To investigate the effects of phosphorus fertilizer on the morphological traits, active ingredients and rhizosphere soil microbial community of Polygala tenuifolia. The phosphorus fertilizer was calculated in terms of P_2O_5. Five treatments were set up: 0(CK), 17(P1), 34(P2), 51(P3), and 68(P4) kg per Mu(1 Mu≈667 m~2). A randomized block design was adopted. Samples of P. tenuifolia and its rhizosphere soil were collected under different superphosphate fertilizer treatments. Illumina high-throughput sequencing was used to analyze the rhizosphere soil microbial community, 9 morphological traits were measured and the content of 11 active ingredients were determined. The results showed that the whole plant weight, shoot fresh weight, root weight, and root peel thickness were the highest under P1 treatment, increasing by 34.41%, 38.80%, 39.21%, and 3.17% respectively compared to CK. Under P2 treatment, the plant height, stem diameter, root thickness, and core thickness were significantly higher than CK. Phosphorus fertilizer had a significant impact on the content of tenuifolin, sibiricose A5, sibiricose A6, arillanin A, 3,6'-disinapoyl sucrose, and polygalaxanthone Ⅲ. Correlation analysis results showed that the relative abundance of Arthrobacter, Bacillus, norank＿f＿Vicinamibacteraceae, norank＿o＿Vicinamibacterales, MND1 and other bacteria, as well as the relative abundance of Neocosmospora, Paraphoma and other fungi were positively correlated with root diameter, wood core diameter, the whole plant weight, root weight, shoot fresh weight of P. tenuifolia. Bacillus, Neocosmospora, Subulicystidium were significantly positively correlated with oligosaccharides such as 3,6'-disinapoyl sucrose, sibiricose A5、sibiricose A6、glomeratose A、arillanin A and tenuifoliside C. Arthrobacter, Humicola, Aspergillus, Paraphoma were positively correlated with tenuifolin and norank＿f＿Vicinamibacteraceae, norank＿o＿Vicinamibacterales, Fusarium were positively correlated with polygalaxanthone Ⅲ. Evidently, appropriate phosphorus application is conducive to the growth and quality improvement of P. tenuifolia, and can increase the abundance of beneficial microorganisms in the soil.
Rhizosphere ; Phosphorus/pharmacology* ; Soil Microbiology ; Polygala/anatomy & histology* ; Fertilizers/analysis* ; Bacteria/metabolism* ; Soil/chemistry* ; Microbiota/drug effects* ; Plant Roots/metabolism*

OBJECTIVE: To investigate the anti-oxidant and anti-inflammatory effects of ethanol extract of Polygala sibirica L. var megalopha Fr. (EEP) on RAW264.7 mouse macrophages. METHODS: RAW264.7 cells were pretreated with 0-200 µg/mL EEP or vehicle for 2 h prior to exposure to 1 µg/mL lipopolysaccharide (LPS) for 24 h. Nitric oxide (NO) and prostaglandin (PGE₂) production were determined by Griess reagent and enzyme-linked immunosorbent assay (ELISA), respectively. The mRNA levels of inducible nitric oxide synthase (iNOS), cyclooxygenase-2 (COX-2), tumor necrosis factor α (TNF-α), interleukin-1beta (IL-1β), and IL-6 were determined using reverse transcription polymerase chain reaction (RT-PCR). Western blot assay was used to determine the protein expressions of iNOS, COX-2, phosphorylation of extracellular regulated protein kinases (ERK1/2), c-Jun N-terminal kinase (JNK), inhibitory subunit of nuclear factor Kappa B alpha (Iκ B-α) and p38. Immunofluorescence was used to observe the nuclear expression of nuclear factor-κ B p65 (NF-κ B p65). Additionally, the anti-oxidant potential of EEP was evaluated by reactive oxygen species (ROS) production and the activities of catalase (CAT) and superoxide dismutase (SOD). The 2,2-diphenyl-1-picrylhydrazyl (DPPH), hydroxyl (OH), superoxide anion (O₂^-) radical and nitrite scavenging activity were also measured. RESULTS: The total polyphenol and flavonoid contents of EEP were 23.50±2.16 mg gallic acid equivalent/100 g and 43.78±3.81 mg rutin equivalent/100 g. With EEP treatment (100 and 150 µg/mL), there was a notable decrease in NO and PGE₂ production induced by LPS in RAW264.7 cells by downregulation of iNOS and COX-2 mRNA and protein expressions (P<0.01 or P<0.05). Furthermore, with EEP treatment (150 µg/mL), there was a decrease in the mRNA expression levels of TNF-α, IL-1β and IL-6, as well as in the phosphorylation of ERK, JNK and p38 mitogen-activated protein kinase (MAPK, P<0.01 or P<0.05), by blocking the nuclear translocation of NF-κ B p65 in LPS-stimulated cells. In addition, EEP (100 and 150 µg/mL) led to an increase in the anti-oxidant enzymes activity of SOD and CAT, with a concomitant decrease in ROS production (P<0.01 or P<0.05). EEP also indicated the DPPH, OH, O₂^- radical and nitrite scavenging activity. CONCLUSION EEP inhibited inflammatory responses in activated macrophages through blocking MAPK/NF-κ B pathway and protected against oxidative stress.
Animals ; Mice ; Antioxidants/pharmacology* ; Lipopolysaccharides/pharmacology* ; Polygala ; Transcription Factor RelA/metabolism* ; Tumor Necrosis Factor-alpha/metabolism* ; Ethanol/chemistry* ; Interleukin-6/metabolism* ; Anti-Inflammatory Agents/chemistry* ; Reactive Oxygen Species/metabolism* ; Cyclooxygenase 2/metabolism* ; Nitrites/metabolism* ; NF-kappa B/metabolism* ; Nitric Oxide/metabolism* ; Superoxide Dismutase/metabolism* ; RNA, Messenger ; Nitric Oxide Synthase Type II/metabolism*

Five compounds (tenuifoliside C, tenuifoliside D, telephiose A, telephiose C and polygalaxanthone III) from polygala tenuifolia wild were incubated together with CYP probe substrate in human liver microsomes to investigate the inhibitory effect towards CYP450 enzyme. Phenacetin (CYP1A2), coumarin (CYP2A6), paclitaxel (CYP2C8), diclofenac (CYP2C9), S-mepheriytoin (CYP2C19), dextromethorphan (CYP2D6), chlorzoxazone (CYP2E1), midazolam (CYP3A) were selected as the isoforfn specific substrate. And the formation of paracetamol, 7-hydroxycoumarin, 6alpha-hydroxy paclitaxel, 4'-hydroxydiclofenac, dextrorphan, 6-hydroxychlorzoxazone, 1'-hydroxymidazolam, 4'-hydroxymephenytoin were detected respectively to measure the effect towards CYP450 by high-pressure liquid chromatography (HPLC). The result shows that five compounds from polygala tenuifolia willd significantly inhibit chlorzoxazone 6-hydroxylation catalyzed by CYP2E1, while showed no effect towards CYP1A2, CYP2A6, CYP2C8, CYP2C9, CYP2C19, CYP2D6, CYP3A. And IC50 value was 38.73, 54.14, 61.77, 62.22, 50.56 micromol x L(-1), respectively.
Cytochrome P-450 Enzyme System ; metabolism ; Esters ; pharmacology ; Glycosides ; pharmacology ; Humans ; Microsomes, Liver ; drug effects ; enzymology ; Oligosaccharides ; pharmacology ; Polygala ; chemistry ; Xanthones ; pharmacology

OBJECTIVETo observe the effect of Polygala tenuifolia Willd YZ-50 on the mRNA expression of brain-derived neurotrophic factor (BNDF) and its receptor TrkB in rats with chronic stress depression.es.

METHODSNormal male Wistar rats were divided in to control group, model group, desipramine (20 mg/kg) group, and low and high-dose (2.8 and 5.6 g/kg) YZ-50 groups. The total RNA was extracted from the rats with chronic stress depression, and the mRNA expression of BDNF and TrkB was detected by RT-PCR.

RESULTSCompared with the model group, YZ-50 at both low and high doses significantly increased the mRNA expression of BDNF and TrkB in the hippocampus of rats with chronic stress depression, and the effect was more obvious in the high-dose group (P<0.01).

CONCLUSIONYZ-50 can up-regulate the expression of BDNF and TrkB mRNA to promote the recovery of the neurons from chronic stress-induced damages and produces anti-depressant effect.

Animals ; Brain-Derived Neurotrophic Factor ; genetics ; metabolism ; Depression ; drug therapy ; etiology ; metabolism ; Drugs, Chinese Herbal ; pharmacology ; Male ; Polygala ; chemistry ; RNA, Messenger ; genetics ; metabolism ; Rats ; Rats, Wistar ; Receptor, trkB ; genetics ; metabolism ; Stress, Physiological ; physiology ; Up-Regulation ; drug effects

OBJECTIVETo observe the effect of tenuigenin (TEN) on aggregated amyloid beta-protein 1-40 (Abeta(1-40)-induced cytotoxicity of primary cultural cortical neurons in vitro.

METHODIn order to establish neurotoxic model, the primary cultural rat cortical neurons were treated with 25 micromol x L(-1) aggregated Abeta(1-40), which were divided into a model group and 3 different dose groups of TEN (50, 100, 200 micromol x L(-1), respectively), and a normal control group with no treatment of Abeta(1-40) was set up. The morphological changes of the neurons before and after administration of TEN were examined under a phase contrast microscope. Neuronal viabilities were detected by MTT colorimetry. Injuring degrees of the neuronal membrane were assessed by lactate dehydrogenase (LDH) colorimetry.

RESULTAs compared with the normal control, treatment of primary cultural neurons with Ap, (25 micromol x L(-1)) for 24 h caused a significant decrease in viabilities and morphological changes of nerve cells, with neurons losing adherent ability or shedding, and the synapse shortening found by microscope. The percentage of apoptotic nerve cells and the LDH leakage were significantly decreased, and the survival rate of neurons was significantly increased in both the TEN high and medium dose groups.

CONCLUSIONThe aggregated Abeta(1-40) has a definite neurotoxicity for cultural cortical neurons, and TEN can significantly protect the neurons from the cytotoxicity of Abeta(1-40).

Amyloid beta-Peptides ; toxicity ; Animals ; Apoptosis ; drug effects ; Cell Survival ; drug effects ; Cells, Cultured ; Cerebral Cortex ; cytology ; Colorimetry ; Dose-Response Relationship, Drug ; Drugs, Chinese Herbal ; isolation & purification ; pharmacology ; Female ; L-Lactate Dehydrogenase ; metabolism ; Neurons ; cytology ; drug effects ; metabolism ; Neuroprotective Agents ; isolation & purification ; pharmacology ; Peptide Fragments ; toxicity ; Plants, Medicinal ; chemistry ; Polygala ; chemistry ; Pregnancy ; Rats ; Rats, Sprague-Dawley

OBJECTIVE: To investigate the effect of reinioside C (RC) on the expression of lectin-like oxidized low density lipoprotein receptor (LOX)-1 mRNA and LOX-1 protein induced by oxidized low density lipoprotein (ox-LDL) in cultured human umbilical vein endothelial cells (HUVEC). METHODS: HUVECs were cultured with ox-LDL (50 mg/L) for 24 h in the absence or presence of RC (1, 3, and 10 micromol/L). The expressions of LOX-1 mRNA and LOX-1 protein were examined by RT-PCR and Western-blot. RESULTS: Incubation with ox-LDL (50 mg/L) significantly raised the expression of LOX-1 mRNA and LOX-1 protein,which was concentration-dependent. CONCLUSION RC can inhibit the increased expression of LOX-1 mRNA and LOX-1 protein induced by ox-LDL in HUVECs.
Cells, Cultured ; Drugs, Chinese Herbal ; pharmacology ; Endothelium, Vascular ; Humans ; Lipoproteins, LDL ; pharmacology ; Polygala ; chemistry ; RNA, Messenger ; biosynthesis ; genetics ; Receptors, LDL ; biosynthesis ; genetics ; Saponins ; pharmacology ; Umbilical Veins ; cytology ; metabolism

Occupational asthma is induced by many agents, including herbal materials, that are exposed in working places. Although there are a few case reports for occupational allergy induced by herbal materials, there is none for that induced by Wonji (Polygala tenuifolia). This study was conducted to evaluate clinical characteristics and immunologic mechanism of Wonji-induced asthma in a exposed-worker. A patient who complained of asthma and rhinitis symptoms, and who had worked in a herbal manufacturing factory for 8 yr, underwent a skin prick test with crude extract of Wonji under the impression of occupational asthma induced by the agent. The patient had a strong positive response to the extract on the skin prick test. Allergen bronchial challenge to the extract demonstrated a typical dual response. Serum specific IgE level to the extract was higher in the patient than in healthy controls, and ELISA inhibition test revealed complete inhibition of IgE binding with the extract, but no inhibition with Der p 2 or mugwort extracts. Six IgE binding components to the extract (10, 25, 28, 36, 50, and 90 kDa) were detected using SDS-PAGE and immunoblot analysis. These findings suggest that Polygala tenuifolia, a herbal material, can induce IgE-mediated bronchoconstriction in exposed workers.
Allergens ; Asthma/*chemically induced ; Dose-Response Relationship, Drug ; Electrophoresis, Polyacrylamide Gel ; Enzyme-Linked Immunosorbent Assay ; Humans ; Immunoblotting ; Immunoglobulin E/chemistry ; Male ; *Medicine, Herbal ; Middle Aged ; Occupational Exposure ; Pollen/chemistry ; Polygala/metabolism ; Protein Binding ; Research Support, Non-U.S. Gov't ; Rhinitis/*chemically induced ; Time Factors

OBJECTIVETo study the chemical constituents in roots of P. fallax and their anti-oxidation activities in vitro.

METHODColumn chromatographic techniques were employed for isolation and purification of chemical constituents of the plant. The structures were elucidated on the basis of the spectral evidence and the physical and chemical character. The isolated compounds were screened with four anti-oxidation models in vitro.

RESULTSeven xanthones, 1,7-dihydroxy-2,3-methylenedioxyxanthone (1), 1-methoxy-2,3-methylenedioxyxanthone (2), 3-hydroxy-1,2-dimethoxyxanthone (3), 1,6,7-trihydroxy-2,3-dimethoxyxanthone (4), 7-hydroxy-1-methoxy-2,3-methylenedioxyxanthone (5), 1,3-dihydroxy-2-methoxyxanthone (6) and 1,3,7-trihydroxy-2-methoxyxanthone (7), were isolated from the roots of P. fallax. And compounds 1 - 7 showed different anti-oxidation activities in the different pharmacological models.

CONCLUSIONCompounds 2, 3, 5 and 7 were isolated from this plant for the first time. Xanthones from this plant showed anti-oxidation activities. The pharmacological activities of the pure compounds from this plant were also reported for the first time.

Animals ; Antioxidants ; isolation & purification ; pharmacology ; Lipid Peroxidation ; drug effects ; Macrophages ; physiology ; Mitochondria, Liver ; metabolism ; Oxidation-Reduction ; drug effects ; Plant Roots ; chemistry ; Plants, Medicinal ; chemistry ; Polygala ; chemistry ; Rats ; Respiratory Burst ; drug effects ; Xanthones ; isolation & purification ; pharmacology