中药厚朴及其类似品的有效成分分析

本文对中药厚朴及其类似品中的有效成分——厚朴酚、和厚朴酚及木兰箭毒碱分别进行了高效液相色谱法和双波长扫描法测定;对其挥发油中化学成分进行了气-质联用分离与鉴定。     

厚朴不同炮制方法有效成份的变化

厚朴未经加工不能入药，而经过不同的加工炮制方法炮制的饮片，由于加工过程中因标准的控制不同，有效成份含量有很大差异，因而对疗效产生影响。笔者对厚朴未经炮制与经过不同炮制方法加工炮制的饮片，通过高效液相色谱法（HPLC）分析其有效成份厚朴酚、和厚朴酚、木兰箭毒碱的变化，为厚朴的加工炮制提供一个理论依据。     

厚朴和大叶木兰中厚朴酚与和厚朴酚的含量测定

本文报道了用高效液相层析法,分别测定厚朴和大叶木兰中的和厚朴酚与厚朴酚的方法。高效液相层析是用YWG-CH填充柱1(250×4 mm i d),用甲醇—水(78:22)作流动相,压力72kg/cm²,和厚朴酚与厚朴酚能完全分离,且样品中其它成分对分离和测定无影响。本法简便、快速、重现性好。用本法测定了不同产地和来源的厚朴中和厚朴酚与厚朴酚的含量。也测定了大叶木兰的干皮、枝皮和叶子中的和厚朴酚与厚朴酚的含量。     

厚朴炮制前后指纹图谱及主要成分含量变化的研究

摘要：目的:建立厚朴及其炮制品的HPLC指纹图谱,并对其所含的6个有效成分进行定量分析。方法:采用HPLC,Inertsil ODS-3 C18色谱柱（250 mm×4.6 mm,5μm）,流动相乙腈-0.1%磷酸水溶液梯度洗脱,检测波长:250 nm,流速:1.0 ml·min-1,建立生厚朴和姜厚朴的HPLC指纹图谱,通过相似度评价、聚类分析对厚朴炮制前后指纹图谱进行分析,并对紫丁香苷、木兰花碱、木兰苷B、木兰苷A、和厚朴酚与厚朴酚进行含量测定。结果:建立了厚朴及其炮制品的指纹图谱,以木兰苷B为参照峰,生厚朴标定了14个共有峰,姜厚朴标定了16个共有峰,与各自对照指纹图谱的相似度均>0.950。紫丁香苷、木兰花碱、木兰苷B、木兰苷A、和厚朴酚和厚朴酚在炮制前的质量分数分别为0.042 3%,0.061 4%,0.081 5%,0.088 1%,1.190 0%,0.969 0%;经炮制后,上述6种成分在姜厚朴中的含量分别为0.034 6%,0.033 1%,0.065 7%,0.070 5%,1.330 0%,1.090 0%。结论:厚朴的HPLC指纹图谱在炮制前后发生了显著变化,通过对比共有峰的保留时间和色谱图,指认出部分成分;并发现峰4和峰5为姜厚朴的特有峰,因此峰4和峰5可用于区别生厚朴和姜厚朴; 6个有效成分含量在炮制前后均发生了一定程度的变化,样品中紫丁香苷、木兰花碱、木兰苷B和木兰苷A含量排序为厚朴>姜厚朴,样品中和厚朴酚和厚朴酚含量排序则为姜厚朴>厚朴。     

基于网络药理学厚朴姜炙的科学内涵研究

目的 通过网络药理学法构建厚朴姜炙前后的差异成分-靶点-通路网络,探讨厚朴姜炙前后药理作用差异的分子机制,揭示厚朴姜炙的科学内涵。方法 查阅文献筛选出厚朴姜炙前后的差异成分,利用Swiss Target Prediction预测差异成分的潜在靶点。利用STRING数据库构建蛋白-蛋白互作网络,然后通过DAVID数据库进行GO注释和KEGG通路富集分析,最后运用Cytoscape 3.8.2软件构建差异成分-靶点-通路网络。结果 通过查阅文献共筛选出8个差异成分并预测得到85个靶点。DAVID数据库筛选出排名前10位的KEGG通路,通过Cytoscape 3.8.2软件得到差异成分-靶点-通路网络。结论 厚朴姜炙后可通过紫丁香苷、厚朴酚、和厚朴酚、木兰花碱、木兰箭毒碱及β-桉叶醇等成分含量的变化来改变厚朴的药效作用。姜炙后,厚朴主要是通过血清素突触、钙信号通路、前列腺癌和膀胱癌等信号通路发挥镇痛、抗胃溃疡、抗肿瘤的作用。     

木兰科药用植物的研究——Ⅱ.厚朴的原植物和资源利用

本文对厚朴进行了本草考证。研究了木兰科厚朴原植物来源,包括厚朴类,姜朴类、枝子皮类和土厚朴类等多种植物。同时用红外、紫外光谱和薄层层桁等方法检查了各类厚朴中厚朴酚(magnolol)(Ⅰ)与和厚朴酚(honokiol)(Ⅱ)以及β-桉油醇(β-cudcsmol)(Ⅲ)的存在。实验证明上述成分厚朴类都含,其余各类厚朴或含少量或不含。但从植物系统位置和植物化学分类观点来看,则Ⅰ,Ⅱ,Ⅲ的分布有一定规律,即属于木兰亚属(Subgen.Magnolia)皱皮木兰组(Seot.Rytidospermum)是Ⅰ,Ⅱ,Ⅲ的分布最集中的植物群。玉兰亚属(Subgen.Pleurochasma)各组除个别种外,均不含Ⅰ,Ⅱ,Ⅲ。未研究过的土厚朴类来源于木莲属(Manglietia),如四川木莲等也含Ⅰ或Ⅱ成分。     

厚朴酚与和厚朴酚含量测定方法

厚朴为胃肠道疾病常用中药,具有温中下气,化湿引滞的功效。厚朴中的主要成分是厚朴酚与和厚朴酚。我国药典已将厚朴酚与和厚朴酚的含量作为厚朴的质量控制标准。据报道,各地商品中有近30种植物皮混作厚朴用。在已发现的厚朴中,对于厚朴酚及和厚朴酚的含量测定方法很多,有薄层色谱法、高效液相色谱法、气相色谱法等,但缺乏对其中厚朴酚或和厚朴酚的含量测定方法进行综合系统的报道,笔者为此对有关文献作一综述。     

木兰科药用植物的研究——Ⅱ.厚朴的原植物和资源利用

本文对厚朴进行了本草考证。研究了木兰科厚朴原植物来源,包括厚朴类,姜朴类、枝子皮类和土厚朴类等多种植物。同时用红外、紫外光谱和薄层层桁等方法检查了各类厚朴中厚朴酚(magnolol)(Ⅰ)与和厚朴酚(honokiol)(Ⅱ)以及β-桉油醇(β-cudcsmol)(Ⅲ)的存在。实验证明上述成分厚朴类都含,其余各类厚朴或含少量或不含。但从植物系统位置和植物化学分类观点来看,则Ⅰ,Ⅱ,Ⅲ的分布有一定规律,即属于木兰亚属(Subgen.Magnolia)皱皮木兰组(Seot.Rytidospermum)是Ⅰ,Ⅱ,Ⅲ的分布最集中的植物群。玉兰亚属(Subgen.Pleurochasma)各组除个别种外,均不含Ⅰ,Ⅱ,Ⅲ。未研究过的土厚朴类来源于木莲属(Manglietia),如四川木莲等也含Ⅰ或Ⅱ成分。     

不同商品规格厚朴提取液中厚朴酚与厚朴酚含量测定

厚朴酚和厚朴酚为厚朴的主要酚性成分，利用气相色谱法对不同商品规格厚朴的厚朴酚与和厚朴酚进行测定。结果厚朴酚含量从高至低依次为：根朴，靴朴，筒朴，枝朴，和厚朴酚含量最高的是根朴，以下依次为筒朴，靴朴，枝朴。     

湖北恩施产厚朴中厚朴酚与和厚朴酚的定量分析

目的：探索厚朴酚类成分的积累与分布规律。方法：采用高效液相色谱法测定厚朴酚、和厚朴酚含量，流动相为甲醇-水（78：22），检测波长为294nm。结果：树龄增长，厚朴酚与和厚朴酚总含量下降；树龄相同，酚含量与胸径、皮厚呈正相关；根皮的酚含量远高于干皮和枝皮，干皮自下而上酚含量逐渐降低，枝皮的酚含量比中上部干皮的高。结论：若单以厚朴酚与和厚朴酚含量为指标，厚朴采割年限应适当缩短；根、干、枝皮应混合用药。     

Magnolol and honokiol account for the anti-spasmodic effect of Magnolia officinalis in isolated guinea pig ileum

Magnolia officinalis is a commonly used traditional Chinese medicine for treating gastrointestinal disorders. HPLC quantification analysis revealed that magnolol and honokiol were the most abundant constituents of M. officinalis extracts, with their contents in the ethanol extract being the highest, the water extract the least and the 50 % ethanol extract in between. In guinea pig isolated ileum, both magnolol and honokiol inhibited contraction to acetylcholine. The herbal extracts also produced inhibitory responses, in an order of decreasing efficacy: ethanol extract > 50 % ethanol extract > water extract. The differences in inhibitory efficacies among the three extracts were similar to the differences in their magnolol and honokiol contents. Further examination demonstrated that two mixtures containing solely magnolol and honokiol at concentrations identical to those determined in the ethanol and water extracts exhibited similar levels of anti-spasmodic effects as their respective extracts while a "blank" ethanol extract free of magnolol and honokiol failed to produce any response. These observations suggest that the magnolol and honokiol contents account for the anti-spasmodic effects of M. officinalis extracts in guinea pig isolated ileum.     

In vitro antibacterial and anti-inflammatory effects of honokiol and magnolol against Propionibacterium sp

Honokiol and magnolol, two major phenolic constituents of Magnolia sp., have been known to exhibit antibacterial activities. However, until now, their antibacterial activity against Propionibacterium sp. has not been reported. To this end, the antibacterial activities of honokiol and magnolol were detected using the disk diffusion method and a two-fold serial dilution assay. Honokiol and magnolol showed strong antibacterial activities against both Propionibacterium acnes and Propionibacterium granulosum, which are acne-causing bacteria. The minimum inhibitory concentrations (MIC) of honokiol and magnolol was 3-4 microg/ml (11.3-15 microM) and 9 microg/ml (33.8 microM), respectively. In addition, the killing curve analysis showed that magnolol and honokiol killed P. acnes rapidly, with 10(5) organisms/ml eliminated within 10 min of treatment with either 45 microg (169.2 microM) of magnolol or 20 microg (75.2 microM) of honokiol per ml. The cytotoxic effect of honokiol and magnolol was determined by a colorimetric (3-(4,5-dimetyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide) (MTT) assay using two animal cell lines, human normal fibroblasts and HaCaT. In this experiment, magnolol exhibited lower cytotoxic effects than honokiol at the same concentration, but they showed similar cytotoxicity when triclosan was employed as an acne-mitigating agent. In addition, they reduced secretion of interleukin-8 and tumor necrosis factor alpha (TNF-alpha) induced by P. acnes in THP-1 cells indicating the anti-inflammatory effects of them. When applied topically, neither phenolic compound induced any adverse reactions in a human skin primary irritation test. Therefore, based on these results, we suggest the possibility that magnolol and honokiol may be considered as attractive acne-mitigating candidates for topical application.     

不同树龄厚朴生物活性成分的比较分析

利用高效液相色谱法(HPLC)测定不同树龄厚朴中绿原酸、芦丁、金丝桃苷、槲皮苷、槲皮素、和厚朴酚和厚朴酚的含量。目的:为了比较分析同一来源不同树龄厚朴中7种活性物质含量变化,为最佳厚朴采集年龄提供参考。结果:绿原酸、金丝桃苷、槲皮素和厚朴酚在13年树龄时含量最高,分别达到0.28、0.033、0.018和5.90%;芦丁和槲皮苷在7年树龄时含量最高,其含量分别为0.34和0.52%;和厚朴酚和厚朴酚在10年树龄时其含量最高为2.40%。厚朴中的厚朴酚和和厚朴酚为主要成分在10年树龄和到26年树龄其成分的变化不大,10、13和26年树龄厚朴中7种活性物质总的含量分别为7.99、8.75和8.73%,总含量变化也不显著,因此,选10年的树为最佳厚朴采集年龄。     

高温致厚朴SFE-CO2萃取物中厚朴酚、和厚朴酚降解

目的了解影响厚朴中有效成分厚朴酚、和厚朴酚的不稳定因素.方法用HPLC法测定在不同实验条件下,厚朴SFE-CO2萃取物中厚朴酚、和厚朴酚的含量;用加速实验测定有效期.结果厚朴酚、和厚朴酚的有效期(25℃)分别为387 d和476 d.结论温度是影响厚朴酚、和厚朴酚有效期的最主要因素, 降解符合一级速度反应.     

高温致厚朴SFE-CO2萃取物中厚朴酚、和厚朴酚降解

目的了解影响厚朴中有效成分厚朴酚、和厚朴酚的不稳定因素。方法用HPLC法测定在不同实验条件下,厚朴SFE-CO₂萃取物中厚朴酚、和厚朴酚的含量;用加速实验测定有效期。结果厚朴酚、和厚朴酚的有效期(25℃)分别为387 d和476 d。结论温度是影响厚朴酚、和厚朴酚有效期的最主要因素, 降解符合一级速度反应。     

厚朴提取物中主要成分的小肠吸收特性

目的从其吸收部位、药物浓度两个方面对厚朴的主要成分和厚朴酚、厚朴酚的吸收特性进行研究。方法采用在体大鼠肠段回流实验。结果和厚朴酚、厚朴酚在小肠上部吸收最佳 ;在小肠中部及下部吸收无明显差异 ,但较小肠上部相比稍差。和厚朴酚、厚朴酚在肠道中的吸收量与其浓度成线性关系 ,随着药物浓度的增大吸收量增加 ,其吸收百分率基本不变。结论和厚朴酚、厚朴酚在肠道中吸收良好 ,吸收机制为被动转运     

Antimicrobial activities of hydroxybiphenyl derivatives (I)

It was revealed that magnolol and honokiol isolated from the stem bark ofMagnolia obovata, had potent antibacterial activity againstBacillus anthracis. A quantitative analytical method of magnolol and honokiol by HPLC has been established, and the amounts of the two components in the dried stem bark ofM. obovata were 1.94% and 0.44%, respectively.

     

重庆产厚朴不同部位厚朴酚与和厚朴酚分布规律的研究

目的 研究厚朴不同部位厚朴酚与和厚朴酚的分布规律,为进一步开发利用厚朴资源并对其进行质量控制提供科学依据。方法 采用HPLC测定厚朴不同部位中厚朴酚与和厚朴酚的含量。结果 厚朴根皮、干皮、枝皮和厚朴叶中厚朴酚与和厚朴酚的总含量分别约为：8.22%,2.30%,2.07%和0.43%,厚朴各部位中厚朴酚与和厚朴酚的总含量差异具有统计学意义(P<0.05)。结论 厚朴不同部位中厚朴酚与和厚朴酚的含量差异较大,其质量控制有待深入研究,厚朴不同部位在临床用药时应有所区别,另外厚朴叶资源具有较大的开发价值。     

Differential inhibitory effects of honokiol and magnolol on excitatory amino acid-evoked cation signals and NMDA-induced seizures

The effects of honokiol and magnolol, two major bioactive constituents of the bark of Magnolia officinalis, on Ca(2+) and Na(+) influx induced by various stimulants were investigated in cultured rat cerebellar granule cells by single-cell fura-2 or SBFI microfluorimetry. Honokiol and magnolol blocked the glutamate- and KCl-evoked Ca(2+) influx with similar potency and efficacy, but did not affect KCl-evoked Na(+) influx. However, honokiol was more specific for blocking NMDA-induced Ca(2+) influx, whereas magnolol influenced with both NMDA- and non-NMDA activated Ca(2+) and Na(+) influx. Moreover, the anti-convulsant effects of these two compounds on NMDA-induced seizures were also evaluated. After honokiol or magnolol (1 and 5 mg/kg, i.p.) pretreatment, the seizure thresholds of NMRI mice were determined by tail-vein infusion of NMDA (10 mg/ml). Data showed that both honokiol and magnolol significantly increased the NMDA-induced seizure thresholds, and honokiol was more potent than magnolol. These results demonstrated that magnolol and honokiol have differential effects on NMDA and non-NMDA receptors, suggesting that the distinct therapeutic applications of these two compounds for neuroprotection should be considered.