林下参化学成分及SD大鼠皮下注射Rh2药理和药动学的研究
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摘要
林下参，又称育山参，是人为地把园参的种子撒播到自然的环境里，任其自然生长，不移栽、不搭棚、不施肥、不打药、不锄草、不动土，10年后才收取的半野生山参。林下参长白山地区蕴藏量的商业价值约50亿元，但有关其基础研究开展的还不是非常充分，本文利用现代的分离纯化手段从林下参的水提物中分离得到25个化合物，依据其理化性质及波谱数据解析(1H-NMR、13C-NMR、DEPT、HMBC等)，鉴定了其中23个化合物的结构，分别为：20(S)-原人参二醇(1)、20(R)-原人参三醇(2)、西洋参皂苷F6(3)、人参皂苷Rb3(4)、人参皂苷Rc(5)、人参皂苷Rd(6)、人参皂苷Re(7)、人参皂苷Rf(8)、人参皂苷Rg1(9)、20(S)-人参皂苷Rg2(10)、20(S)-人参皂苷Rg3(11)、20(R)-人参皂苷Rg3(12)、20(S)-人参皂苷Rh1(13)、20(S)-人参皂苷Rh2(14)、β-谷甾醇(15)、豆甾醇(16)、胡萝卜苷(17)、儿茶酚(18)、蔗糖(19)、十一酸(20)、正三十八烷(21)、琥珀酸(22)、二十四酸(23)。其中化合物3、4、5、6为首次从林下参中分离得到。
首次采用气相色谱-质谱联用(GC-MS)技术对林下参根中分得的油状物成分进行了分析，从气相色谱图中共检测出40个峰，通过标准质谱图数据库检索，鉴定了其中18种成分，并用峰面积归一化法确定了其相对含量。
首次对林下参根、叶片中无机元素的组成及含量进行了对比分析。通过将样品消化后定容，利用电感耦合等离子发射光谱仪（ICP）测定的方法，测出各部位含45种无机元素，其中含量较高的元素为Ca、Mg等常量无机元素，重金属元素含量很低。
首次对20(S)-人参皂苷Rh2在SD大鼠血液中的药物动力学进行了初步研究。皮下注射给予大鼠20(S)-人参皂苷Rh2，剂量为10mg/kg，采用固相萃取法提取大鼠血清中的药物，并应用HPLC-UV检测大鼠血中的药物含量。20(S)-人参皂苷Rh2在浓度0.25-100μg/mL区间内线性良好。给药后0.5h大鼠血浆中的20(S)-人参皂苷Rh2浓度达到峰浓度，为0.79μg/mL，t1/2=2.8h，AUC=3.4μg.h/mL，给药后6h内平均血药浓度为0.50±0.24μg/mL。
首次研究了20(S)-人参皂苷Rh2对异丙肾上腺素造成的大鼠心肌缺血的保护作用。通过给予SD大鼠皮下注射异丙肾上腺素来复制心肌损伤实验动物模型，研究了20(S)-人参皂苷Rh2对异丙肾上腺素致心肌损伤的影响。通过在指定时间点测定大鼠收缩压、舒张压、心率等心脏指标和动物心脏指数、死亡率等指标发现20(S)-人参皂苷Rh2对异丙肾上腺素造成的大鼠心肌缺血具有一定的保护作用，故本研究成果为20(S)-人参皂苷Rh2的心血管方面的药物研发提供了一定的科学依据。
首次探讨了给予大鼠皮下注射不同剂量的20(S)-人参皂苷Rh2后，大鼠红细胞中ATP、ADP、AMP、GDP、GMP等腺苷的浓度的变化，发现ATP的浓度跟大鼠心肌缺血情况存在关联，20(S)-人参皂苷Rh2可以通过降低心肌细胞的能量代谢的途径来降低心肌耗氧量和心肌坏死程度，从而起到对心脏的保护作用，因此ATP可以作为20(S)-人参皂苷Rh2对心血管系统的保护作用的生物标记物。
Panax ginseng C.A.Meyer cv. Silvatica is a new kind of ginseng which was cultivatedartificially in the natural environment and grew freely about10years or more. Being asemi-wild ginseng between cultivated and grew wildly ginseng, the development of Panaxginseng C.A. Meyer cv. Silvatica has developed rapidly in recent years. However, theresearch on its theoretical basis has not carried out yet as much as possible. It is urgent tobegin the study as the the quantity of wild ginseng is reducing abruptly, while the cultivationand application of Panax ginseng C.A.Meyer cv. Silvatica are growing widely.
In order to find more new active constituents from the Panax ginseng C.A.Meyer cv.Silvatica, the chemical study on the ethonal extract of the root of this plant was carried out.By various methods of isolation,25compounds were isolated, and by the means of thephysicochemical properties and spectroscopic methods(1H-NMR、13C-NMR、DEPT、HMBC),23compounds were elucidated as:20(S)-protopanaxadiol(1),20(R)-protopanaxatriol(2),quinquenoside-F6(3), ginsenoside-Rb3(4), ginsenoside-Rc(5), ginsenoside-Rd(6), ginse-noside-Re(7), ginsenoside-Rf(8), ginsenoside-Rg1(9),20(S)-ginsenoside-Rg2(10),20(S)-ginsenoside-Rg3(11),20(R)-ginsenoside-Rg3(12),20(S)-ginsenoside-Rh1(13),20(S)-ginsenoside-Rh2(14), β-sitosterol(15), Stigmasterol(16), Daucosterol(17), Catechol(18),D-sucrose(19), Undecanoic acid(20), Octadecane(21), Succinic acid(22), Isoselachocericacid(23) respectively. Among of them, compound3,4,5and6were obtained from thePanax ginseng C.A. Meyer cv. Silvatica for the first time.
At the same time, the liposoluble components from the root of Panax ginsengC.A.Meyer cv. Silvatica were analyzed by GC-MS computer technology.40peaks wereobtained and18of them were identified by means of indexing the spectrum storeroom forthe first time.
The inorganic elements in the root and leaf of Panax ginseng C.A.Meyer cv. Silvaticawere determined by ICP-AES instrument.45kinds of inorganic elements were detected inthe samples and the contents of Ca and Mg were higher than others. The amount of somekinds of inorganic elements in the root apppeared some difference comparing to the leaf.
The effects of20(S)-Ginsenoside-Rh2which is an active compound separated fromPanax ginseng C.A.Meyer cv. Silvatica on myocardial ischemia induced by isoproterenol and pharmacokinetic were investigated in experimental SD rats. The pharmacology resultsshowed that20(S)-Ginsenoside-Rh2has the effect of protecting the heart from damageinduced from ischemia by means of increasing blood pressure and lowering the heart ratios.The pharmacokinetic results showed that the mean plasma concentration-time data could beadequately described by a one compartment open model with r2=0.9976and t1/2=2.8h, AUC=3.4μg.h/mL.
The concentrations of ATP, ADP, AMP, GTP and GDP in the rat plasma were measuredby means of HPLC-UV detection. The concentrations of ATP have been used as a biomarkerfor reflecting the protection to the cardiovascular system.
The chemical constituents of Panax ginseng C.A. Meyer cv. Silvatica and bioactivities of20(S)-Ginsenoside-Rh2were made a deep investigation in the thesis. The results would fillthe theoretical basis research inadequaty of it as well as supply good methods for qualitycontrol. Further more, they also offered strong, accurate and scientific theoretical basis to thewide application of this kind of ginseng, which might produce even higher theoretical andeconomic value.
首次采用气相色谱-质谱联用(GC-MS)技术对林下参根中分得的油状物成分进行了分析，从气相色谱图中共检测出40个峰，通过标准质谱图数据库检索，鉴定了其中18种成分，并用峰面积归一化法确定了其相对含量。
首次对林下参根、叶片中无机元素的组成及含量进行了对比分析。通过将样品消化后定容，利用电感耦合等离子发射光谱仪（ICP）测定的方法，测出各部位含45种无机元素，其中含量较高的元素为Ca、Mg等常量无机元素，重金属元素含量很低。
首次对20(S)-人参皂苷Rh2在SD大鼠血液中的药物动力学进行了初步研究。皮下注射给予大鼠20(S)-人参皂苷Rh2，剂量为10mg/kg，采用固相萃取法提取大鼠血清中的药物，并应用HPLC-UV检测大鼠血中的药物含量。20(S)-人参皂苷Rh2在浓度0.25-100μg/mL区间内线性良好。给药后0.5h大鼠血浆中的20(S)-人参皂苷Rh2浓度达到峰浓度，为0.79μg/mL，t1/2=2.8h，AUC=3.4μg.h/mL，给药后6h内平均血药浓度为0.50±0.24μg/mL。
首次研究了20(S)-人参皂苷Rh2对异丙肾上腺素造成的大鼠心肌缺血的保护作用。通过给予SD大鼠皮下注射异丙肾上腺素来复制心肌损伤实验动物模型，研究了20(S)-人参皂苷Rh2对异丙肾上腺素致心肌损伤的影响。通过在指定时间点测定大鼠收缩压、舒张压、心率等心脏指标和动物心脏指数、死亡率等指标发现20(S)-人参皂苷Rh2对异丙肾上腺素造成的大鼠心肌缺血具有一定的保护作用，故本研究成果为20(S)-人参皂苷Rh2的心血管方面的药物研发提供了一定的科学依据。
首次探讨了给予大鼠皮下注射不同剂量的20(S)-人参皂苷Rh2后，大鼠红细胞中ATP、ADP、AMP、GDP、GMP等腺苷的浓度的变化，发现ATP的浓度跟大鼠心肌缺血情况存在关联，20(S)-人参皂苷Rh2可以通过降低心肌细胞的能量代谢的途径来降低心肌耗氧量和心肌坏死程度，从而起到对心脏的保护作用，因此ATP可以作为20(S)-人参皂苷Rh2对心血管系统的保护作用的生物标记物。
Panax ginseng C.A.Meyer cv. Silvatica is a new kind of ginseng which was cultivatedartificially in the natural environment and grew freely about10years or more. Being asemi-wild ginseng between cultivated and grew wildly ginseng, the development of Panaxginseng C.A. Meyer cv. Silvatica has developed rapidly in recent years. However, theresearch on its theoretical basis has not carried out yet as much as possible. It is urgent tobegin the study as the the quantity of wild ginseng is reducing abruptly, while the cultivationand application of Panax ginseng C.A.Meyer cv. Silvatica are growing widely.
In order to find more new active constituents from the Panax ginseng C.A.Meyer cv.Silvatica, the chemical study on the ethonal extract of the root of this plant was carried out.By various methods of isolation,25compounds were isolated, and by the means of thephysicochemical properties and spectroscopic methods(1H-NMR、13C-NMR、DEPT、HMBC),23compounds were elucidated as:20(S)-protopanaxadiol(1),20(R)-protopanaxatriol(2),quinquenoside-F6(3), ginsenoside-Rb3(4), ginsenoside-Rc(5), ginsenoside-Rd(6), ginse-noside-Re(7), ginsenoside-Rf(8), ginsenoside-Rg1(9),20(S)-ginsenoside-Rg2(10),20(S)-ginsenoside-Rg3(11),20(R)-ginsenoside-Rg3(12),20(S)-ginsenoside-Rh1(13),20(S)-ginsenoside-Rh2(14), β-sitosterol(15), Stigmasterol(16), Daucosterol(17), Catechol(18),D-sucrose(19), Undecanoic acid(20), Octadecane(21), Succinic acid(22), Isoselachocericacid(23) respectively. Among of them, compound3,4,5and6were obtained from thePanax ginseng C.A. Meyer cv. Silvatica for the first time.
At the same time, the liposoluble components from the root of Panax ginsengC.A.Meyer cv. Silvatica were analyzed by GC-MS computer technology.40peaks wereobtained and18of them were identified by means of indexing the spectrum storeroom forthe first time.
The inorganic elements in the root and leaf of Panax ginseng C.A.Meyer cv. Silvaticawere determined by ICP-AES instrument.45kinds of inorganic elements were detected inthe samples and the contents of Ca and Mg were higher than others. The amount of somekinds of inorganic elements in the root apppeared some difference comparing to the leaf.
The effects of20(S)-Ginsenoside-Rh2which is an active compound separated fromPanax ginseng C.A.Meyer cv. Silvatica on myocardial ischemia induced by isoproterenol and pharmacokinetic were investigated in experimental SD rats. The pharmacology resultsshowed that20(S)-Ginsenoside-Rh2has the effect of protecting the heart from damageinduced from ischemia by means of increasing blood pressure and lowering the heart ratios.The pharmacokinetic results showed that the mean plasma concentration-time data could beadequately described by a one compartment open model with r2=0.9976and t1/2=2.8h, AUC=3.4μg.h/mL.
The concentrations of ATP, ADP, AMP, GTP and GDP in the rat plasma were measuredby means of HPLC-UV detection. The concentrations of ATP have been used as a biomarkerfor reflecting the protection to the cardiovascular system.
The chemical constituents of Panax ginseng C.A. Meyer cv. Silvatica and bioactivities of20(S)-Ginsenoside-Rh2were made a deep investigation in the thesis. The results would fillthe theoretical basis research inadequaty of it as well as supply good methods for qualitycontrol. Further more, they also offered strong, accurate and scientific theoretical basis to thewide application of this kind of ginseng, which might produce even higher theoretical andeconomic value.
引文
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