血根碱酊剂体外抗真菌初步研究

通过分析博落回(Macleaya cordata (wild)R.Br.)提取物—血根碱(制成血根碱酊剂)、总生物碱对须癣毛癣菌(Trichophyton mentagrophytes)、玫瑰色毛癣菌(Trichophyton rosaceum)、许兰黄癣菌(Trichophyton schoenleini)、红色毛癣...     

血根碱抗水霉菌体外活性研究

采用测OD值和琼脂稀释法,以孔雀石绿、氯化钠作为对照,分别进行了3种药物对水霉菌孢子和菌丝的体外抑菌活性研究,其中血根碱以2%DMSO(二甲亚砜)为溶剂,孔雀石绿和NaCl均以水为溶剂。结果表明:血根碱对水霉孢子的MIC80为200μg/mL,对菌丝的MIC为256μg/mL;孔雀石绿对水霉菌孢子的MIC80为6μg/mL,对菌丝的MIC为12μg/mL;氯化钠对水霉菌孢子的MIC80为15 000μg/mL,对菌丝的MIC为20 000μg/mL,说明血根碱具有开发为抗水霉菌药物应用于渔药领域的前景。     

Induction of heme oxygenase-1 by Macleaya cordata extract and its constituent sanguinarine in RAW264.7 cells

Macleaya cordata (plume poppy) is used in traditional Chinese medicine for its anti-inflammatory and antibacterial activities. In this study, we examined whether M. cordata extract and/or its major alkaloid constituents, protopine, allocryptopine, sanguinarine and chelerythrine activate the Nrf2 signalling pathway which regulates the expression of cytoprotective enzymes including heme oxygenase-1 (HO-1) and thioredoxin 1. In murine macrophage RAW264.7 cells, M. cordata extract increased both mRNA and protein levels of HO-1. Of the alkaloids examined, only sanguinarine appeared to be responsible for these effects. At the concentration of 2 μM, sanguinarine induced nuclear accumulation of Nrf2, increased the expression of Hmox1 gene encoding HO-1 and elevated HO-1 protein levels. Sanguinarine-induced Hmox1 mRNA expression was suppressed by SB203580, a pharmacologic inhibitor of p38 mitogen-activated protein kinases (p38 MAPKs). The upregulation of HO-1 in RAW264.7 cells by sanguinarine was, however, accompanied by decrease in cell viability. Nonetheless, sanguinarine at micromolar, non-cytotoxic concentrations elevated protein levels of HO-1 and thioredoxin 1 in primary cultures of human hepatocytes. We conclude that sanguinarine may, under appropriate conditions, increase the capacity of the enzymatic antioxidant defence system via activation of the p38 MAPK/Nrf2 pathway.     

血根碱对菜青虫的生物活性

[目的]进一步明确血根碱的杀虫活性及作用机理。[方法]采用叶蝶法和点滴法测定了血根碱对菜青虫幼虫的触杀活性及对其幼虫生长发育、化蛹及羽化的影响。[结果]血根碱对菜青虫3龄幼虫具有很强的触杀活性,在4 000 mg/L浓度下的校正死亡率为57.14%。血根碱具有很强的生长发育抑制作用,处理后幼虫的体重减轻,化蛹后的蛹重减轻;血根碱无杀卵、杀蛹作用。[结论]为进一步开发研制新型高效杀虫剂提供了依据。     

高通量测序分析血根碱对中华鳖肠道菌群结构的影响

为研究血根碱对中华鳖(Trionyx sinensis)肠道菌群多样性及丰度的影响,选取480只初始体重为(31.75±7.20) g的中华鳖,设置添加0的对照组(RS1)和添加50 (RS2)、100 (RS3)和150 mg/kg (RS4)的3个实验组。实验结束后,提取中华鳖中肠组织的总DNA,进行16S rRNA V3-4区基因组测序。结果显示,实验组的总操作分类单元(Operational taxonomic units, OTUs)数量显著高于对照组,肠道菌群复杂度及丰富度较对照组有明显增加。肠道菌群结构组成分析发现,RS1与RS2结构较为接近,而RS1与RS3、RS4较远。在门分类水平上,变形菌门(Proteobacteria)和衣原体门(Chlamydiae)是优势菌群;在属分类水平上,螺杆菌属(Helicobacter)占绝对优势。通过比较发现,实验组中螺杆菌属丰度较对照组显著下降,而不动杆菌属(Acinetobacter)、乳杆菌属(Lactobacillus)和双歧杆菌属(Bifidobacterium)丰度均有明显上升。研究表明,在饲料中添加适量的血根碱(100~150 mg/kg)能有效提高中华鳖肠道菌群多样性及丰富度,并能改善肠道菌群结构。     

Distinct G-quadruplex structures of human telomeric DNA formed by the induction of sanguinarine and nitidine under salt-deficient condition

The inducing abilities of two alkaloids, Sanguinarine (San) and Nitidine (Nit), to fold human telomeric DNA d[5′-TTGGG(TTAGGG)₃A-3′] (H24A) into G-quadruplex structures has been investigated by using CD and ¹H NMR spectroscopy. It is found that, under salt-deficiency condition, San could induce H24A to fold into a normal anti-parallel G-quadruplex while Nit may induce hybrid or mixed-type G-quadruplex structures. In order to understand the structure of H24A induced by Nit, the interaction between Nit and existed hybrid G-quadruplex H24A in the presence of K⁺ has been examined for comparison. The different behaviors of these two molecules interacted with DNA could be ascribed to distinct binding mode, which may be resulted from the steric hindrance of the molecular frame of these two alkaloids.     

血根碱对6种非靶标生物的安全性评价

为评价血根碱对环境有益生物的风险,测定40%血根碱母药对日本鹌鹑、家蚕、斑马鱼、意大利工蜂、大型溞和斜生栅藻6种非靶标生物的急性毒性。结果显示,血根碱对日本鹌鹑急性经口 7 d-LD₅₀值为556 mg/kg体重,家蚕急性经口 96 h-LC₅₀值为680 mg/L,意大利工蜂急性接触 48 h-LD₅₀ 值为 37.7 μg/蜂;急性经口 48 h-LD₅₀值为 46.2 μg/蜂;对斑马鱼96 h-LC₅₀值为0.64 mg/L,大型溞48 h-LC₅₀值为 0.34 mg/L;斜生栅藻72 h-E_rC₅₀为285 μg/L,72 h-E_yC₅₀为61.1 μg/L。研究表明,血根碱母药对日本鹌鹑、家蚕、蜜蜂均为低毒,而对斑马鱼、大型溞、斜生栅藻表现为高毒。血根碱具有一定风险性,其产品开发应以旱田作物有害生物防治为主。     

Phytochemical and antimicrobial characterization of Macleaya cordata herb

Macleaya cordata (plume poppy) is a source of bioactive compounds, mainly isoquinoline alkaloids which are used in phytopreparations with anti-inflammatory and antimicrobial activities. In this study, the alkaloids sanguinarine, chelerythrine, their dihydro derivatives, protopine and allocryptopine and phenolics, gallic, protocatechuic, p-hydroxybenzoic, m-hydroxybenzoic, gentisic, p-coumaric, caffeic, ferulic and sinapic acids were determined in extracts prepared from M. cordata aerial part, seeds, and seed capsules using HPLC with UV detection and/or LC/MS with electrospray ionization. The highest content of sanguinarine and chelerythrine was found in capsules. Protopine and allocryptopine were major alkaloids in leaves including footstalks. The seed oil contained dihydrosanguinarine, dihydrochelerythrine and twelve fatty acids of which linoleic, oleic, palmitic and stearic acids predominated. In addition, sanguinarine reductase, a key enzyme in sanguinarine/dihydrosanguinarine equilibrium in plants, was found for the first time, in the soluble proteins of leaves. Finally, extracts were tested for antimicrobial activity using the microdilution method on standard reference bacterial strains.     

博落回生物碱提取工艺研究

以博落回血根碱的提取率为考察指标优化博落回生物碱提取工艺,结果表明:酸性醇提取法比酸水法、醇提法对血根碱的提取效率高。采用正交试验方法对酸性醇提取法进行优化,最佳工艺条件为:在pH值为3的75%乙醇溶液中回流提取2次,每次1 h。此条件下血根碱提取率为5.82 mg/g。对最佳工艺条件稳定性试验的结果表明:其稳定性较好,RSD=1.65%。     

血根碱对菜青虫生长发育及几种代谢酶系的影响

为进一步深入了解血根碱的杀虫活性,采用小叶碟添加法测定了血根碱对菜青虫(Peris rapae L.)生长发育及体内羧酸酯酶(CarE)、酸性磷酸酯酶(ACP)、碱性磷酸酯酶(AKP)、谷胱甘肽S-转移酶(GSTs)和乙酰胆碱酶系(AChE)活性的影响。结果表明:血根碱能明显抑制菜青虫3龄幼虫的生长发育;可显著激活羧酸酯酶、酸性磷酸酯酶、碱性磷酸酯酶的活性;明显抑制乙酰胆碱酯酶的活性;对谷胱甘肽S-转移酶表现为先抑制后激活再抑制的趋势,24 h时被明显激活,12 h和36 h时接近于同期对照,而6 h和48 h时被明显抑制。