地黄环烯醚萜苷类成分的制备及血清药物化学研究
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摘要
地黄始载于《神农本草经》,列为上品,是我国著名的的“四大怀药”之一,使用其治疗疾病有着几千年的历史,在我国的多个省市都有栽培,道地药材主产于河南武陟、修武、温县等地。地黄为玄参科植物地黄Rehmannia glutinosa libosch.的新鲜或干燥的块根或是炮制加工品,入药依次为:鲜地黄、生地黄、熟地黄。这三者在药效方面存在着明显的差异:鲜地黄清热生津,凉血,止血;生地黄清热凉血,养阴,生津;熟地黄滋阴补血,益精填髓。在2005年版的《中华人民共和国药典》一部收载的中药方剂中,有1/5都涉及到地黄。
近年来,国内外学者对地黄化学成分的研究较为普遍,对其真正有效成分的研究较少,其确定的“有效成分”多是某种药物中所含的主要成分或指标成分,并无依据证明其为有效成分,这也限制了对中药材的综合开发利用。任何药物都是与人体相互作用后才发挥疗效的,日本学者田代真一在1989年提出了“血清药化学”和“血清药理学”的概念,通过分析口服给药后血清中的成分来确定中药直接进入体内的物质,这成为快速、简便、准确研究药物药效物质基础的有效途径。
本文第一章对地黄的化学成分、提取与分离、药理作用、血清药物化学的研究进展和近年来中药血清药物药物化学的研究进展进行了综述。
第二章提取分离鲜地黄中环烯醚萜苷类成分。药材经粉碎,醇提,大孔吸附树脂,反复硅胶柱层析,葡聚糖凝胶柱纯化,从地黄醇提物中分离得到3个化合物。经TLC检测和光谱分析鉴定结构,这3个化合物分别是梓醇、地黄苷D、益母草苷。
第三章建立一套地黄血清药物化学的实验方法。以含药血清中梓醇、地黄苷D、益母草苷的峰面积的值为指标,确定最佳给药方式及采血时间。以梓醇的峰面积为指标,确定最佳给药量。确定了给药方案是:按浓度为0.7g/ml,1ml/100g,连续给药3d、1次/d,在给药后1h采血,能在含药血清中发现目标成分。这为地黄血清药物化学的进一步研究奠定了基础。
第四章应用血清药物化学的方法,对地黄环烯醚萜苷类入血成分进行分析,为确定地黄的活性成分奠定基础。通过比较对照品、含药血清、空白血清的HPLC指纹图谱,初步确定地黄的入血成分。在给药血清中发现了2个入血成分,且均是原型成分(分别是梓醇、地黄苷D)。这2个入血成分可能是地黄在体内直接作用的物质。
第五章建立地黄醇提物及环烯醚萜苷部位血清药物化学的初步研究。通过比较对照品、含药血清、空白血清的HPLC指纹图谱,确定地黄醇提物及环烯醚萜苷部位的入血成分。在给药血清中发现了3个入血成分,且均是原型成分(分别是梓醇、地黄苷D、益母草苷)。这3个入血成分可能是地黄在体内直接作用的物质,对其进行深入的研究将为进行地黄活性成分的进一步研究奠定基础。
Rehmannia glutinosa libosch was originally recorded in“Shennong bencaojing”and classied as the upper class. It is one of the Four famous herbs produced in Henan HuaiQing. It has been used for medical treatment for several thousands years. It has been cultivated in many provinces, but mainly in Wuzhi, Xiuwu and Wenxian and other areas of Henan province. Radix Rehmannia .is the fresh or dry block root or processed product of the Scrophulariaceae plant Rehmannia glutinosa libosch, medicine followed by: fresh Radix Rehmannia, dry Radix Rehmannia and Radix Rehmannia Praeparata. There are distinct differences between the three: fresh Radix Rehmannia can removing heat to promote salivation, cooling blood, and stop bleeding ; dry Radix Rehmannia can removing heat to cool blood,nourishing Yin,body fluid regeneration;Radix Rehmannia Praeparata can nourishing Yin and blood,tonifying didney jing and filling pith. In the Pharmacopoeia of People's Republic of China (2005 edition),which recorded Chinese medicinal formulae, there were one fifth of it concerned Radix Rehmannia.
In recent years,researching chemical composition of Radix Rehmannia were common among domestic and oversea scholars. Studies on the constituents of its few real active ingredients, its determination of“active ingredients”are mostly some sort of drugs or indicators contained in the main component composition, there is no basis to prove that as an active ingredients, which also limits the Chinese crude drug developed and utilized.Any drug interactions are related to the human body after the therapeutic work. Shinichi Tashiro Japanese scholar in 1989 proposed the concept of“Serum Medicinal Chemistry”and“Serum Pharmacology”.By analyzing the serum after oral administration to determine the composition of traditional Chinese drug directly into the body of material, it become fast, simple and accurate study of the material basis of drug efficacy effective way.
In the first chapter,we summarize the study on the chemical,extraction,pharmacological, serum pharmacochemisty of Rehmannia glutinosa Libosch and serum pharmacochemisty of traditional chinese drug.
In the second chapter,we extraction Iridoid glycosides from Rehmannia glutinosa Libosch .The materia medica by crushing ,ethanol extraction , Separation and purification through the macroporous resin, silica gel column chromatography, Sephadex column,identify the structure of compound through TLC and spectroanalysis.From the ethanol extract of Rehmannia glutinosa Libosch isolated three compounds .There are Catalpol, Rehmannia glycoside D, Motherwort glycosides
In the third chapter,we establish experimental method for Serum pharmacochemisty of Rehmannia glutinosa Libosch. According to peak area value of Catalpol、Rehmannioside D、Leonuride in drug-containing serum catalpol as an indicator to determine the best administration style and blood collection time. According to peak area value of Catalpol in drug-containing serum catalpol as an indicator to determine the best dosage. By comprehensive analysis, the dosage regimen is as following: to continuously dose for 3 days one time a day by the concentration of 0.7g/ml, 1ml/100g. To collect blood after one hour of dosage, we can examine the destination ingredients. Thus, it can lay the foundation of further research on the Serum pharmacochemisty of Rehmannia glutinosa Libosch.
In the fourth chapter,we using the method of serum pharmacochemisty and analysis the compounds of Rehmannia glutinosa libosch absorbed into the blood,lays the foundation for the Rehmannia glutinosa libosch definite active constituent. The constituents absorbed into blood were determined by comparing the HPLC fingerprints of the reference substance、tested serum samples and blank serum sample. Two compounds absorbed into blood were detected, they were original constituents of Rehmannia glutinosa libosch(Catalpol、Rehmannioside D). Two constituents absorbed into blood were possible bioactive components of Rehmannia glutinosa libosch.
In the fifth chapter,we investigate the parameters of serum pharmacochemistry of ethanol extract of Radix Rehamannia and Iridoid glycosides. The constituents absorbed into blood were determined by comparing the HPLC fingerprints of the reference substance、tested serum samples and blank serum sample. Three compounds absorbed into blood were detected, they were original constituents of Rehmannia glutinosa libosch(Catalpol、Rehmannioside D、Leonuride). Three constituents absorbed into blood were possible bioactive components of Rehmannia glutinosa libosch, Its serum pharmacochemistry should be subject to complete investigation so as to lay a foundation for the further study of active ingredients of Rehmannia glutinosa libosch.
近年来,国内外学者对地黄化学成分的研究较为普遍,对其真正有效成分的研究较少,其确定的“有效成分”多是某种药物中所含的主要成分或指标成分,并无依据证明其为有效成分,这也限制了对中药材的综合开发利用。任何药物都是与人体相互作用后才发挥疗效的,日本学者田代真一在1989年提出了“血清药化学”和“血清药理学”的概念,通过分析口服给药后血清中的成分来确定中药直接进入体内的物质,这成为快速、简便、准确研究药物药效物质基础的有效途径。
本文第一章对地黄的化学成分、提取与分离、药理作用、血清药物化学的研究进展和近年来中药血清药物药物化学的研究进展进行了综述。
第二章提取分离鲜地黄中环烯醚萜苷类成分。药材经粉碎,醇提,大孔吸附树脂,反复硅胶柱层析,葡聚糖凝胶柱纯化,从地黄醇提物中分离得到3个化合物。经TLC检测和光谱分析鉴定结构,这3个化合物分别是梓醇、地黄苷D、益母草苷。
第三章建立一套地黄血清药物化学的实验方法。以含药血清中梓醇、地黄苷D、益母草苷的峰面积的值为指标,确定最佳给药方式及采血时间。以梓醇的峰面积为指标,确定最佳给药量。确定了给药方案是:按浓度为0.7g/ml,1ml/100g,连续给药3d、1次/d,在给药后1h采血,能在含药血清中发现目标成分。这为地黄血清药物化学的进一步研究奠定了基础。
第四章应用血清药物化学的方法,对地黄环烯醚萜苷类入血成分进行分析,为确定地黄的活性成分奠定基础。通过比较对照品、含药血清、空白血清的HPLC指纹图谱,初步确定地黄的入血成分。在给药血清中发现了2个入血成分,且均是原型成分(分别是梓醇、地黄苷D)。这2个入血成分可能是地黄在体内直接作用的物质。
第五章建立地黄醇提物及环烯醚萜苷部位血清药物化学的初步研究。通过比较对照品、含药血清、空白血清的HPLC指纹图谱,确定地黄醇提物及环烯醚萜苷部位的入血成分。在给药血清中发现了3个入血成分,且均是原型成分(分别是梓醇、地黄苷D、益母草苷)。这3个入血成分可能是地黄在体内直接作用的物质,对其进行深入的研究将为进行地黄活性成分的进一步研究奠定基础。
Rehmannia glutinosa libosch was originally recorded in“Shennong bencaojing”and classied as the upper class. It is one of the Four famous herbs produced in Henan HuaiQing. It has been used for medical treatment for several thousands years. It has been cultivated in many provinces, but mainly in Wuzhi, Xiuwu and Wenxian and other areas of Henan province. Radix Rehmannia .is the fresh or dry block root or processed product of the Scrophulariaceae plant Rehmannia glutinosa libosch, medicine followed by: fresh Radix Rehmannia, dry Radix Rehmannia and Radix Rehmannia Praeparata. There are distinct differences between the three: fresh Radix Rehmannia can removing heat to promote salivation, cooling blood, and stop bleeding ; dry Radix Rehmannia can removing heat to cool blood,nourishing Yin,body fluid regeneration;Radix Rehmannia Praeparata can nourishing Yin and blood,tonifying didney jing and filling pith. In the Pharmacopoeia of People's Republic of China (2005 edition),which recorded Chinese medicinal formulae, there were one fifth of it concerned Radix Rehmannia.
In recent years,researching chemical composition of Radix Rehmannia were common among domestic and oversea scholars. Studies on the constituents of its few real active ingredients, its determination of“active ingredients”are mostly some sort of drugs or indicators contained in the main component composition, there is no basis to prove that as an active ingredients, which also limits the Chinese crude drug developed and utilized.Any drug interactions are related to the human body after the therapeutic work. Shinichi Tashiro Japanese scholar in 1989 proposed the concept of“Serum Medicinal Chemistry”and“Serum Pharmacology”.By analyzing the serum after oral administration to determine the composition of traditional Chinese drug directly into the body of material, it become fast, simple and accurate study of the material basis of drug efficacy effective way.
In the first chapter,we summarize the study on the chemical,extraction,pharmacological, serum pharmacochemisty of Rehmannia glutinosa Libosch and serum pharmacochemisty of traditional chinese drug.
In the second chapter,we extraction Iridoid glycosides from Rehmannia glutinosa Libosch .The materia medica by crushing ,ethanol extraction , Separation and purification through the macroporous resin, silica gel column chromatography, Sephadex column,identify the structure of compound through TLC and spectroanalysis.From the ethanol extract of Rehmannia glutinosa Libosch isolated three compounds .There are Catalpol, Rehmannia glycoside D, Motherwort glycosides
In the third chapter,we establish experimental method for Serum pharmacochemisty of Rehmannia glutinosa Libosch. According to peak area value of Catalpol、Rehmannioside D、Leonuride in drug-containing serum catalpol as an indicator to determine the best administration style and blood collection time. According to peak area value of Catalpol in drug-containing serum catalpol as an indicator to determine the best dosage. By comprehensive analysis, the dosage regimen is as following: to continuously dose for 3 days one time a day by the concentration of 0.7g/ml, 1ml/100g. To collect blood after one hour of dosage, we can examine the destination ingredients. Thus, it can lay the foundation of further research on the Serum pharmacochemisty of Rehmannia glutinosa Libosch.
In the fourth chapter,we using the method of serum pharmacochemisty and analysis the compounds of Rehmannia glutinosa libosch absorbed into the blood,lays the foundation for the Rehmannia glutinosa libosch definite active constituent. The constituents absorbed into blood were determined by comparing the HPLC fingerprints of the reference substance、tested serum samples and blank serum sample. Two compounds absorbed into blood were detected, they were original constituents of Rehmannia glutinosa libosch(Catalpol、Rehmannioside D). Two constituents absorbed into blood were possible bioactive components of Rehmannia glutinosa libosch.
In the fifth chapter,we investigate the parameters of serum pharmacochemistry of ethanol extract of Radix Rehamannia and Iridoid glycosides. The constituents absorbed into blood were determined by comparing the HPLC fingerprints of the reference substance、tested serum samples and blank serum sample. Three compounds absorbed into blood were detected, they were original constituents of Rehmannia glutinosa libosch(Catalpol、Rehmannioside D、Leonuride). Three constituents absorbed into blood were possible bioactive components of Rehmannia glutinosa libosch, Its serum pharmacochemistry should be subject to complete investigation so as to lay a foundation for the further study of active ingredients of Rehmannia glutinosa libosch.
引文
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