Effects of Different Harvest Periods on Seed Quality of Alpinia oxyphylla Miq.

[Objectives] To clarify the appropriate harvest period of Alpinia oxyphylla Miq. seed and improve the quality of seeds. [Methods] In this experiment,the basic parameters and germination characteristics of A. oxyphylla Miq. seeds during different harvest periods were analyzed and compared. [Results] The results showed that the A. oxyphylla Miq. fruit picking period was closely related to the seed quality,and the seed maturity was highest 100 d after the flowering of A. oxyphylla Miq.,that is,the A. oxyphylla Miq. fruit skin turned from green to yellow( yellow-green),the peel had a small number of brown spots,it was spicy enough,the fruit dry/fresh weight ratio was greater than0. 34,the seed thousand kernel weight reached 11 g or more,and the dried seed moisture content was 13. 01%. Using the TTC method,the seed viability index was determined to be 85%,and in this case,it was the best time of harvesting seed. The water permeability of A. oxyphylla Miq. seed was poor,and the seed imbibition and water absorption time was long,about 72 h. The maximum water absorption rate was about23. 71% at the imbibition and water absorption stage,and it was inferred that the A. oxyphylla Miq. seeds were hard seeds. The germination time of A. oxyphylla Miq. seeds was long,the mature A. oxyphylla Miq. seeds began to germinate on the 12 th d,and the germination rate reached 75. 56% on the 50 th d. [Conclusions] This study determined the optimum harvest period of A. oxyphylla Miq. seeds,clarified the biological characteristics of slow seed water absorption and long germination cycle,and concluded that the seed coat was the main factor leading to slow seed water absorption and long seed germination cycle.

Network pharmacology and verification experiment-based prediction of active components and potential targets of Alpiniae Oxyphyllae Fructus-Saposhnikoviae Radix(Yizhiren-Fangfeng)for treatment of diabetic kidney disease

Background:In this study,we analyzed the potential active components,related crucial targets and possible signaling pathway mechanisms of Alpiniae Oxyphyllae Fructus and Saposhnikoviae Radix(AOF-SR)herb pairs in the treatment of diabetic kidney disease(DKD)using network pharmacology and verification experiments.Methods:The active compounds and potential targets of AOF-SR were derived from the Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform,The Encyclopedia of Traditional Chinese Medicine,and PubChem databases,and the potential therapeutic targets of DKD were derived from the OMIM,Drugbank,and DisGeNET databases.The“compounds-diseases-targets”network was constructed using Cytoscape 3.6.0.ClusterMaker functionality in Cytoscape is being used to screen important targets for AOF-SR treatment of DKD.Gene Ontology and Kyoto Encyclopedia of Genes and Genomes enrichment analysis of important targets were performed using DAVID database.In addition,according to the predicted results of network pharmacology,HK-2 cells were used to construct DKD model for verification experiment.HK-2 cells were divided into control group,high glucose(HG)group and AOF-SR(HG+AOF-SR)group to detect survival rate and expression of key proteins in NF-κB and PI3K/Akt signaling pathways.Results:A total of 38 compounds were selected from AOF-SR,of which 23 were Alpiniae Oxyphyllae Fructus and 15 were Saposhnikoviae Radix.Through enrichment analysis of 82 important targets,88 signaling pathways were identified;some of these pathways,such as the NF-κB,PI3K-Akt,IL-17,and JAK/STAT signaling pathways,regulate the pathological process of DKD.In verification experiment,the HK-2 cells survival rate was higher in the HG+AOF-SR group than in the HG group(P<0.05).Moreover,western blotting results showed that the expression levels of NF-κB,p-PI3K,and p-Akt in HG+AOF-SR group were significantly lower than those in HG group(P<0.05).Conclusion:Overall,this study revealed the active compounds,important targets and possible mechanisms of AOF-SR treatment for DKD,and conducted preliminary verification experiments on its correctness,provided novel insights into the treatment of DKD by AOF-SR.

Oxyphyllones A and B,novel sesquiterpenes with an unusual 4,5-secoeudesmane skeleton from Alpinia oxyphylla

Two novel 4,5-secoeudesmane sesquiterpenoids, oxyphyllones A （1） and B （2） were isolated from the fruits of Alpinia oxyphylla. Their structures were established by spectroscopic methods including 1D and 21） NMR spectra. These two compounds are the first example of naturally occurring sesquiterpenoids with a 4,5-secoeudesmane skeleton in the family of Zingiberaceae and oxyphyllone A （1） is the first 4,5-secoeudesmane type of 13-norsesquiterpenoid. Compounds 1 and 2 exhibited no cytotoxicities against three cancer cell lines at 10 μg/mL. ？ 2009 Ning Hua Tan. Published by Elsevier B.V. on behalf of Chinese Chemical Society. All rights reserved.

Protective effects of organic extracts of Alpinia oxyphylla against hydrogen peroxide-induced cvtotoxicitv in PC12 cells

Alpinia oxyphylla,a traditional herb,is widely used for its neuroprotective,antioxidant and memory-improving effects.However,the neuroprotective mechanisms of action of its active ingredients are unclear.In this study,we investigated the neuroprotective effects of various organic extracts of Alpinia oxyphylla on PC12 cells exposed to hydrogen peroxide-induced oxidative injury in vitro.Alpinia oxyphylla was extracted three times with 95%ethanol(representing extracts 1–3).The third 95%ethanol extract was dried and resuspended in water,and then extracted successively with petroleum ether,ethyl acetate and n-butanol(representing extracts 4–6).The cell counting kit-8 assay and microscopy were used to evaluate cell viability and observe the morphology of PC12 cells.The protective effect of the three ethanol extracts(at tested concentrations of 50,100 and 200μg/mL)against cytotoxicity to PC12 cells increased in a concentration-dependent manner.The ethyl acetate,petroleum ether and n-butanol extracts(each tested at 100,150 and 200μg/mL)had neuroprotective effects as well.The optimum effective concentration ranged from 50–200μg/mL,and the protective effect of the ethyl acetate extract was comparatively robust.These results demonstrate that organic extracts of Alpinia oxyphylla protect PC12 cells against apoptosis induced by hydrogen peroxide.Our findings should help identify the bioactive neuroprotective components in Alpinia oxyphylla.

Exploration of the effective components and mechanism of Alpinia oxyphylla-Saposhnikovia Radix in the treatment of diabetic kidney disease based on network pharmacology

Objective:Network pharmacology was used to predict the main targets and related mechanism pathways of the key compounds and active ingredients of the Alpinia oxyphylla-Saposhnikovia Radix based on the therapeutic strategy of tonifying kidney and dispelling pathogenic wind for the treatment of diabetic kidney disease(DKD).Methods:The effective components of Alpinia oxyphylla-Saposhnikovia Radix were screened through the data platform of Traditional Chinese Medicine(TCM).After screening effective targets,the disease target gene database was used to obtain targets for DKD,and then gene enrichment analysis to predict relevant signalling pathways after obtaining the key target genes through Venn analysis.Results:There were 324 active components of Alpinia oxyphylla-Saposhnikovia Radix through the data platform of TCM.In addition,Venn analysis showed that there were 48 key target genes for the treatment of DKD.The biological pathways for the action of Alpinia oxyphylla-Saposhnikovia Radix in DKD were closely related to many signaling pathways including NF-kappaB,classical MAP kinase,JNK and P38 MAP kinase,and ERK5.Conclusions:Treatment of DKD with Alpinia oxyphylla-Saposhnikovia Radix based on the therapeutic strategy of tonifying kidney and dispelling pathogenic wind has the characteristics of multi-components,multi-targets and multi-pathways,which could provide new ideas and methods for further researches on the treatment of DKD with Alpinia oxyphylla-Saposhnikovia Radix.

Effects of Alpinia oxyphylla on oxidative stress and expression of p47phox in diabetic nephropathy rats

Objective:To explore whether Yizhi(Alpinia oxyphylla,AO)could treat diabetic nephropathy(DN)through the inhibition of p47phox expression which is related to oxidative stress.Methods:Model of DN was established in SD rats with high glucose and fat diet for 4 weeks after STZ intraperitoneal injection.The rats were randomly divided into blank group,model group,AO group and irbesartan(IRB)group.Drugs were administered for 4 weeks.Blood glucose and 24-hour urinary protein were measured regularly.The index of ROS,SOD,MDA and Nox2 were tested by ELISA and the expression of p47phox protein in renal tissues were tested by techniques of RT-PCR and Western blot.Results:The decrease of blood sugar in AO group and IRB group were not obvious(P>0.05)while the drugs could significantly reduce the amount of protein in urine(P<0.05).AO could decrease ROS,MDA and Nox2 and increase SOD(P<0.05).It also could inhibit the expression of p47phox which was increased in model group(P<0.05).The IRB group has the similar effects compared with AO group(P>0.05).Conclusion:AO could reduce the amount of 24-hour urinary protein in DN rats and the mechanism may be related to the inhibition of p47phox expression thus alleviates excessive oxidative stress response.

益智仁-乌药药对调控PI3K/Akt/mTOR通路介导细胞自噬保护肾小球足细胞的作用机制研究

目的研究益智仁-乌药药对通过调控PI3K/Akt/mTOR信号通路促进足细胞自噬治疗糖尿病肾病(Diabetic Nephropathy,DN)的作用。方法60只造模成功的C57BL/KSJ-db/db(以下简称db/db)小鼠随机分为模型组、二甲双胍组、缬沙坦组、益智仁-乌药药对(低、中、高剂量)组,每组10只;另取10只C57BL/KSJ-db/m(以下简称db/m)小鼠为正常组,正常组和模型组给予生理盐水,治疗组小鼠分别给予相应药物,给药8周后检测小鼠肾脏病理学改变,足细胞自噬体数量、结构及相关蛋白表达。结果与模型组相比,益智仁-乌药药对组可显著减轻糖尿病肾病小鼠肾小球基底膜增厚情况,增加足细胞自噬体数量,显著升高自噬相关蛋白表达(P<0.05),降低PI3K/Akt/mTOR信号通路相关蛋白的表达(P<0.05)。其中益智仁-乌药药对高剂量组各指标改善优于益智仁-乌药低、中剂量组。结论益智仁-乌药药对通过抑制PI3K/Akt/mTOR信号通路激活,提高足细胞自噬水平,减轻足细胞损伤,发挥治疗糖尿病肾病的作用。

天然片断生境中山姜(Alpinia japonica)种群遗传结构

理论和实验研究表明片断化是导致生物多样性丧失的主要因素之一，但有关研究基本上集中在人类活动引发的片断化生境体系中。由于长期气候变化以及地形等因素的影响，一些物种的种群之间天然情况下就存在隔离，对这些种群进行研究可以弥补人类活动引起的片断化体系经历时间较短的不足。山姜是一个喜湿耐荫的多年生植物，分布在热带亚热带地区，由于气候和地形原因，该物种在浙江东部呈现天然片断化状态，如在浙江天童森林公园及其周边地区，仅分布在少数几个沟谷中。以这些种群为对象，采用RAPD标记分析经历长期天然片断化山姜种群遗传多样性程度和遗传分化格局。7条RAPD引物获得了69个位点，其中多态位点68个。种群水平遗传多样性较高，多态位点百分比、期望杂合度和Shannon多样性指数分别为78.81％-85.51％、0.3170-0.3430、0.4560-0.4914，这与其异交的繁殖方式有关，同时与山姜克隆生长，每个世代维持的时间长，种群更新降低也有关系。遗传变异大多分配在种群内，但种群问遗传分化达到显著程度，φST为0.297，计算的基因流中等，为0.592，不足以克服长期遗传漂变导致的种群分化，Mantel检验表明遗传距离与空间距离不存在相关关系，显示了长期片断化状态下遗传漂变的作用。

五种不同提取方法益智果实挥发油化学成分分析

目的:分析比较5种不同提取方法所得益智果实挥发油化学成分的差异。方法:采用水蒸气蒸馏、超临界CO_(2)萃取、超声辅助提取、溶剂提取及索氏提取益智果实挥发油,通过气相色谱-质谱联用技术分析其化学成分,以峰面积归一化法计算各成分的相对含量。结果:5种不同提取方法挥发油得率为1.50%~10.37%,从5种提取方法中共鉴定出142个化合物,其中共有化合物5个,从水蒸气蒸馏法提取的挥发油中鉴定出65个化合物,超临界CO_(2)萃取法鉴定出48个化合物,溶剂提取法鉴定55个化合物,超声辅助提取法鉴定出45个化合物,索氏提取法鉴定40个化合物。结论:不同提取方法所得挥发油成分均以萜类及含氧衍生物为主,含量最高的成分均为圆柚酮,但不同提取方法化合物种类和含量存在较大差异,研究结果为益智植物进一步开发利用提供了理论依据。

益智不同种质资源3种主要成分的含量比较

为比较不同种质资源益智中3种活性成分的差异,以资源圃保存的44份益智种质资源为试验材料,采用UHPLC-MS方法对不同种质资源益智果实中圆柚酮、白杨素和原儿茶酸的含量进行检测分析。结果表明,44份益智果实中圆柚酮、白杨素、原儿茶酸平均含量分别为3.013mg/g、0.054mg/g、4.882μg/g,圆柚酮含量范围为0.214mg/g~3.898mg/g,白杨素含量范围为0.001mg/g~0.145mg/g,原儿茶酸含量范围为1.242μg/g~8.015μg/g,种质YZ5-1、YZ1-5圆柚酮含量最高,YZ8-1、YZ6-3白杨素含量最高,YZ1-1原儿茶酸含量最高。益智不同种质资源3种主要成分含量差异较大,以益智圆柚酮、白杨素和原儿茶酸含量为综合评价指标,YZ5-1、YZ6-3、YZ8-1、YZ8-3的药材质量较好,研究结果可为益智品种选育和开发利用提供参考依据。

益智仁在疾病中的研究进展

益智仁为姜科山姜属植物益智的干燥成熟果实,是我国著名的四大南药之一,在临床上常用于治疗腹泻、尿频、炎症以及癌症等疾病。近年来,对益智仁的药理研究主要在其提取物和化学成分上,本文作者综述了其在腹泻、阿尔茨海默病(AD)、癌症、肾病、骨疾病等方面的研究,为以后对益智仁的研究、开发与应用提供了基础。

近5年益智化学成分与药理作用研究进展

查阅中英文文献,综述近5年在益智(Alpiniae oxyphyllae Fructus)化学成分研究、益智提取物及单体化合物的药理作用研究等方面的新进展,以期为益智的临床应用和药物开发提供参考。

益智药材质量标准概述

益智为药食同源药材,应用广泛,其质量的优劣直接影响药效。益智药材质量标准的建立能够促进益智饮片市场规范化,保障用药的安全。目前,不同国家或地区益智标准存在差异。对益智的国内外质量标准、炮制规范、商品规格等现状进行综述,旨在为益智质量标准的提升和规范研究提供参考。

基于GC指纹图谱及多指标成分含量测定的益智仁质量评价

目的:采用GC建立17批益智仁指纹图谱及4个成分含量测定方法。方法:以萘为内标物,建立17批益智仁GC指纹图谱,以MATLAB软件及自编程序建立共有模式,并结合相似度评价、系统聚类分析、主成分分析对不同产地益智仁进行质量评价,并同时测定益智仁中巴伦西亚橘烯、石竹素、α-香附酮、圆柚酮4个成分的含量。结果:17批益智仁指纹图谱相似度>0.930,确定了21个共有峰,不同产地样品中成分差异较小。17批益智仁中上述4个成分的含量分别为0.6563~2.3369 mg/g、0.1230~0.3313 mg/g、0.1318~0.3586 mg/g、1.4994~2.3376 mg/g,样品间成分含量差异较小,与指纹图谱结果吻合。结论:该研究建立的益智仁GC指纹图谱及4个成分含量测定方法稳定、可靠,可为益智仁的质量控制提供参考。

基于GC-MS及主成分综合评价广东不同产地益智挥发油成分差异

[目的]比较广东5个主产地的益智挥发油成分的差异性,为益智的合理利用和资源开发提供参考。[方法]采用水蒸气蒸馏法对不同产地的益智挥发油进行提取,用GC-MS法对所得挥发油组分进行鉴定分析,同时进行谱库检索,并通过SIMCA+P统计软件进行主成分分析。[结果]广东5个主产地的益智果实挥发油含量均大于1.00%;采用GC-MS技术共鉴定出58个化合物,共有化合物53个,其中γ-松油烯的含量最高;利用主成分分析将53个指标简化为4个综合指标,主成分综合得分从高到低排序为高州、阳春、罗定、信宜、化州;聚类分析表明,5份益智样品在欧氏距离d=8.00的水平上,可划分为3大类群。[结论]广东省5个不同产地益智挥发油化学成分在组成和含量上存在一定的差异。

益智离体再生体系建立

为建立益智高效再生体系,以益智笋芽为外植体,研究无菌外植体的获得,及不同培养基组分对丛生芽增殖、生根苗试管姜培养效果的影响。结果表明:无菌外植体获取以饱和漂白粉溶液处理30min+0.1%升汞溶液处理15min最佳;最佳增殖培养基为MS+6-BA 1.0mg/L+TDZ 0.1mg/L,添加蔗糖45g/L,增殖倍数达6.85;最利于生根苗试管姜诱导的培养基为MS+NAA 1.5mg/L+IBA 0.5mg/L+甲派鎓1.0mg/L,添加蔗糖90g/L,生根率99.20%,试管姜发生率97.29%,移栽成活率95.93%。

以黄精、益智仁为主的复方固体饮料健脾渗湿的作用研究

目的:研究以黄精、益智仁为主的复方固体饮料健脾渗湿作用。方法:将小鼠随机分为5组,每组10只,空白组正常饲养,每天经口灌胃给予阴性对照组生理盐水,低、中、高3个剂量组分别给予2 g·(kg bw)^(-1)、4 g·(kg bw)^(-1)、6 g·(kg bw)^(-1)固体饮料,每次灌胃间隔12 h。用药10 d后,测量小鼠脾脏与胸腺重量,计算脾脏指数、胸腺指数、给药前后体重增长率。结果:灌胃复方固体饮料的低、中、高剂量组小鼠体重增长率、脾脏指数、胸腺指数均高于阴性对照组,且呈浓度依赖关系,差异有统计学意义(P<0.05)。结论:以黄精、益智仁为主的复方固体饮料可以增强脾的运化功能,消除水湿。

中药益智化学成分的研究

目的 :研究中药益智AlpiniaoxyphyllaMiq .的化学成分。 方法 :利用各种色谱技术进行分离纯化 ,根据化合物的理化性质和光谱数据进行结构鉴定 ;用GC MS联机技术鉴定挥发油的化学成分。结果 :从益智分得 10种化合物 ,其中 2个二芳基庚酮 :yakuchinoneA ( 1)、益智醇 (oxyphyllacinol 2 ) ,3个eremophilane烷碳架的倍半萜valencene ( 3 )、nootkanone ( 4 )、nootkanol ( 5 ) ,两个黄酮 :杨芽黄酮 (tectochrysin 6)、白杨素 (chrysin 7) ,以及 β 谷甾醇( 8)、胡萝卜苷 ( 9)和脂肪酸 ( 10 )。用GC MS鉴定了挥发油中 64个化学成分 ,其中含量较高的有 :p cymene( 4 4 87% ) ,valencene ( 9 13 % ) ,linalool ( 4 3 9% ) ,myrtenal ( 3 90 % ) ,β pinene ( 3 87% ) ,α pinene ( 2 93 % ) ,furopelargone ( 2 62 % ) ,terpinen 4 ol( 2 5 6% )等。 结论 :益智醇 ( 2 )为新二芳基庚醇 ,其化学结构为 1 ( 4′ hyroxy 3′ methoxy phenyl) 7 phenyl 3 heptanol,3 ,6和

益智仁挥发油对帕金森病模型小鼠脑内纹状体和黑质损伤的影响

目的观察益智仁挥发油(VOA)脑保护作用的机制。方法C57BL小鼠随机分为正常对照组、帕金森病(PD)模型组、司来吉兰10mg·kg-1阳性对照组,VOA0.278,0.833和2.5mL·kg-1组。除正常对照和模型组外,各给药组分别预先ig给予VOA或司来吉兰,每天1次,连续12d。第6天开始,除正常对照组外,各组ip给予1-甲基-4-苯基-1,2,3,6-四氢吡啶(MPTP)30mg·kg-1,每天1次,连续7d,制备PD小鼠模型。末次给药后第2天处死小鼠,荧光分光光度法测定小鼠脑内纹状体高香草酸(HVA)和5-羟色胺(5-HT)含量,比色法检测纹状体内超氧化物岐化酶(SOD)活性、还原型谷胱甘肽(GSH)和丙二醛(MDA)含量,SABC免疫组化法测定黑质致密部神经细胞内Bcl-2和天冬氨酸半胱氨酸蛋白酶3(caspase 3)的表达。结果与正常对照组比较,模型组小鼠脑内纹状体HVA,5-HT和GSH含量显著下降,SOD活性下降,MDA含量明显升高;黑质致密部Bcl-2和caspase 3免疫阳性细胞数增加。预先给予VOA0.833和2.5mL·kg-1可明显抑制MPTP所诱导的HVA和GSH含量及SOD活性的降低,并抑制MDA含量升高,2.5mL·kg-1可明显抑制5-HT含量降低。VOA0.833和2.5mL·kg-1可增加黑质致密部Bcl-2免疫阳性细胞数,减少caspase 3免疫阳性细胞数。VOA2.5mL·kg-1的作用与司来吉兰的作用相近。结论VOA的脑保护作用可能与其增加脑内纹状体单胺类神经递质释放、减轻氧化应激反应以及减少黑质致密部神经元凋亡等因素有关。

益智红外指纹图谱的研究

益智产区的鉴别对于药材市场的质量控制以及提高药材的质量显得极为重要。益智为我国四大南药之一,主要药效成分是诺卡酮和益智酮。红外指纹图谱用于中药质量控制研究有着良好的发展前景,借助红外光谱的指纹特性,以八种益智精油红外指纹图谱的共有峰率和变异峰率为指标,计算了样品相对于该标准品的共有峰率和变异峰率,并按照共有峰率、变异峰率的大小建立了不同的序列。结果表明:广东、广西和海南的益智油红外图谱比较吻合,但也存在一定的差异。益智油的红外图谱特征峰为1 710和1 666cm-1,广东和海南的益智油的红外指纹图谱(400～4 000 cm-1)共有峰率为62%,广西和海南的共有峰率为52%,海南的益智油共有峰率为66%～84%。从多维共有峰率和变异峰率双指标序列分析法可以对不同产地具有准确、精细且非常直观的鉴别能力,该方法可以准确地区别不同产地的益智。