Effect mechanism research of procyanidin on gingivitis rats by regulating the PI3K/Akt/VEGF signal pathway / 中国药房

OBJECTIVE To investigate the potential mechanism of procyanidin on rats with gingivitis by regulating phosphoinositide 3-kinase （PI3K）/protein kinase B （Akt）/vascular endothelial growth factor （VEGF） signaling pathway. METHODS The rat model of gingivitis was constructed by sewing the neck of the first maxillary molar with silk thread+applying maltose on the gum+feeding with 20% sucrose solution and soft food. Forth-eight model rats were randomly divided into model group， procyanidin group （160 mg/kg）， 740Y-P group （PI3K/Akt signaling pathway activator， 0.02 mg/kg）， and procyanidin+ 740Y-P group （procyanidin 160 mg/kg+740Y-P 0.02 mg/kg）， with 12 rats in each group； another 12 rats were selected as control group； each medication group was treated with corresponding drugs intragastrically or/and intraperitoneally， once a day， for 7 consecutive days. Twenty-four hours after the last administration， the gingival index of rats was measured； the levels of interleukin- 18 （IL-18）， inducible nitric oxide synthase （iNOS） and alkaline phosphatase （ALP） in gingival crevicular fluid， as well as the levels of superoxide dismutase （SOD）， catalase （CAT） and reactive oxygen species （ROS） in gingival tissues of rats were detected； the pathological changes in gingival tissues were observed； the expression levels of PI3K/Akt/VEGF signaling pathway- related proteins in gingival tissues of rats were detected. RESULTS Compared with control group， the gingival tissues of rats in the model group had severe pathological damage，which was manifested as local tissue expansion and congestion， new capillaries， degeneration and loss of collagen fibers and disorder of arrangement， and a large number of inflammatory cell infiltration in the gingival sulcus wall. The gingival index， the levels of IL-18， iNOS， ALP in gingival crevicular fluid， the level of ROS in gingival tissues， the phosphorylations of PI3K and Akt， as well as the protein expression of VEGF in gingival tissues were significantly increased； the levels of SOD and CAT in gingival tissues of rats in model group were significantly decreased （P＜0.05）. Compared with model group， the pathological damage to the gingival tissues of rats in procyanidin group was reduced， and all quantitative indicators were significantly improved （P＜0.05）； 740Y-P could reverse the improvement effect of procyanidin on various indicators （P＜0.05）. CONCLUSIONS Procyanidin may alleviate gingival tissue damage， and improve gingival inflammation and oxidative stress in rats with gingivitis by inhibiting PI3K/Akt/VEGF signaling pathway.

Study on the mechanism of procyanidin mediated anti gastric cancer SNU-1 cell line by inducing the production of reactive oxygen species / 国际肿瘤学杂志

Objective:To investigate the effect and molecular mechanism of procyanidin on the proliferation, apoptosis and reactive oxygen species (ROS) level of gastric cancer cell line SNU-1 in vitro. Methods:SNU-1 cells were divided into control group and 12.5, 50.0, 200.0 μg/ml procyanidin groups. The effect of procyanidin on the proliferation of SNU-1 cells was detected by CCK-8 assay. The apoptosis level and ROS positive rate of cells were detected by flow cytometry, and 2 mmol/L glutathione was added to SNU-1 cells added with 200.0 μg/ml procyanidin to detect the apoptosis level and ROS positive rate of cells. The expression of apoptosis-related protein in cells was detected by Western blotting.Results:The results of CCK-8 experiment showed that the proliferation activities of SNU-1 cells in the control group and the 12.5, 50.0, 200.0 μg/ml procyanidin groups were 3.69±0.30, 3.29±0.41, 0.91±0.39, 0.45±0.22 respectively, with a statistically significant difference ( F=279.84, P<0.001) . Compared with the control group, the proliferation activities of SNU-1 cells in the three procyanidin groups were significantly inhibited ( P=0.006, P<0.001, P<0.001) . The results of flow cytometry showed that the early apoptosis rates of SNU-1 cells in the control group and the 12.5, 50.0, 200.0 μg/ml procyanidin groups were (0.00±0.00) %, (0.00±0.00) %, (0.09±0.07) % and (0.45±0.22) % respectively, with a statistically significant difference ( F=7.14, P=0.003) . The 50.0 and 200.0 μg/ml procyanidin groups increased significantly compared with the control group ( P=0.003, P=0.007) . The late apoptosis rates of SNU-1 cells in the four groups were (0.00±0.00) %, (0.01±0.00) %, (6.98±0.77) % and (33.32±2.78) % respectively, with a statistically significant difference ( F=654.28, P=0.003) . The 50.0 and 200.0 μg/ml procyanidin groups increased significantly compared with the control group ( P<0.001, P<0.001) . The positive rates of ROS in SNU-1 cells in the four groups were (0.02±0.01) %, (0.10±0.05) %, (1.15±0.26) % and (1.58±0.22) % respectively, with a statistically significant difference ( F=162.24, P<0.001) . The 50.0 and 200.0 μg/ml procyanidin groups increased significantly compared with the control group ( P<0.001, P<0.001) . The positive rates of ROS in SNU-1 cells in the 200.0 μg/ml procyanidin group and the glutathione intervention group were (1.25±0.63) % and (0.13±0.02) % respectively, with a statistically significant difference ( t=5.39, P=0.001) . The early apoptosis rates of the two groups were (10.56±3.24) % and (2.09±0.24) % respectively, and the late apoptosis rates were (29.65±6.01) % and (23.63±1.52) % respectively, with statistically significant differences ( t=2.61, P=0.048; t=3.97, P=0.012) . The expressions of Bcl-2 protein in SNU-1 cells in the control group and the 12.5, 50.0, 200.0 μg/ml procyanidin groups were 1.00±0.00, 0.83±0.05, 0.60±0.14 and 0.41±0.23 respectively, with a statistically significant difference ( F=10.63, P=0.004) . The 50.0 and 200.0 μg/ml procyanidin groups decreased significantly compared with the control group ( P<0.001, P<0.001) . Conclusion:Procyanidin can inhibit proliferation and promote apoptosis of gastric cancer SNU-1 cells in vitro, which may be achieved by increasing intracellular ROS levels and reducing Bcl-2 protein expression.

Antibacterial substances from leaves of Protium spruceanum (Burseraceae): in vitro and in silico evaluation

Due to the increase of bacterial resistance, the search for new antibiotics is necessary and the medicinal plants represent its most important source. The aim of this study was to evaluate the antibacterial property of extract and fractions from Protium spruceanum leaves, against pathogenic bacteria. By means of diffusion and microdilution assays, the crude extract was active against the nine bacteria tested being the hydromethanolic fraction the most active. During phytochemical procedures, procyanidin (1) and catechin (2) were identified as the main antibacterial constituents of this fraction. In silico results obtained using PASSonline tool indicated 1 and 2 as having good potential to interact with different targets of currently used antibiotics. These results no indicated potential to none DNA effect and indicated the cell wall as mainly target. Electrophoresis result supported that had no DNA damage. Cell wall damage was confirmed by propidium iodide test that showed increased membrane permeability and by cell surface deformations observed in scanning electronic microscopy. The in vitro assays together with the in silico prediction results establish the potential of P. spruceanum as source of antibacterial compounds that acts on important bacterial targets. These results contribute to the development of natural substances against pathogenic bacteria and to discovery of new antibiotics.

Screening of flavonoids in Litchi Semen and its targets of intervention in colonic adenocarcinoma by weighted gene co-expression network analysis and molecular docking / 中草药

Objective: To screen the flavonoid constituents and targets of Litchi Semen in the intervention of progression and metastasis of colon adenocarcinoma (COAD). Methods: Through DRAR-CPI and SWISS database, potential targets of 19 flavonoids in Litchi Semen were searched. COAD gene expression data and clinical characteristic data from TCGA database were downloaded. Weighted gene co-expression network analysis (WGCNA) was used to establish the gene co-expression network and identify the co-expression module of COAD. The common targets of co-expression module and potential targets were used as the compound to interfere with the drug target of COAD. Protein interaction network analysis, KEGG and GO analysis were performed by String database. The Hub gene was extracted as potential biomarkers of COAD by the cytoHubba, and the interaction network of components, targets and pathways was established by the Cytoscape. The expressions of potential biomarkers were verified by HPA database, and the compounds were docked with the potential biomarkers. Results: A total of 18 co-expression modules were identified with seven of them were correlated with clinical features, such as survival time and tumor stage. Turquoise module was related to the development and transfer of COAD. 19 flavonoids in Litchi Semen acted on 380 potential targets. 34 targets repeated with turquoise module were selected as targets. GO analysis showed that the target points were enriched in 304 GO items, including 229 biological processes, 31 cell composition and 44 molecular functions; KEGG analysis showed that target points were enriched in cancer pathways, cell cycle, and progesterone-mediated 40 pathways including oocyte cancer pathway, cell senescence, and p53 signaling pathway. The genes of CDC25A, CDC25C, CCNB2 and AURKB were screened by cytoHubba as potential biomarkers which related to the progress and transfer of COAD. Compared with para-cancerous tissues, immunohistochemistry results obtained from HPA database showed that the protein expressions of CDC25C, AURKB and CCNB2 in COAD were increased significantly (P < 0.05), which were consistent with gene expression in TCGA data set. Narirutin, procyanidin A2, phloridzin and ent-epicatechin which were well combined to CDC25A, CDC25C and AURKB through hydrogen bond were screened. Conclusion CDC25A, CDC25C, CCNB2 and AURKB were the potential biomarkers closely related to the progression and metastasis of COAD. The mechanism of intervention of flavonoids in Litchi Semen on the progression and metastasis of COAD may be related to the regulation of biological processes, such as cell division, G2/M phase transformation of cell cycle, and the regulation of cancer pathway, p53 signaling pathway and other signaling pathways. Narirutin, procyanidin A2, phloridzin, ent-epicatechin and rutin could be treated as potential inhibitors of CDC25A, CDC25C and AURKB.

Protective Effect of Procyanidin B2 on Acute Liver Injury Induced by Aflatoxin B in Rats / 生物医学与环境科学（英文）

Objective@#This study aimed to explore the protective effect of procyanidin B2 (PCB2) on acute liver injury induced by aflatoxin B (AFB ) in rats.@*Methods@#Forty Sprague Dawley rats were randomly divided into control, AFB , AFB + PCB2, and PCB2 groups. The latter two groups were administrated PCB2 intragastrically (30 mg/kg body weight) for 7 d, whereas the control and AFB groups were given the same dose of double distilled water intragastrically. On the sixth day of treatment, the AFB and AFB + PCB2 groups were intraperitoneally injected with AFB (2 mg/kg). The control and PCB2 groups were intraperitoneally administered the same dose of dimethyl sulfoxide (DMSO). On the eighth day, all rats were euthanized: serum and liver tissue were isolated for further examination. Hepatic histological features were assessed by hematoxylin and eosin-stained sections. Weight, organ coefficient (liver, spleen, and kidney), liver function (serum alanine aminotransferase, aspartate aminotransferase, alkaline phosphatase, total bilirubin, and direct bilirubin), oxidative index (catalase, glutathione, superoxide dismutase, malondialdehyde, and 8-hydroxy-2'-deoxyguanosine), inflammation factor [hepatic interleukin-6 (IL-6) mRNA expression and serum IL-6], and bcl-2/bax ratio were measured.@*Results@#AFB significantly caused hepatic histopathological damage, abnormal liver function, oxidative stress, inflammation, and bcl-2/bax ratio reduction compared with DMSO-treated controls. Our results indicate that PCB2 treatment can partially reverse the adverse liver conditions induced by AFB .@*Conclusion@#Our findings indicate that PCB2 exhibits a protective effect on acute liver injury induced by AFB .

Simultaneous liquid chromatography- tandem mass spectrometry method to quantify epicatechin and procyanidin B2 in rat plasma after oral administration of Trichilia catigua (catuaba) extract and its application to a pharmacokinetic study

Abstract Trichilia catigua A. Juss., Meliaceae, known as catuaba in Brazil, is traditionally used for the treatment of stress, sexual impotence and memory deficits. To our knowledge, there is no analytical method described in literature for simultaneous quantification of catuaba extract marker substances in biological matrices. The aim of this study was to develop and validate a bioanalytical method by LC-MS/MS to quantify epicatechin and procyanidin B2 in rat plasma after administration of standardized extract of T. catigua. Chromatographic separation was achieved with a C18 column, methanol and 0.1% aqueous formic acid at a flow rate of 0.25 ml/min. Detection was performed using electrospray ionization in negative mode. The lower limits of quantification were 5 ng/ml and 12.5 ng/ml for procyanidin B2 and epicatechin, respectively. Intra- and inter-day assays variability were less than 15%. The extraction recovery was 104% for epicatechin and 74% for procyanidin B2 using one-step liquid-liquid extraction with ethyl acetate. Epicatechin and PB2 were detected in plasma up to 300 min after oral administration of 400 mg/kg of standardized extract of T. catigua in rats. This rapid and sensitive method for the analysis of the epicatechin and procyanidin B2 in rat plasma can be applied to pharmacokinetic studies.

Effect of procyanidin B2 on inflammatory mediators of human periodontal ligament cells / 上海交通大学学报（医学版）

Objective: To investigate the effects of procyanidin B2 on the expression of inflammatory mediators in human periodontal ligament cells (hPDLCs) induced by Porphyromonas gingivalis lipopolysaccharide (LPS). Methods: hPDLCs were cultured using tissue explant method in vitro. The effect of procyanidin B2 on the cell viability of hPDLCs was detected by MTT assay. hPDLCs were stimulated by P. gingivalis LPS after treatment with procyanidin B2 for 1 h. The expressions of IL-1β, IL-6, and IL-8 mRNA and proteins were detected by real-time PCR and ELISA assay. Reactive oxygen species (ROS) in the cytoplasm was observed under fluorescence microscope. Nitric oxide (NO) in the supernatant was detected by Griess assay. Results: 100.00 µg/mL procyanidin B2 could enhance the cell viability of hPDLCs. Procyanidin B2 could inhibit the expressions of IL-1β, IL-6, and IL-8 mRNA and proteins in hPDLCs. It could also downregulate ROS and NO in hPDLCs induced by P. gingivalis LPS. Conclusion: Procyanidin B2 can play an anti-inflammatory role by inhibiting inflammatory mediators in hPDLCs induced by P. gingivalis LPS.

Protective effect of procyanidin B2 on intestinal barrier and against enteritis in a mouse model of trinitrobenzene sulphonic acid-induced colitis / 南方医科大学学报

OBJECTIVE@#To investigate the protective effect of procyanidin B2 (PCB2) on the intestinal barrier and against enteritis in mice with trinitrobenzene sulphonic acid (TNBS)-induced colitis and explore the possible mechanism.@*METHODS@#A mouse model of TNBS-induced colitis was established in male Balb/c mice aged 6-8 weeks. The successfully established mouse models were randomly divided into PCB2 treatment group (=10) and model group (=10) and were treated with daily intragastric administration of PCB2 (100 mg/kg, 0.2 mL) and 0.2 mL normal saline, respectively. After 4 weeks, the disease symptoms, intestinal inflammation, intestinal mucosal cell barrier function and the changes in PI3K/AKT signaling were evaluated using HE staining, immunofluorescence assay and Western blotting.@*RESULTS@#The disease activity index of the mice was significantly lower and the mean body weight was significantly greater in PCB2 group than in the model group in the 3rd and 4th weeks of intervention ( < 0.05). The levels of colonic inflammation and intestinal mucosal inflammatory mediators IL-1β and TNF-α were significantly lower while IL-10 was significantly higher in PCB2 group than in the model group ( < 0.05). Compared with those in the model group, the mice in PCB2 treatment group showed a significantly lower positive rate of bacterial translocation in the mesenteric lymph nodes and a lower thiocyanate-dextran permeability of the intestinal mucosa ( < 0.05). Western blotting showed that PCB2 treatment significantly increased the expressions of claudin-1 and ZO-1 ( < 0.05) and significantly lowered the expression levels of p-PI3K and p-AKT in the intestinal mucosa as compared with those in the model group ( < 0.05).@*CONCLUSIONS@#PCB2 suppresses intestinal inflammation and protects intestinal mucosal functions and structural integrity by inhibiting intestinal PI3K/AKT signaling pathway, suggesting the potential of PCB2 as a new drug for Crohn's disease.

Virtual screening of Chinese herbal monomer against EV71 3C protease / 中草药

Objective: To search for Chinese herbal monomer with the antiviral activity of enterovirus 71 (EV71) 3C protease by virtual screening. Methods: A total of 1998 monomers from antiviral Chinese herbs downloaded from TCM Database@Taiwan database were chosen as screening ligands, and EV71 3C protease was selected as a target. The binding mode of Chinese herbal monomer and EV71 3C protease was simulated through the molecular docking tools of AutoDock Vina. The monomers were sorted according to the binding free energy. The binding poses of the monomers with docking scores less than -37.673 kJ/mol were visualized by Discovery Studio 2018 Visualizer program. Results: Two monomers, namely, procyanidin B5 and 1,2,3,6-tetra-O-galloyl- β-D-glucose, were obtained with the lowest binding energies. Conclusion: The promising candidate monomers with anti-EV71 3C protease activity were screened out from traditional Chinese medicine, providing an alternative method for further development of therapeutically useful anti-EV71 agents.

Properties of decellularized rabbit carotid artery with two different cross-linked technologies: A comparative study / 中南大学学报(医学版)

Objective:To compare the properties between decellularized rabbit carotid artery with different cross-linked technologies.Methods:The decellularized rabbit carotid arteries were randomly divided into a photo-oxidation group and a procyanidins group.One group was cross-linked with photo-oxidation and the other group was cross-linked with procyanidins.The in vitro or in vivo properties of the two groups were evaluated by testing heat-shrinking temperature,max tensile strength and the max elongation or by testing tissue structure,inflammatory reaction and calcification degree.Results:The heat-shrinking temperature,max tensile strength and the max elongation were similar in the two groups (P＞0.05).The tissue structure and inflammatory reaction were also similar in the two groups.Mthough the result of Von-Kossa calcium salt stain was slightly different,the calcium content was lower in the procyanidins group than that in the photo-oxidation group (P＜0.05).Conclusion:The grafts by two cross-linked technologies show excellent mechanical capability,lower immunogenicity,good biological stability and anti-calcification ability.The procyanidins group shows a better anti-calcification property than the photo-oxidation group.

Benefits of procyanidins on gut microbiota in Bama minipigs and implications in replacing antibiotics

Several studies have reported the effect of absorption of procyanidins and their contribution to the small intestine. However, differences between dietary interventions of procyanidins and interventions via antibiotic feeding in pigs are rarely reported. Following 16S rRNA gene Illumina MiSeq sequencing, we observed that both procyanidin administration for 2 months (procyanidin-1 group) and continuous antibiotic feeding for 1 month followed by procyanidin for 1 month (procyanidin-2 group) increased the number of operational taxonomic units, as well as the Chao 1 and ACE indices, compared to those in pigs undergoing antibiotic administration for 2 months (antibiotic group). The genera Fibrobacter and Spirochaete were more abundant in the antibiotic group than in the procyanidin-1 and procyanidin-2 groups. Principal component analysis revealed clear separations among the three groups. Additionally, using the online Molecular Ecological Network Analyses pipeline, three co-occurrence networks were constructed; Lactobacillus was in a co-occurrence relationship with Trichococcus and Desulfovibrio and a co-exclusion relationship with Bacillus and Spharerochaeta. Furthermore, metabolic function analysis by phylogenetic investigation of communities by reconstruction of unobserved states demonstrated modulation of pathways involved in the metabolism of carbohydrates, amino acids, energy, and nucleotides. These data suggest that procyanidin influences the gut microbiota and the intestinal metabolic function to produce beneficial effects on metabolic homeostasis.

Ameliorative effects of type-A procyanidins polyphenols from cinnamon bark in compound 48/80-induced mast cell degranulation / 대한해부학회지

Allergic diseases are a significant health concern in developing countries. Type-A procyanidin polyphenols from cinnamon (Cinnamomum zeylanicum Blume) bark (TAPP-CZ) possesses antiasthmatic and antiallergic potential. The present study was aimed at the possible anti-allergic mechanism of TAPP-CZ against the compound 48/80 (C48/80)–induced mast cell degranulation in isolated rat peritoneal mast cells (RPMCs). TAPP-CZ (1, 3, 10, and 30 µg/ml) was incubated for 3 hours with isolated, purified RPMCs. The C48/80 (1 µg/ml) was used to induce mast cell degranulation. The mast cell viability was assessed using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay whereas histamine, β-hexosaminidase (β-HEX), and interleukin-4 (IL-4) levels were determined in RPMCs. TAPP-CZ (3, 10, and 30 µg/ml) showed significant and dose-dependent decrease in a number of degranulated cells and levels of markers (histamine, β-HEX, and IL-4) as compared with C48/80 control. In conclusion, TAPP-CZ stabilizes mast cell and cause inhibition of the allergic markers such as histamine, IL-4, and β-HEX in IgE-mediated manner. The present study supports mast cell stabilization as a possible mechanism of action of TAPP-CZ against immune respiratory disorders such as asthma and allergic rhinitis.

Advances of mechanism research on procyanidin in prevention and treatment of type 2 diabetes mellitus / 中国中药杂志

Diabetes mellitus, a chronic disease, is characterized by high blood glucose that could induce various complications. Procyanidin, a kind of polyphenol compounds existing in many plants, have shown to be effective in preventing and treating type 2 diabetes mellitus as they may lower blood glucose, moderate insulin resistance and protect islet β cells. This review focused on the research advances on the preventive and therapeutic application of procyanidin in promoting glucose absorption, protecting islet β cells, modulating intestinal microbiota and regulating diabetic complications of type 2 diabetes mellitus, which should provide useful reference for subsequent studies.

Simultaneous determination of epicatechin and procyanidin A2 markers in Litchi chinensis leaves by high-performance liquid chromatography

ABSTRACT The fruits of Litchi chinensis Sonn., Sapindaceae, are renowned for their biological activities. However, their leaves are poorly explored, although they represent an important source of vegetable raw material with biological properties as antioxidant, anti-inflammatory and antinociceptive. An HPLC method was developed and validated for the simultaneous quantification of epicatechin and procyanidin A2 in the leaf hydroethanolic extract of L. chinensis. The markers and other unidentified components were separated on a Luna Phenomenex C18 column (250 mm × 4.6 mm, 5 µm) with mobile phase composed of acetonitrile: water pH 3.0 (with sulfuric acid), in a gradient run; at 1.0 ml min-1, 30 ºC and 278 nm for detection. The method was linear over an epicatechin and procyanidin A2 concentration range of 10–100 µg ml-1. The Limit of Quantification for epicatechin and procyanidin A2 were 1.7 and 2 µg ml-1, respectively. The Relative Standard Deviation (%) values for markers (intra- and inter-day precision studies) were <4.0% and the accuracy was 100 ± 5%. The method was applied to ten samples collected in the state of Santa Catarina (Brazil), which showed 14.8–44.5 and 44.8–69.6 mg g-1 of epicatechin and procyanidin A2, respectively. The proposed method could be a valuable tool for quality assessment of L. chinenis leaves as well as their herbal derivatives.

Simultaneous determination of procyanidin B1, procyanidin B2, procyanidin B4, rutin, and quercetin in Aronia melanocarpa berry by UPLC / 中草药

Objective: The ultra performance liquid chromatography diode array detection method was used to establish the determination methods for the contents of procyanidin B1, procyanidin B2, procyanidin B4, rutin, and quercetin in Aronia melanocarpa berry. Methods: Waters Acquity UPLC® HSS T3 C18 column (100 mm × 2.1 mm, 1.8 μm) was used. The mobile phase was methanol (A)-0.03% phosphoric acid in water (B) for the gradient elution with flow rate of 0.2 mL/min. The column temperature was 35℃. The determination wavelength for procyanidin B1, procyanidin B2 and procyanidin B4 was 280 nm, and that for rutin and quercetin was 360 nm. Results: There was a linear correlation between the concentration of procyanidin B1 2.2-44.0 μg/mL (r = 0.999 2), procyanidin B2 50-1 000 μg/mL (r = 0.999 5), procyanidin B4 3.6-54.0 μg/mL (r = 0.999 2), rutin 7.4-148.0 μg/mL (r = 0.999 4), and quercetin 2.1-42.0 μg/mL (r = 0.999 7). The average recovery rates of procyanidin B1, procyanidin B2, procyanidin B4, rutin, and quercetin were 101.23% (RSD = 2.13%), 99.64% (RSD = 1.23%), 102.31% (RSD = 2.89%), 98.74% (RSD = 2.96%), and 103.53% (RSD = 2.64%), respectively. The average contents of procyanidin B1, procyanidin B2, procyanidin B4, rutin, and quercetin in Aronia melanocarpa berry were 129.78, 3 834.99, 196.02, 134.40 and 79.10 μg/g, respectively. Conclusion: The analysis method is simple, rapid, reproducible, accurate, and reliable, and can be used to identify and evaluate the quantitative determination of five constituents in A. melanocarpa berry.

Determination of catechin,epicatechin,gaIIic acid and procyanidin B2 in procy-anidins by RP-HPLC / 中国药科大学学报

To develop a RP-HPLC method for the determination of catechin (C),epicatechin (EC),gallic acid (GA)and procyanidin B2 (PCB2 )in procyanidins and compare the contents of C,EC,GA and PCB2 in procyani-dins purchased from different manufacturers.A RP-HPLC method was developed and the determination was car-ried out on a Hypersil ODS2 column (4.0 mm ×200 mm,5 μm).The mobile phase consisted of methanol and 2% acetic acid with gradient elution and the detection wave-length was at 280 nm.There was a good linear rela-tionship between concentration and the peak area in the range of 0.1-50 μg/mL (r =0.998 6)for catechin,0.1-50 μg/mL (r =0.994 5)for epicatechin,0.05-50 μg/mL (r =0.999 9)for gallic acid and 0.1-50 μg/mL (r =0.992 2)for procyanidin B2 ,respectively.The average recoveries of catechin,epicatechin,gallic acid and PCB2 were 98.36%,98.21%,89.60% and 98.47%,respectively and the RSDs were 1.39%,0.84%,2.12% and 2.46%,respectively.The method is simple,accurate,reproducible and can be used for assay of C,EC,GA,PCB2 in procyanidins.There was a great difference in the content of four substance in procyanidins purchased from dif-ferent manufacturers.

Effects of Procyanidin on Myocardial Apoptosis and Related Protein Expressions in Experimental Rats With Myocardial Ischemia Reperfusion Injury / 中国循环杂志

Objective: To observe the effects of procyanidins on myocardial apoptosis and related protein expressions of caspase-3, Bcl-2 and Bax in experimental rats with ischemia reperfusion (I/R) injury and to explore the possible mechanism. Methods: A total of 40 male SD rats were randomly assigned to 4 groups: Sham group, IR group, Low-dose procyanidin (50 mg?kg-1) group, and High-dose procyanidin (100 mg?kg-1) group. n=10 in each group and the rats were pre-treated by intra gastric drug administration once/day for 2 weeks, then left anterior descending artery (LAD) occlusion was conducted for 30 minutes followed by reperfusion for 120 minutes to establish IR model. Blood levels of CK-MB activity and myocardial infarction (MI) size were examined; protein expressions of caspase-3, Bcl-2 and Bax were determined by Western blot analysis; myocardial apoptotic index was measured by TUNEL method. Results: Compared with Sham group, IR group presented the higher CK-MB activity, enlarged MI size, increased index of apoptosis, elevated protein expressions of caspase-3 and Bax, while reduced protein expression of Bcl-2 and the ratio of Bcl-2/Bax, P<0.05. Compared with IR group, both Low-dose and High-dose procyanidin groups had the lower CK-MB activity, smaller MI size, decreased index of apoptosis, reduced protein expressions of caspase-3 and Bax, while elevated protein expression of Bcl-2 and the ratio of Bcl-2/Bax,P<0.05. Conclusion: Procyanidin could reduce myocardial apoptosis index in experimental IR rats, which might be related to decreased protein expressions of caspase-3 and Bax, increased protein expression of Bcl-2 and the ratio of Bcl-2/Bax.

Protective effect of procyanidin single active ingredient B2 on human en-dothelial progenitor cells under high glucose stimulation / 中国病理生理杂志

AIM:To study the protective effect of procyanidin single active ingredient B2 (PC-B2) on human endothelial progenitor cells ( EPCs) stimulated with high glucose.METHODS: The human EPCs were isolated from pe-ripheral blood of healthy people and identified.The EPCs were divided into control group (PBS treatment), hypertonic control group (25 mmol/L mannitol treatment), high glucose (30 mmol/L) group, and different concentrations (2, 10 and 50 mg/L) of PC-B2+30 mmol/L glucose groups.The viability of EPCs was detected by CCK-8 assay.The levels of LDH, MDA, SOD and GSH in the EPCs were detected.The changes of NO, ET-1, ICAM-1 and VCAM-1 in the EPCs cultured medium were measured by ELISA.The cell apoptotic rate and reactive oxygen species ( ROS) in the EPCs were analyzed by flow cytometry.The expression of VEGF and VEGFR-2 in the EPCs were determined by Western blot.RE-SULTS:Compared with control group, the viability of human EPCs was decreased significantly in 30 mmol/L glucose group (P cantly (P<0.05), but SOD and GSH activity and NO production were decreased significantly (P<0.05).The ROS and cell apoptotic rate were increased significantly (P<0.05).The expression of VEGF and VEGFR-2 in the EPCs were de-creased (P<0.05).When human EPCs were treated with different concentrations of PC-B2 and 30 mmol/L glucose, the viability was obviously rebounded (P<0.05), the LDH leakage, MDA content and the releases of ET-1, ICAM-1 and VCAM-1 were decreased gradually (P<0.05), the SOD, GSH activity and NO production were increased significantly (P<0.05), the ROS and cell apoptotic rate were decreased significantly (P<0.05), and the expression of VEGF and VEGFR-2 in the EPCs was increased gradually ( P <0.05 ) .CONCLUSION: PC-B2 enhances the viability of human EPCs under high glucose condition, reduces high glucose-induced oxidative damage, restores the EPCs normal function, and reduces the releases of inflammatory cytokines and apoptosis, thus playing a protective effect on human EPCs through inducing the expression of VEGF and VEGFR-2.

Screening tumor vaccine adjuvants from four kinds of Chinese herbal monomer in vitro / 中国免疫学杂志

Objective:To detect immunological effects of procyanidin ,gastrodin,baicalein and apigenin on antigen presenting cells of RAW264.7 and DC2.4 and search for new tumor vaccine adjuvant.Methods: After different concentrations of four kinds of Chinese herbal monomer and LPS were used to stimulate RAW 264.7 or DC2.4 cells for 48 hours,cells were obtained and stained with CD80,CD86,MHCⅠand MHCⅡ antibody.Then protein expression levels of CD80,CD86,MHCⅠ and MHCⅡ on RAW264.7 and DC2.4 cells were analyzed by flow cytometry.Results: Results showed that after 48 hours stimulated with different concentrations of four kinds of Chinese herbal monomer on RAW 264.7 or DC2.4 cells,the expression levels of CD80,CD86,MHCⅠ,MHCⅡ were all up-regulated in comparison with control , procyanidin and baicalein exhibited stronger immunological stimulating activity on a cellular level.Conclusion:Procyanidin and baicalein were found having potential application value in clinical treatment as tumor vaccine adju -vants.

Procyanidin inhibits multidruy resistance 1 mRNA expression via regulating microRNA-27a and microRNA-451 / 国际药学研究杂志

Objective To investigate the effect of expression of microRNA-27a(miR-27a) and microRNA-451 (miR-451) in A2780/T cells and its relativity to multidrug resistance (MDR); mRNA inhibition by procyanidin. Methods Stem-loop PCR method was performed to evaluate the expression of miR-27a and miR-451 in use of procyanidin (0-40 mmol/L) in 0-48 h in A2780/T cells. Additionally, over-expressing or interfecting microRNAs by using mimics or inhibitor of miR-27a and miR-451, the expression of MDR1 mRNA was assessed by RT-PCR in cells exposing to procyanidin. Results The expression of miR-27a and miR-451 was significant inhibited by procyanidin in both time- and concentration-dependency. Over-expressed MDR1 mRNA associated with miR- 27a or miR-451 mimics was blocked by procyanidin, whereas there was no effect on down-expressed MDR1 mRNA associated with miR-27a or miR-451 inhibitor by procyanidin. Conclusion Procyanidin inhibits MDR1 mRNA expression by inhibiting miR-27a and miR-451 expression in A2780/T cells.