Protective Effects of Danmu Extract Syrup on Acute Lung Injury Induced by Lipopolysaccharide in Mice through Endothelial Barrier Repair / 中国结合医学杂志

OBJECTIVE@#To investigate the effects of Danmu Extract Syrup (DMS) on lipopolysaccharide (LPS)-induced acute lung injury (ALI) in mice and explore the mechanism.@*METHODS@#Seventy-two male Balb/C mice were randomly divided into 6 groups according to a random number table (n=12), including control (normal saline), LPS (5 mg/kg), LPS+DMS 2.5 mL/kg, LPS+DMS 5 mL/kg, LPS+DMS 10 mL/kg, and LPS+Dexamethasone (DXM, 5 mg/kg) groups. After pretreatment with DMS and DXM, the ALI mice model was induced by LPS, and the bronchoalveolar lavage fluid (BALF) were collected to determine protein concentration, cell counts and inflammatory cytokines. The lung tissues of mice were stained with hematoxylin-eosin, and the wet/dry weight ratio (W/D) of lung tissue was calculated. The levels of tumor necrosis factor-α (TNF-α), interleukin (IL)-6 and IL-1 β in BALF of mice were detected by enzyme linked immunosorbent assay. The expression levels of Claudin-5, vascular endothelial (VE)-cadherin, vascular endothelial growth factor (VEGF), phospho-protein kinase B (p-Akt) and Akt were detected by Western blot analysis.@*RESULTS@#DMS pre-treatment significantly ameliorated lung histopathological changes. Compared with the LPS group, the W/D ratio and protein contents in BALF were obviously reduced after DMS pretreatment (P<0.05 or P<0.01). The number of cells in BALF and myeloperoxidase (MPO) activity decreased significantly after DMS pretreatment (P<0.05 or P<0.01). DMS pre-treatment decreased the levels of TNF-α, IL-6 and IL-1 β (P<0.01). Meanwhile, DMS activated the phosphoinositide 3-kinase/protein kinase B (PI3K/Akt) pathway and reversed the expressions of Claudin-5, VE-cadherin and VEGF (P<0.01).@*CONCLUSIONS@#DMS attenuated LPS-induced ALI in mice through repairing endothelial barrier. It might be a potential therapeutic drug for LPS-induced lung injury.

Optimization of ethanol precipitation process of Nauclea officinalis extract based on concept of quality by design(QbD) / 中国中药杂志

The ethanol precipitation process of Nauclea officinalis extract was optimized based on the concept of quality by design(QbD). Single factor tests were carried out to determine the levels of test factors. The ethanol volume fraction, pre-ethanol precipitation drug concentration, and ethanol precipitation time were taken as critical process parameters(CPPs). With the comprehensive scores of strictosamide transfer rate and solid removal rate as the critical quality attributes(CQAs), Box-Behnken design was employed to establish the mathematical models and space design between CPPs and CQAs, and the obtained optimal operating space was validated. The optimal operating space included ethanol volume fraction of 65%-70%, pre-ethanol precipitation drug concentration of 22-27 mg·mL~(-1), and ethanol precipitation time of 12 h. Based on the concept of QbD, this study adopted the design space to optimize the ethanol precipitation process of N. officinalis extract, which provided a reliable theoretical basis for the quality control in the production process of N. officinalis preparations. Moroever, this study provided a reference value for guiding the research and industrial production of traditional Chinese medicines.

Study on HPLC Fingerprints of Nauclea officinalis Extract Syrup and Content Determination of 9 Components / 中国药房

OBJECTIVE： To establish HPLC fingerprints of Nauclea officinalis extract syrup， and to determine the contents of 9 components. METHODS： HPLC method was adopted. The determination was performed on Diamonsil C18（2）column with mobile phase consisted of acetonitrile-0.1％ phosphoric acid solution （gradient elution） at the flow rate of 1.0 mL/min. The detection wavelength was set at 240 nm， and column temperature was 30 ℃. The sample size was 10 μL. Using strictosamide as reference， HPLC chromatograms of 20 batches of N. officinalis extract syrup were drawn. The similarity of HPLC chromatograms were evaluated by using TCM Fingerprint Similarity Evaluation System （2004A edition） to confirm common peaks. The contents of 9 components were determined by standard curves. RESULTS： There were 26 common peaks in 20 batches of HPLC chromatograms， and the similarity was higher than 0.98. Compared with mixed control， 9 chemical components were identified， such as 3，4-dihydroxybenzoic acid， neochlorogenic acid， loganic acid， chlorogenic acid， cryptochlorogenic acid， swertioside， pumiloside， strictosamide and vincosamide. The linear range of 9 components were 17.24-275.84， 7.56-120.96， 15.40-246.40， 7.84-125.44， 8.64-138.24， 7.96-127.36， 8.40-134.40， 48.56-776.96， 4.16-66.56 μg/mL（all r≥0. 999）， respectively. The limits of detection were 0.043 1， 0.126 0， 0.038 5， 0.130 7， 0.144 0， 0.066 3， 0.070 0， 0.012 1， 0.052 0 μg/mL， respectively. The limits of quantitation were 0.215 5， 0.189 0， 0.077 0， 0.196 0， 0.288 0， 0.132 7， 0.105 0， 0.097 6， 0.138 7 μg/mL， respectively. RSDs of precision， stability and reproducibility tests were all lower than 2.0％ （n＝6）. Average recoveries were 99.6％、106.3％、100.1％、102.0％、98.4％、100.0％、99.3％、100.6％ and 101.2％， and RSDs were 1.20％、0.24％、0.59％、1.00％、0.73％、1.30％、1.10％、1.80％、1.90％（n＝6）. CONCLUSIONS： Established HPLC fingerprints and quantitative determination method of N. officinalis extract syrup are accurate， specific and sensitive. It can provides reference for quality control of N. officinalis extract syrup.

Chemical constituents from stems and leaves of Nauclea officinalis / 中草药

Objective: To study the chemical constituents from stems and leaves of Nauclea officinalis. Methods: The chemical constituents from N. officinalis were separated and purified by silica gel, ODS, Sephadex LH-20 gel column chromatographies, and preparative HPLC. Their structures were identified by physicochemical properties, spectroscopic analysis, as well as comparisons with the data in literature. Results: Eighteen compounds were isolated from 85% ethanol extract from N. officinalis, and identified as dihydroactinidiolide (1), loliolide (2), 3,4,5-trimethoxyphenol (3), 4-hydroxy-3,5-dimethoxybenzaldehyde (4), methyl-2,4-dihydroxy- 3,6-dimethyl-benzoate (5), p-methoxy cinnamic acid (6), caffeic acid methyl ester (7), ethyl caffeate (8), methyl isoferulate (9), ethyl ferulate (10), secoxyloganin (11), secologanoside (12), 1H-indole-3-aldehyde (13), 1,2,3,4-tetrahydronorharman-1-one (14), vinmajine I (15), 19-O-methyl-3,14-dihydroangustoline (16), naucleidinal (17), and strictosamide (18). Conclusion: Compounds 1-15 were isolated from the genus Nauclea Linn. for the first time.

Study on Quality Standard for the Leaves of Nauclea officinalis / 中国药房

OBJECTIVE:To establish the quality standard for the leaves of Nauclea officinalis. METHODS:Microscopic identi-fication and TLC methods were used for qualitative identification of N. officinalis. The moisture and ash of medicinal materials were determined. HPLC method was adopted to determine the content of strictosamide in medicinal materials. The determination was per-formed on Lichrospher C18 column with mobile phase consisted of acetonitrile-0.1% phosphoric acid solution(gradient elution)at the flow rate of 1 mL/min,injection volume was 10 μL. The detection wavelength was set at 226 nm,and the column temperature was 30 ℃. RESULTS:Microscopic identification of the leaves of N. officinalis had strong characteristics,and TLC sports were clear and well-separated without interference from negative control. The linear range of strictosamide were 0.0105-0.21 mg/mL(r=0.9995);RSDs of precision,stability and reproducibility tests were all lower than 2.0%. Average recoveries were 96.25-101.82%(RSD=1.86%,n=9). Total ash of medicinal materials was 5.27%-6.44%,acid insoluble ash was 0.23%-0.36%,water content was 9.48%-11.46%,strictosamide was 0.124%-1.003%. CONCLUSIONS:Established standard can be used for quality evaluation of N. officinalis.

Alkaloids from stems of Nauclea officinalis / 中草药

Objective: To study the alkaloids from the stems of Nauclea officinalis. Methods: The chemical constituents were separated and purified by silica gel, ODS column chromatography, and preparative HPLC. Their structures were determined by UV, MS, and NMR spectroscopic analyses. Results: Thirteen compounds were isolated from the stems of N. officinalis, the structures were identified as 3-R-3,4-dihydroangustoline (1), blumenol A (2), naucleofficine D (3), 1,2,3,4-tetrahydro-β-carboline (4), 3-S-3,4-dihydroangustoline (5), latifoliamide D (6), latifoliamide B (7), angustoline (8), 3,14-dihydroangustine (9), 3,14,18,19- tetrahydroangustine (10), 6'-acetyl-strictosamide (11), vincosamide (12), and strictosamide (13). Conclusion: Compounds 2 and 4 are obtained from the plants of Nauclea L. for the first time. Compounds 2, 4, 6, 7, 9, and 10 are obtained from N. officinalis for the first time.

Alkaloidal constituents of Nauclea officinalis / 中草药

Objective To investigate the alkaloidal constituents of Nauclea officinalis, an anti-inflammatory and antibacterial traditional Chinese herb. Methods The alkaloids were isolated and purified by silica gel, Sephadex LH20, and C-18 ODS column chromatography repeatedly, and their structures were identified by spectral analysis. Results Eleven alkaloids were isolated from the stems of N. officinalis and their structures were identified as angustoline (Ⅰ), 19-O-ethylangustoline (Ⅱ), 3-S-3, 4-dihydroangustoline (Ⅲ), 3-R-3, 4-dihydroangustoline (Ⅳ), naucleamide A (Ⅴ), strictosamide (Ⅵ), vincosamide (Ⅶ), 6′-acetyl-strictosamide (Ⅷ), 2′-acetyl-strictosamide (Ⅸ), pumiloside (Ⅹ), 3-epi-pumiloside (Ⅺ). Conclusion Compounds Ⅰ-Ⅺ are isolated from this plant for the first time.