4'-O-ß-D-glucosyl-5-O-methylvisamminol ameliorates imiquimod-induced psoriasis-like dermatitis and inhibits inflammatory cytokines production by suppressing the NF-κB and MAPK signaling pathways

Psoriasis is a chronic inflammatory skin disorder in humans, and the inflammatory reaction plays an important role in development and onset of psoriasis. 4'-O-β-D-glucosyl-5-O-methylvisamminol (4GMV) is one of the major active chromones isolated from Saposhnikoviae divaricata (Turcz.) Schischk, which has been reported to exhibit excellent anti-inflammatory activities. However, the possible therapeutic effect on psoriasis and underlying mechanism has not been reported. Thus, the aim of this study was to investigate the protective effect of 4GMV on the imiquimod (IMQ)-induced psoriasis-like lesions in BALB/c mice and the anti-inflammatory effect on the lipopolysaccharide (LPS)-induced inflammation in RAW264.7 macrophages. The results demonstrated that 4GMV decreased IMQ-induced keratinocyte proliferation and inflammatory cell infiltration. Moreover, 4GMV treatment significantly inhibited the production of NO, PEG 2, and cytokines such as interleukin (IL)-1β, IL-6, interferon (IFN)-γ, and IL-22 in LPS-stimulated RAW264.7 macrophages. 4GMV also suppressed the LPS-upregulated protein expressions of iNOS and COX-2 in a dose-dependent manner. Furthermore, qRT-PCR analysis showed that 4GMV down-regulated the mRNA level of IL-1β and IL-6 expression. Further studies by western blot indicated that 4GMV inhibited the activation of upstream mediator NF-κB by suppressing the expression of TLR4 and the phosphorylation of IκBα and p65. The phosphorylation of JNK, p38, and ERK were also markedly reversed by 4GMV in LPS-treated RAW264.7 macrophages. Taken together, these results demonstrated that 4GMV showed a protective effect in IMQ-induced psoriasis-like mice and inhibited inflammation through the NF-κB and MAPK signaling pathways, indicating that 4GMV might be a potential therapeutic drug for psoriasis.

4′-O-β-D-Glucosyl-5-O-Methylvisamminol Attenuates Pro-Inflammatory Responses and Protects against Oxidative Damages

We attempted to examine anti-inflammatory and anti-oxidant effects of 4′-O-β-D-glucosyl-5-O-methylvisamminol (GOMV), the first epigenetic inhibitor of histone phosphorylation at Ser10. While GOMV did not affect the viability of murine macrophage RAW 264.7 cells, it significantly suppressed lipopolysaccharide (LPS)-induced generation of prostaglandin E₂ (PGE₂) and nitric oxide (NO) through transcriptional inhibition of cyclooxygenase-2 (COX-2) and inducible nitric oxide synthase (iNOS). GOMV also scavenged free radicals in vitro, increased NF-E2-related factor 2 (NRF2), and activated antioxidant response element (ARE), thereby resulting in the induction of phase II cytoprotective enzymes in human keratinocyte HaCaT cells. Finally, GOMV significantly protected HaCaT cells against 12-O-tetradecanoylphorbol-13-acetate (TPA)-induced oxidative intracellular damages. Together, our results illustrate that GOMV possesses anti-inflammatory and anti-oxidant activity.

HPLC fingerprint and determination of multi-components in Chitong Xiaoyanling Granules / 中草药

Objective To establish an HPLC fingerprint and to determine six compounds in Chitong Xiaoyanling Granules (CXG) for reference of the effective quality control. Methods The analysis was carried out on an analytical column Dikma Luster ODS (250 mm × 4.6 mm, 5 μm) with gradient elution by methanol (A)-0.1% phosphoric acid solution (B) (0-15 min, 20%-30% A; 15-30 min, 30% A; 30-40 min, 30%-60% A; 40-55 min, 60% A), at the detection wavelengths of 254, 283, 274, and 300 nm and a flow rate of 1.0 mL/min. The column temperature was 30 ℃. Similarity evaluation software was used to evaluate the fingerprint of 10 batches of CXG, and the six marker components were quantified. Results The common mode of the fingerprint was set up with 18 common peaks, and six of them were identified by comparison with the reference. The similar degrees of 10 batches of samples were over 0.9, they were prim-O-glucosylcimifugin, 4'-O-β-D-glucosyl-5-O-methylvisamminol, pulegone, hesperidin, paeonol, and isoimperatorin. The linear ranges were 0.013-0.505 mg/mL (r = 0.999 8), 0.052-2.097 mg/mL (r = 0.999 2), 0.019-0.772 mg/mL (r = 0.998 9), 0.025-1.003 mg/mL (r = 0.999 1), 0.006-0.251 mg/mL (r = 0.999 5), and 0.014-0.576 mg/mL (r = 0.999 4) for prim- O-glucosylcimifugin, 4'-O-β-D-glucosyl-5-O-methylvisamminol, pulegone, hesperidin, paeonol, and isoimperatorin, respectively. The contents of prim-O-glucosylcimifugin, 4'-O-β-D-glucosyl-5-O-methylvisamminol, pulegone, hesperidin, paeonol, and isoimperatorin were 7.267-7.333, 4.260-4.522, 2.033-2.093, 12.234-12.771, 19.023-19.334, and 11.152-11.291 mg/g in 10 batches of samples, respectively. Conclusion The established method has high sensitivity and specificity, and can be used for the quality control of CXG.

Fingerprint and multi-components quantitative determination of Xiaochuan Granula by fused-core column / 中草药

Objective To establish an HPLC fingerprint of Xiaochuan Granula (XCG), and to make a quantitative analysis of seven components by fused-core column. Methods Kromasil C18 (150 mm × 4.6 mm, 3.5 μm) was used with the mobile phase of Methanol (A)-acetonitrile (B)-water (C), at a flow rate of 1.0 mL/min. The detection wavelength was set at 210 nm for amygdalin and magnolin, 240 nm for morroniside, loganin, prim-O-glucosylcimifugin, 4'-O-β-glucopyranosyl-5-O-methylvisamminol, schizandrin, and fingerprint; And the column temperature was maintained at 40 ℃. Common peaks had been identified by UPLC-Q-TOF/MS and standard compounds. Results The fingerprint chromatography included 20 mutual peaks, and the similarity was more than 0.90. Eleven chemical components were identified by UPLC-Q-TOF/MS and standard compounds, which were 3-morroniside, oxypaeoniflorin, 5-loganin, 6-prim-O-glucosylcimifugin, 9-4'-O-β-glucopyranosyl-5-O-methylvisamminol, 11-magnolin, 15-schisandrin, 16-schisandrol B, 18-schisantherin A, 19-deoxyschizandrin, and 20-γ-schizandrin B. Moreover, seven active components (morroniside, oxypaeoniflorin, loganin, prim-O-glucosylcimifugin, 4'-O-β-glucopyranosyl-5-O-methylvisamminol,magnolin, and schisandrin) were quantified and the average recovery rates ranged from 97.3% to 103.8% with RSDs less than 2.0%; Seven components in 10 batches samples were morroniside 0.51-0.69 mg/g, amygdalin 5.01-5.95 mg/g, loganin 1.02-1.33 mg/g, prim-O-glucosylcimifugin 0.35-0.45 mg/g, 4'-O-β-glucopyranosyl-5-O-methylvisamminol 0.45-0.55 mg/g, magnolin 0.38-0.48 mg/g, schisandrin 0.89-1.08 mg/g, respectively, and RSD of each component was less than 11.0%. Conclusion The method for establishing HPLC fingerprint and quantitative analysis of seven components is rapid, simple, and accurate, and can be used for the quality control of XCG.

Optimization of extraction technology for Xiaochuan Decoction based on information entropy theory / 中草药

Objective: To optimize the extraction technology of Saposhnikoviae Radix, Perillae Folium, Magnoliae Flos, Armeniacae Amarum Semen, and honey-fried Ephedrae Herba in Xiaochuan Decoction by information entropy theory. Methods: With the contents of prem-O-glucosylcimifugin, 4'-O-beta-glucopyranosyl-5-O-methylvisamminol, amygdalin, and the yield of extract as comprehensive evaluation indexes in order to optimize the extraction process parameters of orthogonal test, the weight coefficient of each index was determined by the information entropy weight method. Results: Optimum extraction technology was as follows: reflux extraction for 3 times with 10 fold water, for 1.5 h each time. Conclusion: The optimized method is stable and reliable, and can provide the reference for further development and utilization of the formula.

Optimization of Water-extraction Technology of Compound Kuhuang Formulation by Orthogonal Test / 中国药房

OBJECTIVE:To optimize the water-extraction technology of Compound kuhuang formulation. METHODS:The wa-ter-extraction technology of Compound kuhuang formulation was optimized by orthogonal design,with the total contents of matrine and oxymatrine,the total content of prim-O-glucosylcimifugin and 5-O-methylvisamminol,the content of berberine hydrochloride and extract yield,and with the extraction time,amount of water(3 reflux extractions)and soaking time as the factors;and verifi-cation tests were conducted. RESULTS:The extraction time had significant effect on comprehensive evaluation value (P<0.05). The optimal water-extraction technology was as follows as extraction time of 120 min,water amount of 8,6,and 6 times as the amount of herbs,and soaking time of 20 min. In verification tests,the average total content of matrine and oxymatrine was 3.152 6 mg/g(RSD=1.03%,n=3),that of prim-O-glucosylcimifugin and 5-O-methylvisamminol was 4.977 2 mg/g(RSD=2.27%,n=3),the average content of berberine hydrochloride was 3.345 0 mg/g (RSD=1.19%,n=3) and the average extract yield was 49.23%(RSD=2.43%,n=3). CONCLUSIONS:The optimal water-extraction technology of Compound kuhuang formulation is stable and feasible.

Chomones from fruit of cnidium monnieri and their effects on proliferation of UMR106 cells / 中草药

Objective: To study the chomenes from the fruit of Cnidium monnieri and their effects on proliferation of UMR106 cells. Methods: The constituents were separated by column chromatography, and their structures were elucidated by spectroscopic data analyses. The effects of all isolated compounds on proliferation of osteoblast-like UMR106 cells were determined. Results: Ten compounds were isolated and identified as cnidimoside A (1), cnidimol B (2), peucenin (3), 5,7-dihydroxychromone (4), 5-O-methylvisamminol (5), 4'-O-β-D-glucosyl-5-O-methylvisamminol (6), hamaudol (7), 2,5-dimethyl-7-hydroxychromone (8), cimifugin (9), and 5-hydroxy-chromone-7-O-β-D-glucoside (10). Compounds 1,5,9, and 10 showed the significant proliferative activities against UMR106 cells lines at the concentration of 0.10 nmol/L and the proliferative ratios were 30.23%, 31.56%, 35.29%, and 33.36%. Conclusion: Compounds 3-10 are isolated from species of genus Cnidium Cuss. for the first time. Compounds 1,5,9, and 10 (0.10 nmol/L) could increase the proliferation of UMR106 cells to some extent.

Determination of 4’-O-?-D-glucosyl-5-O-methylvisamminol in Ganmao Qingre Granule by HPLC / 中国中医药信息杂志

Objective To establish a method for determining the content of 4’-O-?-D-glucosyl-5-O methylvisamminol in Ganmao Qingre Granule by HPLC. Method The seperation was performed in Hypersil C18 colunm (4.6 mm?250 mm, 5 ?m) with the mobile phase of methanol-water (44∶56). The flow rate was 1.0 mL/min and the wavelength was set at 292 nm. Results The calibration curves was linear in the range of 4.608～46.08 ?g/mL for 4’-O-?-D-glucosyl-5-O-methylvisamminol (r =0.999 9). The average recovery was 97.9%, RSD=0.71% (n=6). Conclusion The method is simple, reproducible and can be used for the quality control of Ganmao Qingre Granule.

Determination of 5-O-Methylvisamminol in Shufeng Oral Liquid by HPLC / 中药新药与临床药理

Objective To establish a method for the determination of 5-O-methylvisamminol in Shufeng oral liquid.Methods The sample was extracted with methyl and HPLC method was used.The chromatographic conditions were as follows: Alltima-C18 column(150 mm ? 4.6 mm,5 ?m)with a mobile phase of methanol-water(41∶59),the detection wavelength at 293 nm and the flow rate being 1.0mL?min-1.Results A linearity was obtained from 0.12 ?g to 0.84 ?g of 5-O-Methyvisammioside in Shufeng oral liquid with a good correlation(r = 0.99998,n = 7).The average recovery was 99.8 % and RSD = 2.05 %(n = 5).Conclusion This method for determination of 5-O-Methyvisammioside in Shufeng oral liquid is simple,sensitive,specific and accurate.

Determination of 4′-O-?-D-glycosyl-5-O-methylvisamminol in Jingfuzhiyang Granules by RP HPLC / 中成药

AIM: To establish a RP HPLC method for the determination of 4′ O ? D glycosyl 5 O methylvisamminol in Jingfuzhiyang Granules (Herba Schizonepetae, Fructus kochiae, Radix Saposhnikoviae, Fructus Crataegi, etc.). METHODS: The sample was separated on Turnner Kromasil C 18 column (4.6mm?200mm,5?m) with mobile phase of methanol 0.05mol?L -1 potassium dihydrogen phosphate acetic acid isopropyl alcohol (85∶170∶4∶4,v/v). The detection wavelength was set at 254nm and the flow rate was 1.0mL?min -1 . The column temperature was 30℃. RESULTS: The calibration curve was linear in the range of 0.0606～0.606?g (r=0.9994). The average recovery was 92.2%. CONCLUSION: This method proves to be relatively simple and accurate, and available for the quality control of Jingfuzhiyang Granules

Determination of prim-O-glucosylcimifugin and 4′-O-?-D-glucosyl-5-O-methylvisamminol in Ganmaoqingre Granules by RP-HPLC / 中成药

Objective: A reversed phase HPLC method was described for determination of prim O glucosylcimifugin and 4′ O ? D glucosyl 5 O methylvisamminol in Ganmaoqingre Granules. Methods:The sample was separated on ODS column with mobile phase of methanol 40 mmol?L -1 sodium acetate pH 6.9 (35∶65) for prim O glucosylcimifugin and H 2O methanol THF(62∶38∶1) for 4′ O ? glucosyl 5 O methylvisamminol. The flow rate was 0.8 mL?min -1 , and the detection was set at 254 nm. Results: The calibration curves were linear in the range of 0.72 ?g?mL -1 ～6.5?g?mL -1 for prim O glucosylcimifugin and 0.92?g?mL -1 ～16.5?g?mL -1 for 4′ O ? D glucosyl 5 O methylvisamminol( r =0.9999). The average recovery was 100.3% and 94.7%. The content of prim O glucosylcimifugin and 4′ O ? D glucosyl 5 O methylvisamminol in Ganmaoqingre Granules was 0.133 mg?g -1 and 0.167 mg?g -1 , respectively. Conclusion: The method is fast and specific for both constitutents of Ganmaoqingre Granules.