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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.
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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.
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Animals , Humans , Mice , Arthritis , Arthritis, Experimental , Arthritis, Rheumatoid , Blotting, Western , In Vitro Techniques , Macrophages , Myeloid Differentiation Factor 88 , Osteoarthritis , Synovial Membrane , Toll-Like Receptor 4 , VitisABSTRACT
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.
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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.
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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.
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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.
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<p><b>OBJECTIVE</b>To determine the ability of grape seed proanthocyanidin extract (GSPE) in alleviating arsenic-induced reproductive toxicity.</p><p><b>METHODS</b>Sixty male Kunming mice received the following treatments by gavage: normal saline solution (control); arsenic trioxide (ATO; 4 mg/kg); GSPE (400 mg/kg); ATO+GSPE (100 mg/kg); ATO+GSPE (200 mg/kg) and ATO+GSPE (400 mg/kg). Thereafter, the mice were sacrificed and weighed, and the testis was examined for pathological changes. Nuclear factor (erythroid-derived 2)-like 2 (Nrf2), heme oxygenase 1 (HO1), glutathione S-transferase (GST), NAD(P)H dehydrogenase, and quinone 1 (NQO1) expression in the testis was detected by real-time PCR. Superoxide dismutase (SOD), glutathione (GSH), total antioxidative capability (T-AOC), malondialdehyde (MDA), 8-hydroxydeoxyguanosine (8-OHdG), and reproductive indexes were analyzed.</p><p><b>RESULTS</b>ATO-treated mice showed a significantly decreased sperm count and testis somatic index and activity levels of SOD, GSH, and T-AOC than control group. Compared to the ATO-treated group, ATO +GSPE group showed recovery of the measured parameters. Mice treated with ATO+high-dose GSPE showed the highest level of mRNA expression of Nrf2, HO, NQO1, and GST.</p><p><b>CONCLUSION</b>GSPE alleviates oxidative stress damage in mouse testis by activating Nrf2 signaling, thus counteracting arsenic-induced reproductive toxicity.</p>
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Animals , Male , Mice , Antioxidants , Metabolism , Arsenic , Toxicity , Grape Seed Extract , Pharmacology , Lipid Peroxidation , NF-E2-Related Factor 2 , Genetics , Metabolism , Oxidative Stress , Proanthocyanidins , Pharmacology , Signal Transduction , Sperm Count , Testis , Cell Biology , MetabolismABSTRACT
@#Objective To investigate the effect of grape seed proanthocyanidin extract (GSPE) on learning and memory ability after cerebral ischemia/reperfusion injury in rats and its mechanism. Methods 72 healthy male Sprague-Dawley rats were divided into sham group (n=18), model group (n=18) and GSPE groups (20 mg/kg, 200 mg/kg, n=18 for each group). The GSPE groups were administered GSPE orally for 4 weeks, while the sham group and model group were given water 10 ml/kg. Then their middle cerebral arteries were obstructed for 2 h and reperfused, excepted the sham group. 6 rats from each group were selected to test with Morris water maze 12 h, 24 h and 48 h after reperfusion respectively. And then, their brain tissues were stained with Hematoxylin-Eosin staining to observe under optical microscope. The level of superoxide dismutase (SOD) and malondialdehyde (MDA) in the brain tissues were measured. Results Compared with the sham group, the latency significantly prolonged, and the incidence of crossing the area the platform located reduced in the model group in the Morris water maze test, with the SOD decreasing and MDA increasing (P<0.05). Compared with the model group, the latency reduced and the incidence of crossing the area increased in the GSPE 200 mg/kg group, with the SOD increasing and MDA decreasing (P<0.05). Conclusion GSPE may suppress peroxidation after the cerebral ischemia/reperfusion to protect brain and learning and memory ability from injury.
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Objective To investigate the influences of grape seed proanthocyanidin extract (GSPE) on cardiovascular system, nervous system and respiratory system of experimental animals, and provide general pharmacological data for further research and application. Methods The influences of GSPE on blood pressure, heart rate, electrocardiogram, breathing frequency and tidal volume in anesthetic dogs after duodenal administration were observed, the impacts on spontaneous activity, coordinated motion, and the sleep situation with threshold dose and subthreshold dose of pentobarbital sodium in mice after intragastric administration were observed. Results GSPE showed no side effects on blood pressure, heart rate, electrocardiogram, breathing frequency and tidal volume in anesthetic dogs at the dosage of 857.00, 214.29, 42.86 mg/kg (P>0.05). At the dosage of 428.57, 214.29, 42.86 mg/kg, GSPE had no obvious influence on spontaneous activities and coordinated movements in mice (P>0.05). GSPE did not evidently change the number of sleeping animals, the sleep latency and the sleeping duration with subthreshold dose and threshold dose of pentobarbital sodium (P>0.05). Conclusion GSPE has no evident adverse effects on central nervous system, cardiovascular system and respiratory system in animals.
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Objective To investigate the effect of grape seed proanthocyanidin extract (GSPE) on learning and memory ability after cere-bral ischemia/reperfusion injury in rats and its mechanism. Methods 72 healthy male Sprague-Dawley rats were divided into sham group (n=18), model group (n=18) and GSPE groups (20 mg/kg, 200 mg/kg, n=18 for each group). The GSPE groups were administered GSPE orally for 4 weeks, while the sham group and model group were given water 10 ml/kg. Then their middle cerebral arteries were obstructed for 2 h and reperfused, excepted the sham group. 6 rats from each group were selected to test with Morris water maze 12 h, 24 h and 48 h after re-perfusion respectively. And then, their brain tissues were stained with Hematoxylin-Eosin staining to observe under optical microscope. The level of superoxide dismutase (SOD) and malondialdehyde (MDA) in the brain tissues were measured. Results Compared with the sham group, the latency significantly prolonged, and the incidence of crossing the area the platform located reduced in the model group in the Mor-ris water maze test, with the SOD decreasing and MDA increasing (P<0.05). Compared with the model group, the latency reduced and the incidence of crossing the area increased in the GSPE 200 mg/kg group, with the SOD increasing and MDA decreasing (P<0.05). Conclu-sion GSPE may suppress peroxidation after the cerebral ischemia/reperfusion to protect brain and learning and memory ability from injury.
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Background The pathogenesis of diabetic retinopathy (DR) might be related with oxidative stress and its mechanism has not been fully elucidated.Grape seed proanthocyanidin extracts (GSPE),a powerful antioxidant,plays roles in some systemic diseases.However,the effect and mechanism of GSPE in DR has not been illuminated clearly.Objective This study was to investigate whether GSPE has a protective effect on the retinas of diabetic subjects and explore its mechanism.Methods Thirty SPF adult Wistar rats were divided into the control group,diabetic group and diabetes + GSPE group according to the randomized number table.Diabetic models were induced by intraperitoneal injection of 100 mg/kg strebtozotocin (STZ) prepared with citrate buffer,and only the equal amount of citrate buffer was used in the same way in the control group.GSPE solution was intragastrally used 250 mg/kg daily in the rats of the diabetes+GSPE group from injective day through 8 weeks,and distilled water was used in the same way in the rats of the control and diabetic groups.The rats were sacrificed in the eighth week after injection and the retinas were isolated.The morphology of the retina were examined by hematoxylin and eosin stain.Retinal homogenatewas prepared for the assay of superoxide dismutase (SOD),glutathione peroxidase (GSH-Px) activity and malonaldehyde (MDA) content.The expressions and location of nuclear erythroid related factor 2 (Nrf2) was determined by immunochemistry,and the expressions of Nrf2 and heme oxygenase-1 (HO-1) were quantitatively analyzed by Western blot.The apoptosis of retinal cells was detected by TUNEL.Results In the eighth week after modeling,the blood glucose levels were significantly higher and the body weight was lower in the rats of the diabetic group and diabetes+GSPE group compared with the control group (all at P＜0.01).Retinal structure was normal in the rats of the control group.However,loose tissue,irregular arrangement of cells and thickness decrease of retinal fiber layer,retinal ganglion cell layer and inner plexiform layer were exhibited in the rats of the diabetic group,while the morphology abnormality was slight in the rats of the diabetes+GSPE group.The SOD and GSH-Px activities and MDA content were significantly different among the 3 groups (F =11.010,P =0.001 ; F =12.072,P =0.000 ; F =25.224,P=0.000),and activities of SOD and GSH-Px in the retinas were significantly lower,and the MDA level was higher in the rats of diabetic group than those of the diabetes+GSPE group and the control group (all at P＜0.01).The relative expressing levels of Nrf2 were (165.5±29.4) ％ and (134.8 ±7.8) ％ in the diabetes+ GSPE group and diabetic group,with a significant difference between them (t=2.450,P=0.044).Compared with the diabetic group,the expressing level of the HO-1 was sigficantly increased ([170.2±22.0)％ versus [125.3±9.2] ％,t =2.360,P =0.002).TUNEL showed that the retinal apoptotic cells of diabetic rats were mainly located in the retinal fiber layer,RGCs layer,inner and outer nuclear layer,and the number of apoptotic cells was less in the diabetes+GSPE group compared with the diabetic group under the fluorescence microscope.Conclusions GSPE can play a protective role on diabetic rats by activating Nrf2 pathway and therefore improving retinal oxidative stress and decreasing apoptosis.
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Objective To investigate the effects of grape seed proanthocyanidin(GSPE) on mitochondrial injury in hippocampus and learning-memory impairment after obstructive sleep apnea hypoxia in rats.Methods Male SD rats(n=80) were randomly divided into control group,model group,low dose of GSPE treatment group and high dose of GSPE treatment group.Rats in control group were exposed in air,the model group were suffered from intermittent hypoxia conditions (50 ml/L,8-hour-intermittent hypoxia everyday,and the duration of experiment 2 and 6 weeks,respectively).Mitochondrion pathology in hippocampal region was observed using electron microscope;malondialdehyde (MDA) contents and superoxide dismutase activity were detected by colorimetry and apoptotic cells was measured by TUNEL method.The cognitive function of rats in each group was assessed with the Morris water maze (MWM).Results After hypoxia,mitochondrion was significantly injured.The MDA contents were increased(79.86 ± 2.52,88.26 ± 2.86) and SOD level decreased (70.67 ± 6.70,64.26 ± 7.86).The number of neural apoptotic cells was significantly enhanced (9.68 ± 0.79,15.9 ± 2.92).MWM test showed that the escaping latency was prolonged and the frequency of crossing the platform was decreased (P ＜ 0.05).Compared with that in the model group,low dose of GSPE decreased MDA contents (76.38 ± 1.96,82.16 ±2.02),increased SOD level(76.20 ± 6.86,70.58 ± 6.86),and decreased apoptotic cells (6.60 ± 0.69,9.54 ±1.36).MWM test showed that the escaping latency was shortened and the frequency of crossing the platform was increased in GSPE treatment groups(P ＜ 0.05).Compared with low dose of GSPE,high dose of GSPE decreased MDA contents increased SOD level and decreased apoptotic cells.MWM test showed that the escaping latency was shortened and the frequency of crossing the platform was increased (P＜ 0.05).Conclusion GSPE can attenuate mitochondrial injury and improve learning-memory function after obstructive sleep apnea hypoxia.
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0.05) during the experiment. The TC of middle dose GSPE group was marked ly different compared with that of normal group(P
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Objective: To investigate the effects of grape seed proanthocyanidin(GSP) and casein peptide on the liver function and protein nutritional status in rays irradiated rats. Methods: Adult wistar rats were randomized into four groups with 20 rats in each group including normal control, radiation control, GSP and GSP+casein peptide combined groups. The GSP group was fed with basic diet containing 1% GSP, the combined group was fed with basic diet containing 1% GSP plus 5% casein peptide. Animals except normal control were irradiated by 60 Co? rays(7Gy) 10 days later. 6 survivors were randomly selected from each group and killed on the 7 th d after radiation. The levels of the total protein (TP) and the albumin (ALB), the activities of the glutamate pyruvate transaminase(GPT) and the glutamate oxaloacetale transaminase(GOT) in serum were measured. The change of the body weight of the animals was observed during the experiment time. Results: After radiation the body weight of the rats dropped sharply. The loss of the weight in combined group were significantly lower than in radiation control group and in GSP group(P