川鄂山茱萸及其近缘植物的生药学研究
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摘要
目的：通过对川鄂山茱萸(Cornus Chinense Wanger．)的生药学系统研究，以山茱萸的苷类化学组分-熊果酸、马钱苷为指标成分，从川鄂山茱萸的不同器官与细胞组织中筛选含量最高的部位，并通过其近缘植物灯台树、小梾木的相关性研究，探讨此类化学组分分布状况，拟证明宏观的形态特征与微观的细胞特征以及分子水平的化学组分存在一定的相关性，为进一步筛选有效药物组分奠定基础，为创新药物的应用与开发提供实验技术依据。
方法：以文献查阅-整理-调查为基础，综合运用植物学、植物解剖学、中药鉴定学知识与实验技术对川鄂山茱萸及其近缘植物进行分类学处理与解剖学研究，结合天然药物化学、仪器分析、药物分析等学科的知识与实验技术，用TLC定性分析川鄂山茱萸及其近缘植物的苷类化学组分；用HPLC测定川鄂山茱萸不同器官、细胞组织、果实不同采集期的熊果酸、马钱苷的含量并进行色谱条件优化。
结果：(1)川鄂山茱萸(Cornus chinense Wanger．)、灯台树(Bothrocaryum controversum(Hemsl．)Pojark)、小梾木(Swida paucinervia(Hance)Sojak)从同属植物分为山茱萸属(Comus)、灯台树属(Bothrocaryum)、梾木属(Swida)。从解剖学与化学的角度支持了形态分类的正确性，证明宏观与微观在生物种上的鉴定一致。
(2)川鄂山茱萸果实解剖学特征可以区别川鄂山茱萸及其近缘植物，以免相似入药部位产生的混淆。
(3)分别建立了川鄂山茱萸及其近缘植物的熊果酸、马钱苷TLC其定性分析方法。熊果酸以环己烷-氯仿-醋酸乙酯(20:5:8)为展开剂；马钱苷以氯仿-甲醇(4:1)为展开剂。川鄂山茱萸不同器官与细胞组织、小梾木、灯台树果实在不同程度上含有熊果酸，其中小梾木果实与川鄂山茱萸总苞片熊果酸含量较高；川鄂山茱萸不同器官与细胞组织含有环烯醚类化合物，而小梾木、灯台树果实可能不含环烯醚类化合物。
(4)分别建立了川鄂山茱萸的熊果酸、马钱苷HPLC含量测定法。其中熊果酸色谱条件为：乙腈:水:磷酸(80:20:0.1)为流动相，检测波长为206nm，流速为1.000ml／min，柱温为30℃；马钱苷色谱条件为：乙腈:水:磷酸(15:85:0.1)为流动相，检测波长为240nm，流速为1.000ml／min，柱温为30℃。方法学考察结果表明重现性好，精密度高，可以作为川鄂山茱萸中熊果酸、马钱苷的检测方法之一。
(5)对川鄂山茱萸不同器官与细胞组织的熊果酸、马钱苷含量测定，川鄂山茱萸不同器官与细胞组织的熊果酸、马钱苷含量有差异。熊果酸的含量为总苞片＞茎＞叶＞花＞果皮＞果实；马钱苷的含量：花＞总苞片＞叶＞果实＞茎＞果皮。
(6)对不同采集期的川鄂山茱萸果实的熊果酸、马钱苷含量测定，2006年采集和2007年采集的川鄂山茱萸果实中熊果酸含量分别为：0.1757％，0.0889％；马钱苷含量分别为：0.4151％，0.2284％。
结论：川鄂山茱萸含有熊果酸、马钱苷，从化学组分的观点支持了川鄂山茱萸有补肝益肾的功效。单纯以此组分确定功效，川鄂山茱萸以总苞片入药最佳；川鄂山茱萸在熊果酸含量上与山茱萸相似，在环烯醚萜类-马钱苷化学组分上有一定差异，有进一步研究的价值。对川鄂山茱萸及其近缘植物灯台树、小梾木的相关性研究，证明宏观的形态特征与微观的细胞特征以及分子水平的化学组分存在一定的相关性，可做为药物筛选的重要参考依据。
本课题运用了植物分类学、植物解剖学、TLC法、HPLC等方法对川鄂山茱萸药材的分析，可做为川鄂山茱萸药材质量标准的主要参考依据。
Objective:Through the research of pharmacognosy in Cornus chinense Wanger,and according to the content of ursolic acid and loganin in Cornus officinalis Sieb. et Zucc. The highest content of part in Cornus chinense Wanger is screened.By the correlation study of Bothrocaryum controversum(Hemsl.)Pojark and Swida paucinervia (Hance) Sojak and to probe into the distribution of these chemical components,prove that certain correlation were exsited in the macroscopic of morphological characteristics,the microscopic of cells characteristics and the molecular level of chemical components,to lay the foundation for screening effective drug components and to provide experimental evidence of drug application for innovation and technology development.
Method: In the light of looking up -finishing-inquiring into the literature,The subject ntegratedly applied the knowledge of botany and phytotomy and Authentication of Chinese Medicines and the experimental technology to study taxonomy and anatomy of Cornus chinense Wanger and its kindred plant.To combine the knowledge of natural medicinal chemistry instrumental analysis,pharmaceutical analysis and experimental technique,Glycosides chemical components of Cornus chinense Wanger and its kindred plant were qualitatively analysised by using TLC.The content of different organ,cell organization of Cornus chinense Wanger and its fruit of different packing time were determined by HPLC, to optimize chromatographic conditions.
Result: (1) Cornus chinense Wanger,Bothrocaryum controversum (Hemsl.) Pojark,Swida paucinervia (Hance) Sojak were divided into Cornus, Bothrocaryum,Swida from the same genus,so plant taxonomy was proved to be correct by anatomy and chemistry and proved the coincidence of orgamism identification on microscopic and on macroscopic.
(2) The Cornus chinense Wanger and its kindred plant were distinguished by the anatomical characteristics of the fruit of Cornus chinense Wanger,in order to avoid a similar medicine parts confusion.
(3) The TLC method of ursolic acid and loganin in Cornus chinense Wanger and its kindred plant were respectively seted up.The lamellar condition of ursolic acid is hexamethylene-chloroform-ethyl acetate(20:5:8);The lamellar condition of loganin is chloroform-water(4:1);ursolic acid were exsited in different organ and cellular tissue in Cornus chinense Wanger,Swida paucinervia (Hance) Sojak and Bothrocaryum controversum (Hemsl.)Pojark,especially in the Swida paucinervia (Hance) Sojak and in the bract of Cornus chinense Wanger,the different organ and cell organization of Cornus chinense Wanger contain iridoids,but Swida paucinervia (Hance) Sojak and Bothrocaryum controversum(Hemsl.)Pojark couldn't contain it.
(4) The content of ursolic acid and loganin in Cornus chinense Wanger.Its kindred plant were respectively seted up by HPLC.The chromatographic condition of ursolic acid:acetonitrile-water- phosphoric acid(80:20:0.1)was the mobile phase,the detection wavelength was 206nm,the flow rate was 1.0ml/min,the column temperature was 30℃;The chromatographic condition of loganin:acetonitrile-water-phosphoric acid(15:85:0.1) was the mobile phase;the detection wavelength was 240nm,the flow rate was 1.0ml/min and the column temperature was 30℃;Methodological experiments indicated the good reproducibility and the high precision,to be a detection method of ursolic acid and loganin in Cornus chinense Wanger.
(5) Through determining the conten of ursolic acid ,loganinin different organ and cell organization of Cornus chinense Wanger,the content of ursolic acid loganin was different.The content of ursolic acid in bract,stem,leaf,flower,pericarp and fruit gradually became low;The content of loganin in flower,bract,leaf ,fruit,stem,pericarp gradually be came low.
(6) Through determining the conten of loganin,ursolic acid in the fruits of Cornus chinense Wanger in different packing time,the results indicate that the conten of loganin was 0.1757%,0.0889% and the conten of ursolic acid was 0.4151%,0.2284% in 2006 and 2007.
Conclusion ursolic acid,Loganin, were exsited in Cornus chinense Wanger, which prove the effects of novishing kidney and live of its from chemistry.To decide the effects with simple component,the bract of Cornus chinense Wanger was the best medicine part.The content of ursolic acid in Cornus chinense Wanger was similar to Cornus officinalis Sieb.et Zucc,but the component of iridoids in Cornus chinense Wanger was invariably different from Cornus officinalis Sieb.et Zucc,so it has the value of further research.
By the correlation study of Bothrocaryum controversum(Hemsl.)Pojark and Swida paucinervia (Hance) Sojak and to probe into the distribution of these chemical components,prove that certain correlation were exsited in the macroscopic of morphological characteristics,the microscopic of cells characteristics and the molecular level of chemical components,which provide important evidence for screening medicine.
The subjiect applied the method of plant taxonomy,plant anatomy,TLC, HPLC to analysis Cornus chinense Wanger,which establish the initial quantity standard of its.
方法：以文献查阅-整理-调查为基础，综合运用植物学、植物解剖学、中药鉴定学知识与实验技术对川鄂山茱萸及其近缘植物进行分类学处理与解剖学研究，结合天然药物化学、仪器分析、药物分析等学科的知识与实验技术，用TLC定性分析川鄂山茱萸及其近缘植物的苷类化学组分；用HPLC测定川鄂山茱萸不同器官、细胞组织、果实不同采集期的熊果酸、马钱苷的含量并进行色谱条件优化。
结果：(1)川鄂山茱萸(Cornus chinense Wanger．)、灯台树(Bothrocaryum controversum(Hemsl．)Pojark)、小梾木(Swida paucinervia(Hance)Sojak)从同属植物分为山茱萸属(Comus)、灯台树属(Bothrocaryum)、梾木属(Swida)。从解剖学与化学的角度支持了形态分类的正确性，证明宏观与微观在生物种上的鉴定一致。
(2)川鄂山茱萸果实解剖学特征可以区别川鄂山茱萸及其近缘植物，以免相似入药部位产生的混淆。
(3)分别建立了川鄂山茱萸及其近缘植物的熊果酸、马钱苷TLC其定性分析方法。熊果酸以环己烷-氯仿-醋酸乙酯(20:5:8)为展开剂；马钱苷以氯仿-甲醇(4:1)为展开剂。川鄂山茱萸不同器官与细胞组织、小梾木、灯台树果实在不同程度上含有熊果酸，其中小梾木果实与川鄂山茱萸总苞片熊果酸含量较高；川鄂山茱萸不同器官与细胞组织含有环烯醚类化合物，而小梾木、灯台树果实可能不含环烯醚类化合物。
(4)分别建立了川鄂山茱萸的熊果酸、马钱苷HPLC含量测定法。其中熊果酸色谱条件为：乙腈:水:磷酸(80:20:0.1)为流动相，检测波长为206nm，流速为1.000ml／min，柱温为30℃；马钱苷色谱条件为：乙腈:水:磷酸(15:85:0.1)为流动相，检测波长为240nm，流速为1.000ml／min，柱温为30℃。方法学考察结果表明重现性好，精密度高，可以作为川鄂山茱萸中熊果酸、马钱苷的检测方法之一。
(5)对川鄂山茱萸不同器官与细胞组织的熊果酸、马钱苷含量测定，川鄂山茱萸不同器官与细胞组织的熊果酸、马钱苷含量有差异。熊果酸的含量为总苞片＞茎＞叶＞花＞果皮＞果实；马钱苷的含量：花＞总苞片＞叶＞果实＞茎＞果皮。
(6)对不同采集期的川鄂山茱萸果实的熊果酸、马钱苷含量测定，2006年采集和2007年采集的川鄂山茱萸果实中熊果酸含量分别为：0.1757％，0.0889％；马钱苷含量分别为：0.4151％，0.2284％。
结论：川鄂山茱萸含有熊果酸、马钱苷，从化学组分的观点支持了川鄂山茱萸有补肝益肾的功效。单纯以此组分确定功效，川鄂山茱萸以总苞片入药最佳；川鄂山茱萸在熊果酸含量上与山茱萸相似，在环烯醚萜类-马钱苷化学组分上有一定差异，有进一步研究的价值。对川鄂山茱萸及其近缘植物灯台树、小梾木的相关性研究，证明宏观的形态特征与微观的细胞特征以及分子水平的化学组分存在一定的相关性，可做为药物筛选的重要参考依据。
本课题运用了植物分类学、植物解剖学、TLC法、HPLC等方法对川鄂山茱萸药材的分析，可做为川鄂山茱萸药材质量标准的主要参考依据。
Objective:Through the research of pharmacognosy in Cornus chinense Wanger,and according to the content of ursolic acid and loganin in Cornus officinalis Sieb. et Zucc. The highest content of part in Cornus chinense Wanger is screened.By the correlation study of Bothrocaryum controversum(Hemsl.)Pojark and Swida paucinervia (Hance) Sojak and to probe into the distribution of these chemical components,prove that certain correlation were exsited in the macroscopic of morphological characteristics,the microscopic of cells characteristics and the molecular level of chemical components,to lay the foundation for screening effective drug components and to provide experimental evidence of drug application for innovation and technology development.
Method: In the light of looking up -finishing-inquiring into the literature,The subject ntegratedly applied the knowledge of botany and phytotomy and Authentication of Chinese Medicines and the experimental technology to study taxonomy and anatomy of Cornus chinense Wanger and its kindred plant.To combine the knowledge of natural medicinal chemistry instrumental analysis,pharmaceutical analysis and experimental technique,Glycosides chemical components of Cornus chinense Wanger and its kindred plant were qualitatively analysised by using TLC.The content of different organ,cell organization of Cornus chinense Wanger and its fruit of different packing time were determined by HPLC, to optimize chromatographic conditions.
Result: (1) Cornus chinense Wanger,Bothrocaryum controversum (Hemsl.) Pojark,Swida paucinervia (Hance) Sojak were divided into Cornus, Bothrocaryum,Swida from the same genus,so plant taxonomy was proved to be correct by anatomy and chemistry and proved the coincidence of orgamism identification on microscopic and on macroscopic.
(2) The Cornus chinense Wanger and its kindred plant were distinguished by the anatomical characteristics of the fruit of Cornus chinense Wanger,in order to avoid a similar medicine parts confusion.
(3) The TLC method of ursolic acid and loganin in Cornus chinense Wanger and its kindred plant were respectively seted up.The lamellar condition of ursolic acid is hexamethylene-chloroform-ethyl acetate(20:5:8);The lamellar condition of loganin is chloroform-water(4:1);ursolic acid were exsited in different organ and cellular tissue in Cornus chinense Wanger,Swida paucinervia (Hance) Sojak and Bothrocaryum controversum (Hemsl.)Pojark,especially in the Swida paucinervia (Hance) Sojak and in the bract of Cornus chinense Wanger,the different organ and cell organization of Cornus chinense Wanger contain iridoids,but Swida paucinervia (Hance) Sojak and Bothrocaryum controversum(Hemsl.)Pojark couldn't contain it.
(4) The content of ursolic acid and loganin in Cornus chinense Wanger.Its kindred plant were respectively seted up by HPLC.The chromatographic condition of ursolic acid:acetonitrile-water- phosphoric acid(80:20:0.1)was the mobile phase,the detection wavelength was 206nm,the flow rate was 1.0ml/min,the column temperature was 30℃;The chromatographic condition of loganin:acetonitrile-water-phosphoric acid(15:85:0.1) was the mobile phase;the detection wavelength was 240nm,the flow rate was 1.0ml/min and the column temperature was 30℃;Methodological experiments indicated the good reproducibility and the high precision,to be a detection method of ursolic acid and loganin in Cornus chinense Wanger.
(5) Through determining the conten of ursolic acid ,loganinin different organ and cell organization of Cornus chinense Wanger,the content of ursolic acid loganin was different.The content of ursolic acid in bract,stem,leaf,flower,pericarp and fruit gradually became low;The content of loganin in flower,bract,leaf ,fruit,stem,pericarp gradually be came low.
(6) Through determining the conten of loganin,ursolic acid in the fruits of Cornus chinense Wanger in different packing time,the results indicate that the conten of loganin was 0.1757%,0.0889% and the conten of ursolic acid was 0.4151%,0.2284% in 2006 and 2007.
Conclusion ursolic acid,Loganin, were exsited in Cornus chinense Wanger, which prove the effects of novishing kidney and live of its from chemistry.To decide the effects with simple component,the bract of Cornus chinense Wanger was the best medicine part.The content of ursolic acid in Cornus chinense Wanger was similar to Cornus officinalis Sieb.et Zucc,but the component of iridoids in Cornus chinense Wanger was invariably different from Cornus officinalis Sieb.et Zucc,so it has the value of further research.
By the correlation study of Bothrocaryum controversum(Hemsl.)Pojark and Swida paucinervia (Hance) Sojak and to probe into the distribution of these chemical components,prove that certain correlation were exsited in the macroscopic of morphological characteristics,the microscopic of cells characteristics and the molecular level of chemical components,which provide important evidence for screening medicine.
The subjiect applied the method of plant taxonomy,plant anatomy,TLC, HPLC to analysis Cornus chinense Wanger,which establish the initial quantity standard of its.
引文
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