Inhibitory effects of Jasminum grandiflorum L. essential oil on lipopolysaccharide-induced microglia activation-integrated characteristic analysis of volatile compounds, network pharmacology, and BV-2 cell.

Neuroinflammation is considered to have a prominent role in the pathogenesis of Alzheimer's disease (AD). Microglia are the resident macrophages of the central nervous system, and modulating microglia activation is a promising strategy to prevent AD. Essential oil of Jasminum grandiflorum L. flowers is commonly used in folk medicine for the relief of mental pressure and disorders, and analyzing the volatile compound profiles and evaluating the inhibitory effects of J. grandiflorum L. essential oil (JGEO) on the excessive activation of microglia are valuable for its application. This study aims to explore the potential active compounds in JGEO for treating AD by inhibiting microglia activation-integrated network pharmacology, molecular docking, and the microglia model. A headspace solid-phase microextraction combined with the gas chromatography-mass spectrometry procedure was used to analyze the volatile characteristics of the compounds in J. grandiflorum L. flowers at 50°C, 70°C, 90°C, and 100°C for 50 min, respectively. A network pharmacological analysis and molecular docking were used to predict the key compounds, key targets, and binding energies based on the detected compounds in JGEO. In the lipopolysaccharide (LPS)-induced BV-2 cell model, the cells were treated with 100 ng/mL of LPS and JGEO at 7.5, 15.0, and 30 µg/mL, and then, the morphological changes, the production of nitric oxide (NO) and reactive oxygen species, and the expressions of tumor necrosis factor-α, interleukin-1ß, and ionized calcium-binding adapter molecule 1 of BV-2 cells were analyzed. A total of 34 compounds with significantly different volatilities were identified. α-Hexylcinnamaldehyde, nerolidol, hexahydrofarnesyl acetone, dodecanal, and decanal were predicted as the top five key compounds, and SRC, EGFR, VEGFA, HSP90AA1, and ESR1 were the top five key targets. In addition, the binding energies between them were less than -3.9 kcal/mol. BV-2 cells were activated by LPS with morphological changes, and JGEO not only could clearly reverse the changes but also significantly inhibited the production of NO and reactive oxygen species and suppressed the expressions of tumor necrosis factor-α, interleukin-1ß, and ionized calcium-binding adapter molecule 1. The findings indicate that JGEO could inhibit the overactivation of microglia characterized by decreasing the neuroinflammatory and oxidative stress responses through the multi-compound and multi-target action modes, which support the traditional use of JGEO in treating neuroinflammation-related disorders.

Long-Term Effect against Methicillin-Resistant Staphylococcus aureus of Emodin Released from Coaxial Electrospinning Nanofiber Membranes with a Biphasic Profile.

Methicillin-resistant Staphylococcus aureus (MRSA) is a serious and rapidly growing threat to human beings. Emodin has a potent activity against MRSA; however, its usage is limited due to high hydrophobicity and low oral bioavailability. Thus, the coaxial electrospinning nanofibers encapsulating emodin in the core of hydrophilic poly (vinylpyrrolidone), with a hygroscopic cellulose acetate sheath, have been fabricated to provide long-term effect against MRSA. Scanning electron microscopy and transmission electron microscopy confirmed the nanofibers had a linear morphology with nanometer in diameter, smooth surface, and core-shell structure. Attenuated total reflection-Fourier transform infrared spectra, X-ray diffraction patterns, and differential scanning calorimetric analyses verified emodin existed in amorphous form in the nanofibers. The nanofibers have 99.38 ± 1.00% entrapment efficiency of emodin and 167.8 ± 0.20 % swelling ratio. Emodin released from nanofibers showed a biphasic drug release profile with an initial rapid release followed by a slower sustained release. CCK-8 assays confirmed the nontoxic nature of the emodin-loaded nanofibers to HaCaT cells. The anti-MRSA activity of the nanofibers can persist up to 9 days in AATCC147 and soft-agar overlay assays. These findings suggest that the emodin-loaded electrospun nanofibers with core-shell structure could be used as topical drug delivery system for wound infected by MRSA.

Comprehensive analysis of the compound profiles of Folium Camelliae Nitidissimae extract by ultrafast liquid chromatography with quadrupole-time-of-flight mass spectrometry and hepatoprotective effect against CCl4 -induced liver injury in mice.

Folium Camelliae Nitidissimae (jinhuacha in Chinese, JHC) is a kind of caffeine-less tea with antioxidant, antitumor and antibacterial effects. Studies on the chemical profiles and hepatoprotective effects of JHC extracts have not been systematically conducted so far. This study comprehensively investigated the compound profiles of JHC extract by ultrafast liquid chromatography with quadrupole time-of-flight tandem mass spectrometry. We also determined JHC's hepatoprotective effects against CCl4 -induced liver injury in mice. A JHC extract was administered orally to mice at 1.95 and 7.80 g/kg body weight once daily for 14 consecutive days prior to CCl4 treatment. Eighty-four compounds including flavonoids, organic acids, catechins, coumarins, phenylpropanol, amino acids, anthraquinones, saponins and nucleosides in JHC extract were authentically identified or tentatively identified by comparing MS information and retention times with those of authentic standards or available references. JHC administration significantly decreased elevated levels of aspartate aminotransferase and alanine aminotransferase in mouse serum, inhibited hepatic malondialdehyde formation and enhanced glutathione and superoxide dismutase activities in the liver of CCl4 -treated mice. The histological observations also further supported the results. These results demonstrate that JHC contains various chemical compounds and its hepatoprotective effects against CCl4 -induced liver injury correlated with decreasing lipid oxidation are significant.

Simultaneous determination of wogonin, oroxylin a, schisandrin, paeoniflorin and emodin in rat serum by HPLC-MS/MS and application to pharmacokinetic studies.

Wogonin and oroxylin A in Scutellariae Radix, schisandrin in Chinensis Fructus, paeoniflorin in Moutan Cortex and emodin in Polygoni Cuspidate Rhizome et Radix are anti-inflammatory active compounds. A method for simultaneous determination of the five compounds in rat was developed and validated using high-performance liquid chromatography with tandem mass spectrometry (HPLC-MS/MS). The separation was performed on a Symmetry C18 column (4.6 × 50 mm, 3.5 µm) with acetonitrile and 0.1% formic acid aqueous solution as the mobile phases. The detection was performed using multiple-reaction monitoring with electrospray ionization source in positive-negative ion mode. The calibration curves showed good linearity (r ≥ 0.9955). The lower limit of quantification (LLOQ) was 5 ng/mL for wogonin and schisandrin, 10 ng/mL for oroxylin A and emodin, and 15 ng/mL for paeoniflorin, respectively. The relative standard deviations of intraday and interday precisions were <11.49 and 14.28%, respectively. The extraction recoveries and matrix effects were acceptable. The analytes were stable under the experiment conditions. The validated method has been successfully applied to pharmacokinetic studies of the five compounds in rats after oral administration of Hu-gan-kan-kang-yuan capsule. This paper would be a valuable reference for pharmacokinetic studies of Chinese medicine preparations containing the five compounds.

Developing an Absorption-Based Quality Control Method for Hu-Gan-Kang-Yuan Capsules by UFLC-QTOF-MS/MS Screening and HPLC-DAD Quantitative Determination.

Traditional Chinese Medicine Preparations (TCMPs) contain massive numbers of ingredients responsible for their multiple efficacies. An absorption-based quality control method for complicated TCMPs using Hu-gan-kang-yuan Capsule (HGKYC) as an example was developed. To select proper chemical markers for quality control of HGKYC, an ultra-fast liquid chromatography (UFLC) coupled with electrospray ionization quadrupole time-off light mass spectrometry (UFLC-QTOF-MS/MS) method was used for the rapid separation and structural identification of the constituents in the HGKYC extract and the rat serum after oral administration of HGKYC. As a result, one hundred and seven prototype constituents including flavonoids, organic acid, phenylpropanoids, anthraquinones, saponins, alkaloids, terpenes, phenols and amino acids in HGKYC extract, and 43 compounds found in rat serum after oral administration of HGKYC were unambiguously identified or tentatively characterized by comparing retention times and MS information with those of authentic standards or available literature references. Finally, a simple, low-cost and effective method of simultaneous determination for baicalein, wogonin, paeonol and emodin in HGKYC was developed using high performance liquid chromatography coupled with a diode array detector. In conclusion, an absorption-based quality control pattern was developed and successfully used for evaluating HGKYC.

Development and validation of a HPLC-MS/MS method for the determination of phytolaccagenin in rat plasma and application to a pharmacokinetic study.

Radix Phytolaccae (the dried root of Phytolacca acinosa Roxb. or Phytolacca americana L.) is widely used in East Asian countries for the treatment of inflammation-related diseases. The active component of Radix Phtolaccae is Phytolcaccagenin a triterpenoid saponin. Phytolcaccagenin has anti-inflammatory activities that exceed those of Esculentoside A and its derivatives regarding suppression of LPS-induced inflammation, and has a lower toxicity profile with less hemolysis. To date, no information is available about analytical method and pharmacokinetic studies of phytolaccagenin. To explore PK profile of this compound, a HPLC-MS/MS assay of phytolaccagenin in rat plasma was developed and validated. The method was fully validated according to FDA Guidance for industry. The detection was performed by a triple-quadrupole tandem mass spectrometer with multiple reactions monitoring (MRM) in positive ion mode via electrospray ionization. The monitored transitions were m/z 533.2>515.3 for Phytolcaccagenin, and 491.2>473.2 for I.S. The analysis was performed on a Symmetry C18 column (4.6 mm × 50 mm, 3.5 µm) using gradient elution with the mobile phase consisting of acetonitrile and 0.1% formic acid water at a flow rate of 1 ml/min with a 1:1 splitter ratio. The method was validated with a LLOQ of 20 ng/ml and an ULOQ of 1000 ng/ml. The response versus concentration data were fitted with 1/x weighting and the correlation coefficient (r) were greater than 0.999. The average matrix effect and the average extraction recovery were acceptable. This validation in rat plasma demonstrated that phytolaccagenin was stable for 30 days when stored below -20°C, for 6h at room temperature (RT, 22°C), for 12 h at RT for prepared control samples in auto-sampler vials, and during three successive freeze/thaw cycles results at -20°C. The validated method has been successfully applied to an intravenous bolus pharmacokinetic study of phytolaccagenin in male Sprague-Dawley rats (10 mg/kg, i.v.). Blood samples taken from 0 to 24h after injection were collected, and data analyzed with WinNonlin. The half-life and clearance were 1.4±0.9 h and 2.1±1.1 L/h/kg, respectively.

Preparation of lung-targeting, emodin-loaded polylactic acid microspheres and their properties.

Emodin (1,3,8-trihydroxy-6-methylanthraquinone) has been identified to have the potential to improve lung fibrosis and lung cancer. To avoid the liver and kidney toxicities and the fast metabolism of emodin, emodin-loaded polylactic acid microspheres (ED-PLA-MS) were prepared and their characteristics were studied. ED-PLA-MS were prepared by the organic phase dispersion-solvent diffusion method. By applying an orthogonal design, our results indicated that the optimal formulation was 12 mg/mL PLA, 0.5% gelatin, and an organic phase:glycerol ratio of 1:20. Using the optimal experimental conditions, the drug loading and encapsulation efficiencies were (19.0±1.8)% and (62.2±2.6)%, respectively. The average particle size was 9.7±0.7 µm. In vitro studies indicated that the ED-PLA-MS demonstrated a well-sustained release efficacy. The microspheres delivered emodin, primarily to the lungs of mice, upon intravenous injection. It was also detected by microscopy that partial lung inflammation was observed in lung tissues and no pathological changes were found in other tissues of the ED-PLA-MS-treated animals. These results suggested that ED-PLA-MS are of potential value in treating lung diseases in animals.

[High-performance capillary electrophoresis for determining the chemical compositions of Gegen and Fenge extracts].

OBJECTIVE: To develop a method for analyzing the chemical compositions of Gegen and Fenge extracts using high-performance capillary electrophoresis (HPCE). METHODS: Using HPCE/DAD, the chemical composition of the extracts was analyzed with the buffer solution of 40 mmol/L borax containing 16.7% methanol, with injection pressure at 137.9 kPa for 5 s, separation voltage at 25 kV in 0-5 min time range and at 22 kV in 5-25 min time range, and the temperature of the capillary of 20 degrees celsius. RESULTS AND CONCLUSION: The method for analysis of Gegen and Fenge extracts was established, which identified puerarin and daidzein as the two major components. This simple and rapid analysis method can be used for Gegen and Fenge extract fingerprinting.

[Study on the TLC-fingerprint of Xiatianwu].

OBJECTIVE: To develop a method that can well evaluate the quality of Xiatianwu. METHODS: Methods are researched and discussed to construct the TLC-FP (thin-layer chromatography finger-print) of Xiatianwu to evaluate its quality. RESULTS: The areas of six evident peaks in the thin-layer chromatography of Xiatianwu are compared with the area of dl-tetrahydropalmatine peak companied with them in the same chromatography. The Rr of every peak and its relative area were acquired to evaluate the quality of Xiatianwu and identify it. CONCLUSION: The TLC-fingerprint of Xiatianwu can well evaluate the quality of Xiatianwu.

[The study on prescription and preparation of the sustained-released capsule of gegen isoflavones].

OBJECTIVE: To study the preparation of the sustained-released capsule of Gegen isoflavones (SRCGI) and choose the best prescription. METHODS: EC and HPMC were used as sustained-released excipients. Uniform layout test was employed to design the formulation. RESULTS: The best prescription release drug Q0.5h = 50% - 53%, to 100% at 12h and its dissolution behavior belongs to Higuchi Kinetics, Q = 17.19t1/2 +/- 38.75, r = 0.9959. CONCLUSION: SRCGI obtained desirable effect of drug release.

[Influence of decoction methods and compatibility on content of cinnamaldehyde in Mahuang decoction].

OBJECTIVE: To develop a GC-MS method for the determination of cinnamaldehyde and study on the influence of decoction methods and compatibility on the contents of cinnamaldehyde in Mahuang decoction. METHODS: Gas chromatography-mass spectrometry (GC-MS) was used. Chromatography/mass spectrometric conditions were: capillary column DB-17 (30 m x 0.5 mm, 0.25 micron), helium as carrier gas, non-split injection, column primary temperature 120 degrees C, 120 degrees C 8 degrees C/min-->250 degrees C, with 1.0 ml/min runoff, and injection volume 1 microliter. The results were analyzed by one-way ANOVA (SPSS). RESULTS: The calibration curves were linear in the range from 0.0136 mg/ml to 0.136 mg/ml (r = 0.9999). The RSD of precison, stability and reproducibilities were 1.02% (n = 6), 2.57% in 24 h and 4.20%, respectively. The cinnamaldehyde content in the blended Mahuangtang decoction (BMHD) was significantly lower than the one in the separated and mixtured decoction and Ramulus cinnamomi decoction (P < 0.05). The separated and mixtured decoction was higher than Ramulus cinnamomi decoction (P > 0.05). CONCLUSION: The main influencing factor of the cinnamaldehyde content was the compatibility. This method is precise and reliable to study cinnamaldehyde in Mahuang Decoction.