Grape Seed Proanthocyanidin Extract Alleviates Arsenic-induced Oxidative Reproductive Toxicity in Male Mice

Objective To determine the ability of grape seed proanthocyanidin extract （GSPE） in alleviating arsenic-induced reproductive toxicity. Methods 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. Results 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, NO.O1, and GST. Conclusion GSPE alleviates oxidative stress damage in mouse testis by activating Nrf2 signaling, thus counteracting arsenic-induced reproductive toxicity.

Quantitative Mitochondrial Proteomics Study on Protective Mechanism of Grape Seed Proanthocyanidin Extracts Against Ischemia/Reperfusion Heart Injury in Rat

Cardiac ischemia/reperfusion（I/R） injury is a critical condition,often associated with high morbidity and mortality.The cardioprotective effect of grape seed proanthocyanidin extracts（GSPE） against oxidant injury during I/R has been described in previous studies.However,the underlying molecular mechanisms have not been fully elucidated.This study investigated the effect of GSPE on reperfusion arrhythmias especially ventricular tachycardia（VT） and ventricular fibrillation（VF）,the lactic acid accumulation and the ultrastructure of ischemic cardiomyocytes as well as the global changes of mitochondria proteins in in vivo rat heart model against I/R injury.GSPE significantly reduced the incidence of VF and VT,lessened the lactic acid accumulation and attenuated the ultrastructure damage.Twenty differential proteins related to cardiac protection were revealed by isobaric tag for relative and absolute quantitation（iTRAQ） profiling.These proteins were mainly involved in energy metabolism.Besides,monoamine oxidase A（MAOA） was also identified.The differential expression of several proteins was validated by Western blot.Our study offered important information on the mechanism of GSPE treatment in ischemic heart disease.

Inhibitory Effects of Grape Seed Proanthocyanidin Extract on Selenite-induced Cataract Formation and Possible Mechanism

This study investigated the inhibitory effect of grape seed proanthocyanidin extract(GSPE) on selenite-induced cataract formation in rats and the possible mechanism.Eighty 8-day-old Sprague-Dawley rats were divided randomly into 5 groups:control group,model group,three GSPE groups(low dose,medium dose and high dose).Control group received subcutaneous injection of physiological saline.Model group was given subcutaneous injection of sodium selenite(20 μmol/kg body weight) on the postpartum day 10,and once every other day for consecutive three times thereafter.GSPE treated groups were respectively administered GSPE at doses of 50,100,and 200 mg/kg body weight intragastrically 2 days prior to the selenite injection(that was,on the postpartum day 8),and once daily for fourteen consecutive days thereafter.The opacity of lenses was observed,graded and photographed under the slit lamp microscopy and the maximal diameter of the nuclear cataract plaques was measured.The lenses were analyzed for superoxide dismutase(SOD),catalase(CAT),glutathione peroxidase(GSH-PX),malondialdehyde(MDA),calcium(Ca 2+),nitric oxide(NO) and anti-hydroxyl radical ability(anti-OH).The histomorphology of lenses was observed with HE staining under a light microscope.The levels of calpainⅡ,and iNOS protein and mRNA expression in lenses were detected by using immunohistochemistry and real-time quantitative RT-PCR.The results showed subcutaneous injection of sodium selenite led to severe nuclear cataract in model group,and the achievement ratio of model group was 100%.As compared with model group,the degree of lenses opacity and the maximal diameter of nuclear cataract plaques were significantly reduced in GSPE-treated groups.Moreover,we observed selenite treatment caused a significant decrease in the activities of antioxidative enzymes(SOD,CAT,GSH-PX) and anti-OH ability,accompanied by a significant increase in the levels of MDA,NO,Ca 2+ as well as iNOS,and calpainⅡ protein and mRNA expression.Administration of GSPE could dose-dependently preserve the activities of these antioxidative enzymes and anti-OH ability,accompanied by a significant reduction in the levels of MDA,NO,Ca 2+ as well as iNOS,and calpainⅡ protein and mRNA expression.These results suggested that GSPE markedly prevented selenite-induced cataract formation probably by suppressing the generation of lipid peroxidation and free radicals as well as the activation of iNOS,and calpainⅡ in the lenses.

Grape Seed Proanthocyanidin Extract Ameliorates Streptozotocin-induced Cognitive and Synaptic Plasticity Deficits by Inhibiting Oxidative Stress and Preserving AKT and ERK Activities

Progressive memory loss and cognitive impairment are the main clinical manifestations of Alzheimer’s disease(AD).Currently,there is no effective drug available for the treatment of AD.Previous studies have demonstrated that the cognitive impairment of AD is associated with oxidative stress and the inhibition of AKT and ERK phosphorylation.Grape seed proanthocyanidin extract(GSPE)has been shown to have strong antioxidant effect and can protect the nervous system from oxidative stress damage.This study aimed to investigate the protective effect of GSPE on the cognitive and synaptic impairments of AD using a sporadic AD rat model induced by intracerebroventricular(ICV)injection of streptozotocin(STZ)(ICV-STZ).Rats were treated with GSPE(50,100,or 200 mg/kg every day)by intragastrical(ig.)administration for continuous 7 weeks,and ICV-STZ(3 mg/kg)was performed on the first day and third day of week 5.Learning and memory abilities were assessed by the Morris water maze(MWM)test at week 8.After behavioral test,hippocampal long-term potentiation(LTP)was recorded,and the levels of malondialdehyde(MDA),superoxide dismutases(SOD),glutathione(GSH)and the protein expression of AKT and ERK were measured in the hippocampus and cerebral cortex of rats.Our study revealed that ICV-STZ significantly impaired the working learning ability and hippocampal LTP of rats,significantly increased the levels of MDA,and decreased the activity of SOD and GSH in the hippocampus and cerebral cortex.In contrast,GSPE treatment prevented the impairment of cognitive function and hippocampal LTP induced by ICV-STZ,decreased the level of MDA,and increased the level of SOD and GSH.Furthermore,Western blot results showed that GSPE treatment could prevent the loss of AKT and ERK activities in the hippocampus and cerebral cortex induced by ICV-STZ.Our findings demonstrate that GSPE treatment could ameliorate the impairment of cognitive ability and hippocampal synaptic plasticity in a rat model of sporadic AD by inhibiting oxidative stress and preserving AKT and ERK activities.Therefore,GSPE may be an effective agent for the treatment of cognitive deficits associated with sporadic AD.

Beneficial clinical effects of grape seed proanthocyanidin extract on the progression of carotid atherosclerotic plaques

Background Atherosclerotic plaques indicate the occurrence of ischemia events and it is a difficult task for clinical physicians. Grape seed proanthocyanidin extract （GSPE） has been reported to exert an antiatherogenic effect by inducing regression of atherosclerotic plaques in animal experimental studies. In this study, the antiatherogenic effect of GSPE has been investigated in clinical use. Methods Consecu- tive 287 patients diagnosed with asymptomatic carotid plaques or abnormal plaque free carotid intima-media thickness （CIMT） were ran- domly assigned to the GSPE group （n = 146） or control group （n = 141）. The patients in the GSPE group received GSPE 200 mg per day orally, while patients in the control group were only enrolled in a lifestyle intervention program. Carotid ultrasound examination was per- formed at baseline and 6, 12, 24 months during follow-up. Mean maximum CIMT （MMCIMT）, plaque score, echogenicity of plaques and ischemic vascular events were recorded. Results As anticipated, after treatment, GSPE resulted in significant reduction in MMCIMT pro- gression （4.2% decrease after six months, 4.9% decrease after 12 months and 5.8% decrease after 24 months） and plaque score （10.9% de- crease after six months, 24.1% decrease after 12 months and 33.1% decrease after 24 months） for the primary outcome, while MMCIMT and plaque score were stable and even increased with the time going on in control group. The number of plaques and unstable plaques also de- creased after treatment of GSPE. Furthermore, the carotid plaque can disappear after treatment with GSPE. The incidence rate for transitory ischemic attack （TIA）, arterial revascularization procedure, and hospital readmission for unstable angina in GSPE group were statistically significant lower （P = 0.02, 0.08, 0.002, respectively） compared with the control group. Conclusions GSPE inhibited the progression of MMCIMT and reduced carotid plaque size in GSPE treated patients, and with extended treatment, the superior efficacy on MMCIMT and carotid plaque occurred. Furthermore, the GSPE group showed lower rates of clinical vascular events.

Grape Seed Procyanidin Extract Reduces Arsenic- Induced Renal Inflammatory Injury in Male Mice

The aim of the present study is to evaluate the ability and mechanism by which grape seed procyanidin extract （GSPE） relieves arsenic trioxide （As2O3）-induced renal inflammatory injury. Therefore, male Kunming mice were treated with As2O3 and/or GSPE by gavage for 5 weeks. Mice were then sacrificed and inflammatory cytokines of kidneys were examined by ELISA, whereas the expression levels of molecules involved in the nuclear factor （NF）-KB signaling pathway were evaluated by both qRT-PCR and Western blot. Our results indicate that GSPE prevents As2O3-mediated renal inflammatory injury by inhibiting activation of the NF-KB signaling pathway and inflammatory cytokine production, while promoting expression of anti-inflammatory cytokines.

Role of survivin in apoptosis induced by grape seed procyanidin extract in human bladder cancer BIU87 cells

Objects： The aim of this study was to research the effect of grape seed procyanidin extract （GSPE） on cell apotosis in human bladder cancer BIU87 cells and investigate its molecular mechanism. Methods： BIU87 cells were treated with different concentrations of GSPE and cultured for 24 h in vitro while untreated group as control, MTT[3- （4,5-dimethylthiazole- 2-yl） -2, 5-diphenyltetrazolium bromide] assay, Hoechst 33258 stainning, flow cytometry, RT-PCR and Western blot were used to detect the apoptotic induction effect of GSPE on BIU87 cells. Results： We found that GSPE induced cell apoptosis in BIU87 cells by a dose-dependent manner. Semiquantitated RT-PCR and Western blot analyses indicated that GSPE increased the expression of caspase-3 and decreased the expression of survivin （P 〈 0.01）. Conclusion： GSPE induces apoptosis in BIU87 cells in vitro, and the effect maybe related with its down-regulation of survivin.

GSPE通过Nrf2信号通路拮抗砷所致的小鼠肝脏氧化损伤

目的探讨葡萄籽原花青素提取物(grape seed proanthocyanidins extract,GSPE)能否通过Nrf2信号通路拮抗砷所致的小鼠肝脏氧化损伤,为GSPE对抗砷的氧化损伤作用提供理论依据。方法以昆明种小鼠为研究对象,根据As 2O 3(0、4 mg/kg)和GSPE(0、400 mg/kg)的剂量,采用2×2析因设计方差分析,按照体质量分层,随机分为对照组、As 2O 3组、GSPE组和As 2O 3组+GSPE组,使用析因设计分析As 2O 3和GSPE的交互作用和As 2O 3、GSPE的单独效应。采用试剂盒标准的测量方法测量MDA、GSH、ROS、GSH-Px、SOD、CAT、LPO、-SH和T-AOC表达;Western blotting检测Nrf2蛋白的表达;qRT-PCR检测Nrf2 mRNA的表达水平。结果As 2O 3组中,小鼠的体质量有所下降,肝脏重量和脏器系数增加;GSPE可以对抗这一结果;As 2O 3可以增加MDA、ROS、LPO、Nrf2蛋白和mRNA的表达,减少GSH、GSH-Px、SOD、CAT、-SH和T-AOC的表达;在经过GSPE拮抗As 2O 3的作用之后,同组相比,MDA、ROS、LPO、Nrf2蛋白和mRNA的表达减少,而GSH、GSH-Px、SOD、CAT、-SH和T-AOC的表达增加(P均<0.05)。结论GSPE可能通过Nrf2信号通路及相关氧化应激因子拮抗砷所致的小鼠肝脏氧化损伤。

葡萄籽原花青素提取物对心肌梗死大鼠肾素-血管紧张素系统及AQP2蛋白表达的影响

目的:探讨葡萄籽原花青素提取物(GSPE)对心肌梗死大鼠肾素血管紧张素-系统及水通道蛋白2(AQP2)表达的影响。方法:将55只无特定病原体(SPF)级Sprague-Dawley(SD)大鼠按照随机数字表法分为健康组、模型组、辛伐他汀组、GSPE低剂量组及GSPE高剂量组,每组11只。除健康组外其余大鼠均采用冠状动脉结扎法制备心肌梗死模型,建模成功后,GSPE低剂量组、高剂量组大鼠分别灌胃100 mg/kg、400 mg/kg葡萄籽原花青素溶液,辛伐他汀组灌胃40 mg辛伐他汀溶液。苏木精-伊红(HE)染色后观察心肌组织形态;末端脱氧核苷酸转移酶介导的原位缺口末端转移酶标记法(TUNEL)检测心肌细胞凋亡;超声心动图监测心功能;放射免疫法检测血管紧张素Ⅱ(AngⅡ)、血浆肾素活性(PRA)、醛固酮水平;免疫印迹法检测心肌组织中AQP2、B细胞淋巴瘤/白血病2号基因(Bcl-2)及半胱天冬氨酸蛋白酶-3(Caspase-3)蛋白水平。结果:与健康组比较,模型组大鼠左心室收缩末期内径(LVESD)、左心室舒张末期内径(LVEDD)升高,左心室射血分数(LVEF)和左心室短轴缩短分数(LVFS)降低(P<0.05);与模型组比较,GSPE低剂量组、GSPE高剂量组及辛伐他汀组LVEDD、LVESD降低,LVEF、LVFS升高(P<0.05),GSPE低剂量组、GSPE高剂量组间比较差异有统计学意义,而GSPE高剂量组与辛伐他汀组比较差异无统计学意义(P>0.05)。与健康组比较,模型组大鼠AngⅡ、PRA、醛固酮升高(P<0.05);与模型组比较,GSPE低剂量组、GSPE高剂量组及辛伐他汀组AngⅡ、PRA、醛固酮降低(P<0.05)。健康组、模型组、GSPE低剂量组、GSPE高剂量组及辛伐他汀组的心肌细胞凋亡率分别为(5.11±0.33)%、(46.22±3.97)%、(28.46±3.77)%、(15.42±2.33)%及(16.34±2.57)%,各组比较差异有统计学意义(P<0.05)。与健康组比较,模型组大鼠心肌组织中AQP2、Caspase-3蛋白水平升高,Bcl-2蛋白水平降低(P<0.05);与模型组比较,GSPE低剂量组、GSPE高剂量组、辛代他汀组大鼠心肌组织中AQP2、Caspase-3蛋白水平降低,Bcl-2蛋白水平升高(P<0.05)。结论:GSPE对心肌梗死有保护作用,可改善心功能水平,减少心肌细胞凋亡,其作用机制可能与调控AQP2蛋白表达有关。

3种多酚对腐败希瓦氏菌的抑菌效果和机理

为探讨葡萄籽提取物(grape seed extract,GSE)、莲房原花青素(lotus seed proanthocyanidins,LSPC)和莲藕多酚提取物(lotus root polyphenol extract,LRPE)3种植物多酚对腐败希瓦氏菌(Shewanella putrefaciens)的抑菌机理。通过最小抑菌质量浓度(minimal inhibitory concentration,MIC)、生长曲线探究3种多酚对腐败希瓦氏菌的抑菌效果,并通过扫描电子显微镜观察、相对电导率测定、碘化丙啶(propidium iodide,PI)染色观察及碱性磷酸酶(alkaline phosphatase,AKP)活力、胞外蛋白含量、核酸泄漏、Na^(+)K^(+)-ATP酶活性及膜蛋白荧光等测定,探讨3种多酚的抑菌机理。结果表明:GSE、LRPE、LSPC对腐败希瓦氏菌的MIC分别为31.25、62.25、125.00μg/mL;菌体经3种植物多酚处理后,膜蛋白位置发生改变,荧光强度降低,菌体形态发生改变,表面出现凹陷皱缩,且生长明显被抑制,胞外AKP活力、菌悬液中核酸、胞外蛋白含量、相对电导率及PI摄入量显著增加,Na^(+)K^(+)-ATP酶发生一定程度地失活,导致细胞死亡。

Grape seed proanthocyanidin extract induced mitochondria-associated apoptosis in human acute myeloid leukaemia 14.3D10 cells

The important aim in cancer treatment is the selective killing of cancer cells without damaging healthy cells and the most commonly used chemotherapy for eliminating cancer is achieved by activating the mitochondrial apoptotic pathway. Therefore, identification of new agents that can selectively kill cancer cells becomes crucial in cancer clinical treatment.

A novel approach of proteomics to study the mechanism of action of grape seed proanthocyanidin extracts on diabetic retinopathy in rats

Background Diabetic retinopathy （DR） is a leading cause of visual impairment and blindness among the people of occupational age. To prevent the progress of retina injury, effective therapies directed toward the key molecular target are required. Grape seed proanthocyanidin extracts （GSPE） have been reported to be effective in treating diabetic complications, while little is discussed about the functional protein changes. Methods We used streptozotocin （STZ） to induce diabetes in rats. GSPE （250 mg/kg body weight per day） were administrated to diabetic rats for 24 weeks. Serum glucose, glycated hemoglobin and advanced glycation end products （AGEs） were determined. Consequently, 2-D difference gel electrophoresis and mass spectrometry were used to investigate retina protein profiles among control, STZ-induced diabetic rats, and GSPE treated diabetic rats. Results GSPE significantly reduced the AGEs of diabetic rats （P 〈0.05）. Moreover, GSPE significantly suppressed the vascular lesions of central regions, decreased capillary enlargements and neovascularization, similar to those of the control rats under light microscope. Eighteen proteins were found either up-regulated or down-regulated in the retina of STZ-induced diabetic rats. And seven proteins in the retina of diabetic rats were found to be back-regulated to normal levels after GSPE therapy. These back-regulated proteins are involved in many important biological processes such as heat shock, ubiquitin-proteasome system, cell proliferation, cell growth and glucose metabolism. Conclusions These findings might promote a better understanding for the mechanism of DR, and provide novel targets for evaluating the effects of GSPE therapy.

葡萄籽原花青素提取物预灌胃对造影剂诱导糖尿病大鼠急性肾损伤的预防作用观察

目的观察葡萄籽原花青素提取物预灌胃对造影剂诱导糖尿病大鼠急性肾损伤的预防作用,并探讨可能作用机制。方法50只SD肥胖大鼠,腹腔注射1%链脲佐菌素(40 mg/kg),41只成功建成糖尿病大鼠模型,随机分为DM组8只、CM组9只、葡萄籽原花青素提取物低剂量组8只、中剂量组8只、高剂量组8只,另取10只肥胖大鼠为空白对照组(NC组),1 mL/kg腹腔注射柠檬酸缓冲液;低、中、高剂量组大鼠每日分别用50、250、500 mg/kg的葡萄籽原花青素提取物灌胃1次,连续3天,第3天灌胃24 h时尾静脉注射碘海醇(1.8 g I/kg);NC组、DM组、CM组大鼠每日用10 mL/kg生理盐水灌胃1次,第3天灌胃24 h时,NC组、DM组尾静脉注射5 ml/kg生理盐水;CM组尾静脉注射碘海醇(1.8 g I/kg)。末次给药48 h时各组大鼠断尾采血,检测血清肌酐(SCr)和尿素氮(BUN),采血后处死各组大鼠,取肾组织检测肾组织氧化应激指标超氧化物歧化酶(SOD)、丙二醛(MDA),采用原位缺口末端标记法测算各组大鼠肾小管上皮细胞凋亡指数,采用Western Blotting法检测各组大鼠肾组织核因子E2相关因子2(Nrf2)-Kelch样ECH关联蛋白1(Keap1)通路相关醌氧化还原酶1(NQO1)、血红素单加氧酶-1(HO-1)、Nrf2、Keap1蛋白。结果与NC组比较,CM组及低剂量组血清SCr、BUN水平高(P均<0.05)。与CM组比较,NC组、DM组、低中高剂量组血清SCr、BUN水平低(P均<0.05);与低剂量组比较,中、高剂量组大鼠血清SCr、BUN水平低(P均<0.05)。与NC组比较,DM组、CM组、低中剂量组肾组织匀浆SOD水平低,DM组、CM组及低剂量组肾组织匀浆MDA水平高(P均<0.05)。与CM组比较,DM组、低中高剂量组肾组织匀浆组SOD水平高、MDA水平低(P均<0.05)。与CM组比较,中、高剂量组大鼠肾小管上皮细胞凋亡指数小(P均<0.05)。与NC组比较,CM组大鼠肾组织Nrf2、HO-1及NQO1蛋白相对表达量低,中高剂量组大鼠肾组织Nrf2、HO-1及NQO1蛋白相对表达量高(P均<0.05);与CM组比较,低中高剂量组大鼠肾组织Nrf2、HO-1及NQO1蛋白相对表达量高(P均<0.05);与中剂量组比较,低剂量组大鼠肾组织Nrf2、HO-1及NQO1蛋白相对表达量低(P均<0.05)。结论葡萄籽原花青素提取物可改善尾静脉注射造影剂糖尿病大鼠的肾损伤程度,且500 mg/kg时效果最好。葡萄籽原花青素提取物可能通过激活Nrf2—Keap1信号通路,预防造影剂诱导的糖尿病大鼠的急性肾损伤。

葡萄籽原花青素提取物对糖尿病牙周炎大鼠牙周组织炎症的缓解作用及其对牙周组织中TLR4和NF-κB表达水平的影响

目的:探讨葡萄籽原花青素提取物(GSPE)对糖尿病牙周炎大鼠牙周炎症的改善作用,并阐明其作用机制。方法:40只雄性SD大鼠随机分为对照组、糖尿病牙周炎模型组(模型组)、低剂量(100 mg·kg^(-1))GSPE组和高剂量(200 mg·kg^(-1))GSPE组,每组10只,除对照组外,其他各组大鼠采用牙周结扎法联合一次性腹腔注射链脲佐菌素(STZ)建立糖尿病牙周炎模型。模型建立成功后,低和高剂量GSPE组大鼠分别按100和200 mg·kg^(-1)GSPE灌胃给药,对照组和模型组大鼠给予等量生理盐水灌胃,每天1次,连续8周。观察大鼠一般情况和血糖水平变化,采用HE染色观察各组大鼠牙周组织病理形态表现,测量各组大鼠牙槽骨吸收量,采用相关试剂盒检测各组大鼠牙周组织中超氧化物歧化酶(SOD)、过氧化氢酶(CAT)活性和活性氧(ROS)及丙二醛(MDA)水平,酶联免疫吸附试验(ELISA)法检测各组大鼠血清中肿瘤坏死因子α(TNF-α)、白细胞介素1β(IL-1β)和白细胞介素6(IL-6)水平,Western blotting法检测各组大鼠牙周组织中Toll样受体4(TLR4)和核因子κB(NF-κB)蛋白表达水平。结果:与对照组比较,模型组大鼠出现明显“三多一少”症状,血糖水平明显升高(P<0.05),牙周结缔组织中炎性细胞浸润,牙周组织损伤,牙槽骨吸收量明显增加(P<0.05),牙周组织中SOD和CAT活性明显降低(P<0.05),ROS和MDA水平明显升高(P<0.05),血清中TNF-α、IL-1β和IL-6水平明显升高(P<0.05),牙周组织中TLR4和NF-κB蛋白表达水平明显升高(P<0.05);与模型组比较,低和高剂量GSPE组大鼠一般情况均明显好转,血糖水平明显降低(P<0.05),牙周组织损伤明显减轻,牙槽骨吸收量减少(P<0.05),牙周组织中SOD和CAT活性明显升高(P<0.05),ROS和MDA水平明显降低(P<0.05),血清中TNF-α、IL-1β和IL-6水平明显降低(P<0.05),牙周组织中TLR4和NF-κB蛋白表达水平明显降低(P<0.05);与低剂量GSPE组比较,高剂量GSPE组大鼠牙周组织中SOD和CAT活性明显升高(P<0.05),ROS和MDA水平明显降低(P<0.05),血清中TNF-α、IL-1β和IL-6水平明显降低(P<0.05),牙周组织中TLR4和NF-κB蛋白表达水平明显降低(P<0.05)。结论:GSPE可改善糖尿病牙周炎大鼠牙周组织炎症,其作用机制与调节牙周组织中TLR4/NF-κB信号通路有关。

超声辅助提取葡萄籽中原花青素工艺及抗氧化活性研究

本文在单因素试验的基础上,通过设计正交实验优化葡萄籽中原花青素超声提取工艺条件,并研究了原花青素的抗氧化活性。结果表明:最佳工艺条件为乙醇浓度70%,液料比1∶25,超声时间20 min,提取2次,在此条件下原花青素的提取率为4.96%。所得原花青素提取物对ABTS、DPPH、羟基自由基都具有一定的清除率,IC_(50)值分别是0.390、0.242、0.256μg/mL。

基于现行标准的葡萄籽提取物工艺优化研究

目的:为确保葡萄籽提取物(GSE)的安全性和有效性,基于中医药保健品进出口商会和美国药典USP43标准,对GSE的提取工艺进行优化。方法:以原花青素值、儿茶素与表儿茶素含量和作为关键质量属性(CQAs),首先优选提取溶剂,并在单因素实验基础上,通过质量源于设计(QbD)理念建立了葡萄籽提取物的设计空间与优化空间。结果:单因素实验提示,丙酮对原花青素的提取效率最高,且儿茶素、表儿茶素在提取物中的含量显著低于乙醇、甲醇等提取溶剂;方差分析结果表明,通过Box-Behnken设计建立的提取工艺模型具备统计学意义(P<0.0001),葡萄籽提取物的优化提取工艺为丙酮体积分数40%~48%,料液比1∶10 g/mL,提取时间1.5 h,提取1~3次,以优化工艺参数提取,提取物中原花青素值为1.8079%~1.9643%,儿茶素和表儿茶素的含量和不高于0.8131%(RSD<5%),此优化工艺可为GSE的工业化生产提供参考。

原花青素的食用安全性研究进展

目的原花青素是一类在自然界中广泛存在的多酚类化合物,广泛分布于葡萄、松树皮、苹果、荔枝、绿茶、蔓越莓等植物中,具有抗氧化、保护心血管、抗肿瘤、护肤等生物活性,已成为我国保健食品常用的一种原料,有必要对原花青素的食用安全性进行综述。方法详细整理分析国内外文献资料,对原花青素的来源、提取、标准、毒理安全性以及各国允许使用情况等方面进行阐述分析。结果毒理学研究表明,原花青素是安全无毒,健康成年人在大剂量(2500 mg·d^(-1))服用高纯度(80%)葡萄籽原花青素提取物时未有不良事件,富含原花青素的葡萄籽、蔓越莓、越橘、海岸松、辐射松等植物提取物在不同国家被允许作为食品添加剂、膳食补充剂、新食品、保健食品原料或者非处方药使用。结论综合分析世界范围内每日指导用量,建议健康成年人的葡萄籽原花青素每日服用量为100~400 mg,为原花青素作为保健食品原料的食用安全性情况提供参考。

葡萄籽提取物对更年期模型小鼠内分泌代谢的影响

[目的]探讨葡萄籽提取物原花青素对更年期模型小鼠肠道微生物群及内分泌代谢的影响.[方法]将实验小鼠随机分为对照组(对照手术)、模型组(卵巢切除)和模型给药组(卵巢切除加给予葡萄籽提取物),模型给药组于术后当日起灌胃给予葡萄籽提取物每日0.1 mg/g 8周,其他组小鼠给予PBS.术后第6周时进行血糖检测及糖耐量实验;术前、术后第4周及第6周时分别测定各组小鼠体质量;术后第8周时收集小鼠粪便样本,分析厚壁菌和拟杆菌的菌群数量.[结果]与模型组比较,术后第6周时模型给药组小鼠30、60 min时的血糖水平明显降低(P<0.05),第4周及第6周时的体质量显著降低(P<0.01),厚壁菌数量显著减少,拟杆菌数量明显增多(P<0.05).[结论]含原花青素的葡萄籽提取物可改变小鼠肠道厚壁菌及拟杆菌数量,抑制更年期模型小鼠体质量升高,亦可改善糖耐量水平.

葡萄籽原花青素的提取及其抗紫外线活性测定

采用快速溶剂萃取法提取葡萄籽脱脂粉中的原花青素,通过单因素试验和正交试验优化提取工艺,最终获得葡萄籽原花青素提取物(grape seed proanthocyanidins extracts,GSPE)干粉。快速简便地测定不同质量浓度的GSPE乙醇溶液的防晒系数以反映抗紫外线活性,并探究GSPE乙醇溶液质量浓度对其抗紫外线活性的影响。结果表明,快速溶剂萃取法以乙醇做安全提取剂,最佳提取参数为萃取压强1100 psi、萃取时间14 min、萃取温度70℃、乙醇体积分数70%,在此条件下,提取原花青素得率为71.62 mg/g,纯度为41.53%;1000μg/mL的原花青素提取物乙醇溶液的防晒系数值达24.7556±0.7599,具有显著抗紫外线活性;GSPE抗紫外线活性与质量浓度在一定范围内呈显著正相关。

葡萄籽原花青素的毒理学研究

目的:观察葡萄籽提取物原花青素的急性毒性和长期毒性,为其安全应用提供参考。方法:急性毒性实验,对小鼠灌服不同浓度的原花青素混悬液,采用Bliss法计算其LD50。长期毒性实验,将SD大鼠随机分为正常对照组和原花青素高、中、低剂量(428.57、214.29、42.86mg(/kg·d))组,连续灌胃给药26周,停药恢复4周,观察大鼠的一般行为、体重增长、食量消耗,进行血液学及血液生化学指征、尿常规检查、系统尸解及组织病理学诊断。结果:原花青素小鼠LD50为3451.9mg/kg。长期毒性实验中,与同期对照组比较,原花青素高、中、低3个剂量组动物行为活动、进食量等均未见异常;血液学指标、血液生化指标和尿液指标未发现与供试品有关的异常改变,病理组织学检查未发现与原花青素相关的异常改变。结论:在本实验条件下未观察到葡萄籽提取物原花青素有明显毒性反应。