Blueberry anthocyanins extract attenuates oxidative stress and angiogenesis on an in vitro high glucose-induced retinopathy model through the miR-33/GLCCI1 axis

Background:Diabetes retinopathy(DR)is a complication of diabetes that affects patients’vision.Previous studies have found blueberry anthocyanins extract(BAE)can inhibit the progression of DR,but its mechanism is not completely clear.Methods:To study the role of BAE in diabetes retinopathy,we treated human retinal endothelial cells(HRCECs)with 30 mM high glucose to simulate the microenvironment of diabetes retinopathy and used BAE to intervene the in vitro high glucose-induced retinopathy model.HRCEC cell viability and apoptosis rates were examined by Cell Counting Kit 8(CCK-8)assay and flow cytometry assay.The binding sites between miR-33 and glucocorticoid-induced transcript 1(GLCCI1)were assessed by luciferase reporter assay.Retinal neovascularization and oxidative stress contribute to diabetic retinopathy.The tubule formation assay was applied to detect the retinal neovascularization.The oxidative stress in the HRCECs was manifested by the reactive oxygen species(ROS)level,the malondialdehyde(MDA)level,and the superoxide dismutase(SOD)activity.Results:Compared with HRCECs cells cultured under normal conditions,high glucose(HG)can induce oxidative stress in HRCRCs,specifically manifested in the increase of ROS and MDA levels,and the decrease of SOD activity.BAE relieved the tubule formation in n the HRCEC.BAE also relieved the ROS and MDA levels and increased the SOD activity.Luciferase reporter assay revealed that GLCCI1 is a target molecule downstream of miR-33.In HRCEC,BAE significantly inhibited the expression of miR-33 induced by HG.miR-33 mimic inhibited the BAE’s effects on oxidative stress and angiogenesis in an in vitro high glucose-induced retinopathy model.Conclusion:BAE alleviated the oxidative stress and microangiogenesis of HRCEC by regulating the miR-33/GLCCI1 axis.

Characterization of blueberry exosome-like nanoparticles and miRNAs with potential cross-kingdom human gene targets

Edible plant derived exosome-like nanoparticles(ELNs)have been shown to have multiple nutraceutical functions.However,the diversity of plant materials makes the plant derived ELN study challenging.More efforts are still needed to explore the feasible isolation methods of edible plant derived ELNs and the possible roles of food-derived ELNs in improving human health.In this study,a size exclusion chromatography based method was compared with the traditional ultracentrifugation method to isolate blueberry derived ELNs(B-ELNs),and the miRNA profile of B-ELNs was analyzed by high-throughput sequencing.A total of 36 miRNAs were found to be enriched in B-ELNs compared with berry tissue,and their potential cross-kingdom human gene targets were further predicted.Results showed that size exclusion chromatography was effective for B-ELN isolation.The most abundant miRNAs in B-ELNs mainly belonged to the miR166 family and miR396 family.Target gene prediction indicated that B-ELNs could potentially regulate pathways related to the human digestive system,immune system and infectious diseases.

Major Diseases and Pests of Blueberry in Guizhou Province and Progress in Their Prevention and Control

As the blueberry industry continues to evolve,the effective control of its diseases and pests has become an essential component of local agricultural development.This paper provides a comprehensive overview of the principal types of blueberry diseases and pests in Guizhou Province,along with the corresponding control measures,in order to serve as a valuable reference for blueberry growers.

植物乳杆菌发酵蓝莓汁发酵和贮藏过程中理化指标及挥发性风味物质的变化

为了解植物乳杆菌发酵对蓝莓汁在发酵和贮藏过程中理化指标和挥发性风味物质的变化规律的影响,以新鲜巴尔德温为原料,利用植物乳酸菌b-1发酵蓝莓汁,研究发酵和贮藏过程中理化指标和活菌数的变化,并利用HPLC和顶空固相微萃取气质联用(HS-SPME-GC/MS)测定蓝莓汁的有机酸和风味代谢物,分析其发酵和贮藏过程中风味品质的变化规律。结果表明,发酵24 h的蓝莓汁中的活菌数达到了108 CFU/mL;储藏28 d后活菌数为6.4×107 CFU/mL。蓝莓汁发酵期间理化性质变化较大,发酵过程中总酸含量逐渐增加,而pH呈现显著下降趋势(P<0.05)。在蓝莓汁发酵和贮藏过程中,共检测到6种有机酸,其中乳酸是发酵后的产物。贮藏过程中发酵蓝莓汁中挥发性风味物质分布为醇类>酮类>酯类>醛类>烷类>烯类。基于PCoA和OPLS-DA分析结果表明,利用植物乳杆菌发酵的蓝莓果汁在发酵期和贮藏期的风味代谢物差异较大,分别检测到14种、9种差异代谢物,且发酵阶段发酵蓝莓汁中芳樟醇和香叶醇的相对含量有所上升,增强了果汁的柑橘香、花香和玫瑰香气。该研究为植物乳杆菌在蓝莓发酵果汁中的应用提供了科学支撑。

不同乳酸菌发酵蓝莓桑葚原浆营养品质及风味变化分析

为探究不同乳酸菌发酵对蓝莓桑葚原浆营养品质及风味的影响,本研究分别以保加利亚乳杆菌、干酪乳杆菌、植物乳杆菌作为发酵剂,利用电子鼻和顶空微萃取结合气相色谱-质谱法等方法分析乳酸菌发酵后蓝莓桑葚原浆中的功能营养成分和风味。结果表明,3个发酵组的营养品质和抗氧化能力差异显著,其中植物乳杆菌发酵组总酸含量最高(1.52 g·L^(-1))、总糖含量最低(789.59 mg·L^(-1))、总花色苷含量最高(6.04 g·L^(-1)),总抗氧化能力最强;电子鼻分析结果可有效区分3种菌种的发酵风味;气相色谱-质谱共检测出51种挥发性成分,包括醇类6种、酸类14种、醛类9种、酮类7种、酯类8种、其他物质7种,其中2-壬酮在植物乳杆菌发酵组中含量较高,赋予发酵汁果香以及花香。综合而言,植物乳杆菌发酵组在营养成分、气味及风味物质方面均有较大提升,更适用于蓝莓桑葚原浆的发酵,有助于开发出具有抗氧化作用的益生元蓝莓果浆。本研究可为蓝莓桑葚深加工提供一定的借鉴。

蓝莓新品种珍珠蓝的选育

珍珠蓝为北高丛蓝莓新品种,由蓝金自然杂交实生苗选育而成。中等果穗密度,中等厚度且质地均匀的果粉,小而干的果蒂痕,果实蓝色,呈扁圆形,果实较硬(3.82 kg·cm^(-2)),单果平均大小为1.43 cm×1.77 cm,单果平均质量为2.32 g,果实最大可达2.48 g,可溶性固形物含量(w)为17.2%,甜度高、酸度低。该品种的始熟期大约在6月下旬,早熟种。该品种风味好,产量中等,适宜鲜食。

蓝莓转录因子VcICE全基因组鉴定及其在花果发育进程的表达分析

为探究ICE(inducer of CBF expression)转录因子在高丛蓝莓花果发育进程中可能的调控机制,通过对VcICE基因家族进行全基因组鉴定及生物信息学分析,采用实时荧光定量PCR(qRT-PCR)技术分析VcICE基因亚家族成员在蓝莓花芽膨大和果实发育进程中的相对表达水平.结果表明:在高丛蓝莓‘Draper’基因组中鉴定出8个VcICE成员,系统进化分析将其划分为2个亚家族,各亚家族成员均包含S-rich,bHLH和ACT等保守结构域;VcICE基因启动子区域富含光响应、植物激素、生长发育及非生物胁迫相关顺式作用元件;VcICE亚家族基因在高丛蓝莓‘O’Neal’和‘Bluerain’花芽膨大与果实发育进程中的相对表达差异显著.研究结果以期为深入了解VcICE转录因子在蓝莓花果发育进程中的功能提供理论参考.

利用低场核磁共振技术探究机械损伤对蓝莓果实内部水分状态的影响

本研究采用自制损伤装置制作不同损伤等级样本(DI组(撞击1次)和DII组(撞击不同位置2次)),利用低场核磁共振技术,在4℃和25℃两种贮存温度条件下,采集蓝莓损伤发生0、6、24 h的波谱信息和质子密度图像信息。结果表明,损伤显著改变了蓝莓内部水分状态与分布。随着时间的延长,受损蓝莓细胞壁水含量A21减少,而细胞质水含量A22在损伤发生后逐渐上升,特别在损伤程度高的DII组,损伤发生24 h后A22增至0 h的1.45倍以上,而液泡水含量A23和总水分含量A2大幅度下降,表明蓝莓损伤后水分重新分配和向外界环境迅速转移。随着损伤程度上升,液泡水主峰整体呈现显著下降和右移的趋势,同时温度的升高进一步加剧了该变化。在25℃条件下,损伤发生6 h和24 h时,出现了4个弛豫峰,弛豫峰的增加标志着蓝莓品质显著劣变。相比于室温(25℃),4℃低温贮存有利于保持果实的完整性,并减缓受损蓝莓水分流失速率。实验结果可为实现蓝莓机械损伤的快速检测和损伤机理研究提供理论基础和数据参考。

蓝莓真空冷冻干燥工艺优化

为提高蓝莓的真空冷冻干燥速率和品质,对不同隔板温度(40、45、50、55℃)、真空度(65、70、75、80 Pa)及预冻温度(-65、-60、-55、-50℃)的干燥效果进行了探讨,研究了不同干燥工艺对蓝莓干燥特性和品质的影响。在单因素试验的基础上,采用响应面法优化确定了蓝莓真空冷冻干燥的最优工艺参数并进行了验证。结果表明:隔板温度、真空度和预冻温度越高,干燥时间越短,干燥速率越快。适当降低隔板温度和预冻温度可以在一定程度上保持冻干蓝莓的颜色、硬度、花青素含量和抗坏血酸含量,并提高复水比,但真空度的降低会增大冻干蓝莓的色差并降低硬度。蓝莓真空冷冻干燥最佳工艺参数为:隔板温度44℃,真空度71 Pa,预冻温度-55℃。研究结果为优化真空冷冻干燥工艺和提升冻干蓝莓的品质提供了理论依据和参考。

不同蓝莓酒挥发性风味成分和感官品质差异及相关性分析

该研究采用全二维气相色谱-飞行时间质谱(GC×GC-TOFMS)技术结合相对气味活度值(ROAV)法对5款市售商品蓝莓酒挥发性风味成分、重要香气物质、关键香气物质及特征香气物质进行解析,通过感官定量描述分析(QDA)法分析其感官品质,并对挥发性风味成分与感官评价进行Pearson相关性分析。结果表明,5种蓝莓酒共鉴定出挥发性风味成分411种,其中酯类85种、苯环化合物74种、醇类69种、萜类73种、酮类34种、杂环化合物26种、醛类18种、烃类10种、酸类3种和其他类19种。5款商品蓝莓酒中共检出的重要香气化物质(0.1<ROAV<1)38种,关键香气物质(ROAV>1)16种,特征香气物质(ROAV>10)4种。相关性分析结果显示,(E)-β-紫罗兰酮、D-柠檬烯和1-辛烯-3-酮与蓝莓酒呈现的特殊果香和植物香气特征具有显著相关性(P<0.05)。

Adaptability of Some Blueberry Species in Tianjin

[Objective] The aim was to research the adaptability of 8 blueberry species in Tianjin. [Method] With 8 blueberry species as materials, blueberry was explored in terms of plant characters, the type of fruit-bearing branch, phenological phase, fruit economic characters, and yielding capacity. [Result] Al blueberry species were suitable to be grown in Tianjin with improved soils, showing blueberry growth characters, without any virus or diseases. General y speaking, species B, C, D, E, F and G can be grown in soils exposed to the air as fresh food; species A and H are suitable to be cultivation; species C tastes sour, which can be planted for further production. [Conclusion] The research provides references for blueberry introduction, selection, and cultivation in the region with similar ecological characters.

响应面法优化蓝莓酒渣花青素酶法提取工艺

为了高质化利用蓝莓酒渣,以纤维素酶为对照,选择淀粉酶、果胶酶、酸性蛋白酶、中性蛋白酶为实验组,以无水乙醇为提取溶剂提取蓝莓酒渣中的花青素。采用单因素试验考察提取酶添加量、提取时间、提取温度以及初始pH值对花青素提取量的影响,并通过响应面试验优化蓝莓酒渣花青素酶法提取工艺。结果表明,酶法提取花青素的最佳工艺条件为淀粉酶添加量0.7%,蓝莓酒渣与无水乙醇料液比为1∶100(g∶m L),提取温度60℃,提取时间为40 min,初始pH值为5。在此优化条件下,花青素提取量为0.68 mg/g,比未优化前提高了20.8%。

脉冲强光处理对蓝莓汁杀菌效果及品质的影响

为探究脉冲强光(intense pulsed light,IPL)处理对蓝莓汁杀菌效果和品质的影响,以微生物数量、酶活力、营养成分(总花色苷、维生素C、总酚)含量为指标,分析不同脉冲强度和次数的IPL处理对蓝莓汁活菌数和营养品质的影响,并通过气相色谱-质谱(gas chromatography-mass spectrometry,GC-MS)法分析蓝莓汁挥发性风味物质的变化。结果表明:IPL处理后,蓝莓汁中微生物数量降低到20 CFU/mL以下;过氧化物酶完全失活,超氧化物歧化酶活力保留率高于(88.68±0.18)%;蓝莓汁没有发生褐变;总酚含量得以完全保留。400 J强度的IPL处理对蓝莓汁抗氧化能力影响不显著(P>0.05),总花色苷、维生素C含量保留率分别高达(84.83±1.30)%、(95.00±2.59)%。GC-MS分析表明,香气成分中的醛类、醇类、酯类含量有一定的损失,新增异蒲勒醇、柏木烯醇等挥发性风味物质。IPL处理能够有效杀灭蓝莓汁中的微生物,同时保留较多的营养成分。

蓝莓来源的外泌体样纳米颗粒在MNNG诱导人正常胃黏膜上皮细胞恶性转化中的干预效应

目的:提取蓝莓来源的外泌体样纳米颗粒(blueberry derived exosome-like nanoparticles,BELNs)并进行鉴定,探讨BELNs在1-甲基-3-硝基-1-亚硝基胍(N-methyl-N′-nitro-N-nitrosoguallidine,MNNG)诱导人正常胃黏膜上皮细胞恶性转化中的干预效应。方法:采用超速离心法提取BELNs,纳米颗粒跟踪分析仪和透射电镜分别从粒径、Zeta电位、形态对获取的BELNs进行鉴定;激光共聚焦显微镜观察BELNs能否被MNNG诱导的人正常胃黏膜上皮细胞,即胃黏膜上皮细胞转化细胞(transformed gastric mucosal epithelial cells,TGES-1)内吞;用200、400μg/mL的BELNs处理TGES-1细胞72 h后,蛋白质印迹法分析干性基因、上皮细胞间充质转化(epithelial-mesenchymal transition,EMT)及增殖相关蛋白表达的变化。结果:纳米颗粒跟踪分析仪和透射电镜等结果显示BELNs符合外泌体样纳米颗粒的特征,表明成功提取BELNs;激光共聚焦显微镜结果提示BELNs可被TGES-1细胞内吞;蛋白印迹结果表明,400μg/mL的BELNs抑制TGES-1细胞中N-钙黏蛋白(N-cadherin)、波形蛋白(Vimentin)、同源框蛋白(Nanog)和增殖细胞核抗原(proliferating cell nuclear antigen,PCNA)的表达,增强E-钙黏蛋白(E-cadherin)的表达。结论:BELNs在MNNG诱导人正常胃黏膜上皮细胞恶性转化中具有明显的干预效应。

响应面法优化高纤低卡蓝莓果渣司康配方研究

司康配方中使用低热量原料,并添加脂肪替代物蓝莓果渣粉,能够制作出低脂、低糖、高纤的烘焙产品。基于单因素试验、响应面试验、验证试验和感官评价、理化指标检测,最终优化出功能性蓝莓果渣司康的最佳配方:全麦面粉132 g、蓝莓果渣粉18 g、椰子油21 g、牛奶53 g、零卡糖20 g、盐1 g、鸡蛋1个,为蓝莓果渣的废物利用和功能性烘焙食品的开发提供了理论依据和实践借鉴。

Study on Grafting and Cultivation of ‘Spartan'Blueberry

The results of grafting improvement and cultivation trials in the Yangtze River Delta of China showed that the rootstock cultivar had a great influence on plant growth and development, fruit yield and quality of ＂Spartan＂ blueberry. Among the seven rootstock-scion combinations, the northern highbush blueberry （Vaccinium corymbosum） cultivar ＂Croaton＇-＇Spartan＇ combination showed well-healed graft unions, large branch amount, early fruit bearing, high yield, excellent fruit quality and almost no tillers. As a kind of rootstock, ＂Croaton＇ was better than non-grafted ＇Spartan＇ and the two rabbiteye blueberry cultivars ＇Premier＇ and ＇Gardenblue＇. The grafting pattern showed no significant stimulating effect on the plant growth and development, fruit yield and quality of ＇Spartan＇ blueberry in this area, but the overall performance of greenwood cleft grafting was better than that of inlaid bud grafting. Suitable rootstock cultivar and grafting pattern significantly improve plant arowth and development, fruit vield and aualitv of ＇Spartan＇ blueberry.

富硒酵母对蓝莓酒发酵过程中理化性质和风味的影响

为分析富硒酵母对蓝莓酒发酵过程中理化性质和风味的影响,采用外源供硒方式,测定AG1、AG2和LM等4种酵母的富集硒元素能力,筛选出富硒效果最佳的酵母进行蓝莓酒发酵,并对发酵过程中的发酵液氨基酸态氮、总酸和pH等理化指标与主要风味物质进行测定。结果表明,AG2(15 mg/L)的富硒效果最佳;其在蓝莓酒发酵过程中,蓝莓酒液的氨基酸态氮和总酸含量呈上升趋势,在发酵第20天的含量最高,分别为6.2和7.6 g/L;pH先缓慢下降,后趋于稳定;发酵初期(2~6 d)蓝莓酒中的还原糖含量急速下降,后趋于稳定;随着发酵时间的延长,发酵液中酒精度呈先增加后不变的趋势,在第10天酒精度值最大,为13.8%vol。接种该酵母的蓝莓酒与普通酵母发酵的主要风味物质种类和相对含量上既有相似之处,也存在一定差异,二者均以异戊醇、苯乙醇、2-甲基-1-丁醇和2,3-丁二醇等物质发挥风味。本研究为蓝莓酒富硒机理的深度研究提供参考。

Effect of blueberry on hepatic and immunological functions in mice

BACKGROUND: Conventional drugs used in the treatment and prevention of liver diseases often have side effects, therefore research into natural substances are of significance. This study examined the effects of blueberry on liver protection and cellular immune functions. METHODS: To determine the effects of blueberry on liver protective function, male mice were orally administered blueberry (0.6 g/10 g) or normal saline for 21 days. Hepatic RNA was extracted by Trizol reagent, and the expression of Nrf2, HO-1, and Nqol was determined by real-time RTPCR. Superoxide dismutase (SOD) and malondialdehyde (MDA) in liver homogenate were determined, and liver index was measured. To assess the effects of blueberry on cellular immune function, male mice received blueberry (0.4, 0.6, or 0.8 g/10 g) for 35 days, and the percentages of CD3(+), CD41(+), and CDS(+) T lymphocyte subgroups in peripheral blood were detected by flow cytometry, the index of the thymus and spleen was measured, and lymphocyte proliferation in the spleen was determined by MTT assay. RESULTS: Blueberry treatment significantly increased the expression of Nrf2, HO-1, and Nqol, the important antioxidant components in the liver. Hepatic SOD in the blueberry group was higher and MDA was lower than that in the control group (P<0.05). Blueberry also increased the index of the spleen and enhanced the proliferation of lymphocytes of the spleen (P<0.05). The percentages of the CD3(+) and CDA(+) T lymphocyte subsets and the CD4(+)/CD8(+) ratio were also increased by blueberry (P<0.05). CONCLUSIONS: Blueberry induces expression of Nrf2, HO-1, and Nqol, which can protect hepatocytes from oxidative stress. In addition, blueberry can modulate T-cell function in mice. (Hepatobiliary Pancreat Dis Int 2010; 9: 164-168)

Effects of blueberry on hepatic fibrosis and transcription factor Nrf2 in rats

AIM:To investigate the effects of blueberry on hepatic fibrosis and NF-E2-related factor 2(Nrf2) transcription factor in rats.METHODS:Forty-five male Sprague-Dawley rats were randomly divided into control group(A);CCl4-induced hepatic fibrosis group(B);blueberry prevention group(C);Dan-shao-hua-xian capsule(DSHX) prevention group(D);and blueberry + DSHX prevention group(E).Liver fibrosis was induced in rats by subcutaneous injection of CCl4 and a high-lipid/low-protein diet for 8 wk(except the control group).The level of hyaluronic acid(HA) and alanine aminotransferase(ALT) in serum was examined.The activity of superoxide dismutase(SOD),glutathione-S-transferase(GST) and malondialdehyde(MDA) in liver homogenates was determined.The degree of hepatic fibrosis was evaluated by hematoxylin and eosin and Masson staining.Expression of Nrf2 and NADPH quinone oxidoreductase 1(Nqo1) was detected by real-time reversed transcribed-polymerase chain reaction,immunohistochemical techniques,and western blotting.RESULTS:Compared with group B,liver indices,levels of serum HA and ALT of groups C,D and E were reduced(liver indices:0.038 ± 0.008,0.036 ± 0.007,0.036 ± 0.005 vs 0.054 ± 0.009,P<0.05;HA:502.33 ± 110.57 ng/mL,524.25 ± 255.42 ng/mL,499.25 ± 198.10 ng/mL vs 828.50 ± 237.83 ng/mL,P<0.05;ALT:149.44 ± 16.51 U/L,136.88 ± 10.07 U/L,127.38 ± 11.03 U/L vs 203.25 ± 31.62 U/L,P<0.05),and SOD level was significantly higher,but MDA level was lower,in liver homogenates(SOD:1.36 ± 0.09 U/mg,1.42 ± 0.13 U/mg,1.50 ± 0.15 U/mg vs 1.08 ± 0.19 U/mg,P<0.05;MDA:0.294 ± 0.026 nmol/mg,0.285 ± 0.025 nmol/mg,0.284 ± 0.028 nmol/mg vs 0.335 ± 0.056 nmol/mg,P<0.05).Meanwhile,the stage of hepatic fibrosis was significantly weakened(P<0.05).Compared with group A,the activity of GST liver homogenates and expression levels of Nrf2 and Nqo1 in group B were elevated(P<0.05).The expression level of Nrf2 and Nqo1 in groups C,D,and E were increased as compared with group B,but the difference was not significant.CONCLUSION:Blueberry has preventive and protective effects on CCl4-induced hepatic fibrosis by reducing hepatocyte injury and lipid peroxidation.However,these effects may not be related to the activation of Nrf2 during long-term of CCl4.

High-throughput sequencing of highbush blueberry transcriptome and analysis of basic helix-loop-helix transcription factors

The highbush blueberry（Vaccinium corymbosum）,Duke,was used to construct a de novo transcriptome sequence library and to perform data statistical analysis.Mega 4,CLC Sequence Viewer 6 software,and quantitative PCR were employed for bioinformatics and expression analyses of the basic helix-loop-helix（BHLH）transcription factors of the sequencing library.The results showed that 28.38 gigabytes of valid data were obtained from transcriptome sequencing and were assembled into 108 033 unigenes.Functional annotation showed that 32 244 unigenes were annotated into Clusters of Orthologous Groups（COG）and Gene Ontology（GO）databases,whereas the rest of the 75 789 unigenes had no matching information.By using COG and GO classification tools,sequences with annotation information were divided into 25 and 52 categories,respectively,which involved transport and metabolism,transcriptional regulation,and signal transduction.Analysis of the transcriptome library identified a total of 59 BHLH genes.Sequence analysis revealed that 55 genes of that contained a complete BHLH domain.Furthermore,phylogenetic analysis showed that BHLH genes of blueberry（Duke）could be divided into 13 sub-groups.PCR results showed that 45 genes were expressed at various developmental stages of buds,stems,leaves,flowers,and fruits,suggesting that the function of BHLH was associated with the development of different tissues and organs of blueberry,Duke.The present study would provided a foundation for further investigations on the classification and functions of the blueberry BHLH family.