[Synergistic effect and compatibility structure of active anti-inflammatory ingredients from Lamiophlomis rotata based on network pharmacology and component structure theory].

The synergistic effect and compatibility structure of active anti-inflammatory ingredients(iridoid glycosides: shanzhiside methylester and 8-O-acetylshanzhiside methyl ester, flavonoid glycoside: luteoloside, and phenylethanoid glycoside: forsythoside B) from Lamiophlomis rotata were explored based on network pharmacology and component structure theory. In network pharmacology, CTD, SwisseTargetPrediction, and PharmMapper databases were used to collect and screen the targets of all active ingredients. The inflammation-related targets were obtained from CTD and GeneCards databases. The core targets were obtained by Venny 2.1.0, STRING, and Cytoscape 3.9.1. Core targets were annotated by the GO function and enriched by the KEGG pathway based on the DAVID database. In terms of component structure, based on a uniform design method and xylene-induced ear swelling model in mice, tumor necrosis factor-α and interleukin-6 were taken as the dependent variables, and the compatibility relationship among anti-inflammatory ingredients from L. rotata was explored through the quadratic polynomial stepwise regression. In addition, in vivo pharmacological experiments were conducted to verify the results. A network pharmacology study showed that compared with a single ingredient, the combined action of the three ingredients can synergistically exert anti-inflammatory effects through more biological processes, pathways, and targets. Component structure study showed that the optimal structural ratio of shanzhiside methylester and 8-O-acetylshanzhiside methyl ester in the iridoid glycoside ingredient was 1.21∶1. The optimal structural ratio among the three types of ingredients(iridoid glycosides∶phenylethanol glycoside∶flavonoid glycoside) was 4.8∶1.6∶1. In conclusion, each anti-inflammatory ingredient from L. rotata can work synergistically, and there is an optimal compatibility ratio relationship among these ingredients. This work provides a new experimental basis for the intrinsic quality control of L. rotata.

[Two new triterpeneoids from roots of Dolomiaea souliei].

The present study analyzed the chemical constituents in dried roots of Dolomiaea souliei. Chromatographic methods, such as normal-phase, and reversed-phase column chromatography, TLC, and preparative HPLC, were applied to separate and purify the petroleum ether extract of D. souliei. The structures of the purified constituents were identified by multiple spectroscopic methods including 1 D NMR, 2 D NMR, IR, UV, and HR-ESI-MS. Fourteen triterpenoids were obtained and identified as bauer-8-ene-3,11-dione-7α-ol(1), bauer-8-ene-3-one-7α,11α-diol(2), 3-oxo-11α-hydroxy-urs-12-ene(3), 3-oxour-12-ene-1ß,11α-diol(4), 3ß,11α-dihydroxy-urs-12-ene(5), taraxast-20-ene-3ß,30-diol(6), 28-hydroxy-3-oxo-12-ursene(7), 3ß-hydroxytaraxast-20-ene-30-aldehyde(8), urs-12-ene-2α,3ß,28-triol(9), 20-hydroxylupan-3-one(10), monogynol A(11), obtusalin(12), 3-oxo, 11α-hydroxy-olean-12-ene(13), and isocabralealactone(14). Among them, compounds 1 and 2 were new compounds. Compounds 4-10, 12, and 14 were isolated from this genus for the first time. Compounds 3 and 11 were obtained from D. souliei for the first time.

[Network pharmacological analysis and experimental verification of anti-inflammatory and analgesic effect of Zanthoxyli Pericarpium].

To explore the mechanism of anti-inflammatory and analgesic effect of Zanthoxyli Pericarpium based on network pharmacology and inflammatory or pain mouse models. The effective components of Zanthoxyli Pericarpium were screened out by TCMSP database. And their potential corresponding targets were predicted by PharmMapper software. The possible targets relating to inflammation and pain were mainly collected through DrugBank, TTD and DisGeNET databases. The "active ingredient-gene-disease" network diagram was constructed by Cytoscape 3.7.0 software. The network pharmacology results showed 5 potential effective compounds, which were related to 29 targets; 132 targets relating to inflammation and pain were screened out in the DrugBank, TTD and DisGeNET databases. The network analysis results indicated that the phosphatidylinositol 3-kinase catalytic subunit gamma isoform(PIK3 CG) gene may be the key to the anti-inflammatory and analgesic effect of Zanthoxyli Pericarpium. The anti-inflammatory and analgesic effects of essential oil extract and dichloromethane extract of Zanthoxyli Pericarpium were explored through the mouse model of inflammation induced by xylene or carrageenan and the mouse model of pain induced by acetic acid or formalin. The experimental results showed that essential oil extract and dichloromethane extract of Zanthoxyli Pericarpium could reduce xylene-induced ear swelling and carrageenan-induced paw swelling and decrease the number of writhing responses in mice induced by acetic acid and the licking foot time of mice in phase Ⅱ induced by formalin. Western blot results showed that Zanthoxyli Pericarpium extract could inhibit the expressions of PIK3 CG, phosphonated nuclear factor kappaB(p-NF-κB) and phosphonated p38(p-p38 MAPK) protein. The present study showed the anti-inflammatory and analgesic effect of Zanthoxyli Pericarpium through multiple components and targets, so as to provide a pharmacodynamic basis for the study of Zanthoxyli Pericarpium and its mechanism.

[HPLC Fingerprint of Anisodus tanguticus Root].

OBJECTIVE: To establish an HPLC fingerprint of Anisodus tanguticus root for its quality control. METHODS: The analysis was carried out on a Ultimate AQ C18 (250 mm x 4.6 mm, 5 µm) column with the gradient elution of acetonitrile and KH2PO4 buffer soution, whose pH was adjusted to 3.0 with phosphoric acid. The flow rate, column temperature, detection wavelength and injection volume was 1.0 mL/min, 30 degrees C, 210 nm and 10 µL separately. The similarity evaluation and principal component analysis were used to analyze HPLC fingerprint of Anisodus tanguticus root. RESULTS: HPLC fingerprint of Anisodus tanguticus root was established with 15 common peaks by determining 18 batches of Anisodus tanguticus root samples. Four characteristic peaks, anisodine, scopolamine, anisodamine and anisodamine, were confirmed by comparing their retention time and UV spectrum with standard reference substances. The simiarities of 18 batches of Anisodus tanguticus root were between -0.891 and 0.987. Comprehensive evaluation scores of 18 batches of Anisodus tanguticus root were between -0.85 and 0.89 by principal component analysis. CONCLUSION: The established HPLC fingerprint has good precision, repeatability and stability, which can provide more comprehensive information for identification and quality control of Anisodus tanguticus root.

Rapid discrimination of Alismatis Rhizoma and quantitative analysis of triterpenoids based on near-infrared spectroscopy.

This study developed a rapid, accurate, objective and economic method to identify and evaluate the quality of Alismatis Rhizoma (AR) commodities. Traditionally, the identification of plant species and geographical origins of AR commodities mainly relied on experienced staff. However, the subjectivity and inaccuracy of human identification negatively impacted the trade of AR. Besides, liquid chromatographic methods such as ultra-high-performance liquid chromatography (UPLC) and high-performance liquid chromatography (HPLC), the major approach for the determination of triterpenoid contents in AR was time-consuming, expensive, and highly demanded in manoeuvre specialists. In this study, the combination of near-infrared (NIR) spectroscopy and chemometrics as the method was developed and utilised to address the two common issues of identifying the quality of AR commodities. Through the discriminant analysis (DA), the raw NIR spectroscopy data on 119 batches samples from two species and four origins in China were processed to the best pre-processed data. Subsequently, orthogonal partial least squares-discriminant analysis (OPLS-DA) and random forest (RF) as the major chemometrics were used to analyse the best pre-processed data. The accuracy rates by OPLS-DA and RF were respectively 100% and 97.2% for the two species of AR, and respectively100% and 94.4% for the four origins of AR. Meanwhile, a quantitative correction model was established to rapidly and economically predict the seven triterpenoid contents of AR through combining the partial least squares (PLS) method and NIR spectroscopy, and taking the triterpenoid contents measured by UPLC as the reference value, and carry out spectral pre-processing methods and band selection. The final quantitative model correlation coefficients of the seven triterpenoid contents of AR ranged from 0.9000 to 0.9999, indicating that prediction ability of this model had good stability and applicability.

Chemical constituents from the roots of Dolomiaea souliei.

A new benzofuran-type neolignan (1), two new phenylpropanoids (2 - 3), and one new C21 steroid (4) were isolated from the ethyl acetate extract of the roots of Dolomiaea souliei by chromatographic methods, including silica gel, ODS column chromatography, MPLC, and semi-preparative HPLC. Their structures were identified as dolosougenin A (1), (S)-3-isopropylpentyl (E)-3-(4-hydroxy-3-methoxyphenyl) acrylate (2), (S)-3-isopropylpentyl (Z)-3-(4-hydroxy-3-methoxyphenyl) acrylate (3) and dolosoucin A (4) through various spectroscopic techniques including 1D NMR, 2D NMR, IR, UV, HR ESI MS, ORD, and computational ORD methods.

Catalpol Enhances Neurogenesis And Inhibits Apoptosis Of New Neurons Via BDNF, But Not The BDNF/Trkb Pathway.

BACKGROUND: The role of catalpol in brain neurogenesis and newborn neuron survival has not been previously determined in permanent middle cerebral artery occlusion (pMCAO). METHODS: Fifty-four rats were divided into 6 groups: pMCAO (model, n=9); sham operation (NS, n=9); catalpol treatment (5 mg/kg and 10 mg/kg subgroups, n=9 each); K252a (n=9); and K252a+catalpol 5 mg/kg (n=9) with stroke. The effects of catalpol on behavior, neurogenesis surrounding the infarction ipsilateral to pMCAO, and the expression of brain-derived neurotrophic factor (BDNF) and its receptor (TrkB) were evaluated. Vehicle or, K252a (i.p.), an inhibitor of TrkB phosphorylase. RESULTS: Repeated administration of catalpol reduced neurological deficits and significantly improved neurogenesis. Catalpol increased the number of newborn immature neurons, as determined by BrdU+-Nestin+ and BrdU+-Tuj-1+ staining, and downregulated cleaved caspase 3 in Tuj-1+ cells at day 7 following stroke. Moreover, catalpol increased the protein expression of Tuj-1, MAP2, and the Bcl-2/Bax ratio, as determined using Western blot. Catalpol also significantly increased brain levels of BDNF, but not TrkB, resulting in enhanced survival of newborn neurons via inhibition of apoptosis. CONCLUSION: Catalpol may contribute to neurogenesis in infarcted brain regions and help promote the survival of newborn neurons by activating BDNF, but not BDNF/TrkB signaling.

Characterization of Electricity Generated by Soil in Microbial Fuel Cells and the Isolation of Soil Source Exoelectrogenic Bacteria.

Soil has been used to generate electrical power in microbial fuel cells (MFCs) and exhibited several potential applications. This study aimed to reveal the effect of soil properties on the generated electricity and the diversity of soil source exoelectrogenic bacteria. Seven soil samples were collected across China and packed into air-cathode MFCs to generate electricity over a 270 days period. The Fe(III)-reducing bacteria in soil were enriched and sequenced by Illumina pyrosequencing. Culturable strains of Fe(III)-reducing bacteria were isolated and identified phylogenetically. Their exoelectrogenic ability was evaluated by polarization measurement. The results showed that soils with higher organic carbon (OC) content but lower soil pH generated higher peak voltage and charge. The sequencing of Fe(III)-reducing bacteria showed that Clostridia were dominant in all soil samples. At the family level, Clostridiales Family XI incertae sedis were dominant in soils with lower OC content but higher pH (>8), while Clostridiaceae, Lachnospiraceae, and Planococcaceae were dominant in soils with higher OC content but lower pH. The isolated culturable strains were allied phylogenetically to 15 different species, of which 11 were Clostridium. The others were Robinsoniella peoriensis, Hydrogenoanaerobacterium saccharovorans, Eubacterium contortum, and Oscillibacter ruminantium. The maximum power density generated by the isolates in the MFCs ranged from 16.4 to 28.6 mW m-2. We concluded that soil OC content had the most important effect on power generation and that the Clostridiaceae were the dominant exoelectrogenic bacterial group in soil. This study might lead to the discovery of more soil source exoelectrogenic bacteria species.

[Application of Microbial Fuel Cells in Reducing Methane Emission from Rice Paddy].

We aimed to study whether the methane emission from rice paddy with straw return can be alleviated in microbial fuel cells (MFCs). In our study, the soil mixed with 0. 5% ( mass fraction) rice straw was packed into MFCs reactors, then flooded with excess of sterilized water and transplanted with rice seedlings followed by the operation of MFCs. The MFCs were operated for 98 days covering five stages of seeding, tillering, mid-season aeration, rice filling, and ripening. The voltage data were recorded continuously and in real time during the MFCs operation and the methane emitted was collected once a week using the static chamber method and the methane emission flux was determined by gas chromatography. The results showed that the MFCs current increased and reached the peak value in the seeding and tillering stages and the operation of MFCs significantly reduced the accumulative methane emission in these two stages. The possible reason could be that the electrogens competed with methanogens for organic substrates. The height, the above and below ground biomass, and the productivity of rice plants were not significantly affected by the 98-day operation of MFCs. Our study provides a potential green and sustainable technology for the reduction of CH, emission from rice paddy fields.

[Research Progress in Technology of Using Soil Micro-organisms to Generate Electricity and Its Potential Applications].

Microbial fuel cells ( microbial fuel cells, MFCs) are devices in which micro-organisms convert chemical energy into electrical power. Soil has electrogenic bacteria and organic substrates, thus can generate electrical current in MFCs. Soil MFCs can be operated and applied to real-time and continuously monitor soil pollution, remove soil pollutants and to reduce methane emitted from flooded rice paddy, without energy consumption and the application of chemical reagents to the soil. Instead, the operation of soil MFCs generates small amount of electrical power. Therefore, soil MFCs are useful in the development of environment-friendly technology for monitoring and remediating soil pollution, which have potential value for applications in the domain of environmental science and engineering. However, much of advanced technology hasn't been applied into soil MFCs since the studies on soil MFCs was not started until recently. This paper summarized the research progress in related to soil MFCs combining with the frontier of MFCs technology, and brought forward the possible direction in studies on soil MFCs.

Antitumor activity of 4-O-(2″-O-acetyl-6″-O-p-coumaroyl-ß-D-glucopyranosyl)-p-coumaric acid against lung cancers via mitochondrial-mediated apoptosis.

This study was aimed to investigate antitumor activity of 4-O-(2″-O-acetyl-6″-O-p-coumaroyl-ß-D-glucopyranosyl)-p-coumaric acid (4-ACGC) against lung cancer and its mechanisms. The anti-proliferative effects of 4-ACGC on lung cancer cell lines including A549, NCI-H1299, HCC827 were evaluated by MTT method and the IC50 values were calculated, and subsequently a mice xenograft model of A549 was established to investigate the antitumor effect of 4-ACGC in vivo. Furthermore, the apoptosis of the A549 cells was determined by fluorescence microscope by staining with Hoechst 33324 and flow cytometer by staining with FITC conjugated Annexin V/PI, and the further mechanisms were investigated by Western blotting. Our results demonstrated that 4-ACGC possessed notable anti-tumor activity on lung cancer in vivo and in vitro; the mechanisms were involved in inducing mitochondria-mediated apoptosis via up-regulations of caspase-3, caspase-9, Bad and Bax, and down-regulation of Bcl-2. Collectively, our results indicated that the 4-ACGC could be treated as a new candidate for treatment of lung cancer in the future.