莲的非可食用部分中生物碱分布

研究了莲的非可食用部分荷叶、莲子心、莲房、藕节、莲子壳和藕皮中生物碱的组分及含量。采用乙醇水溶液从非可食用部分提取生物碱,然后用高效液相色谱法进行测定。结果表明莲子心提取物中含有5种主要的生物碱,其中4种为莲心碱、异莲心碱、荷叶碱和甲基莲心碱。除莲子心外的其他部分的提取物组分较相似,都含有6～8种主要的生物碱。荷叶碱在荷叶中的含量最高,可达149.64μg/g;莲心碱、异莲心碱和甲基莲心碱在莲子心中的含量最高,分别为457.76、6155.85、1420.90μg/g。莲的六个非可食用部分中都含有5～8种主要的生物碱,有很好的开发利用价值。     

银杏叶有效成分的提取及其镇痛作用研究

采用水煎醇沉法,用正交实验探索银杏叶中黄酮类化合物的最佳提取工艺,并研究复方银杏叶液的镇痛作用。采用煎煮法提取银杏叶药材,醇沉法来纯化,以银杏叶中黄酮含量为指标,选取加水量、煎煮时间、煎煮次数为因素,用正交实验设计方法对银杏叶的提取工艺进行优化,同时在小鼠扭体反应模型上观察其用药后扭体潜伏期和扭体数的变化。银杏叶的最佳提取工艺为14倍加水量,每次煎煮100 min,煎煮3次。复方银杏叶液50、100、200 mg/kg腹腔注射分别显著减少小鼠扭体数,显著延长小鼠扭体潜伏期,并呈一定的剂量依赖性关系。该提取工艺合理可行,与以前文献提取方法比较,减少经济成本,同时袁明复方银杏叶液有明显的镇痛作用。     

丹参酚酸水提取工艺的研究

本文以药典规定的水溶性总酚酸紫外吸收度为检测指标,考察了丹参酚酸的最佳提取工艺。研究结果表明,最佳提取工艺为：采用16倍水煎煮,每次煎煮时间不能超过0．5h,煎煮4次后基本提取完全。另外本文还对水提液的醇沉工艺进行了考察,研究结果表明50％醇沉的效果最好。     

红肉蜜柚汁胞蛋白质双向电泳体系优化研究

为建立适合红肉蜜柚发育过程中汁胞蛋白质组学的研究体系,以着色初始时期的红肉蜜柚汁胞为试材,摸索最适合的蛋白质双向电泳体系参数。结果表明,酚提取法比TCA-丙酮法更适合于红肉蜜柚汁胞总蛋白质的提取,提取得率高,双向电泳图谱清晰。采取18 cm、线性pH 3～10 和18 cm、线性pH 4～7 相结合的分析策略,可获得更为完整的蛋白质组信息,其第一向等电聚焦最佳参数（总伏小时数）分别为30 000 Vhs和43 000 Vhs。     

用电导率法研究低频超声提取小檗碱成分的产额

采用电导率测试法研究了超声提取对药材黄柏中小檗碱成分产额的影响,和煎煮法,浸泡法相比,低频超声辐照可增强从川黄柏中所提得的小檗碱成分的产额,证实了电导率测定法可作为瞬时,动态反馈药材中有效成分提取规律的基本方法。     

正交试验法优选复方四参颗粒剂的提取工艺

本文研究复方四参颗粒剂的提取工艺,采用正交试验法,以丹酚酸B提取率和干膏得率为指标,考察加水量、煎煮时间和煎煮次数对提取效果的影响,确立了复方四参颗粒剂的最佳提取工艺为:加8倍量水,提取2次,第一次时间为2 h,第二次为1 h。在最佳提取工艺条件下,干膏得率为16.58%,丹酚酸B的平均量为22.42mg。与正交试验的结果相符,说明该提取方法合理、稳定、可行。     

正交试验优化水提复方慢肝宁工艺研究

目的优化复方慢肝宁中药材的水提工艺,并建立其提取及含量测定方法。方法采用L_9(3~4)正交试验,以复方提取液中人参皂苷Rb1含量、三七皂苷R1含量、丹参酮I含量、柴胡皂苷A含量的综合评分为评价指标,考察加水量、煎煮时间和煎煮次数对水提工艺的影响,并进行验证试验。结果优化的水提工艺为第1次提取加9倍水,提取90 min;第2次提取加7倍水,提取60 min;第3次提取加7倍水,提取60 min。结论优化的水提工艺稳定、可行,可用于复方慢肝宁中各药材的提取,为复方慢肝宁缓释片的制剂开发提供实验依据。     

双指标优选肠安颗粒中有机酸提取工艺条件(I)

以总有机酸和绿原酸的提取率双指标作为提取条件考核对象,来探讨肠安颗粒中所含有机酸成分的中药材的最佳提取工艺。采用L9(34)正交设计,考察提取溶媒所需加水量、浸泡时间、煎煮时间、煎煮次数四个因素,选择进行3个水平试验。得出结论:以加10倍量的水(第一次煎煮多加1.5倍量),浸泡1 h,煎煮2次,每次1h为最佳提取工艺条件,亦即以A2B2C2D2组合为合理。     

莲心碱对实验性高脂血症大鼠血脂及脂质过氧化的影响

本文研究莲心碱对实验性高脂血症大鼠血脂和抗氧化能力的影响.将32只大鼠随机分为4组,对照组饲喂基础饲料;诱导组饲喂高脂饲料;试验组给予高脂饲料+莲心碱灌胃2.5和5.0 mg·kg-1.测血清中血脂和丙二醛(MDA)水平,以及谷胱甘肽过氧化物酶(GSH-Px)、超氧化物歧化酶(SOD)活性;取肝脏测绝对和相对肝重及MDA含量.结果表明莲心碱可显著降低实验性高脂血症大鼠血清中总胆固醇、甘油三酯、低密度脂蛋白胆固醇水平和动脉粥样硬化指数;显著升高血清高密度脂蛋白胆固醇、GSH-Px和SOD水平;同时还可显著降低血清和肝脏中MDA的含量;其绝对和相对肝重均低于诱导组.     

甲基莲心碱抗乳腺癌 MCF-7细胞的研究

目的:观察甲基莲心碱对乳腺癌细胞系MCF-7增殖和凋亡的影响,并探讨其诱导乳腺癌细胞系MCF-7凋亡的可能作用机制。方法:采用体外培养人乳腺癌细胞系MCF-7,CCK-8实验检测不同浓度甲基莲心碱对MCF-7细胞增殖抑制作用;乳酸脱氢酶(LDH)试剂盒(微板法)检测细胞上清液LDH含量;流式细胞术分析甲基莲心碱对MCF-7细胞周期及凋亡的影响;实时定量PCR(RT-PCR)检测线粒体凋亡相关基因Bax和Bcl-2的表达水平。结果:CCK-8、LDH结果显示甲基莲心碱以时间、浓度依耐性的方式抑制乳腺癌MCF-7细胞的增殖及促进细胞毒性的增加;流式细胞术结果表明不同甲基莲心碱作用下MCF-7的平均凋亡率分别为(15.44±0.52)、(18.81±2.24)、(24.26±2.84)、(36.90±3.15)、(59.27±5.86),且使其周期阻滞于G0/G1期;RT-PCR检测结果证明甲基莲心碱可上调乳腺癌细胞中促凋亡基因Bax的表达,而下调抑制凋亡基因Bcl-2。结论:甲基莲心碱以时间和浓度依赖的方式抑制乳腺癌细胞增殖、细胞毒性增加,导致细胞周期于G0/G1阻滞并促进癌细胞凋亡。甲基莲心碱抗乳腺癌的可能作用机制是激活线粒体凋亡途径。     

莲品种资源的SRAP遗传多样性分析

利用分子标记SRAP技术对国内广泛栽培以及新近育成的39个莲品种进行了DNA多态性分析。选取5对引物扩增基因组DNA,共获得101条带,其中88条为多态性条带,每对引物平均提供20个标记信息。由UPMGA方法得到的聚类分析结果表明了39个品种间的遗传关系,结果表明:花莲、籽莲和藕莲三大类群有明显的界限,花莲和籽莲的遗传距离较近,藕莲与它们的遗传距离较远。分子方差分析结果表明:莲品种间和品种内均存在遗传变异,藕莲品种内的遗传变异略低于品种间的遗传变异,而诱变籽莲、诱变花莲和常规籽莲品种内遗传变异均大于品种间的遗传变异,尤其是诱变籽莲、诱变花莲品种内遗传变异占总变异的分量分别超过了70%和60%,这种品种内的遗传变异,一方面对于莲的高产稳产具有重要的意义,另一方面也说明了从这些国内广泛栽培的品种以及新近育成的莲品种中可以直接进一步选育出更优良的新品种。     

Whole genome re‐sequencing reveals evolutionary patterns of sacred lotus (Nelumbo nucifera)

Sacred lotus (Nelumbo nucifera or lotus) is an important aquatic plant in horticulture and ecosystems. As a foundation for exploring genomic variation and evolution among different germplasms, we re‐sequenced 19 individuals from three cultivated temperate lotus subgroups (rhizome, seed and flower lotus), one wild temperate lotus subgroup (wild lotus), one tropical lotus group (Thai lotus) and an outgroup (Nelumbo lutea). Through genetic diversity and polymorphism analysis by non‐missing SNP sites widely distributed in the whole genome, we confirmed that wild and Thai lotus exhibited greater differentiation with a higher genomic diversity compared to cultivated lotus. Rhizome lotus had the lowest genomic diversity and a closer relationship to wild lotus, whereas the genomes of seed and flower lotus were admixed. Genes in energy metabolism process and plant immunity evolved rapidly in lotus, reflecting local adaptation. We established that candidate genes in genomic regions with significant differentiation associated with temperate and tropical lotus divergence always exhibited highly divergent expression pattern. Together, this study comprehensive and credible interpretates important patterns of genetic diversity and relationships, gene evolution, and genomic signature from ecotypic differentiation of sacred lotus.     

镉在莲各器官中累积规律的研究

莲子是我国重要的特产优质资源,但莲等水生植物容易吸收积累重金属.为了探究莲对重金属镉的吸收积累特点,利用盆栽外施不同浓度的镉(0,1.0 mg·kg-1,2.5 mg·kg-1)培养两个莲子品种,检测分析镉对莲子生物量的影响及莲各器官中镉的含量.结果表明:随着外施镉浓度的增加,两种莲子生物量下降,各器官中镉含量增加,且...     

莲种子蛋白质提取方法比较与双向电泳分析

莲(Nelumbo nucifera Gaertn.)不仅是重要的水生蔬菜作物之一,而且是进行基础研究的好材料。本文采用4种蛋白质提取方法(新型TCA/丙酮法、传统TCA/丙酮法、改良的Tris-HCl法、Tris-饱和酚法)并结合双向电泳技术,对莲子蛋白质提取方法进行筛选与优化。双向电泳实验结果显示,所得蛋白质图谱与莲种子蛋白质组成分布特点一致。通过PDQuest软件分析表明,新型TCA/丙酮法适用于莲子叶和胚芽组织的双向电泳蛋白质提取,而传统TCA/丙酮法则适用于莲胚轴组织双向电泳的蛋白质提取。研究结果为进一步利用质谱进行莲子蛋白质组研究奠定了基础。     

Genetic diversity and differentiation of lotus (Nelumbo nucifera) accessions assessed by simple sequence repeats

Nelumbo nucifera (lotus) is a perennial aquatic crop of substantial economical and ecological importance. Currently, the evaluation of the genetic variation of lotus germplasm accessions using codominant simple sequence repeat (SSR) markers is significant, and it is essential for understanding the population structure of N. nucifera. Here we report the genetic diversity and differentiation of 92 N. nucifera accessions (82 cultivated varieties and 10 wild lotus) using 50 polymorphic SSR markers. A total of 195 alleles were detected, with an average of 3.9 alleles/locus. The mean polymorphic information content (PIC) and the mean expected heterozygosity were 0.43 and 0.50, respectively. The genetic relationships among accessions were estimated using an unweighted pair‐group method with arithmetic average (UPGMA) cluster and principal coordinate analysis (PCA). Both methods revealed that the lotus accessions from China and those from its adjacent Asian countries formed a single cluster, respectively. The cultivated varieties were correlated with their major characteristics in cultivation (the seed, rhizome and flower type) rather than their geographic distribution. On the basis of the Bayesian model‐based analyses, two genetically distinct groups (the seed lotus group and the rhizome lotus group) were generated, with a strong differentiation between them (F_ST = 0.57). The seed lotus group exhibited higher genetic diversity than did the rhizome lotus group. The results herein indicated that the current levels of genetic diversity and differentiation between the lotuses have been greatly influenced by artificial selection.     

Studies on Lotus Seed Protease

A protease from the lotus seed (Nelumbo nucifera Gaertn) was purified by acid-treatment, ammonium sulfate-fractionation, ethylalcohol-fractionation, TEAE-cellulose-treatment and Sephadex G-100 gel-filtration.

The enzyme was purified about 870-fold and was homogeneous in electrophoretic and ultracentrifugal analyses.

Purified lotus seed protease is an acid protease with a pH optimum at 3.8 toward urea-denatured casein. It is active for casein and hemoglobin. But other proteins such as edestin, zein, lotus seed globulin and soybean casein are slightly hydrolyzed and egg albumin is hardly hydrolyzed. This enzyme is most stable at pH 4.0 below 40°C. The enzyme is not a thiol protease, and its activity was completely inhibited by potassium permanganate, remarkably inhibited by sodium dodecylsulfate and accelerated by hydrogen peroxide.