Effects of applying procyanidins on the bond strength of bleached dentin to composite resin / 口腔疾病防治

Objective@#To study the influence of procyanidins on the bonding strength of dentin bleached by carbamide peroxide to composite resin.@*Methods @#By applying different treatments to dentin bonding interfaces, 120 human third molars were randomly divided into 12 groups (n = 10): W group (no bleaching+deionized water), Wa group (no bleaching+deionized water+aging), WT1 group (no bleaching+5% procyanidins for 1 min), WT1a group (no bleaching+5% procyanidins for 1 min+aging), WT2 group (no bleaching+5% procyanidins for 5 min), WT2a group (no bleaching+5% procyanidins for 5 min+aging), C group (carbamide peroxide+deionized water), Ca group (carbamide peroxide+deionized water+aging), CT1 group (carbamide peroxide+5% procyanidins for 1 min), CT1a group (carbamide peroxide+5% procyanidins for 1 min+aging), CT2 group (carbamide peroxide+5% procyanidins for 5 min), and CT2a group (carbamide peroxide+5% procyanidins for 5 min+aging). The bond strength to composite resin was measured by universal mechanical testing machine, microstructure and the nanoleakages were measured by scanning electron microscope immediately or after the thermal cycling aging test.@*Results@#The immediate bond strength of the bleached groups pretreated with procyanidins for 1 min (P<0.001) and 5 min (P<0.001) was higher than that of Group C, and the difference was statistically significant. Meanwhile, there was no statistically significant difference between Group CT1 and Group CT2 (P = 1.000). After the thermal cycles, the bond strength of each group declined. The differences between Group W and Group Wa (P<0.001) and Group C and Group Ca (P<0.001) were statistically significant, but no significant differences between Group CT1 and Group CT1a (P = 0.052) or Group CT2 and Group CT2a (P = 0.053) were found. The main effects of “aging” (P<0.001), “bleaching” (P<0.001) and “procyanidins” (P<0.001) and the second-order interaction effects of “bleaching * procyanidins” (P = 0.008), “bleaching * aging” (P = 0.024), and “aging * procyanidins” (P<0.001) were statistically significant. SEM observations showed that the hybrid layers in Groups C, CT1 and CT2 were not clear, and the hybrid layers in Groups Ca, CT1a and CT2a were partially destroyed and disintegrated. Under backscattering mode, it was observed that there were a large number of silver nitrate particles in the hybrid layer of Group Ca, and the residual silver ions in the hybrid layer of Groups CT1a and CT2a were decreased. @*Conclusion@# Pretreatment with 5% procyanidins for 1 min can improve the immediate bond strength of dentin bleached by carbamide peroxide to composite resin and maintain bonding durability.

In vitro impact of condensed tannins on the caecal metabolites of chickens / Impacto in vitro de taninos condensados nos metabólitos cecais de frangos de corte

Tannins are a diverse group of plant phenolic compounds. Condensed tannins (CTs) represent a major subgroup of tannins and were extracted from tilia (Tilia L.) flowers and black locust (Robinia pseudoacacia) leaves. These extracts were examined for their effects on the metabolic profile of chicken caeca. By using in vitro, a nuclear magnetic resonance (1H-NMR), which was combined with multivariate statistics, the current study was applied for the first time to investigate how three different CT compositions, procyanidins (PC) and/or prodelphinidins (PD) units influenced the metabolic end-products in caecal contents of chickens. In the presence of tannins, glutamate, leucine, lysine, pyroglutamate, phenylalanine, proline, and sarcosine were significantly decreased. CT extracts significantly influenced the fermentation, increasing the concentrations of some fatty acids such as acetate, butyrate, and propionate whereas. In contrast, lactate decreased between the treatments. This study identified the key structural features of CTs that contain either high molar proportions of PD or PC, which might be useful to improve the efficiency of feed utilization in chickens.(AU)

Taninos são um grupo diversificado de compostos fenólicos derivados de plantas. Os taninos condensados (TCs) representam o maior subgrupo de taninos extraídos das flores de tília (Tilia L) e de folhas negras (acácia-bastarda). Estes extratos foram examinados para a avaliação dos seus efeitos no perfil metabólico do ceco de frangos de corte. Com o emprego da ressonância magnética nuclear in vitro (1H-NMR) combinada com estatística multivariada, o presente trabalho foi aplicado pela primeira vez para investigar como três diferentes composições de TCs, unidades de procianidinas (PC) e/ou prodelfinidinas (PD) influenciariam os produtos metabólicos finais dos conteúdos cecais de frangos de corte. Na presença de taninos, houve um significativo decréscimo de glutamato, leucina, lisina, piroglutamato, fenilalanina, prolina e sarcosina. Os extratos de TCs influenciaram significativamente a fermentação, aumentando as concentrações de alguns ácidos graxos, tais como o acetato, butirato e propionato, enquanto em contraste, houve um decréscimo do lactato entre os tratamentos. Este trabalho identificou aspectos estruturais chave que os TCs contêm, tanto as altas proporções molares de PD como as de PC, as quais podem ser úteis para aumentar a utilização de alimentos em frangos de corte.(AU)

Mechanism of Shaoyao Decoction in treatment of ulcerative colitis based on network pharmacology and molecular docking technology / 中草药

Objective: Network pharmacology and molecular docking technology were used to study the material basis and possible mechanism of Shaoyao Decoction in treatment of ulcerative colitis (UC). Methods: TCMSP and TCMID were used to obtain the potential active components and drug targets of Shaoyao Decoction. GeneCard and OMIM were used to search disease targets. The common targets obtained by matching drug targets and disease targets were imported into String to construct a PPI network, and Cyto NCA plug-in was used to screen key targets. The network diagram was drawn by connecting the key targets with the corresponding components, so as to screen the key components. GO and KEGG enrichment analyses were performed on key targets. SYBYL-X2.0 software was used to dock the molecules of the key components with the key targets. The rat UC model was replicated in vivo. After the intervention of Shaoyao Decoction, the disease activity index was observed, the colonic pathological damage was evaluated, and the levels of TNF-α, IL-4 and CXCR4 were detected by ELISA. Results: A total of 424 potential active components were found in Shaoyao Decoction. The key components included quercetin, palmitic acid, catechin, and procyanidins, etc. Its 41 key targets for UC were mainly related to the positive regulation of transcription, the negative regulation of apoptosis process, the signal transduction, and other biological processes. The key targets played a role in treating UC through signaling pathways such as TNF, HIF-1, cancer pathway, TLR, PI3K-Akt, et al. Molecular docking results showed that key components had good binding activity with corresponding targets. Shaoyao Decoction improved colon pathological damage, down-regulated the level of TNF-α and CXCR4, and up-regulated the level of IL-4 in vivo. Conclusion: Shaoyao Decoction has the characteristics of "multi-components, multi-targets, multi-pathways" in treatment of UC, which lays a foundation for further study of its mechanism.

Inhibitory effect of foodborne procyanidins on growth of SH-SY5Y cells and its mechanism / 吉林大学学报(医学版)

Objective: To investigate the inhibitory effect of foodborne procyanidins on the growth of human neuroblastoma SH-SY5Y cells, and to elucidate its mechanism. Methods: The SH-SY5Y cells were cultured and divided into control group, 10 mg · L-1 foodborne procyanidins group, 20 mg · L-1 foodborne procyanidins group and 40 mg · L-1 foodborne procyanidins group; the medium containing different concentrations (0, 10, 20 and 40 mg · L-1) of foodborne procyanidins was added into each group. The proliferation rates of SH-SY5Y cells were measured by MTT method at 24, 48 and 72 h after the drug treatment. Flow cytometry was used to detect the cell cycle of SH-SY5Y cells at 72 h after the drug treatment, and the apoptotic rate of SH-SY5Y cells was detected by Annexin V apoptotic assay kit at 72 h after the drug treatment. Results: Compared with control group, the proliferation rates of SH-SY5Y cells at 24, 48 and 72 h in 10, 20 and 40 mg · L-1 foodborne procyanidins groups were decreased; there were significant differences at 48 and 72 hours in 20 mg · L-1 foodborne procyanidins group (P<0.05 or P<0.01); there were also significant differences at 24, 48 and 72 h in 40 mg · L-1 foodborne procyanidins group (P < 0.01). Compared with control group, the percentage of cells in G0/G1 phase in 40 mg · L-1 foodborne procyanidins group were increased (P<0.01) and the percentage of cells in G2/M phase were decreased (P<0.01). Compared with control group, the apoptotic rates of cells in 10, 20 and 40 mg · L-1 foodborne procyanidins groups were increased significantly (P<0.01). Conclusion: Foodborne procyanidins can inhibit the growth of human neuroblastoma SH-SY5Y cells, and its mechanism is mainly to block the cell cycle and induce the apoptosis.

Antineoplastic Effect of Procyanidin-rich Extract of Lafoensia Pacari in Lung Carcinoma Cells

Abstract Lafoensia pacari A. St. Hill has been used in traditional medicine as an anti-ulcerogenic and anti-inflammatory. Although there is an ethnopharmacological indication for cancer treatment, only a few studies have demonstrated its possible anticancer activity. Thus, the aims of this study were: (1) to evaluate the antineoplastic effect of L. pacari ethanolic extract (LPE) in lung carcinoma cells, (2) to determine the mode of action of LPE and (3) to identify the substances present in LPE. Human and murine lung cancer cell lines were grown in vitro and treated with different concentrations of LPE. Cell cycle and caspase-3 activity assays were performed in order to verify the mode of action. LC-ESI-MS screening was performed to detect the compounds present in LPE. LPE showed a dose-dependent cytotoxic effect, where neoplastic cells were more sensitive than non-neoplastic. The LPE induced sub-G1 cell cycle arrest in cancer cells suggesting cell death, which was confirmed as apoptosis by the activation of caspase-3. The LC-ESI-MS analysis indicated a high level of procyanidins, which could be responsible for the antineoplastic effect of LPE. Thus, we concluded that a Lafoensia pacari extract, rich in procyanidins, is cytotoxic against lung cancer cells through activation of caspase-3-dependent apoptosis.

Protective effect of procyanidins on PC12 cells exposed to Aβ25-35 / 中国病理生理杂志

AIM:To investigate the protective effect of procyanidins on the PC 12 cells exposed to Aβ25-35and the mechanisms.METHODS:Aβ25-35at 25 μmol/L was used to treat the PC12 cells for 48 h, and the PC12 cells were pretreated with procyanidins at 25,50 and 100 mg/L for 24 h.The cell vitality was measured by MTT assay.The content of reactive oxygen species(ROS)was detected by DCFH-DA staining.The change of mitochondrial membrane potential was examined by JC-10 staining.The apoptosis was analyzed by flow cytometry with Annexin V /PI double staining.The protein levels of activated caspase-3 was determined by Western blot.RESULTS:Under the exposure of the PC12 cells to Aβ25-35,procyanidins increased the cell viability,reduced intracellular ROS level, prevented mitochondrial membrane po-tential decline,attenuated the caspase-3 activation and inhibited the apoptosis of PC 12 cells(P<0.05 or P<0.01). CONCLUSION:Procyanidins have a significant protective effect on the PC 12 cells exposed to Aβ25-35.Its mechanism may be related to removing intracellular ROS induced by A β25-35, relieving the damage to the mitochondrial membrane, and thereby inhibiting cell apoptosis.

Calcium mobilizing effect of hawthorn leaf procyanidins in vascular endothelial cells / 中国中药杂志

The hawthorn leaves have the effect of activating blood, removing blood stasis, regulating qi through the veins, dissolving turbidity and lowering lipid. Procyanidinis is one of its main active components and plays an important role in regulating vasoactivity. Previous studies showed that the regulating effect of procyanidins was related to its regulation on nitric oxide secretion from vascular endothelial cells, and this effect was dependent on the extracellular calcium concentration, suggesting that the changes in intracellular calcium ion concentration in endothelial cells may play a key role in this process. However, the research on this issue is still insufficient so far. This study is aimed to observe the effect of hawthorn leaf oligomeric procyanidins (HLP) on calcium mobilization of vascular endothelial cells, and investigate the underlying mechanism. Human umbilical vein endothelial cells (HUVEC) were cultured and labeled with Fura-2. HUVEC were treated with HLP at concentrations of 6.25, 12.5, 25 and 50 mg·L⁻¹, and the intracellular calcium concentrations were measured with a living cell microscope for 30 min. HLP increased the intracellular calcium concentration of HUVEC in a concentration dependent manner; and the intracellular calcium concentrations in 25 and 50 mg·L⁻¹ HLP groups were significantly higher than that in the normal group. With the use of calcium-free incubation buffer, addition of calcium chelating agent EGTA in incubation buffer, or use of inhibitors for sodium calcium exchanger, the effect of HLP was significantly inhibited. On the other hand, the effect of HLP could also be weakened by inhibiting the calcium release from the intracellular storage. In conclusion, these results suggest that HLP can elicit calcium mobilization in vascular endothelial cells, which may be one of the mechanisms for its vascular modulatory activity; and this calcium mobilizing effect may be achieved through promoting both extracellular calcium influx and intracellular calcium release, additionally the former may be related to activating the reverse transport of Na⁺-Ca²⁺ exchangers on the cell membrane.

Effects of proanthocyanidins on cerebral edema and secondary brain injury inflammation following intracerebral hemorrhage / 重庆医学

Objective To investigate the effect of procyanidins on cerebral edema and inflammation of secondary brain injury after intracerebral hemorrhage.Methods Forty-eight C57BL6 mice were randomly divided into 4 groups:sham-operation+ saline group,sham-operation + procyanidins (100 mg/kg) group,intracerebral hemorrhage (ICH) + saline group and ICH + procyanidins (100 mg/kg) group.The Bederson method was used to detect postoperative nerve function score in mice.The brain tissue water content was measured by wet-dry weight method.The variety of morphological in brain tissue was observed by hematoxylin-eosin (HE) staining.The expression of glial fibrillary acidic protein (GFAP) and ionized calcium-binding adapter molecule 1(Iba-1) in mouse brain tissue were observed by western blot.The apoptosis of brain tissue was observed by TUNEL and western blot method.Results Compared with the ICH+saline group after 12h,the nerve function score in the ICH + proanthocyanidins group was significantly decreased (P＜0.05),and the water content of brain tissue was also significantly decreased (P＜0.05).The HE staining results showed that procyanidins decreased the number of neurocytes lesions in cerebral hemorrhage mice and reduced the amount of brain tissue hemorrhage.Western blot results showed that procyanidins promoted the expression of Bcl-2,decreased the expression of Bax,Caspase3,GFAP and Iba-1 in cerebral hemorrhage mice.The TUNEL results showed that procyanidins could inhibit the apoptosis of brain tissue neurons in mouse with intracerebral hemorrhage.Conclusion Procyanidins inhibits the production of neurotoxic substances possibly by reducing the expression of GFAP and Iba-1,and prevents and treats secondary brain injury after intracerebral hemorrhage.

Evaluation of the effect of procyanidin on drug permeability on different intestinal mucosa of rats with Ussing Chamber in vitro / 国际药学研究杂志

Objective To investigate the effect of procyanidin on permeability of rodamin 123(R123)and fluorescein sodium (FS)via different intestinal mucosa in rat. Methods The cumulative permeability and the apparent permeability cofficient(Papp)of R123 and FS via rat intestinal membranes at the procyanidin concentration of 20 mg/L was evaluated by the method of Ussing Chamber. The concentrations of R123 and FS in the receptor samples were determined by fluorospectrophotometry. Results The absorptive directed permeability of R123 across all membranes was increased by co-administration of procyanidins, whereas that of the secretive direct was decreased. Compared with control group, the secretive directed permeability of R123 was significantly decreased in colon (P<0.01). Conclusion Procyanidin could inhibit the secretion of R123 on different intestinal mucosa which might be related to the inhibition of P-gp function.

Evaluation of the effect of procyanidin on drug permeability on different intestinal mucosa of rats with Ussing Chamber in vitro / 国际药学研究杂志

Objective To investigate the effect of procyanidin on permeability of rodamin 123(R123)and fluorescein sodium (FS)via different intestinal mucosa in rat. Methods The cumulative permeability and the apparent permeability cofficient(Papp)of R123 and FS via rat intestinal membranes at the procyanidin concentration of 20 mg/L was evaluated by the method of Ussing Cham?ber. The concentrations of R123 and FS in the receptor samples were determined by fluorospectrophotometry. Results The absorptive directed permeability of R123 across all membranes was increased by co-administration of procyanidins,whereas that of the secretive direct was decreased. Compared with control group,the secretive directed permeability of R123 was significantly decreased in colon (P<0.01). Conclusion Procyanidin could inhibit the secretion of R123 on different intestinal mucosa which might be related to the inhibition of P-gp function.

Effects of Hawthorn Leaf Procyanidins on Over Expressions of ICAM-1 and E-selectin in Human Umbilical Vein Endothelial Cells Induced by TNF-α / 中国中医药信息杂志

Objective To observe the effects of hawthorn leaf procyanidins (HLP) on over expressions of ICAM-1 and E-selectin in human umbilical vein endothelial cells (HUVEC) induced by TNF-α,and clarify the mechanism of HLP's anti-inflammation effect. Methods HUVEC were cultured in vitro. MTT assay was used to detect cell viabilities. The expressions of ICAM-1 and E-selectin in HUVEC were detected by flowcytometry. Results Up to 200 mg/L, HLP showed no significant decrease in cell viabilities; the expression levels of ICAM-1 and E-selectin in the model group significantly increased, compared with that in the normal group; 10, 20, 30, 40, 50 mg/L HLP inhibited the expression elevations of ICAM-1 and E-selectin in concentration-dependent manner; and there were statistical significances in 40, 50 mg/L HLP groups, compared with the model group. Conclusion HLP can inhibit the over expressions of ICAM-1 and E-selectin of vascular endothelial cells induced by TNF-α, which possibly underlies HLP's anti-inflammation effect.

Protective effects and mechanism of pine bark extract on cerebral ischemia reperfusion injury in mice / 中国实验动物学报

Objective To investigate the role of IL-6/STAT3/miR-21 in the treatment of cerebral ischemia reper?fusion injury in mice by pine bark extract. Methods 120 KM mice were randomly divided into normal control group, sham operation group, 14 d and 28 d model groups, 14 d and 28 d treatment groups, each group with 20 mice. The model of cerebral ischemia?reperfusion injury model was made by bilateral common carotid artery occlusion. The mice in the treat?ment group were treated with pine bark extract procyanidins by intragastric administration for 7 days before the execution. The control group and sham operation group were treated with normal saline. The differences in learning and memory abili?ties and histopathological changes in the hippocampus of mice in each group were examined. The levels of interleukin 6, STAT3, p?STAT3 and miR?21 were measured. Results The learning and memory abilities of the mice in the treatment group were higher than that in the model group at the same time points. The treatment group had less irregularity in the ar?rangement of Nissl body, irregular shape of the cells, with relatively lighter cytoplasmic staining, but was better than the model group. The expression level of IL?6 in the 28 d model group was lower than that in the 14 d model group, The IL?6 in the hippocampus of the treatment group was significantly lower than that in the model group at the same time point. Therewere no significant differences in the contents of total STAT3 protein in the hippocampus between the groups (P>0. 05). The expression of p?STAT3 in the treatment group was lower than model group at the same time point. The expression of miR?21 in the hippocampus of 14 d, 28 d treatment groups was lower than that in the model group at the same time point, and decreased with time. Conclusions The procyanidins extracted from pine bark may effectively inhibit IL?6, then the phosphorylation level of STAT3 is significantly decreased and finally decreases the expression of miR?21, so as to alleviate the cerebral ischemia reperfusion injury in mice.

Effect of Grape Seed Procyanidins on Protein Expressions of Caspase-3 and TRAF6 in Renal Tissue of Rats with Renal Ischemia-reperfusion Injury / 中国药房

OBJECTIVE:To study the effect of grape seed procyanidins on protein expressions of cysteine aspartic protease 3 (Caspase-3), tumor necrosis factor-related receptors (TRAF6) in renal tissue of rats with renal ischemia-reperfusion injury (RIRI),and explore the protective mechanism of procyanidins on RIRI. METHODS:50 rats were randomly divided into sham op-eration group,model group,Shenfukang capsules group(positive control,600 mg/kg),procyanidins low-dose,high-dose groups (100,150 mg/kg),10 in each group. All rats were intragastrically administrated once a day,for 7 d. After administration,rats in other groups except for sham operation group were established the RIRI model.After the modeling successed of 24 h,levels of creati-nine(Cr),urea nitrogen(BUN)in serum,and protein expressions of Caspase-3,TRAF6 in renal tissue of rats in each group were detected,and apoptotic rate of renal tubular epithelial cells was determined. RESULTS:Compared with sham operation group,lev-els of Cr,BUN in serum in model group were significantly increased(P<0.05);protein expressions of Caspase-3,TRAF6 in re-nal tissue were obviously enhanced (P<0.05);apoptotic rate of renal tubular epithelial cells was obviously increased (P<0.05). Compared with model group,levels of Cr,BUN in serum in each administration group were obviously decreased(P<0.05);pro-tein expressions of Caspase-3,TRAF6 in renal tissue were obviously weakened(P<0.05);apoptotic rate of renal tubular epithelial cells was obviously decreased(P<0.05);and procyanidins high-dose group showed superior effect to low-dose group and Shenfu-kang granules group (P<0.05). CONCLUSIONS:Grape seed procyanidins can relieve the RIRI of rats,which may be achieved by reducing protein expressions of Caspase-3 and TRAF6 to inhibit the apoptotic rate of renal tubular epithelial cells.

Effects of procyanidins on cell apoptosis and the release of cytochrome C in human trabecular meshwork cells under oxidative stress / 眼科新进展

Objective To investigate the effect of procyanidins (PC) on cell apoptosis and cytochrome C releasing in human trabecular meshwork cells (HTMC) under oxidative stress induced by H2 O2.Methods HTMC were cultured and then divided randomly into 5 groups.Normal cultured HTMC served as normal group without treatment,and normal cultured HTMC in control group were treated with 500 μrnol · L-1 H2O2 for 1 h,as well as normal cells in treatment groups were treated with 500 imol · L-1 H2 O2 for 1 h combined with different concentrations of PC (0.02 g · L-1,0.05 g · L-1,0.10 g · L-1).Then flow cytometry and Western blot were applied to detect the cell apoptosis rate and the release of cytochrome C (CytC) respectively.Results Compared with the normal group,the apoptotic rates of the control group and the PC treatment groups were significantly increased,and the difference was statistically significant (all P ＜ 0.01).Compared with the control group,the apoptotic rates of the 3 PC treatment groups were decreased,and the differences were statistically significant (all P ＜ 0.01).Moreover,the apoptotic rate in 0.02 g · L-1,0.05 g · L-1,0.10 g · L-1 PC treatment groups was decreased as PC concentration increased,and the differences were statistically significant (all P ＜ 0.01).Furthermore,compared with the normal group,the release of cytochrome C in the control group,0.02 g · L-1 and 0.05 g · L-1 PC treatment group enhanced,and the differences were statistically significant (all P ＜ 0.01),but there was no significant difference between 0.01 g · L-1 PC treatment group and the normal group (P ＞ 0.05).Compared with the control group,the release of cytochrome C in the 3 PC treatment group was attenuated with significant differences (all P ＜0.01).Meanwhile,the release of cytochrome C was decreased in the 3 PC treatment groups with a concentration-dependent manner,and the pairwise differences were statistically significant (all P ＜ 0.01).Conclusion Exogenous PC can decrease the apoptotic rate of HTMC under oxidative stress and reduce the release of cytochrome C,as well as it also has an antioxidant effect in a certain concentration-dependent manner.

Effects of procyanidins on cell apoptosis and the release of cytochrome C in human trabecular meshwork cells under oxidative stress / 眼科新进展

Objective To investigate the effect of procyanidins (PC) on cell apoptosis and cytochrome C releasing in human trabecular meshwork cells (HTMC) under oxidative stress induced by H2 O2.Methods HTMC were cultured and then divided randomly into 5 groups.Normal cultured HTMC served as normal group without treatment,and normal cultured HTMC in control group were treated with 500 μrnol · L-1 H2O2 for 1 h,as well as normal cells in treatment groups were treated with 500 imol · L-1 H2 O2 for 1 h combined with different concentrations of PC (0.02 g · L-1,0.05 g · L-1,0.10 g · L-1).Then flow cytometry and Western blot were applied to detect the cell apoptosis rate and the release of cytochrome C (CytC) respectively.Results Compared with the normal group,the apoptotic rates of the control group and the PC treatment groups were significantly increased,and the difference was statistically significant (all P ＜ 0.01).Compared with the control group,the apoptotic rates of the 3 PC treatment groups were decreased,and the differences were statistically significant (all P ＜ 0.01).Moreover,the apoptotic rate in 0.02 g · L-1,0.05 g · L-1,0.10 g · L-1 PC treatment groups was decreased as PC concentration increased,and the differences were statistically significant (all P ＜ 0.01).Furthermore,compared with the normal group,the release of cytochrome C in the control group,0.02 g · L-1 and 0.05 g · L-1 PC treatment group enhanced,and the differences were statistically significant (all P ＜ 0.01),but there was no significant difference between 0.01 g · L-1 PC treatment group and the normal group (P ＞ 0.05).Compared with the control group,the release of cytochrome C in the 3 PC treatment group was attenuated with significant differences (all P ＜0.01).Meanwhile,the release of cytochrome C was decreased in the 3 PC treatment groups with a concentration-dependent manner,and the pairwise differences were statistically significant (all P ＜ 0.01).Conclusion Exogenous PC can decrease the apoptotic rate of HTMC under oxidative stress and reduce the release of cytochrome C,as well as it also has an antioxidant effect in a certain concentration-dependent manner.

Oligomeric Procyanidins (OPCs) Inhibit Procollagen Type I Secretion of Fibroblasts

Wound healing is composed of a complex process that requires harmonies of various cell populations where fibroblasts play the main role. Oligomeric procyanidins (OPC) are main components of grape (Vitis vinifera) seed extracts, and recent studies showed OPC's effects on inflammation, cell migration, and proliferation. We investigated the effect of OPC on fibroblasts to regulate wound healing process. Human dermal fibroblast known as Hs27 cells were treated with various concentrations of OPC (0, 2.5, 5, 10, and 20 µg/µl). Cell cytotoxicity was evaluated by the Cell Counting Kit assay, and the expression levels of secreted procollagen were analyzed. Procollagen levels in OPC treated cells exposed to transforming growth factor beta 1 (TGF-β1) or ascorbic acid were evaluated using Western blot and immunocytochemistry. Relative mRNA expressions of procollagen, molecular chaperone such as HSP47, P4H were determined by real-time PCR in OPC treated cells. OPC showed no cytotoxicity on Hs27 cells at every concentration but inhibited procollagen secretion in a dose-dependent manner. The inhibitory effect also appeared under TGF-β1 induced collagen overproduction. Immunocytochemistry showed that higher levels of intracytoplasmic procollagen were accumulated in TGF-β1 treatment group, whereas ascorbic acid induced a release of accumulated procollagen under OPC treatment. The mRNA expressions of procollagen, molecular chaperone were not affected by OPC, but procollagen level was increased when exposed to TGF-β1. OPC inhibits procollagen secretion from fibroblasts with no effects on cell proliferations even under the environment of TGF-b1-induced collagen overproduction. OPC could regulate the diseases and symptoms of abnormal overabundant collagen production.

Effects of hawthorn leaf polymeric procyanidins on calcium mobilization in human umbilical vein endothelial cells / 中国病理生理杂志

AIM:To observe the effects of hawthorn leaf polymeric procyanidins ( PPC) on calcium mobiliza-tion of vascular endothelial cells , and to study the underlying mechanism .METHODS: Free calcium in cultured human umbilical vein endothelial cells (HUVECs) was labeled with Fura-2.HUVECs were treated with ATP, a positive control drug, and PPC at concentrations of 12.5, 25 and 50 mg/L..The intracellular calcium concentrations were measured with a living cell microscope for 30 min.RESULTS:PPC concentration-dependently increased the intracellular calcium concen-tration of HUVECs .The intracellular calcium concentrations in 25 and 50 mg/L PPC groups were significantly higher than that in normal group (P<0.01).The dynamic manner of calcium concentration elevations elicited by PPC was a slow in -crease which happened after a latency time of several minutes , lasted for several minutes , and reached a plateau finally . This manner was quite different from that elicited by ATP , a typical SOC operator , hinting different mechanisms between them .Inhibiting the intracellular calcium release did not influence the effects of PPC;however , deleting extracellular calci-um, inhibiting the reverse mode of Na +-Ca2+exchange, or deleting extracellular sodium , restrained or even abolished the effects of PPC.CONCLUSION:PPC elicits calcium mobilization in vascular endothelial cells , which may be one of the mechanisms of the vascular modulatory activity of hawthorn procyanidins .This effect may be achieved through inducing the influx of sodium and then activating the reverse mode of Na +-Ca2+exchange.

Protective effects of procyanidins on human trabecular meshwork cells against H2O2 induced oxidative stress / 眼科新进展

Objective To investigate the protection and mechanism of procyanidins (PC) against H2O2 induced oxidative damage of human trabecular meshwork cells (HTMC) in order to provide an experimental foundation for glaucoma clinical treatment.Methods HTMC were cultured and then divided randomly into 5 groups.As untreated group:Normal cultured HTMC;Control group:Normal cultured HTMC + H2O2 (500 μmol · L-1 for 1 hour);Treated group:Normal cultured HTMC + H2O2 (500 μmol ·L-1 for 1 hour) + PC (PC fmal concentrations were 0.02 g · L-1,0.05 g · L-1,0.10 g· L-1).Real-time fluorescence quantitative polymerase chain reaction (PCR) was used to investigate the expression of mitochondrial complex Ⅰ mRNA.Results Compared with untreated group (1.000 0 ± 0.000 0),the differences of mitochondrial complexⅠ mRNA expression in 0.02 g · L-1 PC (0.401 3 ±0.010 3),0.05 g · L-1 PC (0.791 5 ± 0.008 5) groups were statistically significant (all P ＜ 0.01),but the 0.10 g ·L-1 PC group (1.043 0 ± 0.062 2) had no significant differences (P ＞ 0.05).The differences between PC treated groups and control group were statistically significant (P ＜0.01),which showed HTMC treated with PC could increase the expression of mitochondrial complex Ⅰ mRNA.The differences in each PC treated groups were statistically significant (P ＜ 0.01),which showed the expression of mitochondrial complex Ⅰ mRNA were increased along with the concentration of PC gradually increased.Conclusion Exogenetic PC can increase the expression of mitochondrial complex Ⅰ mRNA in the oxidative damaged HTMC,and in a certain range of concentration,the protective effects of PC have the positive relationship of dose-effect,which suggest that PC may be a good candidate for further study of the clinical treatment of glaucoma.

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.

Inhibitory effect of lotus seedpod oligomeric procyanidins on advanced glycation end product formation in a lactose-lysine model system

Background Industrial food processing induces protein glycation modifications and toxic advanced glycation end products (AGEs) which affect human health. Therefore, it is of interest to monitor AGEs in food processing. The present study was carried out to investigate the influence of lotus seedpod oligomeric procyanidin (LSOPC) concentrations, solution pH value and metal ions on AGE formation by heat treatment of lactose-lysine model solutions. Ne-(carboxymethyl) lysine (CML), as one of the common AGEs was also determined by HPLC-MS/MS in this experiment. Results The results showed that LSOPC can inhibit the formation of AGEs effectively at higher concentrations, lower temperature, and it can reverse the promotion function of metal ions because of its high inhibition activity. Also, LSOPC can inhibit CML formation in the Maillard reaction as well. Conclusion These results indicated that LSOPC could be used as functional food ingredients to inhibit AGE formation.