摘要
厚朴,来源于木兰科木兰属植物厚朴Magnolia officinalis Rehd et Wils.和凹叶厚朴Magnolia officinalis Rehd. et Wils. var. biloba Rehd. et Wils的干燥干皮、枝皮和根皮。厚朴始载于《神农本草经》,列为中品。厚朴为常用中药,不少常用处方和中成药制剂中都有厚朴入药,国内外需求量很大。为满足对厚朴不断增大的需求量,目前已在四川、湖北、浙江、福建等主产区建立厚朴药材生产基地,大力发展厚朴种植。据初步统计,全国种植厚朴和凹叶厚朴不少于70万亩,全国厚朴叶的蕴藏量每年可达30万吨以上。历代主要本草中所载正品厚朴其药用部位均为树皮,但古今药学专著中均未见厚朴叶的药用记载。目前,国内大量厚朴叶未加利用,任其凋落腐烂。
本论文在课题组前期研究工作的基础上,进一步探寻厚朴叶的药用价值,以期实现厚朴资源的综合利用。通过对厚朴Magnolia officinalis Rehd et Wils.叶的药效活性和化学成分研究,取得了如下成果并有所创新：
1.首次对厚朴叶乙醇提取物及该提取物不同极性萃取部位进行离体血管环实验研究,并初步讨论其作用机制。其中,厚朴叶乙醇总提取物和该提取物的氯仿萃取部位对基础状态血管环具有收缩作用,该作用能被Ca2+抑制剂和a受体抑制剂阻断,提示在厚朴叶中可能有激活L型钙离子通道和a受体的成分,引起Ca2+内流,从而引起血管平滑肌收缩；厚朴叶乙醇总提取物的正丁醇萃取部位对NE预收缩血管环具有舒张作用,但不同剂量范围舒张效果和舒张机制有差异,表明该部位不同剂量作用时的机制可能不完全相同。
2.系统地研究了厚朴叶血管药效活性部位的化学成分,共分离得到36个化学成分,鉴定出26个化合物,主要包括黄酮类、苯丙素类、以联苯木脂素和双环氧木脂素类为主的木脂素类以及简单芳香族类化合物,其中22个化合物为首次从厚朴M. officinalis Rehd et Wils.叶中分离得到,14个化合物为首次从厚朴M. officinalis Rehd et Wils.植物中分离得到,8个化合物为首次从木兰属(Magnolia L.)植物中分离得到,为厚朴叶的药效研究奠定了物质基础。
3.首次对厚朴叶中分离量较高的8个成分进行离体血管环作用活性追踪,初步明确厚朴叶对离体血管作用的物质基础和作用机制。研究发现8个成分对基础状态血管均无影响；首次发现从舒张血管活性部位分离得到的槲皮苷和阿福豆苷均能使NE预收缩的血管环张力下降,其中槲皮苷非内皮依赖性舒张血管,该作用可能与血管平滑肌细胞有关；阿福豆苷内皮依赖性舒张血管,可能是通过NO-GC-cGMP途径和通过激活环氧合酶途径引发血管舒张效应。此外,芦丁也有报道认为具有体外舒张血管的作用,因此,厚朴叶舒张血管的药效物质基础与槲皮苷、阿福豆苷和芦丁有关。
4.槲皮苷在厚朴叶中含量较高,并且表现出与厚朴叶大极性部位类似的药效活性,提示在厚朴叶舒张血管这一药效中,可将槲皮苷作为其主要活性物质进行研究。本论文首次以槲皮苷为对照品,采用HPLC法测定不同采收季节厚朴叶中的槲皮苷含量,分析其动态变化情况,为综合评价厚朴叶的最佳采收期提供依据。研究发现,不同采收季节的厚朴叶中槲皮苷含量总体上呈现规律性变化,即6月中旬达到最大值,此后含量逐渐下降,11月份以后自然掉落的叶片中槲皮苷含量极低,因此,以槲皮苷为指标,6月中旬为厚朴叶的最佳采收期。
本论文从化学成分、药理作用、含量测定等多方面对厚朴叶进行了比较系统的研究。结果显示厚朴叶有较强的血管调节作用,在心血管系统方面有一定的药用价值,尤其大极性部位的舒张血管活性表明其具有潜在的降压作用,可将厚朴叶作为治疗高血压的血管扩张药进行进一步研究,以便开发利用。本研究为充分合理利用厚朴叶资源,实现厚朴的综合利用和可持续开发奠定理论基础,也为开发价格便宜、获取方便、毒副作用低的厚朴叶中药新资源提供依据。
本论文受教育部博士点基金项目——“厚朴叶药用价值的系统研究”(课题编号200806330006)和科技部“十一五”国家科技支撑项目——“厚朴种植恢复重建关键技术研究与示范”(课题编号2009BAI84803)资助。
Houpo (Magnolia Officinalis Cortex), the bark of stem, root or branch of Magnolia officinalis Rehd et Wils. or Magnolia officinalis Rehd et Wils. var. biloba Rehd et Wils., was first recorded in "Shennong Bencaojing". It has been used in traditional Chinese medicine for the treatment of abdominal distention and dyspepsia, and this crude drug is one of the important components of the prescriptions. At present, the number of the leaves of M. officinalis is so huge for many trees are planted in major producing areas, but most of them has been wasted.
On the foundation of the group's former work, the medicinal value of the leaves of M. officinalis was studied in this paper, including chemical constituents, pharmacological effects and the dynamic variation of quercitrin. The major achievements and creative points were as followed.
1. The paper first reported the contraction activity of alcoholic extract and chloroform extract of the leaves of M. officinalis on rabbit aortic rings with or without endothelium in vitro. They might exert action through the long lasting voltage operated Ca2+channels and alpha-receptors, because the effect could be inhibited by both calcium channel blocker and alpha-blocker. Whereas, the n-Butyl alcohol extract of the leaves of M. officinalis exhibited relaxant effect on the arteries induced by NE, but the mechanism was unclear.
2. From the chloroform extract and n-Butyl alcohol extract of the leaves of M. officinalis,36compounds were isolated and26of which were identified based on spectral techniques. All these compounds were composed of flavonoids, phenylpropanoids, lignans and so on.22compounds were isolated from the leaves of Magnolia officinalis for the first time,14compounds were isolated from Magnolia officinalis for the first time and8compounds were isolated from the genus Magnolia for the first time.
3.8compounds were highly-contained among the identified compounds, they were also assayed on isolated rabbit arteries in vitro in order to initially elucidate efficacious material basis of it. The data indicated that all the8compounds were ineffective on the aortic rings, but quercitrin and afzelin which isolated from the n-Butyl alcohol extract could dose-dependently relax the rabbit arteries induced by NE. The endothelium-independent relaxation effect of quercitrin might be mediated by vascular smooth muscle cell. On the contrary, the excellent vasodilation of afzelin was endothelium-dependent, which was associated with NO-GC-cGMP pathway and cyclooxygenase pathway. In addition, rutin was also been reported of its relaxation on the isolated arteries. Therefore, the efficacious material basis of the leaves of M. officinalis was quercitrin, afzelin and rutin.
4. As the main active ingredient of the leaves of M. officinalis, quercitrin was used as reference standards to analyze the content changes of it in the different harvest time through HPLC method for the first time. The research showed that the content of quercitrin changed regularly, it was highest in the middle of June and the fallen leaves contained little quercitrin after November. Consequently, taking quercitrin as index, the optimum time of the leaves of M. officinalis to harvest was the middle of June.
The findings suggested that the leaves of M. officinalis could modulate the tension of isolated blood vessels, especially vasodilatation, it reminded that the leaves of M. officinalis might be used as cardiovascular drug or antihypertensive drug.
The study was supported by the Ph. D. Programs Foundation of Ministry of Education of China---"Study on the Medicinal Value of the Leaves of M. officinalis'"(No:200806330006). and the National Science and Technology Support Project (2009BAI84B03) from Ministry of Science.
引文
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