苦参生物碱的研究

从苦参(Sophora flavescens Ait)根中分离得到9个生物碱,用波谱等方法确定为槐果碱(sophocarpine)、苦参碱(matrine)、异苦参碱(isomatrine)、槐醇(sophoranol)、N-甲基野靛碱(N-methylcytisine)、槐定(sophoridine)、氧化苦参碱(oxymatrine)、氧化槐果碱(oxysophocarpine)和氧化槐醇(sophoranol N-oxide)。其中氧化槐醇是首次从苦参根中得到的。     

西藏不同生长地砂生槐群体种子生物碱含量及其相关性分析

为了明确不同群体间和群体内砂生槐天然种子富含生物碱含量与组成的差异,该研究选取来自西藏“一江四河”流域、海拔2 900～4 100 m范围内10个群体的砂生槐种子作为研究材料,采用高效液相色谱法,测定10个群体300个单株种子重要生物碱(氧化苦参碱、苦参碱、槐果碱、槐定碱)含量,应用SPSS软件分析了地理分布特征与生物碱含量之间的相关性及群体遗传变异关系,以筛选生物碱含量较高的群体,为砂生槐药物开发利用提供依据。结果表明：(1)10个群体的砂生槐种子中均含有氧化苦参碱、苦参碱和槐果碱,含量依次为：氧化苦参碱(46.18～64.08 mg/g)>苦参碱(1.14～9.82 mg/g)>槐果碱(0.08～1.16 mg/g),其中氧化苦参碱占生物碱总量的90%以上,且群体4中含量最高(64.08±7.37 mg/g);其他微量生物碱在群体3中含量最高;群体间氧化苦参碱、苦参碱和槐果碱均差异极显著(P＜0.01)。(2)氧化苦参碱群体内和群体间变异相对较小,苦参碱和槐果碱群体内和群体间变异系数均较大,而且氧化苦参碱和槐果碱的群体间变异均大于群体内变异,苦参碱的群体间变异小于群体内变异。(3)氧化苦参碱与海拔呈显著正相关关系(r = 0.117^*),苦参碱与海拔、经度和纬度均呈极显著负相关关系(r ＜ -0.326^**),其他关系不显著。氧化苦参碱与苦参碱和槐果碱均呈显著负相关关系(r ＜-0.162^**),而苦参碱与槐果碱呈显著正相关关系(r =0.789^**)。(4)聚类分析将10个群体聚为4类,各类间的距离差异比较大,且大多与地理因素有关。     

苦参碱类生物碱的应用进展

苦参碱是豆科植物苦参、苦豆子、广豆根等中草药的活性成分,是苦参碱类生物碱的代表,这一类生物碱主要还包括氧化苦参碱、槐果碱、氧化槐果碱、槐定碱和槐胺碱等。而对苦参碱和氧化苦参碱的研究最为广泛。苦参碱对中枢神经具有解热、镇痛、抗惊厥、稳定神经等作用;对心血管系统具有明显的负性频率和正性肌力作用、能防治动脉粥样硬化、减轻心肌损伤等作用;对消化系统具有抗肝损伤、抗纤维化、升高白细胞等作用;还具有抗肿瘤、抗肝癌等作用。氧化苦参碱用于治疗白细胞低下和慢性乙型、丙型肝炎、阻止肝纤维化和肝硬化,有抗心律失常、强心、平喘等作用。槐果碱有明显抗柯萨奇B病毒(Coxsackie Virus B,CVB)的作用、免疫调节功能以及抗严重急性呼吸道综合症(Severe Acute Respiratory Syndrome,SARS)病毒的作用。槐定碱有改善心功能、抗炎、抗肿瘤、抗病毒作用。槐胺碱具有免疫抑制、抗心律失常作用。本文综述了这一类生物碱的成分分析及其部分化合物的药理活性和临床应用等方面的研究进展。     

苦豆子生物碱抗烟曲霉、须癣毛癣菌和新生隐球菌的作用研究

目的对比研究苦豆子总碱、槐果碱、苦参碱、槐定碱、氧化苦参碱体外抗烟曲霉、须癣毛癣菌和新生隐球菌的敏感性,为苦豆子进一步开发利用奠定基础。方法分别用管碟扩散法和微量液基稀释法,测定苦豆子总碱、槐果碱、苦参碱、槐定碱、氧化苦参碱对烟曲霉、须癣毛癣菌和新生隐球菌的抗菌活性。结果管碟法显示上述5种生物碱体外对烟曲霉、须癣毛癣菌和新生隐球菌均有不同程度的抗菌活性。前4种生物碱对烟曲霉的最低杀菌浓度(MFC)分别为25.0、4.25、5.0、3.125 mg·mL~(-1),氧化苦参碱对烟曲霉MFC500 mg·mL~(-1);对须癣毛癣菌的MFC分别为3.125、2.125、5.0、1.5625、125 mg·mL~(-1);对新生隐球菌的MFC分别为3.125、4.25、10、6.25、1000 mg·mL~(-1)。结论槐果碱、苦参碱及槐定碱其抗真菌活性与苦豆子总碱相当,氧化苦参碱的作用不显著。     

苦豆子生物碱对萝卜蚜的毒力及其对几种酯酶的影响

苦豆子Sophora alopecuroids(L.)的次生代谢物质为喹诺里西定生物碱类。本研究明确了该生物碱中的野靛碱对萝卜蚜(Lipaphis erysimi)有很高的毒杀作用,对其无翅成蚜的致死中浓度(LC₅₀浸渍法)为(432.59±2.12)mg/L,优于著名的杀蚜生物碱毒黎碱和烟碱,两者对该试虫的LC₅₀分别为(684.70±2.28)mg/L和(1090.65±2.01)mg/L。用小菜蛾(Plutella xylostella)幼虫作试虫,得知苦豆子7种主要生物碱对昆虫的乙酰胆碱酯酶(AChE)有抑制作用,其抑制程度排序为：总碱>野靛碱>槐胺碱>槐定碱>槐果碱>氧化苦参碱>苦参碱>苦豆碱。野靛碱和苦豆碱对a-乙酸萘酯酶、a-乙酸萘酯羧酸酯酶及酯酶同功酶的活性亦表现不同程度的抑制作用。     

黄花棘豆生物碱成分研究

采用硅胶柱层析、Sephadex LH-20和半制备HPLC等色谱手段对草原毒草黄花棘豆(Oxytropis ochrocephala Bunge)生物碱成分进行分离纯化,从全草提取物中分离得到8个生物碱成分。利用MS、1H NMR和13C NMR波谱技术鉴定化合物结构为槐定碱(1)、槐胺(2)、异槐定碱(3)、苦参碱(4)、7,11-去氢苦参碱(5)、槐果碱(6)、羽扇豆碱(7)和苦马豆素(8)。化合物1~6为首次从棘豆属植物中分离得到。     

半夏和苦豆子生物碱的抗线虫活性

首次研究了半夏和苦豆子生物总碱,以及3种主要的苦豆子单体生物碱(槐定碱、氧化苦参碱和氧化槐果碱)的抗线虫活性。半夏生物总碱处理24 h,对松材线虫、南方根结线虫、全齿复合线虫和秀丽隐杆线虫的半抑制浓度(IC50)分别为16.18、20.25、33.24和20.77μg/m L。苦豆子生物总碱处理24 h,对上述4种线虫表现出更强的抑制活性,IC50值分别为0.622、0.383、1.476和1.224μg/m L,抗线虫活性强于或接近阳性对照阿维菌素。3种苦豆子单体生物碱均表现出明显的抗线虫活性,其中氧化槐果碱的抗线虫活性最强。研究结果为植物源杀线虫剂的研究和开发提供了依据。     

PPARα转基因小鼠在药物毒性评价中的应用

目的研究PPARα转基因小鼠在评价PPARα激动剂类药物毒性时,是否比传统动物更敏感。方法8周龄PPARα转基因小鼠(Tg)和C57BL/6J小鼠(WT),雌雄各半,分别随机分成3组,氯贝丁酯高剂量组(400 mg/kg)、氯贝丁酯低剂量组(300 mg/kg)、溶媒对照组(10%羧甲基纤维素钠)。连续灌胃一个月,给药结束后检测血生化指标、心肝肾脏器系数及病理改变,并观察动物的一般生长情况。结果 1血生化:Tg♂高、低剂量组肌酐(CREA)、天门冬氨酸氨基转移酶(AST)分别显著高于各对应的野生型对照组(P0.01,P0.05)。2脏器系数:Tg高、低剂量组肾脏系数与Tg溶媒对照组比较均有显著增加(P0.01,P0.05)。3组织病理:Tg各剂量组肝脏、肾脏病理损伤较WT各剂量组更严重。结论 PPARα转基因小鼠评价PPARα激动剂药物肝脏毒性和肾脏毒性时比常规C57BL/6J野生小鼠更敏感,是一个新的动物模型。     

槐定碱抑制人胃癌MKN45细胞的增殖并促进其凋亡

本研究旨在探讨槐定碱对人胃癌MKN45细胞增殖和凋亡的影响及其可能的作用机制。将人胃癌MKN45细胞分为对照组和槐定碱组(设立6个浓度亚组)。用MTT法检测MKN45细胞增殖情况,用免疫细胞化学染色法和Western blot观察细胞高迁移率组蛋白3(high mobility group-box 3,HMGB3)的蛋白表达,用Hoechst 33342染色法观察细胞形态变化,用流式细胞术检测细胞凋亡。结果显示,槐定碱作用48 h能显著抑制MKN45细胞的增殖,作用呈剂量依赖性。与对照组相比,槐定碱作用48 h后MKN45细胞的HMGB3蛋白表达显著减少(P0.05),细胞凋亡率显著升高(P0.05)。以上结果提示,槐定碱作用48 h可抑制人胃癌MKN45细胞的增殖,促进其凋亡,其机制可能与HMGB3表达下调有关。     

苦豆子生物碱对萝卜蚜的毒力及其对几种酯酶的影响

苦豆子Sophoraalopecuroids（L．）的次生代谢物质为喹诺里西定生物碱类。本研究明确了该生物碱中的野靛碱对萝卜蚜（Lipaphiserysimi）有很高的毒杀作用,对其无翅成蚜的致死中浓度（LC50,浸渍法）为（4325±2．12）mg／L,优于著名的杀蚜生物碱毒黎碱和烟碱,两者对该试虫的LC50分别为（684．70±2．28）mg／L和（1090．65±2．01）mg／L。用小菜蛾（Plutellaxylostella）幼虫作试虫,得知苦豆子7种主要生物碱对昆虫的乙酰胆碱酯酶（AChE）有抑制作用,其抑制程度排序为：总碱＞野靛碱＞槐胺碱＞槐定碱＞槐果碱＞氧化苦参碱＞苦参碱＞苦豆碱。野靛碱和苦豆碱对α-乙酸萘酯酶、α-乙酸萘酯羧酸酯酶及酯酶同功酶的活性亦表现不同程度的抑制作用。     

Selective extraction and separation of oxymatrine from Sophora flavescens Ait. extract by silica-confined ionic liquid

This study highlighted the application of a two-stepped extraction method for extraction and separation of oxymatrine from Sophora flavescens Ait. extract by utilizing silica-confined ionic liquids as sorbent. The optimized silica-confined ionic liquid was firstly mixed with plant extract to adsorb oxymatrine. Simultaneously, some interference, such as matrine, was removed. The obtained suspension was then added to a cartridge for solid phase extraction. Through these two steps, target compound was adequately separated from interferences with 93.4% recovery. In comparison with traditional solid phase extraction, this method accelerates loading and reduces the use of organic solvents during washing. Moreover, the optimization of loading volume was simplified as optimization of solid/liquid ratio.     

Antifeedant activity and acute and residual toxicity of alkaloids from Sophora flavescens (leguminosae) against formosan subterranean termites (Isoptera: Rhinotermitidae)

Ku Shen, Sophora flavescens Aiton (Leguminosae), contains alkaloids that possess broad biological activities. This study showed that total matrines (a mixture of the alkaloids), and matrine and oxymatrine (two major alkaloids) had a strong antifeedant effect against Formosan subterranean termite, Coptotermes formosanus Shiraki (Isoptera: Rhinotermitidae), at all tested concentrations (ranging from 0.01 to 5%). Total matrines and matrine also had significantly toxic effects at 5%. The estimated LD50 of total matrines and matrine was 12.3 and 8.6 microg per insect, respectively, after 24 h; 13.3 and 7.1 microg per insect after 48 h; and 10.1 and 4.8 microg per insect after 1 wk. The antifeedant and toxic properties of total matrines- and matrine-treated wood remained effective at least 12 mo after treatment under both light and dark storage conditions. These chemicals have great potential for commercial development as wood treatment agents.     

Biosynthese und Stoffwechsel der chinolizidinalkaloide von Sophora alopecuroides

Administration of matrine-U-³H and sophocarpine-U-³H to Sophora alopecuroides seedlings shows that these compounds were incorporated into quinolizidine alkaloids such as matrine, sophacarpine, and their N-oxides, but not into sophoridine. It is suggested that there is no stereochemical conversion of alkaloids of matrine configuration into sophoridine by the plant. The incorporation of cadaverine-1,5-¹⁴C was so low that it cannot be regarded with certainty as a physiological precursor of the alkaloids. The N-oxides of matrine and sophocarpine were isolated and identified by their chromatographic and chemical properties.     

苦参碱的提取分离及对小鼠的毒性研究

采用酸性乙醇提取、乙醚萃取、硅胶柱层析分离等方法从苦参中分离到苦参碱单体.以小鼠为实验动物进行毒性测定,小鼠的死亡主要集中在48h内,48h后无小鼠的死亡现象.小鼠对苦参碱的耐受量大于30mg.k-g1,小于140mg.k-g1,致死中量LD50为64.01mg.kg-1,回归方程Y=-3.2370+4.5602X,LD50标准误差SE=6.14.适口性的测定表明,苦参碱对小鼠有较好的适口性,可以作为杀鼠剂使用.     

苦参植株中总生物碱的分布及含量测定

选用酸性染料络合显色法测定了江苏盱眙产苦参的根、茎、叶、种子等部位中苦参总碱的含量 ,为充分利用苦参全植株提供了一定的依据。     

Antiviral effects of sophoridine against coxsackievirus B3 and its pharmacokinetics in rats

Coxsackievirus B3 (CVB3) is a major pathogen for acute and chronic viral myocarditis. The aim of this study was to investigate the antiviral effects of sophoridine, an alkaloid extracted from Chinese medicinal herb, Sophora flavescens, against CVB3, and the underlying pharmacokinetics. First, we determined the antiviral effects of sophoridine against CVB3 in in vitro (primarily cultured myocardial cells), in vivo (BALB/c mice) and serum pharmacological experiments. Then, we determined the pharmacokinetic behavior in serum samples of SD rats after oral administration by HPLC. Finally, we determined the effects of sophoridine on the production of cytokines in a murine viral myocarditis model by measuring mRNA expression of some important cytokines in hearts of infected BALB/c mice by RT-PCR. We found that sophoridine exhibited obvious antiviral effects both in vitro and in vivo, and serum samples obtained from rats with oral administration of sophoridine reduced the virus titers in infected myocardial cells. The serum concentration profile correlated closely with antiviral activity profile. Moreover, sophoridine significantly enhanced mRNA expression of IL-10 and IFN-gamma, but decreased TNF-alpha mRNA expression. In conclusion, sophoridine possesses antiviral activities against CVB3, by regulating cytokine expression, and it is likely that sophoridine itself, not its metabolites, is mainly responsible for the antiviral activities. Therefore, sophoridine may represent a potential therapeutic agent for viral myocarditis.     

苦豆子生物碱对小菜蛾体内部分杀虫剂代谢酶活性的影响

以苦豆子Sophora alopecuroids 7种生物碱和小菜蛾Plutella xylostella幼虫为试材,研究了该生物碱对小菜蛾体内降解杀虫剂的羧酸酯酶、磷酸酯酶、谷胱甘肽-S-转移酶活性的影响。结果表明: 野靛碱和苦豆碱对羧酸酯酶活性有显著的抑制,为可逆抑制类型的非竞争性抑制作用;野靛碱等5种生物碱对酸性磷酸酯酶有明显的抑制,野靛碱对碱性磷酸酯酶有弱抑制作用;其中野靛碱等3种生物碱对谷胱甘肽-S-转移酶有明显的抑制作用。     

Matrine induces Akt/mTOR signalling inhibition‐mediated autophagy and apoptosis in acute myeloid leukaemia cells

Pharmacological modulation of autophagy has been referred to as a promising therapeutic strategy for cancer. Matrine, a main alkaloid extracted from Sophora flavescens Ait, has antitumour activity against acute myelocytic leukaemia (AML). Whether autophagy is involved in antileukaemia activity of matrine remains unobvious. In this study, we demonstrated that matrine inhibited cell viability and colony formation via inducing apoptosis and autophagy in AML cell lines HL‐60, THP‐1 and C1498 as well as primary AML cells. Matrine promoted caspase‐3 and PARP cleavage dose‐dependently. Matrine up‐regulated the level of LC3‐II and down‐regulated the level of SQSTM1/p62 in a dose‐dependent way, indicating that autophagy should be implicated in anti‐AML effect of matrine. Furthermore, the autophagy inhibitor bafilomycin A1 relieved the cytotoxicity of matrine by blocking the autophagic flux, while the autophagy promoter rapamycin enhanced the cytotoxicity of matrine. Additionally, matrine inhibited the phosphorylation of Akt, mTOR and their downstream substrates p70S6K and 4EBP1, which led to the occurrence of autophagy. In vivo study demonstrated that autophagy was involved in antileukaemia effect of matrine in C57BL/6 mice bearing murine AML cell line C1498, and the survival curves showed that mice did benefit from treatment with matrine. Collectively, our findings indicate that matrine exerts antitumour effect through apoptosis and autophagy, and the latter one might be a potential therapeutic strategy for AML.     

Synergistic action between jasmonic acid and nitric oxide in inducing matrine accumulation of Sophora flavescens suspension cells

Secondary metabolites not only play important ecological roles in plants but also are important pharmaceutical and source compounds for derivative synthesis. Production of plant secondary metabolites is believed to be controlled by the endogenous signal network of plants. However, the molecular basis is still largely unknown. Here we show that matrine production of Sophora flavescens Ait. cells treated with low levels of jasmonic acid （JA） and nitric oxide （NO） is significantly increased although treatment with low concentrations of JA or NO alone has no effects on matrine production, showing that JA and NO may act synergistically in triggering matrine production. Moreover, treatment with NO triggers lipoxygenase （LOX） activity and enhances JA levels of the cells, showing that NO may activate the endogenous JA biosynthesis of S. flavescens cells. External application of JA induces nitric oxide synthase-like activities and stimulates NO generation of S. flavescens cells, which suggests that JA may trigger NO generation of the cells. Thus, the results reveal a mutually amplifying reaction between JA and NO in S. flavescens cells. Furthermore, JA and NO inhibitors suppress not only the mutually amplifying reaction between JA and NO but also the synergistic effects of NO and JA on matrine production. Therefore, the data demonstrate that the synergistic action of JA and NO in inducing matrine production might be due to the mutually amplifying reaction between JA and NO in the cells.