【作者】 刘娟；

【导师】 彭成；

【作者基本信息】 成都中医药大学 ， 药理学， 2016， 硕士

【摘要】 目的通过系统的化学成分研究和药理实验,开展益母草对子宫平滑肌影响的药效物质基础研究。方法运用大孔树脂、MCI、硅胶、葡聚糖凝胶Sephadex LH-20、反相C18等柱色谱,以及制备薄层色谱、半制备液相色谱、重结晶等分离方法,对益母草活性部位的化学成分展开系统研究,获得单体化合物,结合MS、IR,1DNMR, 2D NMR等波谱学方法和Marfey’s等化学方法鉴定化合物结构。建立正常大鼠离体子宫模型,观察不同单体化合物对子宫收缩的活动力、张力和频率的影响,用十六通道生理记录系统记录子宫平滑肌收缩曲线,以探寻益母草调节子宫平滑肌的药效物质基础。结果1.益母草化学成分研究本论文采用现代分析技术和色谱方法,从益母草正丁醇部位分离鉴定了21个化合物,包括生物碱10个,环肽5个,黄酮苷3个,木脂素及小分子芳香族化合物3个,其中新化合物4个。2.益母草单体化合物调节子宫平滑肌研究 益母草单体化合物对子宫平滑肌的作用表现为兴奋和舒张双重调节作用。在试验剂量范围内,化合物6、7、9和13对子宫平滑肌具有兴奋作用,具体表现为：化合物6中、高剂量均能明显升高子宫收缩平均值、最小值以及收缩活力；化合物13低、中、高剂量对子宫收缩平均值、最大值以及最小值,高剂量对子宫收缩活力均呈明显升高作用；化合物7低剂量对子宫收缩最小值,中剂量对子宫收缩频率、平均值及活力均有升高；化合物9中、高剂量均能升高子宫收缩平均值。相反,在试验剂量范围内,化合物10、16～18对子宫平滑肌具有舒张作用,具体表现为：化合物18低、中、高剂量对子宫收缩活力,低、高剂量对子宫收缩频率、平均值以及高剂量对子宫收缩最大值均呈明显降低作用；化合物17中、高剂量对子宫收缩频率、平均值、活力,高剂量对子宫收缩最大值均有降低作用。化合物10低剂量对子宫收缩频率、收缩活力,高剂量对子宫收缩频率、收缩活力以及收缩最大值也均有降低作用；化合物16低、中、高剂量组对子宫收缩频率、收缩活力,中剂量组对子宫收缩最大值有降低作用。化合物12低剂量对子宫收缩频率也有降低作用。以上低、中、高剂量的终浓度分别为1μM, 10μM,50μM,所有差异均具有统计学意义(P<0.05或P<0.01)。结论1.从益母草正丁醇部位分离得到21个化合物,有4个为新化合物。在新化合物中,化合物1*和2*为两个新环肽,化合物3*和4*为首次从益母草属中分离得到的特殊N-氧化物型甾体生物碱。2.益母草单体化合物具有兴奋子宫和舒张子宫双重作用。环肽、甾体生物碱苷、胍类生物碱可能为缩宫活性成分类型,黄酮苷、甾体生物碱、亚精胺可能为舒张子宫活性成分类型。3.从益母草中获得具有缩宫活性的环肽,突破了传统观念认为益母草生物碱为缩宫主要药效物质基础的认识。更多还原

【Abstract】 Objective The pharmacodynamic material basis for influencing the activity of uterine smooth muscle of Leonurus japonicus have been studied by systemic chemical constituents and pharmacological experiments.Methods By using column chromatography over macroporous resin, MCI, siliea gel, Sephadex LH-20, and Rp C18, preparative thin layer chromatography, semi-preparative liquid chromatography, and recrystallization methods, the chemical components of bioactive fration of L. japonicus were isolated. Their structures were determined by spectroscopic and chemical techniques, including MS, UV, IR,1DNMR,2DNMR, and Marfey’s methods.The uterus in vitro uterus model of normal rats has been established and applied to detect the effects of the isolated compounds on the mobility, amplitude, tension, and frequency of uterine smooth muscle. The contraction curves of uterine smooth muscle were were recorder with sixteen channels physiological recording system.Results 1. Study on the chemical components of Leonurus japonicus Twenty-one compounds were isolated from L. japonicus, including ten steroidal alkaloid, five cyclic peptides, three flavone glycosides, three phenylpropanoid and aromatic compounds, four of them are new compounds.2. Study on uterine contraction activity of monomeric compound from Leonurus japonicus The effects on uterine smooth muscle of Leonurus japonicus showed dual effects of excitement and relaxation. In the test range, compounds 6,7,9 and 13 showed excitatory effects on the contractile activity of isolated uterus in normal rats. Specifically as follows:both high and medium doses of compound 6 could significantly increase average, minimum, and the contractile activity of uterine contraction; compound 13 could increase the average,maximum, minimum at low, middle, high doses, increasing the contractile activity of uterine contraction at high dose, compound 7 could rise minimum of the uterine contraction at low dose, increasing frequency, average and vitality of the uterine contraction at middle dose; Compound 9 could increase the average of uterine contraction at high dose.On the contrary, in the test range, Compounds 10,16-18 had an inhibition effect on the contractile activity of isolated uterus in normal rats, specifically as follows:compound 18 could reduce the vitality of the uterine contraction at low, middle, high dose, deseasing frequency, average of the uterine contraction at low, high dose, reducing the maximum of the uterine contraction at high dose. Compounds 17 could reduce frequency, average and the vitality of the uterine contraction at middle, high dose, reducing the maximum of the uterine contraction at high dose; Compounds 10 could reduce the frequency and the vitality of the uterine contraction at low dose, reducing frequency, the vitality, the maximum of the uterine contraction at high dose; Compounds 16 could reduce frequency and the vitality of the uterine contraction at low, middle, high dose, reducing the maximum of the uterine contraction at middle dose; Compounds 12 could the frequency of the uterine contraction at low dose, the above low, middle and high doses were respective as the final concentration of 1 μM,10μM,50μM, all the differences were statistically significant (P<0.05 or P＜0.01).Conclusions 1. Twenty-one compounds were isolated from L. japonicus, Among the compounds found, these were four new compounds, including two new cyclic peptide (1* and 2*), two special N-oxide steroidal alkaloid (3* and 4*)were discovered for the first time.2. The in vitro uterus experiment of normal rat indicated that there were two kinds of chemical constituents that had uterine contraction and relaxation effects separately in Leonurus japonicus. Cyclic peptides, steroidal alkaloid glycosides, leonurine were possible the material basis of uterine contraction by analysis of the structures of the compounds, flavone glycosides, steroid alkaloid were the main active constituents of relaxing the uterus.3. Cyclic peptides with uterine contraction activity have been abtained, which break through the traditional concept that motherwort alkaloid was the main pharmacodynamic material basis for uterine contraction.更多还原