【作者】 段雪云；

【导师】 刘焱文；

【作者基本信息】 湖北中医学院 ， 中医基础理论， 2009， 博士

【摘要】 缬草（Valeriana officinalis L.）系败酱科（Valerianace）缬草属（Valerian）植物,根及根茎是主要入药部位。具有镇静安神、解痉止痛之功效,主治心神不安,心悸失眠,癫狂,风湿痹痛,痛经等。现代药理研究表明,缬草具有较好的抗心律失常作用。为了阐释缬草抗心律失常作用的物质基础,在前人研究的基础上,采用学科交叉的研究方法,通过超临界CO₂萃取法和溶剂法,将缬草分为不同提取部位;然后以乌头碱、氯仿两种诱导剂诱导心律失常的模型为指标进行药理筛选,确定了缬草抗心律失常的主要活性部位。缬草水溶性物质部位有稳定的降低室颤发生率,推迟心律失常发生时间,减少死亡率的作用;超临界CO₂萃取物和水液部位对氯仿诱发小鼠心室纤颤具有明显的拮抗作用。氯仿、乌头碱两种诱导剂均能诱导心律失常的发生,但其诱发机理不同。缬草水溶性物质部位对两种诱导剂诱导的心律失常均有明显的对抗作用,说明其抗心律失常机制是多方面的,符合中药多成分多靶点的作用机理。采用色谱分离法和波谱鉴定法等现代科学技术,从缬草抗心律失常有效部位中分离和鉴定了5种化合物,分别为青刺尖木脂醇苷（prinsepiol4-O-β-D-glucoside）、8-羟基松脂醇苷（8-hydroxypinoresinol-4-O-β-D-glucoside）、β-谷甾醇（β-sitosterol）、kanokoside A、绿原酸（Chlorogenic acid）。利用现代电生理学中的膜片钳技术,将上述分离得到的化合物对转基因Kv1.5细胞进行了抗心律失常活性筛选,从中筛选出2个具有抗心律失常活性的化合物,分别为青刺尖木脂醇苷、8-羟基松脂醇苷;并对青刺尖木脂醇苷进行了作用机理探讨,研究结果表明,青刺尖木脂醇苷对Kv1.5钾通道的抑制,说明青刺尖木脂醇苷的抗心律失常可能主要是通过抑制钾通道,延长动作电位和有效不应期而达到治疗目的,初步阐释了缬草抗心律失常的作用机制,所以青刺尖木脂醇苷有望成为治疗抗心律失常药。在抗心律失常物质基础研究的基础上,采用高效液相色谱法对缬草水溶性部位进行了指纹图谱研究,标定了15个共有峰,归属了4个共有峰,分别是青刺尖木脂醇苷、8-羟基松脂醇苷、绿原酸和橙皮苷;并对不同产地、不同采收时间以及缬草近缘品种进行了比较;建立了可靠的分析方法,符合《国家食品药品监督管理局》颁布的中药注射剂指纹图谱技术要求。从而为规范缬草属植物的药用来源及采收季节提供了参考依据。采用高效液相色谱法对缬草中抗心律失常有效成分青刺尖木脂醇苷进行了含量分析研究,建立了灵敏度高、重现性好、可靠性强的含量测定方法。从而为有效控制缬草药材的质量,保证用药安全提供了科学依据。更多还原

【Abstract】 Valeriana officinalis L.belongs to department of Patrinia of valerian,its roots and rhizomes are the main parts of medicine which have sedative and spasmolytic effects, attending uneasy,palpitation insomnia,mania,rheumatism Bitongkang,dysmenorrheal,etc.Modern pharmacological studies have shown that valerian has anti-arrhythmic effect.In order to clarify the material basis of the anti-arrhythmic effects of Valeriana we adopted interdisciplinary research methods based on the previous studies.Through supercritical CO₂ extraction method and solvent method the Valeriana was divided into different valerian extracts,and then using aconitine alkali and chloroform to induce arrhythmia model for pharmacological screening indicators to determine the main active anti-arrhythmic site of Valeriana.The water-soluble parts of Valerian stably decrease the incidence of ventricular fibrillation,postpone arrhythmias time and reduce the mortality.Supercritical CO₂ extract and water-liquid parts have more pronounced antagonism to chloroform-induced ventricular fibrillation in mice.And the other parts also have anti-arrhythmic effects indifferent extent.Although inducing arrhythmia through different mechanisms,the water-soluble part of Valerian have altogether obvious antagonistic effects to the two agent chloroform and aconitine induced arrhythmia, while the other parts have different response.The all above reflect the mechanisms for its anti-arrhythmic is multifaceted and there are multiple active ingredients.Using chromatographic separation and spectral identification method,we can separate and identify five kinds of compounds from the effective anti-arrhythmic parts of Valeriana as follows:Green barbed tip lignan glycosides （prinsepiol-4-0-β-D-glucoside）;8-hydroxy resin glycosides （8-hydroxypinoresinol -4-o-β-D-glucoside）;β-sitosterol; kanokoside A;Chlorogenic acid.Using modern electrophysiological patch-clamp technique to investigate these compounds separated above,there are two compounds that have been proved had anti-arrhythmic activity: Green barbed tip lignan glycosides and 8-hydroxy resin glycosides.We explored the action mechanism of the green lignan glycosides and the results show that the Green barbed tip lignan glycosides through inhibiting potassium channels and prolonging effective refractory period to achieve the anti-arrhythmic therapeutic purposes.Green barbed tip lignan glycosides are expected to become anti-arrhythmic drug.The paper initially explain the mechanism of its anti-arrhythmic. On the basis of basic research of anti-arrhythmic substances we utilized high-performance liquid chromatography to research the fingerprint of water-soluble parts of Valeriana and demarcate 15 common peaks.Valeriana from different original Sources,different harvesting time and Valeriana close related species were also compared,establishing a reliable analytical method in accordance with the "Chinese Pharmacopoeia" and the State Food and Drug Administration issued technical standards.Thereby the research provides a reference for the Valeriana medicinal plants harvesting sources and season.Using high-performance liquid chromatography to analyze active anti-arrhythmic ingredients Green barbed tip lignan glycosides of Valeriana content has high sensitivity,good reproducibility,powerful reliability,through which we can effectively control Valeriana medicinal quality and provide a scientific basis to ensure drug safety.更多还原