博落回的开发利用

博落回提取物主要成份为血根碱、白屈莱红碱等生物碱．国内外已开发为药品、生物农药、兽药等产品。     

博落回生物碱与生物活性及其应用

我国是博落回资源分布最丰富的国家,生物碱是其主要的活性成分。对博落回生物碱的种类、分析和制备方法;抗菌、杀虫、抗肿瘤等生物活性以及临床应用和病虫害防治等方面进行了介绍。     

药用植物博落回形态与发育解剖学研究

博落回(Macleaya cordata)是罂粟科含有异喹啉生物碱的主要药用植物, 其生物碱提取物具有多种药理活性。目前对博落回的研究主要集中在生物碱的成分、活性和药理等方面, 对与其合成、运输、储藏密切相关的发育解剖结构及生物碱组织化学定位研究还未见报道。该文报道了温室中生长的博落回植株形态特征, 并结合徒手切片和改良碘化铋钾沉淀的方法对主要器官(根、茎、叶)的发育结构和生物碱组织化学染色进行了初步分析。结果表明, 博落回发育基本遵循一般双子叶植物的规律。根中生物碱出现在中柱鞘外的薄壁细胞和导管周围。茎和叶柄中的生物碱主要出现在维管束周围, 偶尔也出现在茎的髓细胞中。总体上, 茎中的生物碱积累量少于根。     

小果博落回中2种杀虫活性成分的分离及鉴定

以粘虫(Mythimna separata)3龄幼虫为试虫,采用生物活性示踪法从小果博落回(Macleaya microcarpa)乙醇提取物中分离纯化出2种活性成分,经MS1、H-NMR1、3C-NMR分析鉴定为二氢血根碱和二氢白屈菜红碱。采用小叶碟添加法测试了2种化合物对粘虫3龄幼虫的拒食及毒杀活性。结果表明,2种化合物对粘虫3龄幼虫均具有较高的拒食活性,48 h的拒食AFC50分别为0.168和0.231 mg.mL-1,同时,二氢血根碱对粘虫3龄幼虫具有一定的毒杀活性,96 h的LC50为0.085 mg.mL-1。分析认为二氢血根碱可能是小果博落回的主要杀虫活性成分之一。     

Macleaya cordata extract and Sangrovit genotoxicity. Assessment in vivo

Background: Sanguinarine (SG) has been reported to form DNA adducts in vitro and increase the levels of DNA single strand breaks in the blood and bone marrow of mice treated intraperitoneally with SG. Recently, we showed no genotoxic effects of orally administrated 120 mg/kg feed Macleaya cordata extract (a mixture of sanguinarine and chelerythrine) in pigs or rats in 90-day studies. The goal of this paper was to assess the possible genotoxicity of M. cordata extract when included as a dietary admixture to rodents at concentrations providing 600 mg/kg feed and 100, 7000 or 14000 mg/kg feed Sangrovit (natural feed additive containing M. cordata extract and powdered M. cordata) in a 90-day pilot study. Methods and Results: The rats consumed ad libitum either the standard diet or the diets containing 367 ppm of sanguinarine and chelerythrine in M. cordata extract, and 5, 330, or 660 ppm of total alkaloids in Sangrovit for 90 days. The DNA adducts formation in liver was analyzed by (32)P-postlabeling technique and DNA single strand breaks in lymphocytes were evaluated by Comet assay. The results showed that M. cordata extract and/or Sangrovit induced no DNA damage to rat lymphocytes or hepatocytes after 90-days oral administration. Conclusions: Data from the studies described in this paper and the fact that Sangrovit given to the rats in our experiments were higher than the recommended dose (50 to 100 mg/kg feed), argue strongly in favour of the use of Sangrovit in live stock.     

中药博落回的药理实验研究

博落回对四氯化碳、半乳糖胺所致急性肝损伤模型,均有显著改善肝脏功能作用;对四氯化碳所致慢性肝损伤大鼠模型,博落回可显著降低血清LDH水平。降低动物死亡率。提高血清A/G比值,有效保护肝细胞胰。抑制肝脏纤维化;博落回还可显著增强T和B淋巴细胞功能。     

小果博落回的生物碱分离与鉴定

以D101大孔吸附树脂为吸附剂,将对小果博落回全草的酸水提取物分为50%乙醇洗脱部分和丙酮洗脱部分.对两部分洗脱物分别进行硅胶柱色谱分离,由50%乙醇洗脱部分得到5个生物碱:二氢血根碱(1),二氢白屈菜红碱(2),6-乙氧基血根碱(3),6-丙酮基血根碱(4)和Bocconarborine A (5).由丙酮洗脱部分得到原阿片碱(6),α-别隐品碱(7),β-别隐品碱(8)和小檗红碱(9).采用波谱分析对化合物进行了结构表征,结果表明,化合物3、4、5、9是首次从该植物中分离得到.     

博落回遗传转化体系的初步建立

博落回是一种重要且应用广泛的药源植物,市场前景广阔。现以博落回无菌苗叶片为材料,通过优化卡那霉素和头孢霉素两种抗生素的浓度,初步建立了博落回遗传转化体系,同时利用农杆菌介导法将MtDREB1C基因导入博落回,获得卡那霉素抗性植株。随机抽取50株转基因植株进行PCR鉴定,其中3株成功扩增出了目的条带,经初步统计,获得博落回阳性苗的概率是6%。实验结果表明植物基因工程技术可作为博落回遗传改良的新途径,为博落回分子育种奠定基础。     

博落回SSR引物的开发以及遗传多样性分析

根据前期由本实验室完成的博落回转录组数据设计出65对引物,以浏阳产地的博落回为材料进行筛选,得到了28对有效引物.用这些引物对来自全国4个不同主要产区的博落回进行扩增,共扩增出引物159条带,其中多态性条带121条,占76.10%,共找到特征性条带13条.统计并比较4个不同产地的数据,经研究发现不同地区的博落回的遗传多样性较低,4个不同产地的博落回相似性较高.     

HPLC法测定博落回药材不同器官白屈菜红碱含量

本研究采用高效液相色谱法分离并测定博落回药材不同器官的白屈菜红碱含量。色谱条件:Diamnosil C₁₈柱,以乙腈-0.2%磷酸(22:78)为流动相;检测波长268nm。检测结果表明,其果实的白屈菜红碱含量较高,因此,常法采用博落回果实作为药材具有一定的科学性。     

博落回生物碱对蝗虫成虫毒力试验简报

目的:测定博落回生物碱对蝗虫成虫的毒力作用,为利用药用植物提取物杀灭蝗虫的无公害防治提供参考。方法:用文献法提取博落回果实中的生物碱,制备博落回生物碱溶液;采用喷雾法将博落回生物碱溶液喷施于蝗虫成虫;进行了博落回生物碱溶液对蝗虫成虫的田间小面积试验;用计算机软件SPSS11.5进行统计学分析。结果:在96小时内,博落回生物碱对蝗虫成虫的LD50为29.568mg/L,95%可信限为2.5563～69.473mg/L。田间小面积施药3天后,每平方米内虫口减退达90%左右,持效时间7天。结论:博落回溶液对蝗虫成虫具有一定的杀灭作用。     

博落回杀灭鱼类指环虫和病原菌活性成分的研究

采用常压硅胶柱层析和RP-HPLC制备并结合杀虫、抑菌活性追踪的方法,从博落回中分离获得了杀灭金鱼指环虫和抑制鱼类病原菌的活性成分,通过TLC、UV、m.p.、IR、MS、1H NMR和13C NMR鉴定其为血根碱.杀灭指环虫试验结果表明:血根碱对指环虫有较强杀灭活性,在质量浓度为0.6 mg?L-1时的杀虫率达到100%.血根碱对6株水产致病菌的活性试验结果表明:对嗜水气单胞菌的活性最高,MIC和MBC分别为12.5和25μg·mL-1;对哈维弧菌、鳗弧菌、杀鲑气单胞菌史氏亚种的活性最弱,其MIC和MBC均分别为50和100μg·mL-1.血根碱杀灭鱼类指环虫活性较高,对6株水产病原菌都有明显抑制作用.     

中国科学家完成博落回全基因组测序并解析苄基异奎宁类生物碱途径

博落回(Macleaya cordata)是罂粟属的一种传统中药, 因其富含具有抗菌功能的血根碱(SAN)和白屈菜红碱(CHE), 可以作为牲畜抗菌生长促进剂的安全来源植物。中国科学家以博落回为材料, 利用从头测序手段测定了首个罂粟属植物全基因组序列。研究人员通过组织特异性代谢谱及表达谱分析鉴定了博落回中16个参与血根碱和白屈菜红碱合成的候选基因, 并结合同源克隆及饲喂实验对其中的14个基因进行了体外功能验证。上述研究为进一步探索博落回中血根碱等苄基异奎宁类生物碱(BIAs)合成、调控及后续合成奠定了基础, 也为解析罂粟属其它植物的代谢途径提供了有力工具。     

An Endophytic Sanguinarine-Producing Fungus from Macleaya cordata,Fusarium proliferatum BLH51

Fermentation processes using sanguinarine-producing fungi other than Macleaya cordata may be an alternative way to produce sanguinarine (SA), which is a quaternary benzo[c]phenanthridine alkaloid possessing antibacterial, anthelmintic, and anti-inflammatory properties. In this study, a SA-producing endophytic fungus strain BLH51 was isolated from the leaves of M. cordata grown in the Dabie Mountain, China. Strain BLH51 produced SA when grown in potato dextrose liquid medium. The amount of SA produced by this endophytic fungus was quantified to be 178 μg/L by HPLC, substantially lower than that produced by the host tissue. The fungal SA—which was analyzed by thin layer chromatography and high-performance liquid chromatography—was shown to be identical to authentic SA. Strain BLH51 was identified as Fusarium proliferatum based on the morphological characteristics and nuclear ribosomal DNA ITS sequence analysis. To the best of our knowledge, this is the first report concerning the isolation and identification of endophytic SA-producing fungi from the host plant, which further proved that endophytic fungi are valuable reservoirs of bioactive compounds.     

兽用博落回散对饲用酶制剂酶活性的影响

博落回散能抑菌消炎、改善畜禽肠道内环境,兼具营养和药用双重作用,已经成为一类新型饲料添加剂。酶制剂是目前已在养殖业广泛使用的饲料添加剂,它能提高动物对饲料的消化利用率、提高饲料转化率、降低饲喂成本。博落回散能否和酶制剂同时使用目前尚未有相关的研究报道。本文研究了不同浓度博落回散溶液对8种常用饲用酶制剂活性的影响,实验结果表明,在正常使用浓度(30～50 mg/L)范围内,博落回散不会明显影响淀粉酶、纤维素酶、果胶酶、甘露聚糖酶和木聚糖酶这5种糖基水解酶的活性;对植酸酶、酸性蛋白酶和脂肪酶这3种非糖基水解酶的活性影响不大,这说明博落回散能和酶制剂同时添加到饲料中。本研究为兽用博落回散饲料添加剂在养殖行业的广泛应用提供了实验支持。     

Differential diversifications of South American and Eastern Asian disjunct genera Bocconia and Macleaya (Papaveraceae)

Bocconia (10 species) and Macleaya (2 species) are two disjunct genera between South America and eastern Asia (EAS) in the Papaveraceae offering an opportunity to compare its biogeographic history with that of the well‐known disjunction between EAS and eastern North American (ENA). Our phylogenetic analyses of the chloroplast matK and rbcL gene sequences of Ranunculales including two species of Macleaya and six species of Bocconia supported the monophyly of Bocconia, Macleaya, and Chelidonioideae to which Bocconia and Macleaya belong. Nucleotide sequences of matK, rbcL, and nrDNA ITS supported the sister relationship of Bocconia and Macleaya. Biogeographic analyses of Chelidonioideae using S‐DIVA (statistical dispersal vicariance analysis) and DEC (dispersal extinction cladogenesis) methods inferred Eurasia as the most likely ancestral area of Bocconia and Macleaya and suggested no extinction events in either Bocconia or Macleaya. This agrees with the “Out‐of‐Asia” pattern of the EAS‐ENA disjunction. Molecular dating of Ranunculales with fossil‐based calibrations showed that Bocconia and Macleaya diverged in the late Eocene and early Oligocene, which is much earlier than most EAS‐ENA disjunct taxa. The disjunction may have formed via long distance dispersal or boreotropical connections via the North Atlantic and Bering land bridges. Both Bocconia and Macleaya diversified in the late mid‐Miocene, but Bocconia has apparently experienced a greater diversification probably aided by the evolution of the bird dispersal syndrome in fruit and seed after migration to South America. The greater diversification of Bocconia is also evidenced by the diverse leaf morphology and growth habit in response to colonization in various local habitats in South America.     

Two pairs of new dihydrobenzophenanthridine alkaloid isolated from the root of Macleaya cordata

Two pairs of dihydrobenzophenanthridine alkaloid, named (±)(S)-6-((R)-1-hydroxyethy) dihydrochelerythrine (1a, 1b) and (±)(S)-6-((R)-1-hydroxyethyl)dihydro-sangunarine (2a, 2b) were isolated from the root of Macleaya cordata. Their chemical structures were elucidated by analysis of the spectroscopic data including one-dimensional (1D) and two-dimensional (2D) NMR spectra, and were further confirmed by X-ray crystallographic analysis.     

Modulation of intestinal morphology and microbiota by dietary Macleaya cordata extract supplementation in Xuefeng Black-boned Chicken

Antibiotics are commonly overused to prevent livestock from diseases and to increase production performance. As potential substitutes of antibiotics, plant extracts have attracted the attention of researchers. It was known to all that addition of Macleaya cordata extract (MCE) to the food could advance immunity, intestinal health and animal performance. Thus, it was conducted to investigate the influence of MCE (0, 100, 150 and 200 mg/kg, with six replicate pens/treatment and 24 hens/pen) on intestinal morphology and microbial diversity in different intestinal segments in Xuefeng black-boned chicken in this study. The results showed that MCE supplement (100, 150 and 200 mg/kg) significantly diminished (P < 0.05) the crypt depth of the jejunum as compared to basal diet group. The 100 mg/kg group displayed a marked increase (P < 0.05), compared with 0 and 200 mg/kg group, in ileum microbial diversity as represented by the Shannon's index. In the cecum, treatment of MCE significantly decreased (P < 0.01) the Firmicutes, but Deferribacteres in 200 mg/kg MCE group were significantly raised (P < 0.05). In conclusion, we found that MCE improved intestinal morphology and reduced the crypt depth in jejunum. Together, addition of 200 mg/kg MCE modulated intestinal microbiota, increased beneficial bacteria such as Lactobacillus. Adding 100 mg/kg MCE to diet increased bacterial community diversity and relative abundance in jejunum and ileum, but had no effect on cecum microbial diversity.     

Optimizing the Production of Antioxidant and Antibacterial Alkaloids from Macleaya cordata by Subcritical Water Extraction Technology

In this work, the subcritical water extraction technology was used to extract alkaloids from Macleaya cordata, and the effects of extraction temperature and time on its yield were investigated to find the best extraction conditions. Moreover, the antioxidant capacity and antibacterial activity of Macleaya cordata extract were studied. Furthermore, through the single-factor method, it was found that properly increasing the extraction temperature and prolonging the extraction time was conducive to increasing alkaloid yield. Still, a considerable amount of alkaloids might be decomposed by heat, resulting in a decrease in their yield. The results showed that the optimal extraction temperature of alkaloids from Macleaya cordata with subcritical water is 190 °C, the time is 45 min, and the corresponding maximum yield is 35.19±0.12 mg/g (sanguinarine equivalent in raw materials). In addition, the antioxidation and bacteriostasis abilities of subcritical water extract are better than those of traditional hot water extract, indicating that it is a feasible method to extract alkaloids from Macleaya cordata with subcritical water.     

博落回、虎杖和黄芩提取物对植物病原菌的抑制作用

研究了博落回(Macleaya cordata)、虎杖(Reynoutria japonica)和黄芩(Scutellaria baicalensis)提取物对三种植物病原细菌和六种病原真菌的抑制作用.结果显示:虎杖提取物对根癌土壤杆菌和番茄疮痂病菌表现出较强的抗细菌活性;博落回提取物表现出较强的抗真菌活性,对供试真菌的半抑制浓度(IC50)在0.04～0.76mg/mL之间.