Longxuetongluo capsule alleviates lipopolysaccharide-induced neuroinflammation by regulating multiple signaling pathways in BV2 microglia cells.

BACKGROUND: Longxuetongluo capsule (LTC), derived from the total phenolic compounds of Chinese dragon's blood, is now used in the treatment of ischemic stroke in convalescence. The aim of this study is to explore the neuroprotective effect of LTC from the perspective of neuroinflammation. METHODS: Cell viability and lactate dehydrogenase (LDH) release were measured by 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium (MTS) and LDH assay kit. Proinflammatory mediators and cytokines production including Nitric Oxide (NO), prostaglandin E2, (PGE2), interleukin (IL-ß), IL-6, and tumor necrosis factor-α (TNF-α) were detected by enzyme-linked immunosorbent assay (ELISA) assay. In addition, western blot was used to detect the expression of inflammatory proteins associated with the mitogen-activated protein kinases (MAPKs), janus kinase/signal transducer and activator of tranions (JAK/STAT), nuclear transcription factor κB (NF-κB), and nuclear factor erythroid-2-related actor 2/heme oxygenase 1 (Nrf2/HO-1) signaling pathways. Moreover, immunofluorescence assay and electrophoretic mobility shift assays (EMSA) were performed to determine the Nrf2 translocation and the binding-DNA activity of NF-κB, respectively. RESULTS: LTC at 0.5 to 2 µg/mL significantly increased cell viability and decreased LDH, NO, PGE2, IL-1ß, IL-6, and TNF-α production in oxygen-glucose deprivation/reoxygenation (OGD/R) and lipopolysaccharide (LPS)-induced BV2 microglia cells. Meanwhile, LTC not only decreased the protein expressions of inducible nitric oxide synthase (iNOS), cyclooxygenase-2 (COX-2) but also down-regulated phosphorylation of extracellular signal-regulated kinase (ERK)1/2, p38, and up-regulated HO-1 expression via nuclear translocation of Nrf2. LTC can significantly inhibit the phosphorylation of JAK1/STAT3 and reduce the translocation of NF-κB from cytosol to nucleus as well as the binding-DNA activity. PC12 cell pretreated with LTC-condition medium (CM) significantly alleviated LPS-induced neurotoxicity and increased PC12 cell viability in a dose-dependent manner. CONCLUSION: The present study showed that LTC exhibited a strong antineuroinflammatory activity and neuroprotective effects on LPS-stimulated BV2 microglial cells and PC12 cells.

Ultra-pH-sensitive indocyanine green-conjugated nanoprobes for fluorescence imaging-guided photothermal cancer therapy.

Photothermal therapy (PTT) has been recognized as a promising approach for cancer treatment due to its minimal invasiveness and low systemic side effects. However, developing a photothermal agent with accurate tumor imaging capability is a prerequisite for the efficient PTT. Here, we developed a series of ultra-pH-sensitive indocyanine green (ICG)-conjugated nanoparticles for fluorescence imaging-guided tumor PTT. These nanoparticles exhibited high fluorescence activation ratio (~100-fold) with sharp pH transition (ΔpHon/off <0.25), and superior temperature response than free ICG. The in vivo imaging experiments demonstrated that the nanoparticles generated excellent tumor-to-normal tissue contrast through pH-triggered fluorescence activation in tumor sites, which provided information on tumor mass location, boundaries, and shape. Moreover, comparing to free ICG, the nanosystem had significantly longer blood circulation time and more accurate tumor targeting, providing efficient photothermal therapeutic effect against A549 tumor in living animals. In conclusion, this nanoplatform offers a potential strategy for imaging-guided cancer PTT.

Salvia przewalskii extract of total phenolic acids inhibit TLR4 signaling activation in podocyte injury induced by puromycin aminonucleoside in vitro.

BACKGROUND: TLR4 signaling is known to be involved in podocyte injury. We have previously shown that Salvia przewalskii extract of total phenolic acids (SPE) and its active monomer salvianolic acid B (SalB) and rosmarinic acid (RA) protect podocytes from injury induced by PAN. In the present study, we test whether SPE inhibits TLR4 signaling. METHODS: The conditionally immortalized mouse podocytes were treated with SPE, SalB, RA, SalB + RA or tacrolimus for 30 min, followed by PAN (100 µg/mL) for 24 h. The F-actin staining with phalloidin was used to assess cytoskeletal injury in the podocytes. Western blotting and semi-quantitatives RT-PCR were used to assess the changes of the components in the TLR4 signaling pathway. RESULTS: (1) The F-actin stress fibers of podocytes were almost completely disrupted after PAN treatment for 24 h, and the disruption was significantly alleviated by SPE; (2) the PAN-induced elevation of mRNA levels of TLR4, MyD88 and p65 were inhibited except p65 with high-dose SalB; (3) consistently, the protein levels of TLR4, MyD88 and pp65 were significantly elevated by PAN, and SPE, SalB, RA and admixture, respectively, attenuated the elevations of TLR4 and pp65 proteins; (4) SPE and tacrolimus have a similarly strong effect on inhibition of the expression of TLR4 signaling components. CONCLUSIONS: SPE protects podocytes from PAN-induced injury at least partly through inhibiting TLR4 signaling. SPE is as strong as tacrolimus in inhibiting TLR4 signaling in podocytes.

Ultra-performance liquid chromatography-quadrupole\time-of- flight mass spectrometry with multivariate statistical analysis for exploring potential chemical markers to distinguish between raw and processed Rheum palmatum.

BACKGROUND: The long term use of Rheum palmatum for the treatment of diseases associated with chronic hepatitis and renal failure can lead to liver and kidney damage. To reduce the toxicity of R. palmatum and alleviate any symptoms of decanta and celialgia, the raw material has been subjected to a specific process prior to its use for hundreds of years. Despite its extensive use in medicine, very little is currently known about the nature of the components present in this material in terms of their efficacy and overall toxicity, and the effect that processing has on the levels of these components in the processed material. The aim of this investigation was to explore potential differences in the chemical markers between batches of raw and processed R. palmatum and to develop a deeper understanding of the underlying mechanisms responsible for the enhanced efficacy and reduced toxicity of the processed material. METHODS: Raw and processed R. palmatum samples were analyzed by ultra-performance liquid chromatography-quadrupole time-of-flight mass spectrometry (UPLC/Q-TOF-MS) coupled with multivariate statistical analysis using principal component analysis (PCA) and orthogonal partial least square discriminant analysis (OPLS-DA). RESULTS: The emodin-8-O-glucoside, emodin-O-glucoside, catechin-glucopyranoside, gallic acid-3-O-glucoside, torachrysone, and chrysophanol dimethyl ether were rapidly explored as representative markers to distinguish for the first time between the raw and processed R. palmatum material. Among the potential chemical markers, Emodin-8-O-glucoside and gallic acid-3-O-glucoside were determined to be the best markers for the raw and processed R. palmatum. CONCLUSION: UPLC/Q-TOF-MS with multivariate statistical analysis represents an efficient method for exploring the chemical markers in the raw and processed R. palmatum material, as well as investigating the mechanisms associated with the processing, quality control, and safe application of R. palmatum.

Review of the botanical characteristics, phytochemistry, and pharmacology of Astragalus membranaceus (Huangqi).

Astragalus membranaceus is one of the most widely used traditional Chinese herbal medicines. It is used as immune stimulant, tonic, antioxidant, hepatoprotectant, diuretic, antidiabetic, anticancer, and expectorant. The current paper reviews the botanical characteristics, phytochemistry, and pharmacology of Astragali Radix. Information on Astragali Radix was gathered via the Internet (using Google Scholar, Baidu Scholar, Elsevier, ACS, Medline Plus, CNKI, and Web of Science) as well as from libraries and local books. More than 100 compounds, including flavonoids, saponins, polysaccharides, and amino acids, have so far been identified, and the various biological activities of the compounds have been reported. As an important traditional Chinese medicine, further studies on Astragali Radix can lead to the development of new drugs and therapies for various diseases. The improvement of its utilization should be studied further.

Analysis of the Correlation between Commodity Grade and Quality of Angelica sinensis by Determination of Active Compounds Using Ultraperformance Liquid Chromatography Coupled with Chemometrics.

The contents of ferulic acid, senkyunolide A, butylidenephthalide, ligustilide, and n-butylphthalide were determined by UPLC analytical method; the correlation among the grade, average weight, and content was explored by correlation analysis and analysis of variance (ANOVA); the different commercial grades with average weight and content were revealed by principal component analysis (PCA) and then rationality analysis grade classification of A. sinensis. The results showed that various commercial grades can be distinguished by PCA analysis. And there was significant negative correlation between the commodity grades and average weight, commodity, and the content of bioactive compounds, while the content of senkyunolide A had significant negative correlation with commodity grades (P < 0.01). Average weight had no correlation with chemicals compounds. Additionally, there was significant positive correlation among the bioactive compounds (content of ferulic acid and phthalides) of different grades of A. sinensis. The content of senkyunolide A, butylidenephthalide, and ligustilide had significant positive correlation with the content of ferulic acid. The content of ligustilide and butylidenephthalide had significant positive correlation with the content of senkyunolide A. The content of ligustilide had significant positive correlation with the content of butylidenephthalide. The basis of grades classification is related with the difference levels of the bioactive compounds.

Hypericin: chemical synthesis and biosynthesis.

Hypericin is one of the most important phenanthoperylene quinones extracted mainly from plants of the genus Hypericum belonging to the sections Euhypericum and Campylosporus of Keller's classification. Widespread attention to the antiviral and anti-tumor properties of hypericin has spurred investigations of the chemical synthesis and biosynthesis of this unique compound. However, the synthetic strategies are challenging for organic and biological chemists. In this review, specific significant advances in total synthesis, semi-synthesis, and biosynthesis in the past decades are summarized.

UPLC-QTOF/MS Analysis of Alkaloids in Traditional Processed Coptis chinensis Franch.

The processing technology employed in traditional Chinese medicine (TCM) is significant and distinct. Meanwhile, the processed Coptis chinensis Franch. are significant in clinic based on clinical practice and literature. The current study used ultraperformance liquid chromatography method (UPLC) coupled with quadrupole time of flight mass spectrometry (qTOF/MS) and Marklynx software to analyze the chemical profiles of crude and processed C. chinensis Franch. 13 compounds in these samples are identified, including 3 compounds that are detected in C. chinensis Franch. for the first time. Moreover, the results of the experiment show significant chemical differences between crude and processed C. chinensis Franch. with principal component analysis (PCA). The obvious separation in PCA confirms the traditional processing theory in TCM.

Hydrolysis of flavanone glycosides and degradation of the corresponding aglycones from dried immature Citrus fruit by human fecal flora in vitro.

The hydrolysis of the flavanone glycosides contained in dried immature Citrus fruit, originating from Citrus aurantium L (Family Rutaceae), and degradation of their aglycones in human fecal flora have been analyzed. High-performance liquid chromatography was used to determine the flavanone glycosides and their corresponding aglycones in human fecal flora. The separation of compounds was performed with an ODS column by isocratic and stepwise gradient elution with 0.5 % (v/v) acetic acid-acetonitrile. As a result, the hydrolysis rate of hesperidin and narirutin (flavanone rutinoside) was faster than that of naringin and neohesperidin (flavanone neohesperioside). When the half-life time of each flavanone glycoside was carefully calculated (under the mixed conditions with the human fecal flora), hydrolysis of the flavanone rutinoside turned out to be approximately two times faster than of flavanone neohesperioside. The observed degradation rates of both aglycones was found not to be different. Therefore, it seems that the hydrolysis rate of flavanone glycosides in dried immature citrus fruit with human fecal flora is closely related to the steric hindrance of the sugar. This finding might be effectively used for further pharmacokinetics research on the flavanone glycosides of dried immature Citrus fruit.

Pharmacokinetics of cimifugin in rat plasma after oral administration of the extract of Saposhnikovia divaricatae root. Determination of cimifugin by high performance liquid chromatography coupled with solid phase extraction.

High performance liquid chromatography (HPLC) coupled with the solid phase extraction method was developed for determining cimifugin (a coumarin derivative; one of Saposhnikovia divaricatae's constituents) in rat plasma after oral administration of Saposhnikovia divaricatae extract (SDE), and the pharmacokinetics of cimifugin either in SDE or as a single compound was investigated. The HPLC analysis was performed on a commercially available column (4.6 mm x 200 mm, 5 pm) with the isocratic elution of solvent A (Methanol) and solvent B (Water) (A:B=60:40) and the detection wavelength was set at 250 nm. The calibration curve was linear over the range of 0.100-10.040 microg/mL. The limit of detection was 30 ng/mL. At the rat plasma concentrations of 0.402, 4.016, 10.040 microg/mL, the intra-day precision was 6.21%, 3.98%, and 2.23%; the inter-day precision was 7.59%, 4.26%, and 2.09%, respectively. The absolute recovery was 76.58%, 76.61%, and 77.67%, respectively. When the dosage of SDE was equal to the pure compound calculated by the amount of cimifugin, it was found to have two maximum peaks while the pure compound only showed one peak in the plasma concentration-time curve. The pharmacokinetic characteristics of SDE showed the superiority of the extract and the properties of traditional Chinese medicine.