【作者】 刘永松；

【导师】 潘玲； 蒋立科；

【作者基本信息】 安徽农业大学 ， 基础兽医学， 2006， 硕士

【摘要】 百蕊草(Thesium Chinense Turcz.)为檀香科(Santalaceae)植物,系草本或灌木,稀小乔木植物,以寄生或半寄生为其主要生物学特征。百蕊草全草均可入药,具有抗菌消炎,清热解毒等作用,是一种广谱的抗菌中草药。但由于传统中药研究思路和方法的局限性,严重限制了该中药在临床上的应用。鉴于中药现代发展趋势,其药效的作用机制是当前中药药理研究的热点,所以有效提取百蕊草的主要成分,搞清楚其主要成分的作用机制对百蕊草的临床应用具有重要意义。通过采用超声波萃取(UE)和微波萃取技术(MAE)对百蕊草的有效成分进行提取研究,并与中药研究中最常采用的水和乙醇作为萃取剂萃取靶物质的抑菌检测效果相比较,结果表明:在波长为216nm和276nm的双波长紫外光下检测提取效果,利用微波水萃取技术能够较好地提取出百蕊草的主要成分。鉴于百蕊草对不同细菌的抗菌作用,本项研究采用牛津杯法将百蕊草的提取物对七种常见致病细菌(标准株)进行了体外抗菌检测,并对抑菌试验的条件和参数进行了摸索。结果表明,百蕊草对G+和G-细菌均有作用,对G+细菌的作用强于G-细菌,几种试验菌的敏感顺序由铜绿假单胞菌(P.aeruginosa)、藤黄八叠球菌(B.subtilis)、蜡状芽胞杆菌(S.lutea)、枯草芽胞杆菌(B.cereus)、嗜水气单胞菌(A.hydrophila)和金黄色葡萄球菌(S.aureus)依次递增,其中以金黄色葡萄球菌、嗜水气单胞菌最为敏感,大肠埃希氏菌(E.coli)对百蕊草素敏感性最差。与此同时,试验还测定了各种较敏感细菌的最小抑菌浓度(MIC),这对百蕊草作为一种广谱的抗菌中草药在临床应用上具有指导性作用。在有效提取百蕊草素的基础上,对百蕊草的抗菌作用机制作了初步研究。采用多种生化检测方法跟踪检测金黄色葡萄球菌对糖和蛋白质代谢结果的影响,表明百蕊草素能够显著抑制金黄色葡萄球菌对葡萄糖和蛋白质的代谢,提示百蕊草素改变细菌的代谢机制,破坏或抑制了其对周围物质选择性通透性,干扰或破坏金黄色葡萄球菌内氧化还原状态,从而影响了其新陈代谢;此外,对百蕊草有效成分还进行再分离纯化试验。利用聚酰胺吸附柱层析技术再分离百蕊草有效成分,将分离得的五种组分进行相关抗菌试验检测发现,百蕊草抗菌作用不是单一的药物作用,而是多种成分联合作用的结果,为更深入研究百蕊草药理学奠定了重要的基础。更多还原

【Abstract】 Thesium Chinense Turcz. is a Santalaceae plant, belongs to the herbage or the frutex, is also a rare arbor plant, takes autoeciousness or semiparasite as its main biology characteristic. The whole grass may be used as medicine, has the antibiotic action and diminish inflammation effects, be provided with clearing away heat and detoxifcation function, et al. Thesium Chinense Turcz. has the high medical value as one kind of a broad-spectrum antibiotic Chinese herbal medicine. As a result of being restricted by the thoughts and the methods of the traditional Chinese herbal medicine research, the function of Thesium Chinense Turcz. and its mechanism is not confirmed, which limited its’application in the clinical. In view of the fact of the developing characteristic of modern Chinese herbal medicine, that the functional mechanism of Chinese native herbal medicine is the hot spot in the current pharmacology research. Therefore, effectively extracting the active components, and making clear its effect mechanism is the vital significance to study Thesium Chinense Turcz.The active components of Thesium Chinense Turcz were extracted by Ultrasonic-wave extraction (UE) and Mro-wave-assisted (MAE) technology in the research, Through bacterial susceptibility test, two extraction reagent, water and alcohol were compareted, which mostly be used to extract the components of Chinese herbal medicine. The results indicated that using double wave 216nm and 276nm of ultraviolet ray to examine the extracting can judge availably the effect of the active components,and water-extract in Mro-wave-assisted is the better method to extract the active components in Thesium Chinense Turcz..In order to verified the susceptibility of Thesium Chinense Turcz to different bacterium, Seven kinds of common nosogenesis-bacteria were used to do bacterial examination test by the the method of Oxford-cup tests in the experiment. The antibacterial experiment conditions and the parameter were also studied. The results showed that:Thesium Chinense Turcz. is sensitive to both the G+ and the G-- bacterium, and the effect on the G+ was stronger than that on the G--, the sensitive order of seven kinds of experimental bacterium was S.aureus, A.hydrophila, B.cereus, S.lutea, B.subtilis, and P.aeruginosa. And, the S.aureus and A.hydrophila were the most sensitive bacterium, but E.coli is the worst to Thesium Chinense Turcz.. Each sensitive bacteriums’MIC was also tested in the experiment. The result confirmed that Thesium Chinense Turcz. is available to use as a broad-spectrum antibiotic Chinese herbal medicine in clinic. The antibacterial mechanism of Thesium Chinense Turcz was studied using its active components which were extracted in the former experiment. The sugar and the protein metabolism on the S.aureus were tested by biochemistry examination methods. The result indicated that the active components of Thesium Chinense Turcz. could change the bacterium metabolism mechanism. The effect disturbs or destroys the oxidation reduction condition of the S.aureus, thus affect its metabolism. In addition, chromatographic analysis technology was used to separate active components of Thesium Chinense Turcz, and five kinds of components were obtained to do antibacterial experiment to acquire their correlation. In conclusion, the antibacterial function of Thesium Chinense Turcz. is not the single medicine’s action, but the joint action of its several kinds of ingredients. The test laid an important foundation to continue advanced research on the Chinese herbal medicine Thesium Chinense Turcz.更多还原