葡萄属植物白藜芦醇研究进展

根据有关文献分别从白藜芦醇穴Res雪的发现、化学结构、物理特性、对葡萄属植物及人体健康的作用、作用机理方面作了简要概述,总结了其诱导、取样、测定方法,在葡萄植物体内的分布、动态变化、器官之间含量的相关性,葡萄属植物种间、品种间及葡萄加工品之间的含量差异,并介绍了白藜芦醇的生物合成、几种合成酶,影响代谢的内部、外部因素,对白藜芦醇的研究意义及应用前景也进行了讨论。     

Resveratrol protects cortical neurons in infant rats with bacterial meningitis through miR-223-3p/NLRP3 pathway

AIM To investigate the protective effect of resveratrol (Res) on cortical neurons in rat bacterial meningitis (BM) model. METHODS Group B hemolytic Streptococcus was injected via the posterior cistern to establish a BM model. Resveratrol was administered intranasally and microRNA-223-3p (miR-223-3p) antagomir was administered by intracerebroventricular injection. HE staining was used to observe the pathological changes of the brain tissue. Loeffler scoring method was used to evaluate the neurobehavioral functions. TUNEL staining was used to detect neuronal apoptosis. The expression of interleukin-1β (IL-1β), IL-18, glial fibrillary acidic protein (GFAP) and ionized calcium-binding adaptor molecule 1 (Iba1) was detected by immunofluorescence staining. The protein levels of nucleotide-binding oligomerization domain-like receptor protein 3 （NLRP3）, cleaved caspase-1, IL-1β and IL-18 were determined by Western blot. The expression level of miR-223-3p was detected by RT-qPCR. Online software TargetScan was used to search for the complementary nucleotide sequences between miR-223-3p and NLRP3 mRNA. RESULTS Compared with sham group, the thickness of meninges in BM model was increased, the neurological score was decreased (P<0.05), and the number of TUNEL positive neurons was increased significantly (P<0.05). Astrocytes and microglia were activated, the fluorescence intensity of IL-1β and IL-18 was increased (P<0.05), and the expression levels of NLRP3, cleaved caspase-1, IL-1β, IL-18 and miR-223-3p were increased (P<0.05). Compared with BM group, after treatment with resveratrol, the neurological score was increased (P<0.05), the number of TUNEL positive neurons was decreased significantly (P<0.05), and the inflammatory response of astrocytes and microglia was suppressed. The fluorescence intensity of IL-1β and IL-18 was decreased (P<0.05), the protein levels of NLRP3, cleaved caspase-1, IL-1β and IL-18 were decreased (P<0.05), and the expression level of miR-223-3p was increased (P<0.05). A nucleotide sequence in the 3'-UTR of NLRP3 mRNA might be targeted by miR-223-3p. In the brain of rat BM model, compared with antagomir control group, the expression of NLRP3 was increased in miR-223-3p antagomir group with resveratrol treatment (P<0.05). CONCLUSION Resveratrol may reduce the inflammatory death of cortical neurons in BM model of infant rats through miR-223-3p/NLRP3 pathway, thus playing a protective role for the neurons.     

UV-C辐射对盆栽葡萄北丰叶片和邻近果穗果实白藜芦醇及其糖苷质量分数的影响

以1a生北丰(Vitis thunbergii×V.vinifera)葡萄盆栽结果树为试材,在果实始熟期采用紫外线C(Ultraviolet C,UV-C)辐射结合韧皮部环剥的方法,研究了UV-C辐射对葡萄叶片和邻近果穗果实白藜芦醇(Resveratrol,Res)及其糖苷质量分数的影响。结果表明,UV-C辐射诱导葡萄果实Res合成积累的敏感性低于叶片,辐射葡萄叶片对邻近果穗上果皮Res的影响远远小于UV-C直接辐射葡萄果穗,且辐射诱导葡萄果皮中Res质量分数与叶片中Res质量分数存在着一定的相关性。当叶片位于果穗下方时,叶片中反式白藜芦醇(trans-Resveratrol,trans-Res)、反式白藜芦醇苷(trans-Piceid,trans-PD)和顺式白藜芦醇苷(cis-Piceid,cis-PD)质量分数表现出环剥处理显著高于不环剥处理,然而果皮中的结果正好相反。当叶片位于果穗上方时,果皮中trans-Res、trans-PD和cis-PD质量分数表现出不环剥处理显著高于环剥处理,暗示葡萄Res可能存在韧皮部运输途径,且运输方向是双向的。此外,UV-C直接辐射葡萄果穗处理果皮trans-Res和cis-PD质量分数显著高于对照和UV-C辐射葡萄叶片处理。种子中仅检测到trans-Res,且仅环剥处理和辐射下方叶片不环剥处理显著高于对照。     

白藜芦醇的研究进展

白藜芦醇是一种植物抗毒素。该文主要介绍了几种能够产生工业价值且无毒无害符合食品安全的白藜芦醇提取方法:乙醇水溶液溶剂法、酶化学法和生物发酵法,从防癌、抗肿瘤、保护心血管、抗衰老和美容健体4个方向挖掘我国白藜芦醇的利用现状,阐述我国白藜芦醇生物功效的研究进展,为开发具有药用、保健功效的白藜芦醇产品提供依据。     

白藜芦醇浸提及纯化过程在线检测方法研究

[目的]比较并筛选出适用于白藜芦醇在线检测的方法。[方法]通过试验对比了高效液相色谱(HPLC)和紫外分光光度(UV)法测定花生根白藜芦醇的异同。[结果]从线性相关性看,R(UV)=0.999 6,R(HPLC)=0.999 8,2种方法都有良好的线性度,都适用于花生根白藜芦醇的检测;从精密度[RSD(HPLC)=1.87%,RSD(UV)=2.52%]和回收率[HPLC 98.8%～101.2%,RSD为1.25%;UV法97.8%～103.5%,RSD为2.25%]看,HPLC法优于紫外分光光度法;从平均测得结果看,紫外分光光度法测得白藜芦醇含量为0.699 1%、RSD=2.52%(n=4),HPLC法测得白藜芦醇含量为0.690 4%,RSD=1.87%(n=4),2种方法结果相近。[结论]本着在线监测和仪器的普适性,紫外分光光度法更有优势,更适用于花生根白藜芦醇的检测。     

Resveratrol inhibits chondrosarcoma via mitochondrial and PI3K/Akt signaling pathways

AIM：To investigate the inhibitory effects of resveratrol on chondrosarcoma and the relation with mitochondrial and PI3K/Akt pathways. METHODS：Chondrosarcoma SW1353 cells were treated with resveratrol at concentrations of 25, 50 and 100 μmol/L for the time intervals of 24 h, 48 h and 72 h. The viability and apoptosis of the SW1353 cells in the presence or absence of resveratrol were analyzed by CCK8 assay and Hoechst 33258 staining, respectively. The protein levels of Bcl-2, Bax, activated caspase-3, Akt and p-Akt were detected by Western blotting. The cell migration ability was determined by wound scratch assay. RESULTS：Exposure of the cells to resveratrol resulted in a decrease in the cell viability in a dose- and time-dependent manner (P<0.05). visible nuclei with apoptotic characteristics in resveratrol group were observed. The protein levels of activated caspase-3 and Bax were increased, and Bcl-2 and p-Akt were decreased compared with control group. The total Akt were not significantly changed. Resveratrol also significantly reduced the migration of tumor cells. CONCLUSION：Resveratrol induces apoptosis of chondrosarcoma, which plays a role of part through mitochondrial and PI3K/Akt signaling pathways.     

芪合酶基因的克隆及其酵母表达载体的构建

应用基因工程技术将从中国野生属葡萄种高抗白粉病的华东葡萄白河-35-1中分离出的芪合酶基因cDNA片段（Stilbene synthase，STS）进行PCR扩增；把回收的目的基因PCR产物定向插入克隆载体pGEM-TEasy中进行测序与序列分析；采用定向克隆的方法将芪合酶基因克隆到大肠杆菌-酵母菌穿梭型酿酒酵母表达载体pYES6／CT上，转化大肠杆菌，提取阳性重组质粒转化酿酒酵母菌（Saccharomyces cerevisiae）菌株INVScl，酵母菌落PCR电泳结果显示，在约1200bp处有一条亮带，证明芪合酶基因酵母表达载体构建成功，为后期工作的开展奠定了基础，并为开发一种含有白藜芦醇生物活性成分的酵母微生态制剂提供了重要的实验素材。     

花生种子白藜芦醇含量与黄曲霉产毒抗性的关系

选用抗黄曲霉产毒的花生品种(系)和高产毒品种(系), 采用强产毒菌株AF2202人工接种水分吸涨的花生种子, 培养7 d后,测定接种和未接种的花生种子中白藜芦醇及黄曲霉毒素含量,探讨白藜芦醇与花生种子黄曲霉产毒抗性之间的关系。结果表明,抗黄曲霉产毒花生品种(系)的白藜芦醇含量较高,平均为37.3 µg kg^–1,高产毒品种含量相对较低,平均为13.3 µg kg^–1,抗、感品种之间存在显著差异。吸胀处理后抗产毒花生品种(系)白藜芦醇含量提高2.0倍,感病品种(系)仅提高1.6倍,对不同品种(系)处理后的种子二次接种,令黄曲霉毒素含量下降37.6%~75.8%,但吸胀处理后抗产毒品种(系)的黄曲霉毒素含量仍低于感病品种(系)。相关分析表明,花生白藜芦醇含量与黄曲霉毒素含量呈显著负相关,并且在离体培养基中添加浓度大于3.0 μg mL^–1的白藜芦醇可导致黄曲霉菌产毒量大幅下降,说明白藜芦醇对黄曲霉产毒具有抑制作用。     

白藜芦醇对HA-VSMC细胞膜电位和钙离子的影响

[目的]研究白藜芦醇对人主动脉血管平滑肌细胞(T/G HA-VSMC)膜电位和游离钙浓度的影响。[方法]用荧光染料DiBAC4(3)和Fluo-3-AM分别标记细胞,在流式细胞仪上分别测定细胞膜电位及胞内游离钙浓度的变化。[结果]给予Res后,HA-VSMC膜电位去极化,胞内游离钙浓度升高。[结论]Res可去极化HA-VSMC细胞膜电位,促进钙内流。     

酶法提取虎杖中白藜芦醇的工艺优化

[目的]对酶法提取虎杖中白藜芦醇的工艺条件进行优化。[方法]采用单因素试验与正交试验相结合的方法,确定用酶法从虎杖中提取白藜芦醇的最佳工艺条件。[结果]从虎杖中提取白藜芦醇的最佳工艺为:料水比1∶15(V/V),漆酶含量0.30%,纤维素酶含量0.50%,木聚糖酶含量0.40%,β-葡聚糖酶含量0.20%,pH 4.4,在此条件下白藜芦醇的得率达0.592%。[结论]该研究为白藜芦醇的开发及综合利用提供了研究基础。