不同因素对丹参等药用植物化学成分的影响研究
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摘要
中药在我国的应用有着悠久的历史，在预防及治疗某些常见病、慢性病中有着不可替代的作用。但是近年来随着环境因素的变化，中药退化严重，品质下降，直接影响到临床的治疗效果。中药药效的优劣与其所含的化学成分中的活性物质（药用成分）关系密切，因此考察环境因素对药用植物生长的影响及其与药用成分的相关性分析，有助于我们科学的种植、生产药材，从而为提高中药临床疗效提供理论参考。本课题研究了两种不同环境下的药用植物相关因素对其主要药用成分的影响：有着未来良好发展潜力的黄顶菊（自然环境）中的总黄酮、传统药物丹参（盆栽环境）中的丹参酮和酚酸，主要取得如下成果：
1、本研究条件下，建立了黄顶菊中主要成分总黄酮的一种提取分离方法，最佳工艺条件为：80%乙醇，料液比1:20（w/v），每次回流75min，回流2次。黄顶菊干粉浸膏收率可达17.62％。色谱分离条件为：色谱柱Apollo C18（150mm×4.6mm），流动相A为甲醇溶液，B为φ=0.05％磷酸水溶液，洗脱梯度（0~8min，φ=65％B→φ=55％B；8~25min，φ=55％B→φ=50％B；25~30min，φ=50％B→φ=45％B；30~50min，φ=45％B；50.01~55.01min，φ=65％B），流速1.0ml/min，检测波长360nm，柱温10℃。
2、不同产地黄顶菊间的相同成分存在差异。从理论上验证了地道药材讲究地域的说法。这种情况的产生，可能由土壤因子等环境因素差异引起。
3、黄顶菊样地与空地土壤因子差异明显。其中土壤pH、速效P、碱解N、脲酶增高，而碱性磷酸酶、酸性磷酸酶、有机C降低，提示其抑制其他植物生长的机理有可能是其极强的吸附营养能力使土地变贫瘠。
4、研究发现土壤因子与黄顶菊主要药用成分黄酮类的相关性。土壤速效P与槲皮素糖苷、异鼠李素硫酸酯和紫云英苷显著正相关，相关系数分别为0.701*、0.670*、0.681*。土壤碱性磷酸酶与万寿菊素糖苷和6-甲氧基山奈酚糖苷显著负相关，相关系数分别为-0.740*、-0.777*。由此可见，土壤速效P和碱性磷酸酶对黄顶菊主要成分积累有显著作用。
5、研究发现丹参的生长和养分吸收可以通过接种AM真菌来调节，但AM真菌接种效果会受土壤含水量和N含量的影响。综合各方面因素，本试验表明，在70%水分、施N量为0.16g/kg时接种AM真菌丹参可利用价值较高。
6、相同水分和施N条件下，接种AM能够提高植物K、Ca等元素含量，从而提高丹参生长量。不同水分条件下，接种AM真菌改善了丹参逆境生存能力。施N量会影响丹参药用成分含量，当施N量高于一定数值时，丹参药用成分占植物含量比降低。
本课题考察了不同因素与黄顶菊黄酮类物质间的关系、不同水分和N肥对接种AM真菌丹参生长量和药用成分的影响，为提高药用植物品质及合理应用提供了依据。
The application of traditional Chinese medicine in china has a long history. It takes anirreplaceable role in the disease prevention and treatment of some common diseases, chronicdiseases. But recent years, with the changes of environmental factors, traditional Chinesemedicine has seriously degraded, the quality of which directly affects the clinical treatment.The merits of the traditional Chinese medicine efficacy has a close related with its chemicalcomposition of the active substances-medicinal ingredients, therefore investigating theinfluence of environmental factors on the growth of medicinal plants and correlation analysisof their medicinal ingredients can help us scientific planting medicinal materials production, itcan provide theoretical reference for improving the clinical curative effect of traditionalChinese medicine. The research of this article focus on the effect of two different conditionsof medicinal plant related factors on the main medicinal ingredients: flavonoids in Flaveriabidentis which has a good future potential development (natural environment)、tanshinone andphenolic acid in traditional medicine Salvia miltiorrhiza plants (Potted plant environment),the main results as follows：
1、Under the conditions of this study, establishing a method of extraction and separationof flavonoids which is the main components in Flaveria bidentis, the optimum conditions:80%ethanol, Material liquid than1:20(w/v), Backflow75minutes at a time, Back2times. F.bidentis powder extract yield can reach17.62%. Chromatographic conditions: Apollo C18Column (150mm×4.6mm), Methanol solution for mobile phase A, B for φ=0.05%phosphoric acid aqueous solution, A gradient elution (0~8min, φ=65%B→φ=55%B;8~25min, φ=55%B→φ=50%B;25~30min, φ=50%B→φ=45%B;30~50min, φ=45%B;50.01~55.01min, φ=65%B), a flow rate of1.0ml/min, detection wavelength at360nm, column temperature is10℃.
2、The same composition of Flaveria bidentis are differences among different plots. Ittheoretically verified authentic ingredients attention geographical argument. For this situation,the difference may be caused by environmental factors such as soil factors.
3、The soil factors of F.bidentis of and the blank plots have significant difference. Wheresoil pH, available P, alkali solution N and urease increased, but soil alkaline phosphatase, acidphosphatase and organic carbon decreased. The results suggest that the mechanism of inhibition of the growth of other plants might be its strong capacity of adsorption which could
make the land barren.
4. The study found a correlation between soil factors and F.bidentis’s main medicinalingredient flavonoids. There was a positive correlation among soil available P, quercetinglycosides, isorhamnetin sulfate and AST. The correlation coefficients were0.701*0.670*,0.681*, respectively. The significantly negative correlation was found among soil alkalinephosphatase, marigold glucosides and6-methoxy kaempferol glycosides. The correlationcoefficient were-0.740*,-0.777*, respectively. Thus, soil available P and alkaline phosphatasehad a distinct role in the accumulation of main component of F.bidentis.
5. The study found that the growth and nutrient absorption of the Salvia miltiorrhizacould be adjusted by arbuscular mycorrhizal fungi, but the effect of arbuscular mycorrhizalfungal inoculation was influenced by soil water and nitrogen fertilizer. Taking into all thesefactors, the growth effect was best under70%of soil water, N0.16g/kg.
6. AM inoculation can improve the content of K, Ca and other elements, which promotethe growth of Salvia miltiorrhiza under the same soil water and nitrogen fertilizer conditions.AM inoculation can improve the survival ability of Salvia miltiorrhiza adversity underdifferent soil water conditions. The content of medicinal components of Salvia miltiorrhizaare influenced by nitrogen fertilizer. When the amount of nitrogen is higher than a certainvalue, the content of medicinal components of Salvia miltiorrhiza as a proportion of the totalfalls.
This paper examines the relationship between the different conditions of flavonoids inthe Flaveria bidentis, and the effects of growth and medicinal components of AM inoculationof Salvia miltiorrhiza under different soil water and nitrogen fertilizer conditions It provides atheoretical basis in improving the quality of medicinal plants.
1、本研究条件下，建立了黄顶菊中主要成分总黄酮的一种提取分离方法，最佳工艺条件为：80%乙醇，料液比1:20（w/v），每次回流75min，回流2次。黄顶菊干粉浸膏收率可达17.62％。色谱分离条件为：色谱柱Apollo C18（150mm×4.6mm），流动相A为甲醇溶液，B为φ=0.05％磷酸水溶液，洗脱梯度（0~8min，φ=65％B→φ=55％B；8~25min，φ=55％B→φ=50％B；25~30min，φ=50％B→φ=45％B；30~50min，φ=45％B；50.01~55.01min，φ=65％B），流速1.0ml/min，检测波长360nm，柱温10℃。
2、不同产地黄顶菊间的相同成分存在差异。从理论上验证了地道药材讲究地域的说法。这种情况的产生，可能由土壤因子等环境因素差异引起。
3、黄顶菊样地与空地土壤因子差异明显。其中土壤pH、速效P、碱解N、脲酶增高，而碱性磷酸酶、酸性磷酸酶、有机C降低，提示其抑制其他植物生长的机理有可能是其极强的吸附营养能力使土地变贫瘠。
4、研究发现土壤因子与黄顶菊主要药用成分黄酮类的相关性。土壤速效P与槲皮素糖苷、异鼠李素硫酸酯和紫云英苷显著正相关，相关系数分别为0.701*、0.670*、0.681*。土壤碱性磷酸酶与万寿菊素糖苷和6-甲氧基山奈酚糖苷显著负相关，相关系数分别为-0.740*、-0.777*。由此可见，土壤速效P和碱性磷酸酶对黄顶菊主要成分积累有显著作用。
5、研究发现丹参的生长和养分吸收可以通过接种AM真菌来调节，但AM真菌接种效果会受土壤含水量和N含量的影响。综合各方面因素，本试验表明，在70%水分、施N量为0.16g/kg时接种AM真菌丹参可利用价值较高。
6、相同水分和施N条件下，接种AM能够提高植物K、Ca等元素含量，从而提高丹参生长量。不同水分条件下，接种AM真菌改善了丹参逆境生存能力。施N量会影响丹参药用成分含量，当施N量高于一定数值时，丹参药用成分占植物含量比降低。
本课题考察了不同因素与黄顶菊黄酮类物质间的关系、不同水分和N肥对接种AM真菌丹参生长量和药用成分的影响，为提高药用植物品质及合理应用提供了依据。
The application of traditional Chinese medicine in china has a long history. It takes anirreplaceable role in the disease prevention and treatment of some common diseases, chronicdiseases. But recent years, with the changes of environmental factors, traditional Chinesemedicine has seriously degraded, the quality of which directly affects the clinical treatment.The merits of the traditional Chinese medicine efficacy has a close related with its chemicalcomposition of the active substances-medicinal ingredients, therefore investigating theinfluence of environmental factors on the growth of medicinal plants and correlation analysisof their medicinal ingredients can help us scientific planting medicinal materials production, itcan provide theoretical reference for improving the clinical curative effect of traditionalChinese medicine. The research of this article focus on the effect of two different conditionsof medicinal plant related factors on the main medicinal ingredients: flavonoids in Flaveriabidentis which has a good future potential development (natural environment)、tanshinone andphenolic acid in traditional medicine Salvia miltiorrhiza plants (Potted plant environment),the main results as follows：
1、Under the conditions of this study, establishing a method of extraction and separationof flavonoids which is the main components in Flaveria bidentis, the optimum conditions:80%ethanol, Material liquid than1:20(w/v), Backflow75minutes at a time, Back2times. F.bidentis powder extract yield can reach17.62%. Chromatographic conditions: Apollo C18Column (150mm×4.6mm), Methanol solution for mobile phase A, B for φ=0.05%phosphoric acid aqueous solution, A gradient elution (0~8min, φ=65%B→φ=55%B;8~25min, φ=55%B→φ=50%B;25~30min, φ=50%B→φ=45%B;30~50min, φ=45%B;50.01~55.01min, φ=65%B), a flow rate of1.0ml/min, detection wavelength at360nm, column temperature is10℃.
2、The same composition of Flaveria bidentis are differences among different plots. Ittheoretically verified authentic ingredients attention geographical argument. For this situation,the difference may be caused by environmental factors such as soil factors.
3、The soil factors of F.bidentis of and the blank plots have significant difference. Wheresoil pH, available P, alkali solution N and urease increased, but soil alkaline phosphatase, acidphosphatase and organic carbon decreased. The results suggest that the mechanism of inhibition of the growth of other plants might be its strong capacity of adsorption which could
make the land barren.
4. The study found a correlation between soil factors and F.bidentis’s main medicinalingredient flavonoids. There was a positive correlation among soil available P, quercetinglycosides, isorhamnetin sulfate and AST. The correlation coefficients were0.701*0.670*,0.681*, respectively. The significantly negative correlation was found among soil alkalinephosphatase, marigold glucosides and6-methoxy kaempferol glycosides. The correlationcoefficient were-0.740*,-0.777*, respectively. Thus, soil available P and alkaline phosphatasehad a distinct role in the accumulation of main component of F.bidentis.
5. The study found that the growth and nutrient absorption of the Salvia miltiorrhizacould be adjusted by arbuscular mycorrhizal fungi, but the effect of arbuscular mycorrhizalfungal inoculation was influenced by soil water and nitrogen fertilizer. Taking into all thesefactors, the growth effect was best under70%of soil water, N0.16g/kg.
6. AM inoculation can improve the content of K, Ca and other elements, which promotethe growth of Salvia miltiorrhiza under the same soil water and nitrogen fertilizer conditions.AM inoculation can improve the survival ability of Salvia miltiorrhiza adversity underdifferent soil water conditions. The content of medicinal components of Salvia miltiorrhizaare influenced by nitrogen fertilizer. When the amount of nitrogen is higher than a certainvalue, the content of medicinal components of Salvia miltiorrhiza as a proportion of the totalfalls.
This paper examines the relationship between the different conditions of flavonoids inthe Flaveria bidentis, and the effects of growth and medicinal components of AM inoculationof Salvia miltiorrhiza under different soil water and nitrogen fertilizer conditions It provides atheoretical basis in improving the quality of medicinal plants.
引文
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