Tetrahydroxy stilbene glycoside ameliorates neuroin-flammation for Alzheimer's disease via cGAS-STING / 中国药理学与毒理学杂志

OBJECTIVE Alzheimer's disease(AD)is the most common neurodegenerative disease worldwide.Neuroinflammation is a potential target for the patients with AD.It is attributed to activated microglia and the release of various inflammatory mediators from infec-tion,ischemia and toxin accumulation.Accumulating evi-dence has indicated that the cGAS-STING pathway driven neuroinflammation in neurological disease.TSG is a main natural active ingredient that derived from polyg-onum multiflorum.Previous research from our group found that TSG has beneficial effects of anti-aging,anti-inflammatory action and improving memory function in APP/PS1 transgenic AD mice.Here,we investigated the effects of TSG on cognitive impairment and neuroinflam-mation in APP/PS1-AD mice and explore the underly-ing mechanism by which TSG ameliorates memory func-tion in the cGAS-STING-mediated inflammatory response.METHODS The Morris water mace test and the novel object recognition test were performed to test the effects of TSG on spatial learning and cognitive and memory abil-ity in APP/PS1 double transgenic AD mice model.In addi-tion,real-time quantitative PCR,Western blotting,ELISA analysis,and flow cytometry to examine gene and pro-tein expression of cGAS-STING related pro-inflammatory cytokines and chemokines.Statistical analyses were ana-lyzed using the SPSS 25.0 package by analysis of vari-ance(ANOVA).Neuman-Keuls or Tukey's multiple-com-parisons test were conducted as ANOVA justified post hoc comparisons between group means.RESULTS We demonstrated that AD transgenic mice exhibited cognitive deficits accompanied by the elevated serum and brain inflammation.The expressions of serum inflammatory cytokines and the activation of microglia in cerebral cor-tex and hippocampus were suppressed after TSG treat-ment,which was probably attributable to the decrease of cyclic GMP-AMP synthase(cGAS)and stimulator of interferon genes(STING)triggered immune response.Additionally,the data showed that TSG treatment reduced the expression level of inflammatory cytokines(IL-1β,TNF-α,IFN-β,IFN-α)in microglial cells BV2 primed with LPS and IFN-γ.CONCLUSION TSG implicated the health benefits in preventing cognitive disorders by inhib-iting neuroinflammation via cGAS-STING signalling path-way in AD.

Qualitative and quantitative analysis of chemical compounds in 3 species of wild Veratrum from Inner Mongolia / 中国药房

OBJECTIVE To conduc t qualitative and quantitative analysis for the chemical compounds in 3 species of wild Veratrum（V. nigrum ，V. maackii ，V. dahuricum ）from Inner Mongolia. METHODS HPLC-Q-Exactive-MS/MS technology was used to identify the chemical components of V. nigrum ，V. maackii and V. dahuricum by consulting SciFinder ，ChemSpider database and related literatures and comparing with the reference substance. The contents of polydatin ，oxyresveratrol and resveratrol in 3 species of wild Veratrum were determined by HPLC. RESULTS A total of 31 compounds were identified ，including 13 stilbenes， 11 flavonoids，4 organic acids ，2 glycosides，1 brasilin. Most of the compounds were shared by 2 or 3 species of wild Veratrum， only 2 flavonoids kaempferol and luteolin were owned by V. dahuricum . The total contents of polydatin ，oxyresveratrol and resveratrol in 3 species of wild Veratrum were in the range of 6.618-11.292 mg/g，and the total contents of them in V. nigrum were the highest ，followed by V. maackii and V. dahuricum . The contents of polydatin and resveratrol in V. maackii were the highest ，and the content of oxyresveratrol in V. nigrum was highest. CONCLUSIONS Most of the components of 3 species of wild Veratrum are similar，only kaempferol and luteolin are unique to V. dahuricum . The contents of polydatin ，oxyresveratrol and resveratrol are significantly different among 3 species of wild Veratrum.

Effect of TSG on GSK3β，PKA and PP2A of APP/PS1/Tau Triple-transgenic Mice Dementia Model / 中国实验方剂学杂志

Objective:To observe the effect of tetrahydroxy stilbene glycoside （TSG） on the expression of glycogen synthase kinase 3<italic>β </italic>（GSK3<italic>β</italic>）， cyclic adenosine monophosphate-dependent protein kinase （PKA） and Serine/threonine phosphatase 2A（PP2A） in the brain of amyloid precursor protein/presenilin-1/Tau （APP/PS1/Tau） triple-transgenic mice dementia model. Method:A total of forty-five 8-month-old APP/PS1/Tau transgenic mice were randomly divided into model group， positive control group （Huperzine-A， 0.15 mg·kg^-1）， low， medium and high dose TSG groups （TSG， 0.033，0.1，0.3 g·kg^-1）， with 9 mice in each group， and another nine C5B7L/6J mice of the same age were selected as normal control group. After 60 days of intragastric administration， the general structure of hippocampal neurons was observed by hematoxylin-eosin （HE） staining， immunohistochemical （IHC） was used to detect the expression of PKA protein in the brain of mice in each group， the mRNA expression levels of GSK3<italic>β</italic>， PKA and PP2A were detected by real time quantitative reverse transcription polymerase chain reaction （Real-time PCR）， and protein expression levels of GSK3<italic>β</italic> and PP2A were detected by Western blot. Result:Compared with the normal control group， the apoptosis level of neurons in the model group was significantly increased， the protein and mRNA expression levels of GSK3<italic>β</italic> and PKA were significantly increased （<italic>P</italic><0.05， <italic>P</italic><0.01）， and the protein and mRNA expression levels of PP2A were significantly decreased （<italic>P</italic><0.05， <italic>P</italic><0.01）. Compared with the model group， the apoptosis level of neurons in each treatment group was significantly down-regulated， the protein and mRNA expression levels of GSK3<italic>β</italic> and PKA were significantly down-regulated （<italic>P</italic><0.05， <italic>P</italic><0.01）， and the protein and mRNA expression levels of PP2A were significantly increased （<italic>P</italic><0.05， <italic>P</italic><0.01）. Conclusion:The mechanism of TSG in the treatment of Alzheimer's disease （AD） may be related to lowering the transcription and expression of GSK3<italic>β</italic> and PKA， increasing the transcription and expression of PP2A.

Hepatotoxicity of Main Components of Polygonum multiflorum and Its Effects on Drug Metabolism Enzyme / 中国药房

OBJECTIVE：To study the hepatotoxicity of main components of Polygonum multiflorum ，and investigate its toxic mechanism based on metabolic enzymes. METHODS ：ADMETlab 2.0 platform was used to forecast the toxic or carcinogenic effects of emodin ，physcion，rhein，stilbene glycoside and gallic acid on liver ，skin and heart. The effects of those components on cytochrome P 450 enzyme system （CYP1A2，CYP2C9，CYP2C19，CYP2D6，CYP3A4）were evaluated. The effects of different concentrations of emodin ，rhein，stilbene glycoside and gallic acid （10，20，40，80 μmol/L）on the survival rate of normal hepatocyte L 02 were detected. The effects of major components of P. multiflorum on the activity of UGT 1A1 enzyme were studied by in vitro reaction system ，using bilirubin as substrate. RESULTS ：Main components of P. multiflorum ，ie. emodin ，physcion， rhein and gallic acid ，showed strong toxic effects on the liver ，while stilbene glycosides possessed weak toxic effects on the liver. Emodin and physcion had strong inhibitory effects on CYP 1A2 and medium inhibitory effects on CYP 2C9，CYP2D6 and CYP3A4；rhein showed medium inhibitory effects on CYP 1A2 and CYP 2C9，while stilbene glycoside and gallic acid possessed weak inhibitory effects on the above enzymes. Emodin （40，80 μmol/L）and gallic acid （40，80 μmol/L）could significantly reduce the survival rate of L 02 cells（P＜0.01）. The inhibition rate of 5，10，20，40，80 μmol/L emodin and gallic acid（except for 5 μmol/L emodin）on UGT 1A1 enzyme increased significantly （P＜0.01），and the inhibition effect of emodin on UGT 1A1 enzyme was reversible competitive inhibition. CONCLUSIONS ：The main components of P. multiflorum ，ie. emodin ，rhein and physcion ， are hepatotoxic ；the mechanism of it may be associated with inhibiting the activity of CYP 1A2 and CYP 2C9 and competitively blocking rate-limiting enzyme UGT 1A1 in the process of bilirubin metabolism.

Effects of Stilbene Glycoside on the Phosphorylation of Thr 205 and Ser 404 Sites of Tau Protein in AD Model Mice / 中国药房

OBJECTIVE：To study the e ffects of stilbene glycoside c（TSG）on phosphorylation of Thr 205，Ser404 sites of Tau protein in Aizheimer ’s disease （AD）model mice ，and to investigate the potential anti-AD mechanism of TSG. METHODS ：APP/ PS1/Tau three transgenes （3×Tg-AD）mice were randomly divided into model group ，positive control group （huperzine，0.15 mg/kg），TSG low-dose ，medium-dose and high-dose groups （0.033，0.1，0.3 g/kg），with 6 mice in each group. In addition ，6 C57BL/6J mice were chosen as normal control group. Administration groups were given relevant medicine intragastrically. Model group and normal control group were given equal volume of normal saline intragastrically ，once a day ，for consecutive 60 days. After last medication ，immunofluorescence staining was used to detect Tau protein and phosphorylated Tau protein （Thr205， Ser404 sites） distribution and expression in brain tissue of mice in each group. Western blotting assay was used to detect phosphorylated Tau protein （Thr205，Ser404 sites）expression level in brain tissue of mice in each group. RESULTS ：Compared with normal control group ，the expression of Tau protein，phosphorylated Tau protein （Thr205，Ser404 sites）in 729011126@qq.com the brain tissue of mice were increased in model group ，which were easy to aggregate and distributed more widely ；theirrelative expression were increased significantly （P＜0.01）. Results of Western blotting assay showed that the expression levels of phosphorylat ed Tau protein （Thr205，Ser404 sites）were increased significantly （P＜0.01）. Compared with model group ，the expression of Tau protein ，phosphorylated Tau protein （Thr205，Ser404 sites） in the brain tissue of mice were decreased in positive control group and TSG groups ；aggregation decreased，distribution narrowed and their relative expression were decreased significantly （P＜0.01）. Results of Western blotting assay showed that the expression levels of phosphorylated Tau protein （Thr205，Ser404 sites）were decreased significantly （P＜ 0.01）. Compared with positive control group ，There was no significant difference in the distribution of Tau protein ，phosphorylated Tau protein （Thr205，Ser404 sites）in the brain tissue of mice in TSG groups ；the relative expression were not statistically significant（P＞0.05）；but Western blotting assay showed the expression levels of phosphorylated Tau protein （Thr205 site）in TSG medium-dose and high-dose groups as well as the expression levels of phosphorylated Tau protein （Ser404 site）in TSG groups were decreased significantly （P＜0.05 or P＜0.01）. CONCLUSIONS ：TSG can play an anti-AD effect on AD model mice by down-regulating the expression of phosphorylated Tau protein （Thr205，Ser404 sites）in brain tissue.

Preliminary study on hepatotoxic components in Polygoni Multiflori Radix based on different adjuvants processing for detoxification / 中草药

Objective: To provide references for optimizing adjuvants with Polygoni Multiflori Radix (PMR), we compared the effects of detoxification by different adjuvants processing according to Chinese medicine’s records of past dynasties. Methods: The chemical information of all samples including crude and processed PMR with different adjuvants was characterized by UPLC-Q-TOF-MS, and the normal human hepatocytes (L02 cell line) was cultured in vitro to evaluate the cytotoxicity, then we gave synthetic analyses on effects of processed PMR with different adjuvants for toxicity-decreasing and variations of chemical contents. The difference of toxicity reducing effect and the rule of composition change of PMR processed with different adjuvants were compared comprehensively. Results: Different adjuvants had different level of effects on chemical fingerprint, index component and cytotoxicity of PMR under the same conditions of pressure and time. More specifically, black bean, jujube and rice-rinsing water had greater impact on PMR main components including gallic acid, catechins, cis-stilbene glycoside, trans-stilbene glycoside, emodin-8-O-β-D-glucoside, physcion and emodin as well as hepatotoxicity. The three adjuvants with the best toxicity-decreasing effects were in sequence of rice-rinsing water > jujube > black bean. Furthermore, comprehensive analysis of simple correlation and multiple correlation suggested that cis-stilbene glycoside might be the main chemical component contributed to hepatotoxicity of PMR, and emodin-8-O-β-D-glucoside might be the potential toxicity component. Conclusion: Different adjuvants traditionally recorded can attenuate the toxicity of PMR. In addition to black beans, rice-rinsing water and jujube can also be used as candidate adjuvants for the toxicity-decreasing of PMR.

Regulatory Effects of Stilbene Glucoside on JNK and PP 2B in APP/PS1/Tau Transgenic Dementia Mice / 中国药房

OBJECTIVE：To study the regulatory effects of stilbene glucosid e（TSG）on c-Jun N-terminal kinase （JNK）and protein phosphortase 2B（PP2B）in APP/PS1/Tau transgenic dementia （3×Tg-AD）mice，and to explore its potential mechanism of anti-Alzheimer’s disease （AD）. METHODS ：Totally 45 male 3×Tg-AD mice were randomly divided into model group ，positive control group （huperzine A ，0.15 mg/kg），TSG low-dose ，medium-dose and high-dose groups （0.033，0.1，0.3 g/kg），with 9 mice in each group. Another 9 normal male C 57BL/6J mice were included into normal control group. Administration groups were given relevant medicine intragastrically ，once a day ，for consecutive 60 d. Normal control group and model group were given constant volume of normal saline intragastrically. After medication ，Morris water maze experiment was used to test the spatial learning and memory ability of mice in each group ；Nissl staining was used to observe the changes of Nissl bodies in cerebral cortex and hippocampus ；mRNA and protein expressions of JNK and PP 2B were detected by qRT-PCR and Western blotting assay. RESULTS：Compared with normal control group ，the escape latency was significantly prolonged （P＜0.01），the retention time of the original platform quadrant was significantly shortened （P＜ and the times of crossing the platform was significantly reduced in model group （P＜0.01）；the number of Nissl bodies in cerebral cortex and hippocampus was significantly 729011126@qq.com reduced，the staining was slight ；the relative expressions of JNK mRNA and protein were significantly increased （P＜ 0.01），and the relative expressi ons of PP 2B mRNA and protein were significantly decreased （P＜0.01）. Compared with model group ，the escape latency was significantly shortened in positive control group and TSG groups （P＜0.01）；the retention time of the original platform quadrant was significantly prolonged （P＜0.01）；the times of crossing the platform was significantly increased （P＜0.01）；the number of Nissl bodies in cerebral cortex and hippocampus was increased significantly ，the staining was heavy ；the relative expression of JNK protein was significantly decreased（P＜0.05 or P＜0.01），the relative expressions of PP 2B mRNA and protein were significantly increased （P＜0.01）， while the relative expression of JNK mRNA was significantly decreased in TSG high-dose group （P＜0.05）. CONCLUSIONS ：TSG can improve the learning and memory ability and neuronal damage of 3 × Tg-AD mice. The mechanism may be related to down-regulating the transcription and expression of protein kinase JNK ，up-regulating the transcription and expression of protein phosphatase PP 2B.

Effects of Stilbene Glucoside on the Proliferation and Estrogen Receptor Expression of Human Breast Cancer T-47D Cells / 中国药房

OBJECTIVE：To study the effects of stilbene glucoside （TSG）on the proliferation and estrogen receptor （ER）of human breast cancer T- 47D cells ，and to explore its estrogen-like effect and potential mechanism. METHODS ：Taking ER positive human breast cancer T- 47D cells as subjects ，using β-estradiol（β-E2，1×10－8 mol/L）as positive control ，CCK-8 assay was used to detect the cell proliferation after treated with different concentrations of TSG （1×10－8，1×10－7，1×10－6，1×10－5，1×10－4 mol/L）for 24，48，72 h；the cell proliferation rate was calculated. Western blotting assay and RT-PCR methods were adopted to detect the protein and mRNA expression of ER-α and ER-β in cells after treated with low，medium and high concentrations of TSG （1×10－8， 1×10－6，1×10－4 mol/L）for 48 h. RESULTS ：After treated with different concentrations of TSG for 24，48，72 h，the cell proliferation rate of each administration group at each time point （except for β-E2 group at 48 h）increased significantly ，compared with blank group ；those of TSG groups （1×10－5，1×10－6，1×10－7 mol/L）were significantly higher than β-E2 group（P＜0.05 or P＜0.01）. After treated with low ，medium and high concentrations of TSG for 48 h，protein and mRNA expression of ER-α and ER-β in cells were increased significantly，compared with blank group （P＜0.05 or P＜0.01）；protein expression of ER-β in TSG low concentration group ，mRNA expression of ER-α in TSG groups as well as mRNA expression of ER-β in TSG low and high concentration groups were significantly higher than β-E2 group（P＜0.05 or P＜0.01）. CONCLUSIONS ：TSG can induce the in vitro proliferation of T- 47D cells and exert estrogen-like effects by promoting protein and mRNA expression of ER-α and ER-β， which is stronger than that of β-E2 at a certain concentration.

Exploring Research on Processing Method of Polygoni Multiflori Radix Praeparata / 中国实验方剂学杂志

Objective::To investigate the effects of black bean juice with different stewing times on the appearance character and the content changes of effective components of Polygoni Multiflori Radix Praeparata. Method::HPLC was employed with Agilent ZORBAX Extend-C18 column (4.6 mm×250 mm, 5 μm), a gradient mobile phase of methanol (A)-water (B) was eluted (0-30 min, 5%-100%A; 30-40 min, 100%A), the flow rate was 1.0 mL·min-1, the injection volume was 10 μL and the column temperature was 35 ℃, detection wavelength was set at 280 nm. The contents of stilbene glycoside, emodin, emodin methyl ether, emodin-8-O-β-D-glucoside and emodin methyl ether-8-O-glucoside in samples prepared at different processing times were simultaneously determined by HPLC. Result::The content of stilbene glycoside decreased gradually with the increase of stewing time, compared with 8 h, its content decreased by 76% at 64 h. The contents of emodin-8-O-β-D-glucoside and emodin methyl ether-8-O-glucoside increased first, and then decreased, reaching the highest value at 24 h, and then decreased to the level similar to the content of 8 h after 40 h, and then fluctuated slightly. The contents of emodin and emodin methyl ether increased first, and then decreased, reached the maximum when stewed for 32 h, then decreased slowly and tended to be stable. Conclusion::The stewing time has significant influence on the content of various components in Polygoni Multiflori Radix Praeparata, and the changing trend is different, the processing time of Polygoni Multiflori Radix Praeparata shall be standardized. At the same time, it is not sufficient to take stilbene glycoside and anthraquinones as the indicator ingredients for this decoction pieces, the quality control indicators such as polysaccharides shall be considered.

Developing spectrum-toxicity relationship with rough set theory for hepatotoxicity material basis of Polygonum multiflorum / 中国中药杂志

Idiosyncratic hepatotoxicity of Polygonum multiflorum has attracted a great attention in the world. The most toxic part of idiosyncratic hepatotoxicity was screened by MTT assay and flow cytometry, which was the 50% ethanol elute by macroporous adsorptive resins from alcohol-extraction of P. multiflorum. The fingerprints were collected by HPLC from 50% ethanol elute of crude and processed P. multiflorum from different habitats, then 14 common peaks were determined. Spectrum-toxicity relationship was analyzed by rough set theory(RST). Two main chemical components were predicted for idiosyncratic hepatotoxicity, in which TSG was the greater contributor. Idiosyncratic hepatotoxicity of TSG was tested in vitro, and the results indicated that TSG was the most important constituent contributed to idiosyncratic hepatotoxicity of P. multiflorum. The study showed the discovery of the main chemical components for idiosyncratic hepatotoxicity, and RST was effective for analyzing the spectrum-toxicity relationship, which could be a new method used in the effective/toxic constituents field of traditional Chinese medicine.

Flavestin K, An Isoprenylated Stilbene from the Leaves of Macaranga recurvata Gage

A new isoprenylated stilbene, flavestinK (1) together with two known isoprenylated stilbenes, flavestin B (2), flavestin G (3), and two isoprenilated flavanones, 4-O-methyl-8-isoprenylnaringenin (4) and 8-isoprenyl-5,7-dihydroxyflavanone (5) were isolated from the leaves of Macaranga recurvata Gage. All of the structures have been determined based on HRESIMS, 1D and 2D NMR spectral data. All of the isolated compounds were evaluated for their cytotoxicity against three human cancer cells (HeLa, T47D and WiDr). Compound 1 showed higher activity than doxorubicin against HeLa cells with IC₅₀ value of 13.1 µg/mL.

Analysis of HPLC Characteristic Chromatogram of Standard Pieces of Polygoni Multiflori Radix / 中国实验方剂学杂志

Objective: To establish HPLC characteristic chromatogram of standard pieces of Polygoni Multiflori Radix, and compare with the HPLC characteristic chromatograms of original pieces, raw materials and control materials of Polygoni Multiflori Radix. Method: HPLC analysis was carried out on a Waters BEH-C18 column (4.6 mm×250 mm, 5 μm) with the mobile phase of methanol(A)-0.1%phosphoric acid(B) for gradient elution (0-20 min, 15%-30%A; 20-35 min, 30%-40%A; 35-55 min, 40%-75%A; 55-75 min, 75%-100%A) at a flow rate of 0.8 mL·min-1. The detection wavelength was set at 270 nm and the column temperature was maintained at 30℃. The injection volume was 10 μL. Moreover, similarity and cluster analysis of HPLC characteristic chromatograms of four samples of Polygoni Multiflori Radix were performed. Result: HPLC characteristic chromatogram of standard pieces of Polygoni Multiflori Radix composed of seven peaks was established. The similarity was 0.999 between characteristic chromatograms of standard pieces and original pieces, which was better than similarities among characteristic chromatograms of raw materials, control materials and original pieces. There was no obvious difference on number of peaks between characteristic chromatograms of standard pieces and original pieces, while an obvious difference on number of peaks between chromatograms of control materials and original pieces was found. In addition, standard pieces, original pieces and control materials could generally gather into one class, while raw materials could generally gather into another class. Conclusion: Compared with the raw materials and control materials, the established HPLC characteristic chromatogram of standard pieces can better reflect the internal quality of original pieces, which can be used for the quality control of decoction pieces of Polygoni Multiflori Radix.

Neuroprotective Effect of Tetrahydroxy Stilbene Glucoside on Aβ25-35-induced Neuron Synapses via CREB/BDNF Signaling Pathway / 中国实验方剂学杂志

Objective: To investigate the neuroprotective effect and mechanism of tetrahydroxy stilbene glucoside (TSG) on β-amyloid protein 25-35 (Aβ25-35)-induced neuron synapses damage. Method: Primary neurons were isolated and purified from cerebral cortex of suckling mouse. Then neurons were divided into control group, model group (incubation with Aβ25-35) and TSG groups (after incubation with Aβ25-35, add 6.25, 12.5, 25, 50, 100 μmol·L-1 TSG). Cell counting kit-8 (CCK-8) and Lactate dehydrogenase (LDH) methods were used to observe the viability of neuron, immunocytochemical staining was performed to determine the expressions of synapsin-1 (SYN-1), and the concentration of postsynaptic density-95 (PSD-95) and synaptophysin (SYP) were detected by enzyme-linked immunosorbent assay (ELISA) method. The level of cyclic adenosine monophosphate response element binding protein (CREB) and brain-derived neurotrophic factor (BDNF) mRNA were determined by reverse transcription-polymerase chain reaction (RT-PCR) and the level of CREB, Phosphorylated CREB (p-CREB) and BDNF proteins were determined by immunocytochemical staining or Western blot (WB). Result: Compared with normal group, the cell survival rate of model group was significantly reduced, LDH release was significantly increased (PPPPPPP-1,25 μmol·L-1 TSG can significantly enhance the expression of SYN-1(PPPPConclusion: TSG possesses the neuroprotective effect on Aβ25-35-induced neuron synapses, the mechanism may be associated with the activation of CREB/BDNF signaling pathway.

Identification of Soluble Epoxide Hydrolase Inhibitors from the Seeds of Passiflora edulis Cultivated in Vietnam

Soluble epoxide hydrolases (sEH) are enzymes present in all living organisms, metabolize epoxy fatty acids to 1,2-diols. sEH in the metabolism of polyunsaturated fatty acids plays a key role in inflammation. In addition, the endogenous lipid mediators in cardiovascular disease are also broken down to diols by the action of sEH that enhanced cardiovascular protection. In this study, sEH inhibitory guided fractionation led to the isolation of five phenolic compounds trans-resveratrol (1), trans-piceatannol (2), sulfuretin (3), (+)-balanophonin (4), and cassigarol E (5) from the ethanol extract of the seeds of Passiflora edulis Sims cultivated in Vietnam. The chemical structures of isolated compounds were determined by the interpretation of NMR spectral data, mass spectra, and comparison with data from the literature. The soluble epoxide hydrolase (sEH) inhibitory activity of isolated compounds was evaluated. Among them, trans-piceatannol (2) showed the most potent inhibitory activity on sEH with an IC₅₀ value of 3.4 µM. This study marks the first time that sulfuretin (3) was isolated from Passiflora edulis as well as (+)-balanophonin (4), and cassigarol E (5) were isolated from Passiflora genus.

Rapid screening of potential analgesic ingredients from Draconis Resina by live cell immobilized chromatography coupled with HPLC-DAD-TOF-MS / 中草药

Objective: To rapidly screen the potential analgesic ingredients from Draconis Resina by live cell immobilized chromatography coupled with HRMS. Methods: An HPLC-DAD-ESI-TOF-MS technique was used to rapidly identify the main chemical constituents from Draconis Resina. Based on the bio-specific affinity adsorption of bioactive compounds with receptors or channels on cells, the potential bioactive components in Draconis Resina could be selectively bound to the target cells-mice dorsal root neurons cells, then the chemical constituents with cell target affinity were identified by LC-HRMS. Results: A total of 21 compounds with various structures were tentatively identified and characterized by HPLC-DAD-ESI-TOF-MS, and among them, 10 potentially analgesic active ingredients in Draconis Resina extract combined with dorsal root neurons cells were successfully detected and identified, including two stilbene, two homoisoflavones, one homoisoflavone, two dihydrochalcone, and three flavonoid oligomers. Conclusion: Live cell immobilized chromatography coupled with LC-DAD-HRMS analysis could provide a rapid and efficient tool for finding the potential bioactive components in Draconis Resina for the next pharmacology studies, which provide the reference for exploring of effective materials basis in Chinese medicines.

Effects of Stilbene Glycoside on Okadaic Acid-induced Tau Protein Phosphorylation in NG 108-15 Cells / 中国药房

OBJECTIVE： To observe the effects of stilbene glycosidec （TSG） on okadaic acid （OA）-induced Tau protein phosphorylation in NG108-15 cells， and to investigate the potential anti-Alzheimer’s disease （AD） mechanism of this compound. METHODS： AD model of NG108-15 cells was induced by OA. The survival rate of NG108-15 cells was observed by MTT assay after pretreated with low-dose， medium-dose and high-dose of TSG （50， 100， 200 μmol/L）. The apoptosis of NG108-15 cells was detected by AO/EB double fluorescence staining. The protein and mRNA expression of CDK5 and GSK3β， and the protein expression of Tau and p-Tau were detected by Western blotting assay and RT-PCR. The distribution of CDK5， GSK3β and Tau protein were detected by immunofluorescence. RESULTS： The normal morphology of NG108-15 cells was observed in normal control group， but CDK5， GSK3β and Tau protein were not found or few was found. Contracted or globular early apoptotic cells were observed in model gorup； the distribution of CDK5， GSK3β and Tau protein was increased， while survival rate of the cells was decreased； protein and mRNA expression of CDK5 and GSK3β as well as ratio of the relative expression of p-Tau to that of Tau （p-Tau/Tau） were all increased significantly （P＜0.05 or P＜0.01）. After pretreatment of TSG， the distribution of early apoptotic cells as well as CDK5， GSK3β and Tau protein were all decreased to some extent in administration groups， while survival rates of the cells were increased significantly. Protein expression of CDK5 and p-Tau/Tau in medium-dose group and high-dose group as well as mRNA expression of CDK5， protein and mRNA expression of GSK3β in administration group were decreased significantly （P＜0.05）. CONCLUSIONS： TSG can protect against AD model cells， the effects of which may be associated with improving survival rate of the cells， down-regulating the protein expression and gene transcription level of phosphokinase CDK5 and GSK3β， inhibiting Tau protein phosphorylation.

Antifungal activity of extracts and phenolic compounds from Deguelia duckeana

ABSTRACT Candida spp. is associated with almost 80% of all nosocomial fungal infections and is considered a major cause of blood stream infections. In humans, Cryptococcosis is a disease of the lungs caused by the fungi Cryptococcus gattii and Cryptococcus neoformans. It can be potentially fatal, especially in immune-compromised patients. In a search for antifungal drugs, Deguelia duckeana extracts were assayed against these two fungi and also against Candida albicans, which causes candidiasis. Hexane branches and CH2Cl2 root extracts as well as the substances 4-hydroxylonchocarpine, 3,5,4′-trimethoxy-4-prenylstilbene and 3′,4′-methylenedioxy-7-methoxyflavone were assayed to determine the minimal inhibitory concentration. Phytochemical study of CH2Cl2 root and hexane branch extracts from D. duckeana A.M.G. Azevedo, Fabaceae, resulted in the isolation and characterization of nine phenolic compounds: 4-hydroxyderricine, 4-hydroxylonchocarpine, 3′,4′,7-trimethoxy-flavonol, 5,4′-dihydroxy-isolonchocarpine, 4-hydroxyderricidine, derricidine, 3,5,4′-trimethoxy-stilbene, 3′,4′,7-trimethoxyflavone and yangambin. The only active extract was a CH2Cl2 root showing minimal inhibitory concentration 800 µg/ml against C. gattii, and the investigation of compounds obtained from this extract showed that 4-hydroxylonchocarpine was active against all three fungi (C. neoformans, C. gattii and C. albicans). These results suggest that D. duckeana extracts have potential therapeutic value for the treatment of pathogenic fungi.

Increasing pinosylvin production in Escherichia coli by reducing the expression level of the gene fabI-encoded enoyl-acyl carrier protein reductase

Background: The plant secondary metabolite pinosylvin is a polyphenol from the stilbene family, which have positive effects on human health. Biotechnological production is an attractive alternative for obtaining this stilbene. In Escherichia coli, malonyl-CoA is the precursor for both stilbene and fatty acid syntheses. In this study, with the aim of increasing pinosylvin production, we evaluated a novel approach that is based on reducing the expression of the gene fabI, which encodes the enzyme enoyl-acyl carrier protein reductase that is involved in fatty acid synthesis. Results: A recombineering method was employed to eliminate the chromosomal -35 promoter sequence and the upstream region of the gene fabI in E. coli strain W3110. Analysis, employing RT-qPCR, showed that such modification caused a 60% reduction in the fabI transcript level in the mutant strain W3110Δ-35fabI::Cm compared to the wild type W3110. Synthetic genes encoding a mutant version of 4-coumaroyl-CoA ligase from Streptomyces coelicolor A3 with improved catalytic activity employing cinnamic acid as substrate and a stilbene synthase from Vitis vinifera were cloned to generate the plasmid pTrc-Sc4CL(M)-VvSTS. The production performance of strains W3110Δ-35fabI::Cm/pTrc-Sc4CL(M)-VvSTS and W3110/pTrc-Sc4CL(M)- VvSTS was determined in shake flask cultures with Luria-Bertani medium supplemented with 10 g/L glycerol and 3 mM cinnamic acid. Under these conditions, the strain W3110Δ-35fabI::Cm/pTrc-Sc4CL(M)-VvSTS produced 52.67 mg/L pinosylvin, a level 1.5-fold higher than that observed with W3110/pTrc-Sc4CL(M)-VvSTS. Conclusion: A reduction in the transcript level of fabI caused by the elimination of the -35 and upstream promoter sequences is a successful strategy to improve pinosylvin production in E. coli.

Five New Stilbenes from the Stem Bark of Artocarpus communis

Five new prenylated stilbenes (1 – 5), along with the known compounds cudraflavone C, trans-4-isopentenyl-3,5,2′,4′-terahydroxystilbene, trans-4-(3-methyl-E-but-1-enyl)-3,5,2′,4′-tetrahydroxystilbene, pannokin G, cycloartobiloxanthone, artonin P, morusin, artocarpin, artonin E, kuwanon C, artobiloxanthone, and artoindonesianin C (6 – 17) were isolated from the stem bark of the tropical tree Artocarpus communis. The structures were established by NMR spectroscopic analysis, MS studies, and comparison with spectral data reported in the literature.

Simultaneous Determination of Stilbene Glycosides, Ferulic Acid and Paeonol in Qubai Granules by a Dual-wavelength HPLC Method / 中国药师

Objective: To establish an HPLC determination method for the quantitative indicators in Qubai granules( stilbene gly-cosides, ferulic acid and paeonol). Methods: A dual-wavelength HPLC assay was used with the following conditions: a chromatogra-phy column ODS (150 mm×4. 6 mm, 5 μm) was used, the mobile phase was methanol: 0. 1% phosphoric acid with gradient elution, the column temperature was 35℃, and the injection volume was 10 μl. Results: The linear ranges of the three active constituents were 1.56-156.00 μg·ml-1(r stilbeneglycoside=0.999 8)、1.00~100.00 μg·ml-1(rferulicacid=0.999 8),1.61~161.00 μg·ml-1(rpaeonol=0. 999 7), respectively, and the average recoveries of the three constituents were between 100. 33% and 100. 76% with the RSDS less than 2% (n=6). Conclusion: The method is simple, stable and reliable, which can be used for the quality control of Qubai granules.