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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<sup>-1</sup>）， low， medium and high dose TSG groups （TSG， 0.033，0.1，0.3 g·kg<sup>-1</sup>）， 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.
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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.
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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.
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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.
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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.
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OBJECTIVE:To establish the method for the rapid determination of stilbene glycoside in Shouwu pills. METH-ODS:HPLC method was used to determine the content of stilbene glycoside in Shouwu pills(as measured value). The determina-tion was performed on ODS-C18 column with mobile phase consisted of acetonitrile-water(25:75,V/V)at the flow rate of 0.8 mL/min. The detection wavelength was set at 320 nm,and the column temperature was 30 ℃. The sample size was 10 μL. The partial least square method-near infrared diffuse reflectance spectrophotometry was used to establish quantitative calibration model for pre-dicting the content of stilbene glycoside in Shouwu pills. According to measured value,76 calibration set samples and 24 validation set samples were collected. Standard normalization method and first-order derivative method combined with Savitzky-Goly filter method were used for spectrum pretreatment. The optimal band ranged 9000-4150 cm-1,and main component factor was 12. RE-SULTS:The content determination method of stilbene glycoside in Shouwu pills was in line with the requirements. The correlation coefficients,the root-mean-square error of calibration,the root-mean-square error of predication and the root-mean-square error of cross-validation(RMSECV)of the quantitative calibration model were 0.99190,0.0201,0.0236 and 0.07629. There was no sta-tistical significance between predicted value and measured value(P>0.05). CONCLUSIONS:The method is accurate,stable and simple,and can be used for rapid determination of stilbene glycoside in Shouwu pills.
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Objective: To explore the main toxic components induced hepatotoxicity based on the spectrum-toxicity correlation analysis of processed Polygoni Multiflori Radix (PMR), and to provide reference of PMR for promoting the quality control as well as the safe clinical treatment. Methods: The UPLC-Q-TOF-MS characterized chemical information of all samples including crude and processed PMR which were black soybean steamed with high pleasure in different time, and the main components were identified after referring to the literatures. With the normal human hepatocytes (Lo2 cell line) as in vitro model and cell inhibitory rate as testing index, the simple correlation analysis and multivariate linear correlation analysis were used to screen the main components of PMR hepatotoxicity. Result: Seven main same components of crude and processed PMR were identified as trans-stilbene glycoside, gallic acid, emodin, physcion, emodin-8-O-β-D-glucoside, cis-stilbene glycoside, and catechin. And it was found that trans-stilbene glycoside, physcion, emodin-8-O-β-D-glucoside, cis-stilbene glycoside, and catechin were closely related to PMR hepatotoxicity. Furthermore, physcion and cis-stilbene glycoside were found greatly contributing to hepatotoxicity induced by PMR in the principle component regression analysis, which indicated these two might be the main toxic components. In addition, the results demonstrated apparent effects in detoxification could not be achieved until black soybean steamed with high pleasure at least 36 h. Conclusion: This study will provide data support for further rational use of PMR and deep hepatotoxicity research.
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AIM To establish an ultra-performance liquid-chromatography tandem mass spectrometry (UPLC-MS/MS) method for the simultaneous content determination of five constituents in Jiangzhi Huoxue Tablets (Polygoni multiflori Radix,Astragali Radix,Chuanxiong Rhizoma,etc.).METHODS The 50％ methanol extract of this drug was performed on a 40 ℃ thermostatic Restek UItra BiPh column (100 mm × 2.1 mm,5 μm),with the mobile phase comprising of acetonitrile (containing 0.1％ formic acid)-0.1％ formic acid flowing at 0.4 mL/min in a gradient elution manner.RESULTS Stilbene glycoside,tanshinone Ⅱ A,emodin,ferulic acid and puerarin showed good linear relationships within the ranges of 4.01-1 027,0.7-187,1.48-380,3.98-1 020 and4.285-1 097 ng/mL (r ＞0.994 0),whose average recoveries were 98.57％-101.0％ with the RSDs (n =6) of less than 4.79％.CONCLUSION This specific and sensitive method can be used for the quality control of Jiangzhi Huoxue Tablets.
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OBJECTIVE:To establish a method for the contents determination of harpagoside and stilbene glycoside in Shuang-shen xiaolong granule. METHODS:HPLC of harpagoside was performed on the column of Kromasil 100-5 C18 with mobile phase of acetonitrile-1% acetic acid solution (gradient elution) at flow rate of 1.0 ml/min,detection wavelength was 278 nm,column temperature was 25 ℃ and volume injection was 20 μl. HPLC of stilbene glycoside was performed on the column of Kromasil 100-5 C18 with mobile phase of acetonitrile-water(19∶81,V/V)at flow rate of 1.0 ml/min,etection wavelength was 320 nm,column temperature was 25 ℃ and volume injection was 10 μl. RESULTS:The linear range was 0.555 8-8.893 4 μg for harpagoside(r=0.999 9)and 0.010 6-0.340 2 mg for stilbene glycoside(r=0.999 6);RSDs of precision,stability and reproducibility tests were no more than 1.80%;recoveries were 97.30%-101.35%(RSD=1.43%,n=6) and 96.67%-100.83%(RSD=1.48%,n=6),respec-tively. CONCLUSIONS:The method is simple,accurate and reproducible,and can be used for the contents determination of harpa-goside and stilbene glycoside in Shuangshen xiaolong granule.
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Objective To investigate stilbene glycosides(TSG) and PNS concomitantly on PC12 cell survival rate of Alzheimer's disease.Methods The nerve cells that were seeded on the two culture plates were cultured for 1 day after the removal of primary culture fluid.In addition to the blank group, the model group and drug compatibility group were added 5 μl Aβ25-35 perpore to induced PC12 cell damage.To established AD cell damage model after exposure to the circumstances for 24 hours.Uniform design and factorial design were used respectively.After 1 d, using MTT method in ELISA analyzer measured the OD value of each pore, and calculating the survival rate of cells.Results The uniform design results showed that the cell survival rate was significantly linear with TSG and PNS (P＜0.05).From the equation, The higher the dose, the higher the cell survival rate.In this experimental condition, TSG and PNS respectively 50 mg/L, 200 mmol/L achieved the highest cell survival rate.2×2 factorial design experiments showed that, compared with the model group, the cell survival rate of TSG-PNS group (74.46％ ± 2.06％ vs.65.42％ ± 1.42％) increased (P＜0.05), but there was no interaction between the two groups (P=0.053).This showed that the combination of the two drugs in this dose has a protective effect on AD damage.Conclusion The compatibility of total saponins of two stilbene glucoside and three seven combination has the synergistic effect of anti AD damage.
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Objective: To explore the effects of different drying methods on the transformation of bioactive constituents, such as stilbene glycosides and anthraquinones, in Polygoni Multiflori Radix (PMR), and to provide a scientific evidence for the suitable drying method for PMR. Methods: Using crude PMR as materials, six drying methods including shade-drying, sun-drying, oven-drying, freeze-drying, microwave-drying, and far-infrared-drying were used to process the samples of PMR. HPLC was used to simultaneously determine the contents of stilbene glycosides, free and conjugated anthraquinones with different drying methods and drying time. Results: The contents of stilbene glycosides in the samples processed with the above methods were as follows in descending order: sun-drying > far-infrared-drying > freeze-drying > shade-drying > oven-drying > microwave-drying; The shade-drying and sun-drying methods were more suitable for the conjugated anthraquinones transforming into the free ones and the contents of conjugated anthraquinones in the sample processed with far-infrared-drying method were the highest. The comprehensive evaluation index obtained with principal component analysis showed that the far-infrared-drying method is significantly higher than those with other drying methods for PMR. Conclusion: Far-infrared-drying method is the suitable approach for the primary drying processing of PMR.
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Objective To screen and optimize the preparation process of Radix Ginseng Capsules.This paper compares the reflux extraction and effects of infiltration percolation extraction to the renshenshouwu capsules.Methods Based on the Orthogonal test,the content of ginsenosides Rg1 and Stilbene glycoside were measured by HPLC.The transferring rates of ginsenosides Rg1 and Stilbene glycoside (1:1 )were regarded as the index to evaluate extracting conditions ,such as the ethanol concentration (%),consumption of alcohol (times),extraction time and extraction times (times). Results The optimum extract conditions of ginsenosides Rg1 and Stilbene glycoside are:with 8 times volume of 50%concentration ethanol as the extract liquid,to refluxing extract 1 times,1.5 h every time.Conclusion The preparation method is simple,practical,high stability and provides the basis for industrial production.
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Stilbene glycoside (TSG) has been shown to have many beneficial properties. It is therefore essential to understand the absorption and metabolism of TSG in detail. We determined the recovery of TSG and its metabolites (TSG sulfate/glucuronides) in rat gastric contents, gastric mucosa, portal vein plasma, celiac arterial plasma, bile, and urine after administration of 15mg of TSG in 0.5mL physiological saline or incubation for 20min in situ in the stomach of rats. Within 20min, (64.0±9.8)% of the administered TSG disappeared from the stomach; later, TSG was recovered in both free and conjugated forms in plasma and bile, but not in urine. On the other hand, only free TSG was detected in the gastric contents and mucosa; it was also detected in the portal vein plasma as (48.1±3.5)% of the total TSG (all forms of TSG). However, the proportion of free TSG in the celiac arterial plasma and bile decreased to 4%-10%. In addition, the proportion of free TSG to total TSG in the liver microsome incubation mixture after TSG was incubated in liver microsome at 37℃ for 30min was very low [(10.6 ± 2.6)%]. These results indicate that TSG could be quickly absorbed from the rat stomach, conjugated in liver and excreted in bile. Such novel information would be helpful for the use of TSG as a beneficial natural product which may improve its proposed efficacy in preventing chronic diseases.
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Stilbene glycoside (TSG) has been shown to have many beneficial properties. It is therefore essential to understand the absorption and metabolism of TSG in detail. We determined the recovery of TSG and its metabolites (TSG sulfate/glucuronides) in rat gastric contents, gastric mucosa, portal vein plasma, celiac arterial plasma, bile, and urine after administration of 15 mg of TSG in 0.5 mL physiological saline or incubation for 20 min in situ in the stomach of rats. Within 20 min, (64.0±9.8)% of the administered TSG disappeared from the stomach) later, TSG was recovered in both free and conjugated forms in plasma and bile, but not in urine. On the other hand, only free TSG was detected in the gastric contents and mucosa; it was also detected in the portal vein plasma as (48.1±3.5)% of the total TSG (all forms of TSG). However, the proportion of free TSG in the celiac arterial plasma and bile decreased to 4%-10%. In addition, the proportion of free TSG to total TSG in the liver microsome incubation mixture after TSG was incubated in liver microsome at 37°C for 30 min was very low [(10.6 ± 2.6)%]. These results indicate that TSG could be quickly absorbed from the rat stomach, conjugated in liver and excreted in bile. Such novel information would be helpful for the use of TSG as a beneficial natural product which may improve its proposed efficacy in preventing chronic diseases.
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OBJECTIVE: To improve the quality standards of Yishen wufa oral solution. METHODS:Polygonum multiflorum, Psoraleae corylifolia and Lycium barbarum in the formulation were identified by TLC qualitatively. The content of stilbene glycoside was determined by HPLC. RESULTS: TLC identification was specific. The TLC spots were clear and well-separated. The linear range of stilbene glycoside was 0.03～0.60 ?g(r=0.999 9)with an average recovery of 102.4%(RSD=1.2%,n=6).CONCLUSION: Established method can be used for the quality control of Yishen wufa oral solution.
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Objective To study on purication of stilbene glycoside and icarrin with macroporous resin of radix polygoni multiflori praeparata cum succo glycines sotae and herba epimedii of Fufang Bushenshengjing Capsule. Methods To select the best type of resin from NKA-9, AB-8, D101, X-5 and DM130, to observe the factor of the concentration of water-extraction, the volume of resin, pH of water-extraction, the concentration and volume of ethanol. Results Using AB-8 resin, the concentration of water-extraction is 0.3 g/mL, the volume of resin (mL) to the mass of medicinal materials is 2, pH of water-extraction need't change, eluant is 70% ethanol and its volume (mL) is 5 times of the mass of medicinal materials (g). Conclusion This research can provide reference for the extraction of active component from Fufang Bushenshengjing capsule in industrial production.
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Aim To study the chemical constituents of Dryopteris sublaeta Ching et Hsu. Methods Fresh plant of Dryopteris sublaeta Ching et Hsu was extracted twice with boiling water, concentrated to small volume under reduced pressure at 50 ℃. The concentrated material was partitioned with ether, ethyl acetate, and n-butanol. The fraction of ethyl acetate extract was chromatographed over macroporous adsorption resin (Diaion HP-20) eluted with a mixture of H2O and MeOH in increasing MeOH content.Their fractions from resin were repeatedly chromatographed over Toyopearl HW-40, Sephadex LH-20 and silica gel column chromatography. The compounds were identified on the basis of their physiochemical and spectral data. Results Four compounds were obtained and identified as 3,5-dihydroxy-stilbene-3-O-neohesperidoside ( 1 ), 3,5-dihydroxy-stilbene-3-O-β-D-glucoside ( 2 ), polydotin peceid (3) and 3,5,4'-trihydroxy-bibenzyl-3-O-β-D-glucoside (4). Conclusion Compound 1 is a new compound, the others were isolated from Dryopteris for the first time.
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OBJECTIVE:To establish an HPLC method for the determination of Stillbene glycoside (THPG) in Zhipingk-ang misture.METHODS:The chromatographic column was Hypersil C18(250 mm?4.6 mm,5 ?m).The mobile phase consisted of acetonitribl-water(25∶75) at a flow rate of 1.0 mL?min-1 with detection wavelength set at 320 nm. RESULTS: There was a good linear relationship within the concentration range of 0.5～8.0 ?g for THPG. The average recovery of THPG was 100.03%(RSD=1.07, n=6).CONCLUSION: The adoptd method is accurate, reproducible and suitable for the quality con-trol of Zhipingkang misture.
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Objective To observe the effect of 2,3,5,4'-tetrahydroxy stilbene-2-?-D-glycoside (TSG)on learning and memory and free radicals metabolism in mice with dementia induced by ?-amyloid(A?).Methods The mice models were established by injection of A?1-40 into the right lateral ventricle,and the treatment group was administered with TSG for 8 weeks by gastrogavage.Morris water maze and step-through test were performed in all the mice and then the mice were killed and radioimmunoassay was used to detect the content of interleukin-6(IL-6).Results The learning and memory in model mice were worse and the cortical IL-6 content increased compared to the normal control mice.TSG improved the learning and memory of A?-induced model mice and reduced cortical IL-6 content.Conclusion TSG could improve the learning and memory of dementia mice and decrease cortical IL-6 content,indicating a promising prospect in the treatment of dementia disease such as Alzheimer's disease.