Application of proanthocyanidin in the treatment and repair of deep caries / 口腔疾病防治

@#Proanthocyanidin (PA), as a kind of natural plant polyphenol, have a variety of biological functions, such as promoting remineralization, inducing collagen cross-linking, inhibiting protease activity and inhibiting bacteria. Therefore, PA could be broadly used in the clinical application of treatment and repair of deep caries in the future; for example, PA could promote dentin remineralization, improve resin-dentin bonding durability and improve the dentin acid erosion effect. This application potential of PA arises from several features, firstly, PA can not only promote dentin remineralization on its own or with other remineralizers but also exhibits antibacterial effects, which can inhibit acid production while reducing the formation of cariogenic pathogens and their biofilms. Based on the above features, PA can reduce the incidence of caries disease; thus, PA improves deep caries and long-term effects after treatment. In addition, PA added to adhesives or etch agents can improve the etching and bonding effect of dentin by inducing collagen cross-linking and inhibiting protease activity, thus achieving the ultimate goal of improving the bonding performance of deep caries. This paper summarizes recent progress of research on PA for the treatment and repair of deep caries, including the promotion of dentin remineralization and antibacterial activity as well as the improvement in dentin bonding and acid etching effect, to provide a more comprehensive reference for treating and restoring deep caries in clinical practice.

Protective effect of proanthocyanidins on neurotoxicity of benzoapyrene / 环境与职业医学

Background Benzo[a]pyrene (BaP) is neurotoxic and can cause neuronal damage by oxidative stress. Proanthocyanidin (PC) has antioxidant activity, and its mechanism may related to nuclear factor-erythroid 2-related factor 2 (Nrf2)-heme oxygenase-1 (HO-1) signaling pathway. Objective To explore potential protective effect of PC on hippocampal neuron injury induced by BaP oxidative stress. Methods Hippocampal neurons of neonatal SD rats delivered within 24 h were isolated and cultured, and cell activity was detected by cell counting kit-8 (CCK-8) method. According to the pre-experimental results, a control group and three BaP groups of 10, 20 and 40 µmol·L−1 were set up for Stage I experiment. The length of neurites and number of branches of hippocampal neurons in each group were observed by immunofluorescence method. Reactive oxygen species (ROS) fluorescence probe method was used to measure ROS levels in cells. Real-time quantitative fluorescent polymerase chain reaction (qRT-PCR) and Western blotting were used to detect the mRNA and protein expression of Nrf2, Kelch-like epichlorohydrin associated protein-1 (Keap1), HO-1, B-cell lymphoma-2 (Bcl-2), and Bcl-2-associated X (Bax) in hippocampal neurons of each group, respectively. According to the results of Stage I experiment, three group were set up, including control group, BaP alone treatment group (BaP 20 µmol·L−1), and PC intervention group (BaP 20 µmol·L−1 + PC 2.5 µg·mL−1) for Stage II experiment, with the same protocol as Stage I. Results For Stage I experiment, compared with the control group, the 10, 20, and 40 µmol·L−1 BaP groups showed gradually shortened length of neurites [(177.60±3.49), (142.40±6.52), and (100.50±19.40) µm] (P<0.05) and decreased number of branches (8.00±1.00, 6.33±1.53, 4.33± 0.58) of hippocampal neurons (P<0.05); increased ROS production (2.38±0.33, 8.08±0.26, 9.86±0.19) (P<0.05); the qRT-PCR results showed that the mRNA expression levels of Nrf2 (0.35±0.03, 0.25±0.01, 0.13±0.03), Keap1 (0.70±0.01, 0.47±0.03, 0.15±0.02), HO-1 (0.77±0.02, 0.60±0.02, 0.32±0.01), and Bcl-2 (0.65±0.03, 0.47±0.02, 0.18±0.02) gradually decreased, and the mRNA expression level of Bax gradually increased (1.24±0.01, 2.25±0.15, 4.98±0.30) (P<0.05); the Western blotting results showed that the protein expression trends of Nrf2, Keap1, HO-1, Bcl-2, and Bax were consistent with the mRNA results. For Stage II experiment, compared with the BaP alone treatment group, the length of neurites in the PC intervention group became longer, (149.90±3.01) μm vs (202.00±4.45) μm (P<0.05), the number of branches increased, (4.67±0.58) vs (8.33±0.58) (P<0.05); the ROS production reduced, (10.81±0.63) vs (7.31±0.70) (P<0.05); the mRNA expression levels of Nrf2, Keap1, HO-1, and Bcl-2 increased (P<0.05), and the mRNA expression levels of Bax decreased (P<0.05); the Nrf2, Keap1, HO-1, and Bcl-2 protein expression levels increased (P<0.05), and Bax protein expression level decreased (P<0.05). Conclusion PC may exert neuroprotective effects by activating the Nrf2-HO-1 signaling pathway, inhibiting BaP-induced oxidative stress in neuronal cells, and reducing cytotoxicity.

CONTENIDO DE POLIFENOLES, CAPACIDAD ANTIOXIDANTE Y TOXICIDAD DE Solanum ferrugineum (SOLANACEAE) CON POTENCIAL MEDICINAL / Polyphenol content, antioxidant capacity and toxicity of Solanum ferrugineum (Solanaceae) with medicinal potential

RESUMEN Solanum ferruginium es una planta que crece en lugares perturbados como maleza, es de uso medicinal poco utilizada que presenta un gran potencial como fuente de antioxidantes debido a su alto contenido en polifenoles. Debido a esto se analizó el contenido de polifenoles, flavonoides, proantocianidinas y capacidad antioxidante in vitro (ensayo de captación de radicales DPPH) en hojas, tallos y planta completa de tres localidades (Las agujas, Parque el Nabo y Bosque la Primavera) de Zapopan, Jalisco, así como su toxicidad subcronica en hojas. En general se observó diferencia significativa (p≤ 0,05), en las muestras de las tres localidades, las hojas presentaron el mayor contenido de polifenoles (15,3±0,7 a 22±0,4 mg expresado como equivalente de ácido gálico (EAG/g) en muestras de Parque el Nabo, flavonoides (7,8±0,3 a 13,3±0,3 mg EC/g) y proantocianidinas (3,4±0,1 a 4,2±0,05 mg expresado como equivalente de catequina (EC/g) en el Bosque la Primavera. La capacidad antioxidante fue similar en todas las muestras, con valores de 8,3 a 17 μg/mL de concentración media inhibitora (CI50). En la prueba toxicológica, los ratones no mostraron signos de toxicidad a ninguna dosis por efecto de la administración de la planta en estudio, por lo que la dosis letal media (DL50) es > 15 000 mg/kg de peso corporal. El contenido de polifenoles y actividad antioxidante en S. ferruginium sobre todo en hojas indican un alto potencial con propiedades farmacológicas además de su inocuidad, por lo que es importante realizar estudios de sus compuestos fenólicos individuales, antes de ser utilizada en farmacología.

ABSTRACT Solanum ferrugineum is a plant that grows like a weed. It is a new medicinal plant with great potential as an antioxidant source due to its high polyphenol content. Because of this, polyphenol, flavonoid, proanthocyanidin, and antioxidant capacity in vitro (radical scavenging test by DPPH) were analyzed in leaves, stems, and whole plants from three localities (Las agujas, Nabo Park, and La Primavera Forest) as well as the subchronic toxicity evaluation in leaves. In general, there was a significant difference (p≤ 0.05) in all samples from the three localities. The leaves showed the highest polyphenol content (15.3±0.7 to 22±0.4 mg AGE/g) in samples from the Nabo Park, flavonoid (7.8±0.3 to 13.3±0,3 mg CE/g) and proanthocyanidins (3.4±0.1 to 4.2±0.05 mg CE/g) in La Primavera Forest. Antioxidant capacity was similar across all samples, showing values of 8.3 to 17 μg/mL of IC50. During the toxicology assay, animal specimens showed no signs of toxicity to the doses resulting from the administration of the plant under study so that LD50 > 15 000 mg/kg Bodyweight. The polyphenol content and antioxidant capacity obtained from S. ferruginium leaves, together with its safety, indicate a high pharmacological potential of this plant. Therefore, it is important to carry out studies of its phenolic compounds before being used in pharmacology.

Comparative analysis on immunity of volleyball players before and after taking grape procyanidins sports supplement / Análise comparativa da função de voleibol em suplemento extra-corpóreo para vegetarianos desportivos / Análisis comparativo de la función de inmunodeficiencia del voleibol utilizado por el suplemento deportivo vitro vegetariano

ABSTRACT Grape proanthocyanidin is a good health product, without side effects and excellent biological activity, but research in the field of sports tonic is still relatively slow. Currently, the technology of preparation and extraction of grape proanthocyanidins is relatively mature. This fact laid the groundwork for sports tonic proanthocyanidin research. This study first described the biological structure of proanthocyanidin in grapes, and built the immune system of volleyball players before and after taking proanthocyanidin sports supplements. He then analyzed the factors that influence immunity. The results show that the primary index subsystem is consistent with the total system in each phase, but there are still few differences over time, which can be divided into four phases: development, recession, recovery and stability; at the level of scientific training it is reasonable. Male and female athletes take exercise supplements containing proanthocyanidin at each level of training. Regarding humoral immunity and cellular immunity, there was no adverse reaction. This study may offer some reference value for other athletes before and after taking proanthocyanidin as a sports supplement.

RESUMO A proantocianidina da uva é um produto bom para a saúde, sem efeitos colaterais e excelente atividade biológica, mas a pesquisa no campo do tônico esportivo ainda é relativamente lenta. Atualmente, a tecnologia de preparação e extração das proantocianidinas de uva está relativamente madura. Este fato lançou as bases para a investigação da proantocianidina desportiva tónica. Este estudo descreveu, em primeiro lugar, a estrutura biológica da proantocianidina das uvas, e construiu o sistema imunitário dos jogadores de voleibol antes e depois de tomar suplementos desportivos de proantocianidina. Em seguida analisou os fatores que influenciam a imunidade. Os resultados mostram que o subsistema de índice primário é coerente com o sistema total em cada fase, mas ainda há poucas diferenças no tempo, que podem ser divididas em quatro fases: desenvolvimento, recessão, recuperação e estabilidade; no plano de formação científico e razoável. Os atletas do sexo masculino e feminino tomam suplementos de exercício contendo proantocianidina em cada estágio de treinamento. Com respeito à imunidade humoral e à imunidade celular não houve reação adversa. Este estudo pode oferecer algum valor de referência para outros atletas antes e depois de tomar proantocianidina como suplemento desportivo.

RESUMEN La proantocianidina de la uva es un producto bueno para la salud, sin efectos colaterales y excelente actividad biológica, pero la investigación en el campo del tónico deportivo aun es relativamente lenta. Actualmente, la tecnología de preparación y extracción de las proantocianidinas de uva está relativamente madura. Este hecho lanzó las bases para la investigación de la proantocianidina deportiva tónica. Este estudio describió, en primer lugar, la estructura biológica de la proantocianidina de las uvas, y construyó el sistema inmunitario de los jugadores de voleibol antes y después de tomar suplementos deportivos de proantocianidina. Enseguida analizó los factores que influencian la inmunidad. Los resultados muestran que el subsistema de índice primario es coherente con el sistema total en cada fase, pero aun hay pocas diferencias en el tiempo, que pueden ser divididas en cuatro fases: desarrollo, recesión, recuperación y estabilidad; en el plano de la formación científica es razonable. Los atletas del sexo masculino y femenino toman suplementos de ejercicio conteniendo proantocianidina en cada nivel de entrenamiento. Con respecto a la inmunidad humoral y a la inmunidad celular no hubo reacción adversa. Este estudio puede ofrecer algún valor de referencia para otros atletas antes y después de tomar proantocianidina como suplemento deportivo.

Chemical characterization of the antioxidant, antihyperglycemic and antihypertensive capacities of red rice (Oryza sativa L. ) whole flour / Caracteristicas físico-químicas, capacidad antioxidante, anti-hiperlucémica y antihipertensiva de la harina de arroz rojo (Oryza sativa L. )

Oryza sativa L. rice has large amounts of proteins and minerals, besides presenting several pigmented varieties. Red rice is distinguishable due to its great nutritional value compared to the regular white variety. Its red pericarp pigmentation is due to the bioactive compounds that are responsible for its health benefits. The objective of this study was to evaluate the physical-chemical characterization, antioxidant, antihyperglycemic and antihypertensive capacity of flours of three different red rice cultures (Rubi, Virgínia and Pequeno). All samples presented specific levels of carbohydrates for cereals with low fat content and excellent levels of protein and resistant starch. In addition, the samples had a high antioxidant, antihyperglycemic and antihypertensive capacity. Antihyperglycemic capacities were measured as percent inhibition for amylase (56.7-76.5%) and glycosidase (81.0-76.6%), respectively, and antihypertensive capacity as the percentage inhibition of the angiotensin converting enzyme (38.4-34.7%). In addition, Pequeno flour presented the best results for antioxidant and antihyperglycemic capacity in comparison to the two flours tested. Thus, all red rice flours can be a source of functional compounds when added to food.

El arroz integral (Oryza sativa L.) posee importantes cantidades de proteínas, vitaminas, minerales y fitoquímicos. El arroz rojo se destaca por su gran valor nutricional. La pigmentación roja del pericarpio está asociado al contenido de compuestos bioactivos, que están directamente relacionados a los beneficios de salud humana. Teniendo en cuenta lo antes expuesto se propuso evaluar las caracteristicas físico-químicas, capacidad antioxidante, anti-hiperglucémica y antihipertensiva de las harinas de tres diferentes cultivos de arroz rojo (Rubí, Virginia y Pequeño). Todas las muestras presentaron niveles específicos de carbohidratos para cereales con bajo contenido de grasa y altos contenidos de proteína y almidón resistente. Además, las muestras presentaron una alta capacidad antioxidante, anti-hiperglucémica y antihipertensiva. La capacidad anti-hiperglicémica se midió en porcentaje de inhibidores de α-amilasa (56.7-76.5%) y α-glucosidasa (81.0-76.6%), respectivamente; y capacidad antihipertensiva como el porcentaje de inhibición de la enzima convertidora de la angiotensina (38.4-34.7%). El cultivar Pequeño presentó mayor capacidad antioxidante y anti-hiperglucémica en comparación a los demás cultivares. Así, todas las harinas de arroz rojo pueden ser vehículos de compuestos funcionales en los alimentos.

Evaluation the Effect of Different Antioxidants Applied After Bleaching on Teeth Color Stability / Avaliação do efeito de diferentes antioxidantes aplicados após o clareamento na estabilidade da cor dos dentes

Objective: Vital bleaching is a popular treatment option for discolored teeth; but at post-treatment stage, loss of adhesion is highly reported. Literature focused on antioxidant application for the answer of this issue. The aim of this study was to compare the effects of six different antioxidants on color stability of bleached teeth. Material and Methods: This study included total of 84 extracted intact non-carious lower incisors. 35% hydrogen peroxide was applied on the labial surfaces of specimens in accordance with manufacturer's instructions. The bleached teeth were divided into 7 groups. No antioxidants were applied to the control group. For the experimental groups, the following antioxidants were applied for 10 minutes each: 5% proanthocyanidin, 5% sodium ascorbate, 5% lycopene, %5 green tea, %5 white tea and %5 α-tocopherol. CIE L*, a* and b* values of the teeth were measured by a spectrophotometer. One-way ANOVA was used to determine the differences among the groups. Multiple comparisons were examined with Tukey HSD. Results: The one-way ANOVA test revealed a statistically significant difference between the groups (p < 0.005). Highest color change was observed in lycopene group and the lowest in green tea group. Conclusion: Proanthocyanidin, white tea and green tea could be considered as post-bleaching antioxidant alternatives based on their herbal nature. (AU)

Objetivo: O clareamento vital é uma opção popular de tratamento para dentes descoloridos, mas na fase pós-tratamento, a perda de adesão é altamente relatada. A literatura enfocou a aplicação de antioxidantes para a resposta desta questão. O objetivo deste estudo foi comparar os efeitos de seis diferentes antioxidantes na estabilidade da cor de dentes clareados. Material e Métodos: Este estudo incluiu um total de 84 incisivos inferiores extraídos, intactos e não cariados. Peróxido de hidrogênio a 35% foi aplicado nas superfícies labiais dos espécimes de acordo com as instruções do fabricante. Os dentes clareados foram divididos em 7 grupos. Nenhum antioxidante foi aplicado ao grupo controle. Para os grupos experimentais, os seguintes antioxidantes foram aplicados por 10 minutos cada: proantocianidina a 5%, ascorbato de sódio a 5%, licopeno a 5%, chá verde a 5%, chá branco a 5% e α-tocoferol a 5%. Os valores CIE L *, a * e b * dos dentes foram medidos por um espectrofotômetro. ANOVA um fator foi usada para determinar as diferenças entre os grupos. As comparações múltiplas foram examinadas com Tukey HSD. Resultados: O teste ANOVA revelou uma diferença estatisticamente significativa entre os grupos (p <0,005). A maior mudança de cor foi observada no grupo do licopeno e a menor no grupo do chá verde. Conclusão: Proantocianidina, chá branco e chá verde podem ser considerados como alternativas antioxidantes pós-clareamento com base em sua natureza fitoterápica. (AU)

Effect of feeding grape seed Proanthocyanidin extract on production performance, metabolic and anti-oxidative status of dairy cattle / Efeito da alimentação do extrato de semente de uva Proanthocyanidin sobre o desempenho da produção e o status metabólico e antioxidante de gado leiteiro

Aims to investigate the effects of grape seed proanthocyanidin extract (GSPE) on production performance, metabolism, and anti-oxidative status of Holstein dairy cattle in early lactation. Forty-eight multiparous Holstein dairy cattle were assigned to four groups (CON, G20, G40 and G80) and supplied with 0, 20, 40, and 80mg GSPE/kg of body weight/day. G20 significantly increased milk yield compared with other groups. Milk protein and non-fat-solids were increased in G20, G40 and G80 groups compared with the control group only at the 7th day during the experiment. No significant difference was observed in milk fat and somatic cell count, nor on parameters of energy metabolism in blood, liver function and kidney function between the four groups. There was no significant difference in glutathione peroxidase, superoxide dismutase, total antioxidant capacity, and hydrogen peroxide between the groups; but the malondialdehyde content of G20 significantly increased at day 14 in comparison with CON, and tended to increase at the 28th day. In conclusion, feeding 20mg GSPE/kg of body weight/day was associated with a significant increase in milk yield without detrimental effects on liver or kidney function and with substantial energy metabolism and antioxidant parameters improvement in early lactation dairy cattle.(AU)

O presente trabalho visa investigar os efeitos do extrato de semente de uva Proanthocyanidin (GSPE) sobre o desempenho da produção, o metabolismo e o status antioxidante de gado leiteiro Holstein em lactação precoce. Quarenta e oito vacas leiteiras multíparas Holstein foram divididas em quatro grupos (CON, G20, G40 e G80) e receberam 0, 20, 40 e 80mg de GSPE/kg de peso corporal/dia, respectivamente. O G20 aumentou significativamente o rendimento do leite em comparação com os outros grupos. A proteína e os sólidos não gordurosos do leite foram aumentados nos grupos G20, G40 e G80 somente no sétimo dia durante a experiência. Não foi observada diferença significativa na gordura do leite e na contagem de células somáticas, bem como nos parâmetros de metabolismo energético no sangue, na função hepática e na função renal entre os grupos em relação ao grupo controle. Não houve diferença significativa na glutationa peroxidase, na dimutase de superóxido, na capacidade antioxidante total e no peróxido de hidrogênio entre os grupos, mas o conteúdo malondialdeído do G20 aumentou significativamente no dia 14 em comparação com o CON, e tendia a aumentar no dia 28. Em conclusão, a alimentação de 20mg de GSPE/kg de peso corporal/dia foi associada a um aumento significativo no rendimento do leite, sem efeitos nocivos sobre a função hepática ou a renal, com o metabolismo de energia substancial e a melhoria dos parâmetros antioxidantes de gado leiteiro no início da lactação.(AU)

Study on purification of total flavonoids from Litchi Semen by macroporous resin / 中草药

Objective To screen the macroporous adsorption resin suitable for the separation and purification of total flavonoids from Litchi Semen, and the purification process parameters were established to prepare the total flavonoids of Litchi Semen in accordance with the requirements of effective parts of Chinese materia medica, which laid the foundation for the development of the total flavonoids of Litchi Semen into five new Chinese medicines. Methods The macroporous adsorption resin for purifying the total flavonoids of Litchi Semen by static adsorption-elution test was used. Based on the single factor test, the comprehensive score of adsorption rate was used as the index to investigate the volume fraction of ethanol, the mass concentration of the sample, and the sample solution pH, diameter to height ratio, upper column volume, upper column flow rate, eluent concentration, eluent volume and elution flow rate on the purification process, and determine the optimal purification process parameters. Results The best process condition for separating the total flavonoids of Litchi Semen by AB-8 macroporous adsorption resin were as follows: the mass ratio of resin to medicinal material was 3:1, the concentration of the upper column sample solution was 4—6 mg/mL, sample flow rate was 1 mL/min, and the upper column volume was 2 BV, diameter to height ratio was 1∶12, pH of the sample solution was 2, first impurity removal by 20% ethanol 3 BV, and using 60% ethanol 3 BV for elution, elution flow rate was 4 mL/min. Conclusion AB-8 macroporous resin can be used to purify the total flavonoids of Litchi Semen under the established technological conditions. The mass fraction of total flavonoids in Litchi Semen increased from 29.22% to an average of 67.37%, and the solid content decreased from 1.25 g to 0.40 g. It indicates that the established purification process is stable and feasible, and can be used as a purification process condition for total flavonoids of Litchi Semen.

Isolation and identification of chemical constituents from seeds of Litchi chinensis / 中草药

Objective: To study the chemical constituents from Litchi Semen. Methods: Different column chromatographic techniques were used to separate and purify the chemical constituents and their structures were elucidated by spectral analysis. Results: A total of 15 compounds were isolated and identified as 5-p-trans-coumaroylquinic acid (1), 3-O-trans- coumaroylquinic acid (2), phlorizin (3), naringin (4), rutin (5), naringenin-7-O-rutinoside (6), kaempferol 3-O-(6-O-caffeoyl)-β-D- glucopyranosyl-(1→2)-α-L-rhamnopyranosyl-7-O-α-L-rhamnopyranoside (7), proanthocyanidin A-2 (8), p-hydroxybenzoic acid (9), protocatechuic aldehyde (10), p-hydroxybenzaldehyde (11), protocatechuic acid (12), Z-p-hydroxy-cinnamic acid (13), E-p-hydroxy-cinnamic acid (14), and 3,6-dihydroxy-5,11-epoxy-7Z-megastigmaen-9-one (15). Conclusion: Compounds 1, 2, 7, 13-15 are isolated from the genus Litchi for the first time, and they are also isolated from this plant for the first time.

Grape seed proanthocyanidin extract inhibits platelet-derived growth factor induced proliferation of airway smooth muscle cells / 中国小儿急救医学

Objective To explore the effect of grape seed proanthocyanidin extract(GSPE) on the proliferation of airway smooth muscle cells( ASMCs) induced by platelet-derived growth factor( PDGF) and the underlying molecular mechanism. Methods ASMCs of primary rat were cultured. MTT and flow cytom-etry were used to detect the cell proliferation activity and cell cycle distribution of ASMCs which were treated with PDGF and GSPE respectively. The expression levels of cyclin D1,extracellular regulated protein kinases ( ERK)1/2,p-ERK1/2 and β-actin protein in each group ASMCs were analyzed using western blotting assay after ERK1/2 inhibitor PD98059 intervention. Results Compared with control group,cell proliferative activ-ity,S phase fraction and the expression of cyclin D1 and p-ERK1/2 protein increased in PDGF induced group (P<0. 05). These effects induced by PDGF could be reversed by GSPE. PD98059 also could block PDGF induced higher expression of p-ERK1/2 and cyclin D1 proteins in rat ASMCs. Conclusion GSPE can inhib-it PDGF induced cell proliferation and via ERK1/2 signaling pathway in rat ASMCs,which provide a new way for treatment of bronchial asthma.

Grape seed proanthocyanidin extract ameliorates murine autoimmune arthritis through regulation of TLR4/MyD88/NF-κB signaling pathway

BACKGROUND/AIMS: Grape seed proanthocyanidin extract (GSPE) has been reported to have a beneficial effect on regulating inf lammation. However, the anti-inflammatory mechanism of GSPE remains unclear. The aim of this study was to verify the influence of GSPE on the Toll-like receptor 4 (TLR4)-mediated signaling pathway in the regulation of murine autoimmune arthritis. METHODS: Collagen-induced arthritis (CIA) was induced in dilute brown non-agouti (DBA)/1J mice. The mice were treated with GSPE (0 or 100 mg/kg) intraperitoneally. The severity of arthritis was assessed clinically, biochemically, and histologically. Immunostaining for TLR4 was performed. The expressions of TLR4 and downstream signaling molecules were analyzed by Western blot. The effect of GSPE on lipopolysaccharide (LPS)-induced TLR4 activation was also evaluated using RAW264.7 cells and fibroblast-like synoviocytes (FLSs) from patients with rheumatoid arthritis and from those with osteoarthritis. RESULTS: GSPE attenuated the clinical severity of arthritis and decreased histological damage. GSPE treatment reduced the number of TLR4-stained cells in the synovium of mice with CIA. GSPE also downregulated the expression of TLR4, myeloid differentiation factor 88 (MyD88) and phosphorylated IκBα synovial protein in CIA mice. Concurrently, GSPE inhibited the nuclear translocation of nuclear factor-κB (NF-κB) subunits (p65 and p50). LPS-induced TLR4 activation was suppressed by GSPE in human FLS as well as in murine macrophages in vitro. CONCLUSIONS: Our results demonstrated that GSPE ameliorated CIA by regulating the TLR4-MyD88-NF-κB signaling pathway.

Influence of light on gene expression of key synthesis enzyme genes FtANR and FtLAR about proanthocyanidin in seeds of homologous plant of food and medicine Fagopyrum tataricum / 中国中药杂志

Tartary buckwheat Fagopyrum tataricum is an important medicinal and functional herb due to its rich content of flavonoids in the seeds. F.tataricum exhibited good functions for free radicals scavenging， anti-oxidation， anti-aging activities. Although much genetic knowledge of the synthesis， regulation， accumulation of rutin， the genetic basis of proanthocyanidins(PAs) in tartary buckwheat and their related gene expression changes under different lights(blue， red， far red， ultraviolet light) remain largely unexplored. In this study， we cloned one anthocyanidin reductase gene(ANR) and two leucocyanidin reductase gene(LAR) named FtANR，FtLAR1，FtLAR3 involved in formation of(+)-catechin and(-)-epicatechin precusor proanthocyanidin by digging out F. tataricum seed transcriptome data. The expression data showed that the opposite influence of red light on these gene transcript level compared to others lights. The expression levels of FtANR and FtLAR1 decreased and FtLAR3 appeared increment after exposed in the red light， while the expression levels of those genes appeared opposite result after exposed in the blue and far red light.

Effect of grape seed proanthocyanidins on phenotype marker protein of renal cells in db/db mice / 临床与实验病理学杂志

Purpose To investigate the effect of grape seed proanthocyanidins on the phenotype-transforming marker protein expression of db/db renal cells in mice model of type 2 diabetes,and to explore the protective mechanism of grape seed extract on diabetic renal injury in db/db mice.Methods Male db/db diabetic mice were randomly divided into two groups:diabetic group (db/db group) and diabetic + grape seed proanthocyanidin extract group (db/db + GSPE).The same week-old male db/m mice was used as normal controls (db/m) and grape seed proanthocyanidin extract gavage treatment group (db/m +grape seed proanthocyanidin extract group,db/m + GSPE).The mice of db/db + GSPE group and db/m + GSPE group were administered daily with grape seed proanthocyanidin extract (5mg/kg) by gavage.Results Renal tissues of db/db diabetic mice showed increased expression of α-SMA,p-p38MAPK,pERK1/2 and 8-OHdG level,and down-regulation in E-cadherin expression compared with db/m group (P ＜ 0.05).However,the alternations of α-SMA,p-p38,p-ERK1/2,E-cadherin protein levels,and 8-OHdG level,in db/db group were reversed by addition of grape seed proanthocyanidin extract (P ＜ 0.05).Conclusion Grape seed proanthocyanidin extract inhibits the epithelial to mesenchymal transition (EMT) associated protein,by decreasing ROS production,and activating p38 MAPK and ERK1/2.These findings suggest that grape seed proanthocyanidin extract provides a treatment option for diabetic nephropathy.

Effects of Grape Seed Proanthocyanidin on the Transport of Bile Salts in Colon Glandular Cell Caco-2 / 中国药房

OBJECTIVE:To study the effects of grape seed proanthocyanidin(GSP)on the transmembrane transport of sodium glycocholate (GA) and sodium taurocholate (TA) in colon glandular cell Caco-2. METHODS:Caco-2 model was used,and RP-HPLC was conducted to determine the contents of GA and TA in cell culture medium. The test was divided into GSP group, GA group,TA group,GSP+GA group and GSP+TA group,the transport volumes of transporting GA and TA from Transwell apical (AP)side to basolateral(BL)side by Caco-2 cell at 0,2,4,8 h were detected,respectively. RESULTS:The linear ranges of GA and TA were 0.05-1.2 mmol/L(R2=0.9999). With the time passing,transport volumes of GA and TA in BL site in GA group and TA group were sharply increased;while the transport volumes were obviously decreased after adding GSP,with statistical signifi-cance(P<0.01). CONCLUSIONS:GSP has inhibitory effect on the transmembrane transport of GA and TA in Caco-2 cell.

Effects of grape seed proanthocyanidin extract on airway inflammation and Treg/Th17 balance in asthmatic mice / 国际儿科学杂志

Objective To explore effect of grape seed proanthocyanidin extract (GSPE)on airway inflammation and hyperresponsiveness of ovalbumin-induced murine asthma model and the associated regulatory effect on Treg/Th17 imbalance.Methods A total of 40 mice were randomly assigned to four experimental groups:control,asthma model,low dose GSPE (40 mg/kg),and high dose GSPE (80 mg/kg).Acute asthma model was established with OVA;airway responsiveness of mice in each group was measured with a noninvasive pulmonary function instrument;lung inflammation changes were observed by pathological HE staining;Treg/Th17 cytokines levels in bronchoalveolar lavage fluid were evaluated by ELISA;the expressions of forkhead/winged helix transcription factor(Foxp3) mRNA and retinoid-related orphan receptor gammat (RORγt) mRNA in lung tissue of each group were gained by Real-time PCR method.Results GSPE inhibited ovalbumin-induced increases in airway responsiveness(P ＜ 0.05).Histological studies demonstrated that GSPE substantially inhibited OVA-induced airway inflammation in lung tissue.GSPE decreased IL-17A levels and increased IL-10 levels in bronchoalveolar lavage fluid (P ＜ 0.05).In the asthma model group,RORγt mRNA expression in lung tissue was significantly higher than that in the control group(P ＜ 0.05)and Foxp3 mRNA expression was significantly lower than that in the control group (P ＜ 0.05).In the GSPE group,RORγt mRNA expression was lower than that in asthma model group (P ＜ 0.05),however the Foxp3 mRNA expression was higher than that in asthma model group(P ＜ 0.05).Conclusion GSPE could alleviate airway hyperresponsiveness and airway inflammation of in asthmatic mice.It can modify the asthmatic mice Treg/Th17 imbalance by decreasing IL-17A and increasing IL-10 concentration at the level of cytokines;and also by increasing Foxp3 mRNA expression and inhibiting the expression of RORγt mRNA at the transcriptional level,which provide a new way for treatment of bronchial asthma.

A case control study of grape seed proanthocyandin extract on the aged patients with obstructive sleep apnea-hypopnea syndrome / 中国综合临床

Objective To observe the effect of grape seed proanthocyandin extract(GSPE) on aged patients with obstructive sleep apnea-hypopnea syndrome(OSAHS).Methods One hundred and one cases aged patients with OSAHS who were treated in the Affihated Hospital of North China University of Science and Technology from December 2012 to December 2014 were randomly divided into control group,GSPE group A and GSPE group B,36 cases of each group.The apnea hypopnea index (AHI),rapid eye movement (REM) and micro-arousal index(MAI) were observed by polysomnography (PSG);the fatigue,sleepiness of patients were conducted with fatigue severity scale (FSS) and epworth sleepiness scale (ESS);the peripheral blood malondialdehyde(MDA) levels and superoxide dismutase (SOD) level of before and after treatment were observed by enzyme-hnked immunosorbent (ELISA) method.The control group received continuously positive airways pressure (CPAP) treatment,while GSPE group A and GSPE group B received low and high dose of GSPE treatment oral besides CPAP respectively.Results Before the treatment,there were no significant differences in the terms of AHI,REM,MAI,FSS,ESS,MDA and SOD among the three groups(P＞0.05).After treatment,the scores of FSS,ESS,MAI and MDA in GSPE group B were (2.27±0.84)points,(6.20± 1.16)points,(8.42± 3.27) times/h,(69.40 ± 13.70) nmol/L respectively,lower than that of GSPE group A ((3.84 ± 1.20) points,(8.14± 1.26) points,(10.34± 3.48) times/h,(85.38 ± 12.22) nmol/L respectively) and control group((5.02 ± 1.14) points,(9.40 ± 1.14) points,(13.84 ± 4.08) times/h,(97.96 ± 13.24) nmol/L respectively),the differences were significant(P=0.000).The REM in GSPE group B was (18.28±2.54)％,higher than that of GSPE group A ((15.74 ± 4.32) ％) and control group ((12.38 ± 3.77) ％),there were significant differences among the three groups (P =0.000).While there were no significant differences on SOD levels among the three groups(P＞0.05).The rate of effectiveness in control group,GSPE group A and GSPE B were 70.5％,79.4％ and 90.9％ respectively,the rate of effectiveness in GSPE B was significant higher than GSPE group A and control group,and the difference was significant(P＜0.05).Conclusion GSPE can improve the sleep quality and weaken oxidative stress reaction,and has a good clinical effects for aged patients with OSAHS.

Effects of proanthocyanidin, a crosslinking agent, on physical and biological properties of collagen hydrogel scaffold

OBJECTIVES: The purpose of the present study was to evaluate the effects of proanthocyanidin (PAC), a crosslinking agent, on the physical properties of a collagen hydrogel and the behavior of human periodontal ligament cells (hPDLCs) cultured in the scaffold. MATERIALS AND METHODS: Viability of hPDLCs treated with PAC was measured using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. The physical properties of PAC treated collagen hydrogel scaffold were evaluated by the measurement of setting time, surface roughness, and differential scanning calorimetry (DSC). The behavior of the hPDLCs in the collagen scaffold was evaluated by cell morphology observation and cell numbers counting. RESULTS: The setting time of the collagen scaffold was shortened in the presence of PAC (p < 0.05). The surface roughness of the PAC-treated collagen was higher compared to the untreated control group (p < 0.05). The thermogram of the crosslinked collagen exhibited a higher endothermic peak compared to the uncrosslinked one. Cells in the PAC-treated collagen were observed to attach in closer proximity to one another with more cytoplasmic extensions compared to cells in the untreated control group. The number of cells cultured in the PAC-treated collagen scaffolds was significantly increased compared to the untreated control (p < 0.05). CONCLUSIONS: Our results showed that PAC enhanced the physical properties of the collagen scaffold. Furthermore, the proliferation of hPDLCs cultured in the collagen scaffold crosslinked with PAC was facilitated. Conclusively, the application of PAC to the collagen scaffold may be beneficial for engineering-based periodontal ligament regeneration in delayed replantation.

Anti-inflammatory effects of proanthocyanidin-rich red rice extract via suppression of MAPK, AP-1 and NF-κB pathways in Raw 264.7 macrophages

BACKGROUND/OBJECTIVES: Several pharmacological properties of red rice extract have been reported including anti-oxidant, anti-tumor, and reduced cancer cell invasion. This study was conducted to evaluate the anti-inflammatory effects of red rice extract on the production of inflammatory mediators in lipopolysaccharide (LPS)-induced Raw 264.7 macrophages. MATERIALS/METHODS: Pro-inflammatory cytokines including tumor necrosis factor-α and interleukin-6 were determined by ELISA and cyclooxygenase-2 and inducible nitric oxide synthase expression was evaluated using western blot analysis. In addition, the signaling pathway controlling the inflammatory cascade such as nuclear factor kappa B (NF-κB), activator proteins-1 (AP-1), and mitogen-activated protein kinase (MAPK) was determined. RESULTS: Our results showed that red rice polar extract fraction (RR-P), but not non-polar extract fraction, inhibited interleukin-6, tumor necrosis factor-α, and nitric oxide production in LPS-induced Raw 264.7 cells. RR-P also reduced the expression of inflammatory enzymes, inducible nitric oxide synthase, and cyclooxygenase-2. In addition, activation of AP-1 and NF-κB transcription factor in the nucleus was abrogated by RR-P. RR-P inhibited the phosphorylation of extracellular signaling-regulated kinase 1/2, c-Jun NH2-terminal kinase, and p38 MAPK signaling responsible for the expression of inflammatory mediators in LPS-stimulated Raw 264.7 cells. Based on chemical analysis, high amounts of proanthocyanidin and catechins were detected in the RR-P fraction. However, only proanthocyanidin reduced NF-κB and AP-1 activation in LPS-activated Raw 264.7 cells. CONCLUSION: These observations suggest that the anti-inflammatory properties of RR-P may stem from the inhibition of pro-inflammatory mediators via suppression of the AP-1, NF-κB, and MAPKs pathways.

Anti-inflammatory effects of proanthocyanidin-rich red rice extract via suppression of MAPK, AP-1 and NF-κB pathways in Raw 264.7 macrophages

BACKGROUND/OBJECTIVES: Several pharmacological properties of red rice extract have been reported including anti-oxidant, anti-tumor, and reduced cancer cell invasion. This study was conducted to evaluate the anti-inflammatory effects of red rice extract on the production of inflammatory mediators in lipopolysaccharide (LPS)-induced Raw 264.7 macrophages. MATERIALS/METHODS: Pro-inflammatory cytokines including tumor necrosis factor-α and interleukin-6 were determined by ELISA and cyclooxygenase-2 and inducible nitric oxide synthase expression was evaluated using western blot analysis. In addition, the signaling pathway controlling the inflammatory cascade such as nuclear factor kappa B (NF-κB), activator proteins-1 (AP-1), and mitogen-activated protein kinase (MAPK) was determined. RESULTS: Our results showed that red rice polar extract fraction (RR-P), but not non-polar extract fraction, inhibited interleukin-6, tumor necrosis factor-α, and nitric oxide production in LPS-induced Raw 264.7 cells. RR-P also reduced the expression of inflammatory enzymes, inducible nitric oxide synthase, and cyclooxygenase-2. In addition, activation of AP-1 and NF-κB transcription factor in the nucleus was abrogated by RR-P. RR-P inhibited the phosphorylation of extracellular signaling-regulated kinase 1/2, c-Jun NH2-terminal kinase, and p38 MAPK signaling responsible for the expression of inflammatory mediators in LPS-stimulated Raw 264.7 cells. Based on chemical analysis, high amounts of proanthocyanidin and catechins were detected in the RR-P fraction. However, only proanthocyanidin reduced NF-κB and AP-1 activation in LPS-activated Raw 264.7 cells. CONCLUSION: These observations suggest that the anti-inflammatory properties of RR-P may stem from the inhibition of pro-inflammatory mediators via suppression of the AP-1, NF-κB, and MAPKs pathways.

Redução da atividade proteolítica da dentina após curtos períodos de aplicação de proantocianidina / Reduction of the dentin proteolytic activity after short periods of application of proanthocyanidin

Introdução: Agentes promotores de ligações cruzadas têm sido investigados como inibidores da atividade enzimática da dentina, o que favoreceria a longevidade das restaurações adesivas. Objetivo: Avaliar o efeito do tratamento da dentina com proantocianidina (PA), em curtos períodos de tempo, na inibição da atividade de MMPs in situ. Material e método: Quarenta espécimes de dentina (1×1×6 mm) foram obtidos de molares hígidos e divididos em quatro grupos (n=10). Os espécimes foram condicionados com ácido fosfórico por 15 s, seguido de lavagem em água deionizada. A dentina condicionada foi tratada com: água, 5% PA por 5 s, 15 s ou 30 s. A atividade de MMP foi analisada colorimetricamente (SensoLyte®) e os dados de absorbância (412 nm) foram submetidos aos testes de ANOVA e Tukey (alfa =0,05). Resultado: Todos os períodos de tratamento foram capazes de reduzir a atividade de MMPs, sendo que os melhores resultados foram observados para a dentina tratada com PA por 15 s (63,1% redução) e 30 s (70,2%). O tratamento por 5 s foi capaz de inibir 39,9% das MMPs. Conclusão: A aplicação de PA sobre a dentina condicionada foi capaz de reduzir a atividade de MMPs mesmo em períodos de tempo extremamente curtos, como 5 s. No entanto, melhores resultados foram obtidos com os maiores períodos de tratamento.

Introduction: Collagen cross-linkers have been investigated as inhibitors of the enzymatic activity of dentin, therefore improving the longevity of adhesive restorations. Purpose: To evaluate the effect of etched dentin treatment with proanthocyanidin (PA) in short periods of time on the inhibition of dentin metalloproteinases (MMP) activity in situ. Material and method: Forty dentin specimens (1x1x6mm) were obtained from sound third molars and divided into 4 groups (n=10). The specimens were etched with 37% phosphoric acid for 15s and rinsed in deionized water. Then they were treated with the following solutions: water, 5% PA for 5s, 15s or 30s. The total MMP activity was analyzed by a colorimetric test (SensoLyte®). Absorbance data (412nm) were submitted to ANOVA and Tukey tests (alfa =0.05). Result: All treatment periods were able to reduce the total activity of MMPs. The best results were observed for dentine treated with PA for 15s (63.1% reduction) and 30s (70.2%). Treatment for 5s was capable of inhibiting only 39.9% of the total MMP activity. Conclusion: Application of PA on the etched dentin in extremely short periods of time reduced the MMPs activity of dentin, even after 5s. However, the best results were obtained for the longer periods.