 and 6 post‐race.ResultsDue to large variations in ROM between and within subjects, there were no significant differences in ROM between supplementation groups. However, multiple measures of pain and stiffness were significantly lower for days 1–3 post‐race in the AmLexin group compared to the Placebo group, including WOMAC scores for overall pain at day 1 and 3 post run and a clear trend of less stiffness and better activities of daily living in the AmLexin group. The AmLexin group also showed statistically significant less pain/stiffness than Placebo based on VAS scores for multiple body parts (quadriceps, calves, hamstrings, and knees) at several time points after the race. Redox balance assessment indicated the AmLexin group to show a strong trend toward improved status at day 1, that reached statistical significance for lower oxidative stress and higher oxidative capacity at day 6 post‐race compared to the Placebo group.ConclusionAmLexin has previously been reported to protect joint cartilage from damage, reducing the amount of glycosaminoglycan released by articular cartilage (1 &2), and to be an effective quencher of super oxide anion, a free radical generated by tissue wear and tear (3). These clinical results further demonstrate AmLexin's effectiveness to reduce oxidative stress, improve oxidative capacity, and prevent the development of post‐exercise subjective pain and stiffness in a population of healthy non‐arthritic adults. Therefore, AmLexin may be viewed as an effective dietary supplement to facilitate healthy pain‐free participation in regular physical activity.Support or Funding InformationFunding provided by UnigenThis abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.

Anti-inflammatory and preventive activity of white mulberry root bark extract in an experimental model of pancreatitis

Pancreatitis is characterized by highly morbid inflammation in the pancreas. Currently, there is no specific drug available for pancreatitis except supportive medicines. The present study assessed the pancreato-protective effect of Morus alba root bark extract by using alcohol and cerulein-induced model of pancreatitis. The study also investigated the phytochemical profile through GC-MS and HPLC. Methanolic extract of Morus alba root bark extract (MEMARB) was subjected to GC-MS and HPLC studies. Male albino Wistar rats were administered ethanol (0%–36%) and cerulein (20 μg/kg b.wt. i.p.) with or without MEMARB. Serum lipase, amylase, caspase-1, lipid peroxidation products, glutathione and enzymatic antioxidants were determined. Histological changes in the pancreas were assessed. Cudraflavone B in MEMARB was quantified by HPLC. Significant amount of Cudraflavone B was detected by quantitative HPLC. Marked increase in the levels of serum amylase, lipase, caspase-1, IL-18 and IL-1β were observed in ethanol and cerulein administered rats than in MEMARB co-administered rats. In MEMARB co-administered rats, the antioxidant status was restored to near normal levels. Histological examinations showed that MEMARB significantly reduced the inflammatory and fibrotic changes. The results reveal the potent pancreato-protective effects of Morus alba root bark. The anti-inflammatory effect of Morus alba root bark extract might be due to the presence of various phytonutrients including Cudraflavone B.

Eco- and genotoxicity profiling of a rapeseed biodiesel using a battery of bioassays

Biodiesel is considered an important renewable energy source but still there is some controversy about its environmental toxicity, especially to aquatic life. In our study, the toxicity of water soluble fraction of biodiesel was evaluated in relatively low concentrations using a battery of bioassays: Vibrio fischeri bioluminescence inhibition, Sinapis alba root growth inhibition, Daphnia magna immobilization, boar semen live/dead ratio and DNA fragmentation and Unio pictorum micronucleus test. While the S. alba test indicated nutritive (stimulating) effect of the sample, the biodiesel exerted toxic effect in the aquatic tests. D. magna was the most sensitive with EC50 value of 0.0226%. For genotoxicity assessment, the mussel micronucleus test (MNT) was applied, detecting considerable genotoxic potential of the biodiesel sample: it elucidated micronuclei formation already at low concentration of 3.3%. Although this test has never been employed in biodiesel eco/genotoxicity assessments, it seems a promising tool, based on its appropriate sensitivity, and representativity.

Vascular Protection by Ethanol Extract of Morus alba Root Bark: Endothelium-Dependent Relaxation of Rat Aorta and Decrease of Smooth Muscle Cell Migration and Proliferation.

Morus alba (white mulberry) is native to the northern part of Korea and popularly used as a traditional medicine due to its numerous health benefits against human's disease. However, the possibility that M. alba may also affect the cardiovascular system remains unexplored. This study sought to investigate the vascular protective effects of the root bark extract of M. alba (MAE). Vascular reactivity was performed in organ baths using isolated rat thoracic aorta, while platelet derived growth factor (PDGF) induced proliferation and migration of vascular smooth muscle cells (VSMCs) were studied by 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium (MTS) and wound healing assay, respectively. MAE evoked a concentration dependent vasorelaxation following endothelium-dependent pathway. However, vessel relaxations in response to MAE were markedly reduced after endothelium removal; treatment of endothelial nitric oxide synthase inhibitor, guanylyl cyclase inhibitor, and nonspecific potassium channel inhibitor, however, was not altered by cyclooxygenase inhibitor. Furthermore, MAE also significantly blunted contractile response to vasoconstrictor agent, phenylephrine. Taken together, the current evidence revealed that MAE is a potent endothelium-dependent vasodilator and this effect was involved in, at least in part, nitric oxide cyclic-guanosine monophosphate (NO-cGMP) pathway in combination with potassium (K+) channel activation. Moreover, MAE inhibited proliferation and migration of VSMCs induced by PDGF. Therefore, MAE could be a promising candidate of natural medicine for preventing and controlling cardiovascular diseases linked with endothelial dysfunction.

Ionic Liquid-based Microwave-Assisted Extraction (Il-MAE) of Oxyresveratrol from Morus alba Roots

Ionic Liquid-based Microwave-Assisted Extraction (Il-MAE) of Oxyresveratrol from Morus alba Roots

A strategy for screening of α-glucosidase inhibitors from Morus alba root bark based on the ligand fishing combined with high-performance liquid chromatography mass spectrometer and molecular docking

A new method based on ligand fishing combined with high-performance liquid chromatography quadrupole-time-of-flight mass spectrometer and molecular docking was established to screen α-glucosidase inhibitors from a traditional Chinese medicine Morus alba root bark. α-Glucosidase was immobilized on magnetic nanoparticles, used as a solid support to incubate with crude extract. After ligand fishing, the eluates were analyzed by high-performance liquid chromatography quadrupole-time-of-flight mass spectrometer, obtaining eleven ligands (1–4, 6–12) eventually. In order to discriminate the non-specific binders and discover powerful enzyme inhibitors, molecular docking was further performed and three of the eleven ligands were optimized to be excellent α-glucosidase inhibitors by the confirmation of isolation and bioassay of individual compounds. These three ligands, sanggenons G (6), O (7) and sanggenol G (12) exhibited striking inhibitory activities with extremely low IC50 values. The results suggest that established method will be applied to a wide range of target protein to screen potential bioactive constituents from herbal medicines.

Synergism of prenylflavonoids from Morus alba root bark against clinical MRSA isolates

BackgroundClinical methicillin-resistant Staphylococcus aureus (MRSA) is a thorny problem in current anti-infective therapeutics and a challenge of new drug development. Plant prenylflavonoids possess anti-MRSA activity, but few of the prenylflavonoids have been reported the synergistic anti-MRSA effect when they are used in combination with conventional antibacterial agents. PurposeThis study deals with anti-MRSA activity of four prenylflavonoids from the root bark of Morus alba and their synergism with 11 conventional antibacterial agents. MethodsChromatographic methods and spectral analysis were used to isolate and identify the prenylflavonoids. The antibacterial activity and synergism were assessed by the broth microdilution method, checkerboard dilution test, and time-kill curve assay, respectively. ResultsFour prenylflavonoids, i.e., cyclocommunol (Cy, 1), morusinol (Ml, 2), morusin (Mi, 3) and kuwanon E (Ku, 4), were isolated from Morus alba bark ethanol extract. Compounds 1, 3 and 4 showed high antimicrobial activity on both methicillin-susceptible S. aureus (MSSA) and MRSA strains with MICs/MBCs at 4–16/32–64 and 4–32/16–128 µg/ml, respectively. Ml (2) was not active. Compound 2 showed synergy with amikacin (AK) and streptomycin (SM) against all the ten MRSA isolates. Ml (2) and Ku (4) also showed synergy with ciprofloxacin (CI), etimicin (EM) and vancomycin (VA) against 7–9 isolates. The fractional inhibitory concentration indices (FICIs) ranged 0.09–1.00 and the dose reduction indices (DRIs) of these antibacterial agents ranged 2–128. Cy (1) and Mi (3) showed synergy with the tested antibacterial agents against only 1–3 MRSA isolates except VA. Furthermore, the MRSA resistance could be reversed in the combinations of AK with Cy, Ml, Mi and Ku; EM with Mi and Ku; and SM with Ml by the criteria of MIC interpretive standards for Staphylococcus spp. of CLSI. All the combinations showed only indifference in the 1 × MIC time-killing experiments. The prenylated substitutions play an important role in the activity of the compounds used alone and combined with the tested antibacterials. ConclusionsThe study revealed for the first time the anti-MRSA synergism of prenylflavonoids 1–4 with eleven antibacterial agents and the reversal of MRSA resistance to aminoglycosides, especially amikacin. The results might be valuable for the development of new antibacterial drugs and synergists against MRSA infection.

Anti-Alzheimer’s disease activity of compounds from the root bark of Morus alba L.

The inhibition of acetylcholinesterase (AChE), butyrylcholinesterase (BChE), and β-site amyloid precursor protein cleaving enzyme 1 (BACE1) plays important roles in prevention and treatment of Alzheimer's disease (AD). Among the individual parts of Morus alba L. including root bark, branches, leaves, and fruits, the root bark showed the most potent enzyme inhibitory activities. Therefore, the aim of this study was to evaluate the anti-AD activity of the M. alba root bark and its isolate compounds, including mulberrofuran G (1), albanol B (2), and kuwanon G (3) via inhibition of AChE, BChE, and BACE1. Compounds 1 and 2 showed strong AChE- and BChE-inhibitory activities; 1-3 showed significant BACE1 inhibitory activity. Based on the kinetic study with AChE and BChE, 2 and 3 showed noncompetitive-type inhibition; 1 showed mixed-type inhibition. Moreover, 1-3 showed mixed-type inhibition against BACE1. The molecular docking simulations of 1-3 demonstrated negative binding energies, indicating a high affinity to AChE and BACE1. The hydroxyl group of 1-3 formed hydrogen bond with the amino acid residues located at AChE and BACE1. Consequently, these results indicate that the root bark of M. alba and its active compounds might be promising candidates for preventive and therapeutic agents for AD.

Neuroprotection by Paeoniflorin in the MPTP mouse model of Parkinson's disease

Paeoniflorin (PF) is a major bioactive ingredient in Radix Paeonia alba roots that has low toxicity and has been shown to have neuroprotective effects. Our in vitro experiments suggested that PF affords a significant neuroprotective effect against MPP+-induced damage and apoptosis in PC12 cells through Bcl-2/Bax/caspase-3 pathway. The objectives of the present study were to explore the potential neuroprotective effect of PF in 1-methyl-4-phenyl-1, 2, 3, 6-tetrahydropyridine (MPTP)-treated mouse model of Parkinson's disease (PD). Our results demonstrated that PF treatment ameliorated the behavioral deficits of “in spontaneous motor activity and latency to fall of the rotarod test”, and reduced dopaminergic cell loss that were induced by MPTP in a dose-dependent manner in an in vivo model of PD. In addition, we found that treatment of PF protected dopaminergic neurons by preventing MPTP-induced decreases in striatal and substantia nigra dopaminergic transporter (DAT) and tyrosine hydroxylase (TH) protein levels, and by changing dopamine catabolism and inhibiting dopamine turnover. Furthermore, it was also associated with up-regulation of the Bcl-2/BAD ratio, and inhibition of the activation of caspase-9 and caspase-3. These results showed that PF promoted dopamine neuron survival in vivo due to the MAO-B inhibition, and the PI3K/Akt signaling pathway may have mediated the protection of PF against MPTP, suggesting that PF treatment might represent a neuroprotective treatment for PD.

Application of backpropagation artificial neural network prediction model for the PAH bioremediation of polluted soil

The backpropagation (BP) artificial neural network (ANN) is a renowned and extensively functional mathematical tool used for time-series predictions and approximations; which also define results for non-linear functions. ANNs are vital tools in the predictions of toxicant levels, such as polycyclic aromatic hydrocarbons (PAH) potentially derived from anthropogenic activities in the microenvironment. In the present work, BP ANN was used as a prediction tool to study the potential toxicity of PAH carcinogens (PAHcarc) in soils. Soil samples (16 × 4 = 64) were collected from locations in South-southern Nigeria. The concentration of PAHcarc in laboratory cultivated white melilot, Melilotus alba roots grown on treated soils was predicted using ANN model training. Results indicated the Levenberg-Marquardt back-propagation training algorithm converged in 2.5E+04 epochs at an average RMSE value of 1.06E-06. The averagedR2 comparison between the measured and predicted outputs was 0.9994. It may be deduced from this study that, analytical processes involving environmental risk assessment as used in this study can successfully provide prompt prediction and source identification of major soil toxicants.

Evaluation of the anti-inflammatory activities of tanshinones isolated from Salvia miltiorrhiza var. alba roots in THP-1 macrophages

Ethnopharmacological relevanceSalvia miltiorrhiza var. alba roots are used as the Chinese traditional medicine Danshen for the treatment of cardiovascular diseases in local clinical practice. Tanshinones are the major effective constituents of S. miltiorrhiza var. alba roots, but only tanshinone IIA, tanshinone I, cryptotanshinone, and 15,16-dihydrotanshinone have been investigated for their anti-inflammatory activities. Materials and methodsEleven known compounds were isolated from S. miltiorrhiza var. alba roots, and the structures of all compounds were elucidated by spectroscopic analysis and comparisons with reported data. Immune anti-inflammatory activities were assessed by the ability to inhibit the production of tumor necrosis factor (TNF)-α, interleukin (IL)-1β, and interleukin (IL)-8 using enzyme-linked immunosorbent assay. Quantitative real-time polymerase chain reaction (qRT-PCR) was also used to compare the inhibitory effects of the compounds on TNF-α, IL-1β, and IL-8 mRNA expression with that of tanshinone IIA in lipopolysaccharide-stimulated THP-1 macrophages. ResultsAll tanshinones, except for compound 5, significantly inhibited the mRNA and protein expression of TNF-α, IL-1β, and IL-8, and their anti-inflammatory activities were stronger than that of tanshinone IIA. Compound 9 (5μM) showed the highest inhibitory effects for TNF-α, IL-1β, and IL-8, at 56.3%, 67.6%, and 51.7%, respectively. ConclusionsTen of the 11 tanshinones were shown to have anti-inflammatory properties superior to those of TSIIA, and which significantly inhibited the expression of TNF-α, IL-1β, and IL-8. The present results provided a referential basis for explaining the use of S. miltiorrhiza var. alba root as a Chinese folk medicine for treating cardiovascular diseases associated with inflammation, and show the importance of trace constituents of this herb.

Protective Effects of Paeoniflorin Against MPP(+)-induced Neurotoxicity in PC12 Cells.

In the present study, we investigated the protective mechanism of paeoniflorin (PF), a monoterpene glycoside extracted from Radix Paeoniae alba roots, on MPP(+)-induced neurotoxicity in cultured rat pheochromocytoma cells (PC12). Our work included examination of cell viability assessment, amounts of released lactic dehydrogenase (LDH), intracellular Ca(2+) concentration, cell apoptosis, mitochondrial membrane potential, caspase-3 activity, and expression profiling of two apoptosis-related genes (Bcl-2 and Bax). It was shown that, PF functioned as an MPP(+) antagonist, being able to suppress apoptosis, decrease LDH release and Ca(2+) concentration, attenuate membrane potential collapse and, inhibit caspase-3 activation, decrease in Bax/Bcl-2 ratio. These observations suggest that PF could protect PC12 cells against MPP(+)-induced injury and the mechanism PF's neuroprotective effect was closely associated with Bcl-2 up-regulation and Bax down-regulation. PF has neuroprotective effects on MPP(+)-induced apoptosis in PC12 cells via regulating mitochondrial membrane potential and Bcl-2/Bax/caspase-3 signaling pathways, and this new insight will help develop a PF-based therapeutic strategy for treatmenting neurodegenerative diseases and injury.

Copper excess promotes propagation and induces proteomic change in root cultures of Hyoscyamus albus L.

Hyoscyamus albus L. seedlings respond positively to copper (Cu) excess. In the present study, to understand how roots cope with Cu excess, propagation and proteome composition in the presence of Cu were examined using a root culture system. When H. albus roots were cultured in a medium without Cu, root growth deteriorated. However, in the presence of Cu, root growth increased in a concentration-dependent manner, and vigorous lateral root development was observed at 200 μM Cu. Cu accumulation in the roots increased with the Cu supply. Subcellular fractionation revealed that the highest amount of Cu was present in the cell wall-containing fraction, followed by the soluble fraction. However, the highest specific incorporation of Cu, in terms of fresh weight, was in the mitochondria-rich fraction. High Cu levels enhanced respiration activity. Comparative proteomic analysis revealed that proteins involved in carbohydrate metabolism, de novo protein synthesis, cell division, and ATP synthesis increased in abundance, whereas the proteasome decreased. These results indicate that Cu promotes propagation of H. albus roots through the activation of the energy supply and anabolism. Newly propagated root tissues and newly generated proteins that bind to Cu may provide space and reservoirs for deposition of additional Cu.

Phenolic profile, antioxidant and anti-inflammatory potential of herb and root extracts of seven selected legumes

Herb and root parts of legumes are not commonly used in human consumption, since they are traditionally considered waste material. Although inedible, these plant parts have a complex chemical composition, with potential biological activities. They are especially rich in polyphenols, such as flavonoids, isoflavonoids and phenolic acids. In this paper, phenolic profile of the seven selected legumes (soybean (Glycine max L.), common bean (Phaseolus vulgaris L.), pea (Pisum sativum L.), broad bean (Vicia faba L.), chickpea (Cicer arietinum L.), white lupin (Lupinus albus L.) and grass pea (Lathyrus sativus L.)) was examined using LC–MS/MS method. In the examined herb and root extracts 33 phenolic compounds were identified and quantified. High levels of isoflavones genistein and daidzein were found in G. max, P. vulgaris and L. albus root extracts (1.02–1.53, 0.309–0.648 and 2.81mg/g, respectively). C. arietinum, L. albus and P. vulgaris herb extracts were found to be the richest sources of flavonoid compounds (13.1–19.4, 5.98 and 2.36–3.64mg/g, respectively). Since polyphenols have redox properties, antioxidant activity of the abovementioned plants was investigated using DPPH, NO, and FRAP assay. All samples demonstrated the ability to scavenge both DPPH and NO radical, and exhibited a good FRAP reducing power. V. faba exhibited the highest antioxidant activity (DPPH IC50 11.7–20.7μg/mL, NOIC30 0.080–0.34 mg/mL, FRAP 32.6–66.9 AAE/g dw). Anti-inflammatory potential was examined using in vitroCOX-1/12-LOX pathway inhibition assay, resulting in moderate activity of all samples (IC50 2.09–8.00 for all metabolites). The obtained results indicate a strong potential of waste legume material as a promising source for isolation of bioactive antioxidant and anti-inflammatory compounds for their use as therapeutic and dietary supplements in pharmaceutical and food industry.