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Polysaccharide is one of the functional components of the raspberry, which has various pharmacological effects such as anti-inflammatory, antioxidant, anti-fatigue, hypoglycemic and immunomodulatory. However, whether raspberry polysaccharides have protective effects on UV-induced photodamage to skin cells has not been reported. This study aims to investigate the protective effect of Raspberry Crude Polysaccharide on Ultraviolet B (UVB) -induced photodamage of human immortalized keratinocytes (H a C a T). The photodamage model of HaCaT cells was established by UVB irradiation. To evaluate the anti-UVB activity of R C P, the cell viability was determined by the CCK-8 method, and the Enzyme-linked immunosorbent assays (ELISA) and microplate method were used to measure the contents of matrix metalloproteinase, inflammatory and antioxidant factors in the photodamaged HaCaT cells. The antioxidant activity of RCP was detected by radical scavenging assays against D P P H radical (D P P H •) and ABTS radical (ABTS •
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ObjectiveTo investigate the synergistic effect of Coptidis Rhizoma crude polysaccharide （CCP） and berberine （BBR） in treating ulcerative colitis （UC） model mice. MethodThirty male BALB/c mice were randomized into five groups. Except the 6 mice in the normal group， the rest were given 5% dextran sodium sulfate in their daily drinking water to establish the UC model. After modeling， the mice were administrated with corresponding agents by gavage once daily for 4 days： BBR （100 mg·kg-1） group， BBR （100 mg·kg-1） + low-dose （22.8 mg·kg-1） CCP group， BBR （100 mg·kg-1） + high-dose （45.6 mg·kg-1） CCP group. The mice in the model group and normal group were administrated with the same volume of normal saline. At the end of the experiment， the mice were sacrificed for the collection of colon， and the expression of tight junction proteins zonula occluden-1 （ZO-1）， Claudin-1， and Occludin in colon tissue was detected by Western blot. With the normal group as the control， the disease activity index （DAI） score， colon length， colon histomorphology， and expression levels of tight junction proteins in other groups were evaluated. ResultCompared with the normal group， the modeling down-regulated the protein levels of ZO-1， Claudin-1， and Occludin （P<0.01）. Compared with the model group， BBR did not significantly change the protein level of Claudin-1 and up-regulated those of ZO-1 and occludin （P<0.01）. The expression levels of Claudin-1， ZO-1， and Occludin were up-regulated in BBR + CCP groups （P<0.01）. The expression levels of tight junction proteins in BBR + CCP groups were significantly higher than those in the BBR group （P<0.05）. ConclusionThe administration of CCP combined with BBR can effectively ameliorate intestinal mucosal barrier damage in the mice with UC.
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Objective:Aiming at the residue of Shaoyao Gancaotang， the extraction， qualitative and quantitative study of the small molecule resource components were carried out to clarify the residual small molecule chemical components in the residue and explore the ways of its resource utilization. Method:The ultra-performance liquid chromatography-quadrupole-time-of-flight tandem mass spectrometry （UPLC-QTOF-MS/MS） was used to qualitatively identify the residual small molecule substances in the dregs of Shaoyao Gancaotang. Agilent C<sub>18</sub> reversed-phase chromatographic column （3.0 mm×100 mm， 2.7 µm） was used at the flow rate of 0.4 mL·min<sup>-1</sup>， the injection volume was 5 µL， and the mobile phase was gradient eluted with 0.05% formic acid aqueous solution （A）-acetonitrile （B） （0-1 min， 14%-17.5%B； 1-3 min， 17.5%-19%B； 3-4 min， 19%-20%B； 4-5 min， 20%B； 5-6 min， 20%-21%B； 6-9 min， 21%B； 9-22 min， 21%-36%B； 22-23 min， 36%B； 23-32 min， 36%-43%B）， electrospray ionization （ESI） was employed with negative ion mode scanning and scanning range of <italic>m</italic>/<italic>z</italic> 50-1 200. A high performance liquid chromatography （HPLC） was established for the quantitative analysis of its main components with Agilent C<sub>18</sub> reversed-phase chromatographic column （4.6 mm×150 mm， 5 µm）， the detection wavelength was set at 235 nm， the flow rate was 0.8 mL·min<sup>-1</sup>， and the injection volume was 5 µL. Mobile phase was 0.05% phosphoric acid （A）-acetonitrile （B） for gradient elution （0-1 min， 14%-19%B； 1-4 min， 19%B； 4-18 min， 19%-50%B）. The content changes of main components in the residue of Shaoyao Gancaotang were compared before and after two different techniques of organic solvent extraction and enzymatic extraction. Result:A total of 16 chemical components in the residue of Shaoyao Gancaotang were qualitatively analyzed， and quantitative analysis found that there were many chemical components in the residue， among which the residues of 6 index components such as paeoniflorin and liquiritin reached more than 70% in the original decoction piece. After enzymolysis by cellulase， liquiritin in the residue could be converted into liquiritigenin. The content of crude polysaccharide in enzymatic extract of the residue was 6 times higher than that in the blank group， and the content was up to 12%. Conclusion:There are still many small molecule resource components in the residue of Shaoyao Gancaotang， which has great development potential. Organic solvents can be used to re-extract the target components in the residue， and liquiritin can be converted into liquiritigenin by biological fermentation technology， and the crude polysaccharide from the residue can be extracted by enzymatic method to develop animal feed. This study can provide reference basis and approach for reusing the residues of Shaoyao Gancaotang preparations and dispensing granules， so as to realize the high-value utilization of Shaoyao Gancaotang.
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OBJECTIVE：To screen the components with better anti-inflammatory effect from Codonopsis Radix polysaccharide and explore the anti-inflammatory mechanism. METHODS ：Using Codonopsis Radix as sample ，the crude polysaccharide （CP） was obtained by water extraction and alcohol precipitation. After CP was separated and purified by Fast Flow DEAE Sepharose ， SephadexG-150 gel column and so on ，the components CP 1-2-1 and CP 3-1-1 were obtained and then characterized. The survival rate of RAW 264.7 cell was determined by MTT method after cultured with CP ，CP1-2-1 and CP 3-1-1（0.01，0.1，1，10，100 μg/mL）for 24 and 48 h respectively. The cells were divided into blank group （blank culture medium ），model group （1 μg/mL LPS）and low-，medium- and high-concentration groups of 3 kinds of Codonopsis Radix polysaccharide （1 μg/mL LPS+25，50， 100 μg/mL CP，CP1-2-1，CP3-1-1 solution）. After cultured for 24 h，NO content in cell culture solution ，mRNA expressions of TLR4，IL-6，NF-κB and TNF-α in cells were determined. RESULTS：The sugar content of CP reached （92.20±0.73）％. CP 1-2-1 was a single neutral polysaccharide composed of fructose with a relative molecular weight of 25.8 kDa. CP 3-1-1 was an acidic heteropolysaccharide composed of arabinose ，rhamnose，galactose and galacturonic acid and so on ，with a relative molecular weight of 49.5 kDa. Cell survival rates were higher than 99％，after cultured with 3 kinds of polysaccharide for 24 and 48 h. Compared with blank group ，NO content in cell culture solution and mRNA expressions of TLR 4，IL-6，NF-κB and TNF-α in cells were increased significantly in model group （P＜0.05 or P＜ . Compared with model group ， NO content in cell 2016CFB412） culture solution of each administration group was significantly reduced，mRNA expressions of TLR 4，NF-κB，TNF-α，IL-6 in cells in 50，100 μg/mL CP group were significantly decreased ong1991@163.com （P＜0.05 or P＜0.01）；mRNA expressi ons of TLR 4，NF-κB in cells in 25 µg/mL CP 1-2-1 group and mRNA expressions of TLR 4，NF-κB，TNF-α，IL-6 in cells in 50，100 µg/mL CP 1-2-1 group were significantly decreased （P＜0.05 or P＜0.01）；mRNA expressionsof IL- 6 in cells in 25 µg/mL CP 3-1-1 group，mRNA expressions of NF-κB，TNF-α，IL-6 in cells in 50 µg/mL CP 3-1-1 group and mRNA expressions of TLR 4，NF-κB，TNF-α，IL-6 in cells in 100 µg/mL CP 3-1-1 group were significantly decreased （P＜0.05 or P＜0.01）. CONCLUSIONS ：CP，CP 1-2-1 and CP 3-1-1 all have certain anti-inflammatory activities. The mechanism may be related to inhibiting the generation and releasing of inflammatory cytokines such as TNF-α，IL-6 by inhibiting the activation of TLR 4/NF-κB pathway.
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Objective: To investigate the effects of phytohormones and lignin on the growth of Antrodia cinnamomea mycelium, and to determine the content of crude polysaccharide and crude triterpenoid and antioxidant activity of cultured products. Methods The product cultured by solid-state fermentation in petri dish was ultrasonically extracted. The phenol-sulfuric acid method was selected to determine the content of crude polysaccharide of the extract, and the vanillin-glacial acetic acid method was used to determine the content of crude triterpenoids of the extract. The half-clearing concentrations (IC50) of DPPH free radicals and ABTS free radicals were indexes for evaluating the anti-oxidant activity of the culture product. Results Sampling near the outer edge of the mycelium layer after 20 d of culture was performed by the activation method to obtain inoculated raw materials with better growth activity; The basal medium with good growth, high content of crude polysaccharide and crude triterpenoids was modified PCA medium; On the basis of this medium, the mycelia with 0.5 g/L lignin added into the medium grew fastest, and the diameter of mycelium layer increased to 1.19 times of the control group; When the concentration of IBA was 0.5 mg/L, the dry weight of mycelium was increased by 89.51% compared with the control group, and the yield of crude polysaccharide was increased by 130.57% compared with the control group, which was much higher than other groups. The content and yield of triterpenoids were also increased significantly, 61.31% higher than the control group, and crude triterpenoids yield reached 133.24 mg/L; The content of crude triterpenoids in mycelium was the highest (5.62%) when adding 0.5 g/L powder of Cinnamomum kanehirai, which was 84.26% higher than that of the control group; In addition, the addition of plant hormones and lignin to the culture medium has a certain effect on the anti-oxidant capacity of the Antrodia cinnamomea mycelium, on the whole, the experimental group had good anti-oxidant activity after adding different substances. Conclusion By adding phytohormone and lignin to the modified PCA medium, the growth of the mycelium of Antrodia cinnamomea can be effectively promoted, the content of the active ingredient can be increased, and the anti-oxidant activity can be enhanced.
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Obejctive:To study the extraction, separation, physical and chemical properties and antioxidant activity of the crude polysaccharide from Alhagi sparsifolia Shap.stem-branch.Methods: The crude polysaccharide from Alhagi sparsifolia Shap.stem-branch was extracted by ethanol subsiding method .The total sugar content was determined by phenol-sulfuric acid method and the pro-tein content was determined by coomassie brilliant blue method .The content of uronic acid was determined by carbazole sulfuric acid method and the monosaccharide composition and the relative molar ratio were determined by GC .The antioxidant activities in vitro were evaluated by determining the reducing power of polysaccharide from Alhagi sparsifolia Shap stem-branch and its removal ability to 1,1-diphenyl-2-trinitrophenylhydrazine ( DPPH) radicals and hydroxyl radicals .Results: The total sugar content of crude polysaccharide from Alhagi sparsifolia Shap.stem-branch was 73.2％, and uronic acid accounted for 27.2％of the total sugar content of crude poly-saccharide.The content of protein was 17.6％.The crude polysaccharide from Alhagi sparsifolia Shap .stem-branch was composed of Rha, Ara, Xyl, Man, Glc, Gal, GlcA and GalA, and the relative molar ratio was 1.05:1.00:1.25:0.52:3.05:1.31:0.47:4.78. The reducing power of the polysaccharide from Alhagi sparsifolia shap.stem-branch and the clearance rate on DPPH radicals and hy-droxyl radicals increased with the increase of polysaccharide concentration .Conclusion:The crude polysaccharide from Alhagi sparsi-folia Shap.stem-branch was extracted , and the physical and chemical properties and antioxidant activity in vitro were studied, which provide foundation for the further investigation and comprehensive utilization of Alhagis parsifolia Shap.stem-branch.
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OBJECTIVE:To study the effects of 3 extracts of Acanthopanax sessiliflorus fruits on the proliferation and apoptosis of human hepatocarcinoma cells SMMC-7721,and to provide reference for confirming the mechanism of anti-tumor effect. METHODS:MTT assay was adopted to investigate the effects of low-mass concentration,medium-mass concentration and high-mass concentration of ethanol extract(0.92,1.84,3.68 mg/mL),crude polysaccharide extract(0.06,0.12,0.24 mg/mL)and refined polysaccharide extract (0.04, 0.08, 0.16 mg/mL) from A. sessiliflorus fruits on the proliferation and apoptosis of SMMC-7721 cells after treated for 24,36,48 h,respectively. Flow cytometry was used to investigate the effects of 1.84 mg/mL ethanol extract,0.24 mg/mL crude polysaccharide extract and 0.16 mg/mL refined polysaccharide extract on cell cycle and cell apoptosis after treated for 24 h. The above tests were all negative control(only adding cells without drugs). RESULTS:Compared with negative control,3 extracts of A. sessiliflorus fruits could significantly inhibit the proliferation of SMMC-7721 cells (P<0.01),could significantly decrease the percentage of SMMC-7721 cells in G0/G1 and G2/M phase(P<0.01),could significantly increase the percentage of SMMC-7721 cells in S phase (P<0.01) and the apoptosis rate of SMMC-7721 cells (P<0.05);especially the effects of ethanol extract from A. sessiliflorus fruits were the most obvious. CONCLUSIONS:Three extracts of A.sessiliflorus fruits can inhibit the proliferation of human hepatocarcinoma SMMC-7721 cells,block SMMC-7721 cells in S phase and induce the apoptosis of SMMC-7721 cells.
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[Objective] To investigate the protective effects of different extraction of Agaricus blazei Murrill on HaCaT cells damaged by UVB. [Methods] The cell viability which was detected by MTT method was used to determine the impact of different dose of UVB and different extraction of Agaricus blazei Murrill on normal HaCaT cells and HaCaT cells damaged by UVB. [Results] ①The HaCaT cells viability declined after irradiation with UVB, and it was a dose-dependent manner. The cell viability was 53.23%when the dose of UVB was 20mj/cm2. ②The water extraction at the concentration of 0.2~4mg·mL-1, aqueous extraction at the concentration of 0.1~2mg·mL-1 and crude polysaccharide at the concentration of 0.2~1mg·mL-1 could promote the proliferation of HaCaT cells, while aqueous extraction and crude polysaccharide inhibited the proliferation of HaCaT cells at higher concentrations.③The water extraction at the concentration of 1~4mg·mL-1, aqueous extraction at the concentration of 1~2mg·mL-1 and crude polysaccharide at the concentration of 0.5~1mg·mL-1 could enhance the cells viability which was irradiated by 20mj/cm2 UVB (P<0.01). [Conclusion] The different extraction of Agaricus blazei Murrill all could relieve the photo-damage caused by UVB, and it has the potential of anti-photoaging.
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OBJECTIVE:To optimize the comprehensive extraction technology of crude polysaccharide and total saponins from Fuling kugua capsule. METHODS:Based on single factor test,using liquid ratio,extraction time and extraction times as response factors,the parameters of extraction technology of crude polysaccharides and total saponins from Fuling kugua capsule were opti-mized by using Box-Behnken design of response-surface method. RESULTS:The optimal extraction technology was as follows as liquid ratio of 1:12,extraction time for 1.4 h,extraction times for 2 times. Under this technology,the yield of crude polysaccha-ride was 14.44%(RSD=0.14%,n=3),and that of total saponins was 1.33%(RSD=0.75%,n=3). They were close to predict-ed value 14.69% and 1.36%,and the deviation were 1.70% and 2.21%. CONCLUSIONS:Measured value is close to predicted value,which indicates comprehensive extraction parameters optimized by response-surface methodology can be sued for the extrac-tion of crude polysaccharide and total saponins from Fuling kugua capsule.