非洲山毛豆中鱼藤酮的超临界流体萃取技术研究

采用超临界流体萃取-反相高效液相色谱法,以鱼藤酮萃取率和萃取产物总得率为评价指标,研究提出非洲山毛豆Tephrosia vogelii Hook.f中鱼藤酮的超临界流体萃取较优工艺条件为:萃取物料粒径30目,静态萃取20 min,萃取温度45℃,压力27.58 MPa,流体总流量30 mL/g,萃取前向萃取池中加入3 mL甲醇作夹带剂。采用该法对非洲山毛豆中鱼藤酮的萃取率(质量分数)达0.41%,相当于氯仿浸提法的98%;萃取产物总得率(质量分数)为16.62%,高于氯仿浸提法。与常规溶剂法相比,该方法具有萃取效率高、精确、无毒、经济、操作简便、选择性可调等优点。     

印楝素、鱼藤酮防治茶丽纹象甲成虫室内试验初报

通过室内试验研究了印楝素、鱼藤酮对茶丽纹象甲的杀虫效果。结果表明,药后2d,0.5%印楝素乳油1370倍液对茶丽纹象甲成虫的防治效果即达95.56%,药后3d达到100%,与2.5%联苯菊酯乳油1000倍液的差异不显著。0.5%印楝素乳油可作为有机茶园茶丽纹象甲防治的选择药剂。而7.5%鱼藤酮乳油750倍液,药后5d防效才达51.16%,防治效果较差。     

植物离体培养物中鱼藤酮的分离与HPLC测定

毛鱼藤（Ｄｅｒｒｉｓ ｅｌｌｉｐｔｉｃａ （Ｒｏｘｂ．）Ｂｅｎｔｈ．）和西非灰毛豆（Ｔｅｐｈｒｏｓｉａ ｖｏｇｅｌｉｉ Ｈｏｏｋ．ｆ．）离体培养物经９：１ＣＨＣｌ３／ＭｅＯＨ溶剂提取,Ｃ－１８柱层析分离后,在室温条件下以６６：３４ＭｅＯＨ／Ｈ２Ｏ或４２：５８ＣＨ３ＣＮ／Ｈ２Ｏ为流动相,支样品进行反相柱ＨＰＬＣ分析。结果表明,毛鱼藤和西百灰毛豆的愈伤组织均能合成鱼藤酮,但含量相当低,分别为１７．４μｇ／ｇ,１．８μｇ／ｇ干重。经诱导生根分化的毛鱼藤愈伤组织鱼藤酮含量达１３６．２μｇ／ｇ干重,为愈伤组织含量的７．８倍。西非灰毛豆悬浮培养物的鱼藤酮含量虽比愈伤组织有很大提高,但还不及毛鱼藤愈伤组织含量。结果表明,植物的基因类型在次生代谢上起重要作用,同时离体组织或细胞的分化有助于鱼藤酮的生物合成。     

The effects of phytochemical pesticides on the growth of cultured invertebrate and vertebrate cells

A range of cultured cells of invertebrate and vertebrate origin was grown in the presence of a number of phytochemical pesticides to test the effect of the latter on cell proliferation. The main observation was that azadirachtin was a potent inhibitor of insect cell replication, with an EC50 of 1.5 x 10(10) M against Spodoptera cells and of 6.3 x 10(9) M against Aedes albopictus cells, whilst affecting mammalian cells only at high concentrations (> 10(-4) M). As expected, the other phytochemical pesticides, except for rotenone, had little effect on the growth of the cultured cells. Rotenone was highly effective in inhibiting the growth of insect cells (EC50:10(-8) M) but slightly less toxic towards mammalian cells (EC50:2 x 10(-7) M). Neem terpenoids other than azadirachtin and those very similar in structure significantly inhibited growth of the cell cultures, but to a lesser degree. The major neem seed terpenoids, nimbin and salannin, for example, inhibited insect cell growth by 23% and 15%, respectively.     

绿僵菌与3种杀虫剂混用对沙葱萤叶甲的协同作用

采用点滴法,于室内测定了阿维菌素、茚虫威和鱼藤酮3种杀虫剂单独使用及与绿僵菌混用对沙葱萤叶甲3龄幼虫的协同致死作用,以期为协调使用化学药剂和生防制剂防治沙葱萤叶甲提供参考。3种杀虫剂单独使用时的毒力测定结果表明,阿维菌素对沙葱萤叶甲3龄幼虫的毒力最强,LD50值为4.80 ng/头,其次是茚虫威和鱼藤酮,LD50值分别为10.47和53.21 ng/头。采用不同浓度的3种杀虫剂分别处理绿僵菌24和48 h后,发现药剂对绿僵菌分生孢子萌发均有一定的抑制作用,但抑制率低于20%,且抑制率随着药剂浓度的降低和作用时间的延长而降低,其中茚虫威的平均抑制率最小,分别仅为1.89%±0.57%和0.89%±0.39%。将3种杀虫剂分别与绿僵菌孢子悬液混配使用,对防治沙葱萤叶甲3龄幼虫均具有协同增效作用:其中茚虫威与绿僵菌混用增效作用最强,比单独施用绿僵菌时LT50值缩短了8.12 d;阿维菌素与绿僵菌混用时LT50值缩短了7.60 d;鱼藤酮与绿僵菌混用时LT50值缩短了6.45 d。     

鱼藤酮对小鼠脑神经递质多巴胺及其代谢产物的影响

研究了注射鱼藤酮对C57BL/6N小鼠神经递质多巴胺的影响。给小鼠皮下注射鱼藤酮(10 mg/kg)两次,间隔2 h,在第2次注射8 h后分离脑组织样品,并应用高效液相色谱仪配电化学检测器,测定了小鼠脑内多巴胺(DA)及其主要代谢物3,4-二羟苯酰乙酸(DOPAC)和高香草酸(HVA)的水平。结果表明,第2次注射8 h后,鱼藤酮处理组的脑内DA、DOPAC和HVA水平分别比对照组高40.5%、544.4%和168.2%,差异显著(P<0.01)。实验显示,鱼藤酮对多巴胺能神经系统具有毒性作用。     

Dopamine Release Suppression Dependent on an Increase of Intracellular Ca2+ Contributed to Rotenone-induced Neurotoxicity in PC12 Cells

Rotenone is an inhibitor of mitochondrial complex I that produces a model of Parkinson’s disease (PD), in which neurons undergo dopamine release dysfunction and other features. In neurons, exocytosis is one of the processes associated with dopamine release and is dependent on Ca²⁺ dynamic changes of the cell. In the present study, we have investigated the exocytosis of dopamine and the involvement of Ca²⁺ in dopamine release in PC12 cells administrated with rotenone. Results demonstrated that rotenone led to an elevation of intracellular Ca²⁺ through Ca²⁺ influx by opening of the voltage-gated Ca²⁺ channel and influenced the soluble N-ethylmaleimide attachment protein receptor (SNARE) proteins expression (including syntaxin, vesicle-associated membrane protein 2 (VAMP₂) and synaptosome-associated protein 25 (SNAP-25)); pretreatment with a blocker of L-type voltage-activated Ca²⁺ channels (nifedipine) decreased the intracellular dopamine levels and ROS formation, increased the cell viability and enhanced the neurite outgrowth and exocytosis of synaptic vesicles. These results indicated that the involvement of intracellular Ca²⁺ was one of the factors resulting in suppression of dopamine release suppression in PC12 cells intoxicated with rotenone, which was associated with the rotenone-induced dopamine neurotoxicity.     

鱼藤酮及六种哒唑、咪唑类医用驱虫剂对松材线虫的作用

松材线虫是一种毁灭性的松林病害,发现有效的松材线虫杀虫剂是一项长期而艰巨的任务。测定了鱼藤酮及六种哒唑、咪唑类驱虫药对松材线虫的作用,发现鱼藤酮对松材线虫的杀灭效果远强于以乙酰胆碱酯酶为主要靶标的克线磷,鱼藤酮及阿苯哒唑、甲苯哒唑、氟苯哒唑、噢吩哒唑、磷酸左旋咪唑、盐酸左旋咪唑处理松材线虫120 h后的LC50分别为1.86μg/mL,2.11μg/mL,9.77μg/mL,5.38μg/mL,6.66μg/mL,1.14μg/mL和1.40μg/mL,其中线粒体呼吸链抑制剂鱼藤酮及呼吸代谢抑制剂阿苯哒唑、磷酸左旋咪唑、盐酸左旋咪唑对松材线虫表现出了出色的杀灭效果,松材线虫的呼吸代谢系统是松材线虫的敏感靶标。     

鱼藤酮在不同溶剂中的紫外吸收研究

测定了鱼藤酮在甲醇、三氯甲烷、乙醇、四氯化碳等溶剂中的吸收情况。结果表明，鱼藤酮在不同的溶剂中，其紫外吸收峰和吸收图谱是不完全相同的，它在上述4种溶剂中的最大吸收波长分别是218．0、294．0、211．0和290．0nm。此外，确定了鱼藤酮的紫外检测波长为293．0nm。     

鱼藤酮在3种土壤中的吸附-解吸附特性

为了综合评价鱼藤酮在土壤环境中的吸附-解吸附特性,采用批量平衡法,系统研究了鱼藤酮在砂壤土、黏壤土及壤土3种农业土壤中的吸附-解吸附行为。结果表明,鱼藤酮在3种土壤中的吸附-解吸附行为符合Freundlich模型 (R2≥0.946 8),吸附常数 (K_f-ads) 在1.52~11.39之间,吸附能力为:黏壤土 > 砂壤土 > 壤土;而解吸附常数 (K_f-des) 在1.02~4.55之间,解吸附强弱次序为壤土 > 黏壤土 > 砂壤土。鱼藤酮在砂壤土、黏壤土和壤土3种土壤中有机碳吸附常数 (K_OC) 分别为982、101 7和219,而滞后系数 (H) 分别为0.687 3、0.556 9和0.892 3,表明鱼藤酮在黏壤土及砂壤土中移动性较弱,有正迟滞作用,而在壤土中移动性较强,无迟滞作用。该研究将对鱼藤酮的环境风险评估具有一定的理论指导意义。