CN100340255C - Moutan bark effective constituent and preparation method thereof - Google Patents
Moutan bark effective constituent and preparation method thereof Download PDFInfo
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- CN100340255C CN100340255C CNB2005100604375A CN200510060437A CN100340255C CN 100340255 C CN100340255 C CN 100340255C CN B2005100604375 A CNB2005100604375 A CN B2005100604375A CN 200510060437 A CN200510060437 A CN 200510060437A CN 100340255 C CN100340255 C CN 100340255C
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- Prior art keywords
- paeoniflorin
- mobile phase
- paeonol
- extract
- preparation
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Landscapes
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Abstract
本发明提供一种丹皮有效组分，其活性成分的百分含量为芍药苷10～25%，没食子酰芍药苷40～55%，牡丹皮苷h30～45%；制备方法是将丹皮药材粉碎后加热提取，将提取液浓缩成浸膏，与硅胶进行拌样，用正相硅胶柱对其进行分离，将洗脱液浓缩干燥后得样品，用制备液相色谱继续分离得到的有效组分。本发明的丹皮有效组分加入药用添加剂按照药剂学允许的制备方法制成药用制剂并在制备治疗、预防心血管疾病药物中的应用。本发明方法使用了正相硅胶柱，能有效地除去糖、蛋白质、氨基酸等杂质，提高有效成分的含量，能快速准确的得到有效成分，在生产中更易于药物的质量控制。The invention provides an effective component of paeonol, the percentage content of which is 10-25% of paeoniflorin, 40-55% of galloyl paeoniflorin, and 30-45% of paeoniflorin h; the preparation method is to prepare paeoniflorin After pulverization, heat extraction, concentrate the extract into an extract, mix the sample with silica gel, separate it with a normal phase silica gel column, concentrate and dry the eluent to obtain a sample, and continue to separate the effective group obtained by preparative liquid chromatography point. The effective components of paeonol of the present invention are added with pharmaceutical additives to make pharmaceutical preparations according to the preparation method allowed by pharmacy, and are used in the preparation of medicines for treating and preventing cardiovascular diseases. The method of the invention uses a normal-phase silica gel column, can effectively remove sugar, protein, amino acid and other impurities, increase the content of active ingredients, can quickly and accurately obtain active ingredients, and is easier to control the quality of medicines in production.
Description
Technical field
The present invention relates to Chinese medicine extract, relate in particular to active component that extracts and preparation method thereof from Cortex Moutan, this active component mainly contains peoniflorin, galloylpaeoniflorin, three kinds of chemical compounds of Cortex Moutan Saponin h (Mudanpioside h).
Background technology
Coronary heart disease and angina pectoris are harm humans health " first killers ", in recent years, along with the variation of China's population senescence and people's work, life, dietary structure and environment etc., the incidence rate of cardiovascular and cerebrovascular diseases such as coronary heart disease also increases year by year, and the people's physical and mental health in serious threat.In the natural product many active substances have resist myocardial ischemia, anoxia functions, some of them have been developed to treatment coronary heart disease and anginal new drug, as treat the medicine DIAOXINXUE KANG of coronary heart disease, it is by 8 kinds of pure Chinese medicinal preparations that steroid saponin is made that extract from Chinese medicine; Flavonihippophae is to be that raw material extracts the pure natural medical that processes with the Fructus Hippophae, and its effective ingredient is a Fructus Hippophae total flavones.Thereby from natural product, seek have resist myocardial ischemia, the active substance of anoxia physiologically active, be find, one of effective way of developing new drug.China's medicinal organism resource is very abundant, and its biological active substances is research and finds new drug guide chemicals, the natural treasure-house of developing new drug.At present, China extracts active substance from natural product, be used to be developed to treatment coronary heart disease, safety is good, toxicity is low new drug also seldom, from natural product, extract active substance, be developed to have and resist myocardial ischemia, be used for the treatment of coronary heart disease and anginal new drug, have significant application value and wide development prospect.
Cortex Moutan has another name called Cortex Moutan, is the root bark of dicotyledon medicine ranunculaceae peony (Paeoniasuffruticosa Andr.).Ground such as distribution Hebei, Henan, Shandong, Sichuan, Shaanxi, Gansu.All there is cultivation all parts of the country, ground such as medical material main product Anhui, Sichuan, Gansu, Shaanxi, Hubei, Hunan, Shandong, Guizhou, and in addition, also produce in Yunnan, Zhejiang.Its cold nature, the acrid in the mouth hardship, the heart, liver, kidney channel are gone in cold, have heat clearing away, removing heat from blood, and blood, the effect of repercussive.Cortex Moutan is concocted and to be seen China's beam Dai Taohong scape the earliest and write in " variorums ", thinks: " all urge to break, remove the heart." in the medical book such as Compendium of Material Medica record " the Cortex Moutan core is not used as medicine, and need go it." studies show that: Cortex Moutan contains paeonol (Paeonol), paeoniflorin (Paeoniflorin), hydroxypaeoniflorin, benzoylpaeoniflorin (Benzoylpaeonifforin), benzoyl hydroxypaeoniflorin, Radix Paeoniae aglycon (Paeoniflorigenone), 6-Hydroxycoumarin (6-hydroxycoumarin) and gallic acid (GallicAcid), and benzoic acid, plant sterol, sucrose, glucose, arabinose etc.Contain benzoic acid (Benzoic Acid), tetradecane hydrocarbon (Tetradecane), 2-hydroxyl-4-methoxyl group-1-Phenylethanone. (2-hydroxy-4-methoxy acetophenone), 2 in the volatile oil, the two tertiary butyl 1,4-benzoquinone (2 of 6-, 6-ditert-butyl-p-Benzoquinone), diethyl phthalate (Diethyl Phthalate), isopropyl myristate chemical compound (Chen Yuejiao etc. such as (Isopropyl Myristate), Guangzhou food industry science and technology, 2002,18 (4): 36-37).People such as Wu Shaohua separate from Cortex Moutan first and have obtained following five kinds of chemical compounds: betulic acid (Betulinic Acid), betulin (Betulin), oleanolic acid (Oleanolic Acid), 3 β, 23-dihydroxy-30-norolean-12,20 (29)-dien-28-oic acid, 3-O-methylpaeonisuffral (Wu Shaohua etc., Chinese herbal medicine, 2002,33 (8): 679-680).
Cortex Moutan has the degree of injury of alleviating effect to expeirmental myocardial ischemia, and can reduce myocardial oxygen consumption, increases coronary flow; think that Cortex Moutan has that to regulate blood capable, the effect of mediation blood vessels, thereby myocardial ischemia had protective effect (Ma Yuling etc.; the Shanxi medical magazine, 1984,13 (4): 212-214).Peoniflorin has antiplatelet aggregative activity, and antithrombotic effect is to the protective effect and the antitumor action of liver.Peoniflorin has blocking effect (Zhang Guangxin etc., Chinese Pharmacological circular, 2003,19 (8): 863-866) to myocardial cell Ica, L.Peoniflorin can increase neurocyte survival quantity, reduces mortality rate, excitatory neuron damage (Wu Yumei etc., Chinese J Pharmacol Toxicol, 2002,16 (3): 172-175) due to the antagonism KA.Peoniflorin has significant protective effect (Yang Jun etc., Chinese Journal of New Drugs, 2001,10 (6): 426-428) to PC12 cell injury due to the calcium overload.The colony that peoniflorin strengthens syndrome of deficiency of blood mouse bone marrow cells hematopoietic stem/progenitor cells forms ability; Peoniflorin can promote marrow stromal cell secretion Hemopoietic factor (G-CSF, GM-CSF, PDGF-α) etc., suppresses secretion (Gao Yue, Tianjin Chinese medicine, 2003,20 (6): 47-51) of hemopoietic inhibitive factor (M-CIP).
Summary of the invention
The purpose of this invention is to provide a kind of moutan bark effective constituent, active component has peoniflorin, galloylpaeoniflorin, three kinds of chemical compounds of Cortex Moutan Saponin h (Mudanpioside h).
In the moutan bark effective constituent that the inventive method provides, content of paeoniflorin is 10～25%, and the content of galloylpaeoniflorin is 40～55%, and the content of paeonoside h (Mudanpioside h) is 30～45%.
In the preferred moutan bark effective constituent, content of paeoniflorin is that the content of 12～21% galloylpaeoniflorins is 42～51%, and the content of paeonoside h (Mudanpioside h) is 32～4 1%.
In the best moutan bark effective constituent, content of paeoniflorin is 14～18%, and the content of galloylpaeoniflorin is 44～48%, and the content of paeonoside h (Mudanpioside h) is 34～38%.
Another object of the present invention provides the preparation method of moutan bark effective constituent, is achieved through the following technical solutions:
Ethyl acetate and ethanol will be added after the Cortex Moutan pulverizing medicinal materials, heating extraction gets extracting solution, extracting solution is condensed into extractum, and itself and silica gel are mixed sample, it is separated with normal phase silicagel column, at first use petroleum ether and ethyl acetate as mobile phase, get eluent I, abandon it, change chloroform and methanol then as mobile phase, get eluent II, will get sample behind the eluent II concentrate drying; Continue to separate the active component that obtains with preparative liquid chromatography; The separation condition of preparative hplc: chromatographic column is a semi-preparative column, and mobile phase is water and acetonitrile, and gradient elution is collected solution, and solution obtains active component behind concentrate drying.
Preferred moutan bark effective constituent preparation method realizes by following steps: will add ethyl acetate and ethanol (1: 0.8～1.2) after the Cortex Moutan pulverizing medicinal materials, reflux 0.8～1.2 hour, extract 1～3 time, merging filtrate gets extracting solution, extracting solution is condensed into extractum, and itself and silica gel mixed sample, with normal phase silicagel column it is separated, at first use petroleum ether and ethyl acetate (47～53: 1) as mobile phase, get eluent I, abandon change then chloroform and methanol (8～13: 1) as mobile phase, eluent II, will be behind the eluent II concentrate drying sample; Continue to separate the active component that obtains with preparative liquid chromatography; The separation condition of preparative hplc: chromatographic column is a semi-preparative column, and mobile phase is water and acetonitrile, and gradient elution, flow velocity are 2.8～3.2ml/min, and column temperature is a room temperature.Collect solution, solution obtains active component behind concentrate drying.
Best moutan bark effective constituent preparation method realizes by following steps: will add ethyl acetate and ethanol (1: 1) after the Cortex Moutan pulverizing medicinal materials, reflux 1 hour is extracted 2 times, and merging filtrate gets extracting solution; Extracting solution is condensed into extractum, and itself and silica gel mixed sample, with normal phase silicagel column it is separated, at first use petroleum ether and ethyl acetate (50: 1) as mobile phase, get eluent I, abandon it, change chloroform and methanol (10: 1) then as mobile phase, get eluent II, will get sample behind the eluent II concentrate drying; Continue to separate the active component that obtains with preparative liquid chromatography; The separation condition of preparative hplc: chromatographic column is semi-preparative column (Lichrospher C18; 10.0mm * 250mm, 5 μ m), mobile phase is water (A) and acetonitrile (B), and the gradient elution program is as follows: in the time of 0 minute, mobile phase A is that 80% aqueous solution, Mobile phase B are 20% acetonitrile solution; In the time of 40 minutes, mobile phase A is that 20% aqueous solution, Mobile phase B are 80% acetonitrile solution; In the time of 50 minutes, mobile phase A is that 5% aqueous solution, Mobile phase B are 95% acetonitrile solution; Flow velocity is 3ml/min, and column temperature is a room temperature; Sample 100% dissolve with ethanol separates through preparative liquid chromatography, collects solution at time period 1 4.2～18.4min, and solution obtains active component behind concentrate drying.
Another object of the present invention provides this moutan bark effective constituent adding medical additive and makes pharmaceutical formulation according to the preparation method that pharmaceutics allows.
Another purpose of the present invention provides this moutan bark effective constituent and the application of preparation in preparation treatment, angiocardiopathy preventing medicine thereof.
Usefulness of the present invention is: the method for extraction separation from Cortex Moutan, use normal phase silicagel column, can remove impurity such as desaccharide, protein, aminoacid effectively, improved content of effective, simultaneously in preparation method, adopt preparative hplc, can obtain effective ingredient fast and accurately.Moutan bark effective constituent provided by the invention has protective effect to myocardial ischemia, and its chemical constituent is simply clear and definite, is easier to illustrate its mechanism of action on pharmacological research, is easier to the quality control of medicine aborning.
Description of drawings
Fig. 1 is the HPLC analysis chart of moutan bark effective constituent of the present invention.
The specific embodiment
Further describe flesh and blood of the present invention below in conjunction with embodiments of the invention, this embodiment only is used to the present invention is described and the present invention is not limited.
The preparation technology of embodiment one moutan bark effective constituent
Ethyl acetate and ethanol will be added after the 200g Cortex Moutan pulverizing medicinal materials, heating extraction gets extracting solution, extracting solution is condensed into extractum 16.8g, and itself and silica gel mixed sample, with normal phase silicagel column it is separated and at first uses petroleum ether and ethyl acetate as mobile phase, eluent I, abandon it, change chloroform and methanol then as mobile phase, get eluent II, will get sample 3.9g behind the eluent II concentrate drying; Continue to separate the sample that obtains with preparative liquid chromatography; The separation condition of preparative hplc: chromatographic column is a semi-preparative column, and mobile phase is water and acetonitrile, gradient elution.Sample 100% dissolve with ethanol separates through preparative liquid chromatography, collects solution at time period 14.2～18.4min, obtains active component 0.40g behind the concentrate drying.
The preparation technology of embodiment two moutan bark effective constituents
Ethyl acetate and ethanol (1: 0.8～1.2) will be added after the 500g Cortex Moutan pulverizing medicinal materials, reflux 0.8～1.2 hour, extract 1～3 time, merging filtrate gets extracting solution, extracting solution is condensed into extractum 42g, and itself and silica gel mixed sample, with normal phase silicagel column it is separated, at first use petroleum ether and ethyl acetate (47～53: 1) as mobile phase, get eluent I, abandon it, change chloroform and methanol (8～13: then 1) as mobile phase, get eluent II, will get sample 11.8g behind the eluent II concentrate drying; Continue to separate the sample that obtains with preparative liquid chromatography; The separation condition of preparative hplc: chromatographic column is a semi-preparative column, and mobile phase is water and acetonitrile, and gradient elution, flow velocity are 2.8～3.2ml/min, and column temperature is a room temperature.Sample 100% dissolve with ethanol separates through preparative liquid chromatography, collects solution at time period 14.2～18.4min, obtains active component 1.04g behind the concentrate drying.
The preparation technology of embodiment three moutan bark effective constituents
Get Cortex Moutan medical material 250g, it is pulverized the back add ethyl acetate and ethanol (1: 1), reflux 1 hour is extracted 2 times, filtrate merge extracting solution.Extracting solution is condensed into extractum gets 22g, with extractum and silica gel mixed sample, separate with normal phase silicagel column, at first use petroleum ether and ethyl acetate (50: 1) as mobile phase, get eluent I, change chloroform and methanol (10: 1) then as mobile phase, get eluent II, get the 6.2g sample behind the concentrate drying.Continue to separate the sample that obtains with preparative liquid chromatography.The separation condition of preparative hplc: chromatographic column is semi-preparative column (the Lichrospher C18 of Chinese nation; 10.0mm * 250mm, 5 μ m), mobile phase is water (A) and acetonitrile (B), and gradient sees Table 2, and flow velocity is 3ml/min, and column temperature is a room temperature.Sample 100% dissolve with ethanol separates through preparative liquid chromatography, collects solution at time periods 14.2～1 8.4min, obtains active component 0.58g behind the concentrate drying.
The analysis of embodiment four moutan bark effective constituents
(1) peoniflorin in the above-mentioned Cortex Moutan extract, galloylpaeoniflorin, paeonoside h (Mudanpioside h), assay (high performance liquid chromatography)
Chromatographic condition chromatographic column Agilent Zorbax SB-C18 post (4.6mm * 150mm, 5 μ m); Adopt gradient elution, mobile phase A is 0.2% glacial acetic acid aqueous solution mutually, and Mobile phase B is mutually for containing the acetonitrile solution of 0.2% glacial acetic acid; The gradient elution program is as follows: in the time of 0 minute, mobile phase A is that 90% 0.2% glacial acetic acid aqueous solution, Mobile phase B are the acetonitrile solution of 10% 0.2% glacial acetic acid; In the time of 10 minutes, mobile phase A is that 50% 0.2% glacial acetic acid aqueous solution, Mobile phase B are the acetonitrile solution of 50% 0.2% glacial acetic acid; Mobile phase A is that 5% 0.2% glacial acetic acid aqueous solution, Mobile phase B are the acetonitrile solution of 95% 0.2% glacial acetic acid in the time of 30 minutes; Flow velocity 0.5mLmin
-1It is long to detect the wavelength all-wave; 30 ℃ of column temperatures; ELSD condition: 100 ℃ of drift tube temperatures; Nitrogen flow rate 1.4L/min
The preparation of need testing solution takes by weighing this product, in volumetric flask, is diluted to scale with dissolve with methanol solution, shakes up, promptly.
The accurate need testing solution of drawing of assay method injects chromatograph of liquid, measures, promptly.
(2) above-mentioned Cortex Moutan extract HPLC-ELSD finger printing
Assay method is referring to the peoniflorin in (1) above-mentioned Cortex Moutan extract, galloylpaeoniflorin, paeonoside h (Mudanpioside h) assay (high performance liquid chromatography).The record chromatograph time is 30 minutes.
The HPLC-ELSD analysis of spectra of analysis result moutan bark effective constituent is seen Fig. 1, and 1,2, No. 3 peak among Fig. 1 is respectively peoniflorin, Mudanpioside h and galloylpaeoniflorin.The relative retention time at three peaks is followed successively by 9.8 (peoniflorins), 10.67 (galloylpaeoniflorins), 10.97 (paeonoside h).
Among the present invention, the structural formula of peoniflorin is:
Among the present invention, the structural formula of galloylpaeoniflorin is:
Among the present invention, the structural formula of paeonoside h (Mudanpioside h) is:
Embodiment five moutan bark effective constituent preparations
Get moutan bark effective constituent 0.5g and 10.5g Polyethylene Glycol-6000 mix homogeneously of embodiment three, heating and melting moves in the drop pill drip irrigation behind the change material, and in ℃ liquid paraffin of medicine liquid droplet to 6～8, oil removing makes 400 of drop pill.
Embodiment six moutan bark effective constituent preparations
Get moutan bark effective constituent 0.5g, glucose 4.5g, sodium thiosulfate 0.9g and the distilled water 1ml of embodiment three, behind the said components mix homogeneously, lyophilization, 500 of packing, promptly.
Embodiment seven moutan bark effective constituent preparations
Get Lignum Dalbergiae Odoriferae oil 1.5g, join in the saturated hydroxypropyl of 13ml, stirring and dissolving filters, the filter leaf cold drying, Lignum Dalbergiae Odoriferae oil and the clathrate powder of hydroxypropyl.Except that above-mentioned Lignum Dalbergiae Odoriferae oil closes the clathrate powder of hydroxypropyl, get Cortex Moutan extract 0.5g, mannitol 5.5g, calcium disodium edetate 0.9g and the distilled water 2ml of embodiment three again, behind the said components mixing, lyophilization, 300 of packing, promptly.
The pharmacological evaluation of embodiment eight moutan bark effective constituents
Pharmacological model: neonatal rat myocardial cell hypoxic-ischemic model
After former generation, myocardial cell was cultivated birth SD neonatal rat execution in 1～3 day, it was dirty to core, and subtracts into 1mm after PBS liquid cleans
3About fragment.With 0.125% trypsin Sigma, USA)+0.05% (Gibco is USA) in 37 degree digestion 3～4 times for collagenase II.The differential adherent method was separated into fibrocyte after 1.5 hours, and cell suspension is transferred to (every hole about 4 * 10 in 48 orifice plates
5Cell).(Gibco, (Falcon, USA) cell of cultivating 3-6 days supplies medicine efficacy screening USA) to add 10% hyclone through DMEM.Screen and replaced with serum-free medium in preceding 1 day.Cultivate 3～6 days myocardial cell, PBS washing back adds the pastille culture fluid.Place 95%N
2And 5%CO
2Anoxia is 6 hours in the incubator.Then at CO
2Reoxygenation is 3 hours in the incubator.Get cell conditioned medium liquid and measure LDH content.
Dosage regimen is extracted the moutan bark effective constituent that obtains and is made into 10 with sugar-free Hanks liquid
-4G/mL test liquid, liposoluble constituent can add a small amount of DMSO hydrotropy (the DMSO final concentration is controlled at below 0.2%).The final concentration of each component in culture fluid is controlled at 10
-5G/mL.
Lactic acid dehydrogenase content can be represented the cell injury degree in the lactic acid dehydrogenase assay cell culture fluid.The lactic acid dehydrogenase content that bleeds in the supernatant is high more, shows that cell injury is also remarkable.The determination of lactate dehydrogenase test kit builds up bio-engineering research institute available from Nanjing.Assay method is: get 30 microlitre cell conditioned medium liquid, add 50 microlitre substrate buffer and 10 microlitre lactic dehydrogenase enzyme cofactors, 37 degree vibrations 15 minutes add 50 microlitre 2,4 dinitrophenyl hydrazines again, 37 degree vibrations 15 minutes.Parallel blank.Extract reaction solution 50 microlitres and add 200 microlitre 0.4mol/L sodium hydroxide solutions, leave standstill 3～4 minutes after, measure shading value with microplate reader in 450nm.
Drug effect result
All data are represented with the mean value variance.Adopt monolateral variance analysis and T check carrying out statistical analysis.Compare with model group, P＜0.05 is thought significantly effectively.Drug effect the results are shown in Table 1.According to myocardial cell lactic acid dehydrogenase (LDH) testing result, moutan bark effective constituent has the highly significant effect to reducing LDH release.
Table 1
| Mean | Std | P | |
| Moutan bark effective constituent | 0.1355 | 0.031522 | 0.003716 |
| Model group | 0.22575 | 0.032887 |
Another pharmacological evaluation of embodiment nine moutan bark effective constituents
Pharmacological model: huve cell anoxia reoxygenation model
The healthy newborn umbilical cord is got in former generation huve cell cultivation, cleans the interior residual bloodstain of vein blood vessel with 30～50mlHanks liquid.Look the 0.1% collagenase I that umbilical cord length adds respective amount, change incubator digestion 15～20 minutes over to.Subsequently Digestive system in the umbilical cord is carefully collected in the beaker, and with Hanks liquid with beaker rinse twice, collect in the lump and be sub-packed in centrifuge tube in the beaker, centrifugal 10 minutes of 900rpm.Supernatant is removed in suction, and (contain 10% hyclone, 100U/ml is two anti-, 0.15mg/ml Heparin, 10uMVC) abundant dissolution precipitation with the M199 culture fluid.Gained cell suspension is sub-packed in 5cm
2Culture bottle places in the incubator and cultivates.Changed in second day and adopt the M199 culture fluid that contains 30ug/ml ECGS, changed a culture fluid and be paved with the bottom surface in per afterwards three days until cell.Used cell is former generation endotheliocyte.The culture bottle inner cell is seeded to 96 orifice plates before the modeling and cultivated one day, used cell concentration is 3.5～40,000/hole.During modeling, inhale and abandon old M199 culture fluid, adding pastille does not have phenol red M199 culture fluid, and every hole total amount is 100ul.Place 95%N
2And 5%CO
2Anoxia is 12 hours in the incubator, then at CO
2Reoxygenation is 2 hours in the incubator.Get cell conditioned medium liquid and measure LDH content and NO content.
Dosage regimen is extracted the moutan bark effective constituent that obtains and is made into 10 with sugar-free Hanks liquid
-4G/mL test liquid, liposoluble constituent can add a small amount of DMSO hydrotropy (the DMSO final concentration is controlled at below 0.2%).The final concentration of each component in no phenol red M199 culture fluid is controlled at 10
-5G/mL.
Lactic acid dehydrogenase content can be represented the cell injury degree in the lactic acid dehydrogenase assay cell culture fluid.The lactic acid dehydrogenase content that bleeds in the supernatant is high more, shows that cell injury is also remarkable.The determination of lactate dehydrogenase test kit builds up bio-engineering research institute available from Nanjing.Assay method is: get 30 microlitre cell conditioned medium liquid, add 50 microlitre substrate buffer and 10 microlitre lactic dehydrogenase enzyme cofactors, 37 degree vibrations 15 minutes add 50 microlitre 2,4 dinitrophenyl hydrazines again, 37 degree vibrations 15 minutes.Parallel blank.Extract reaction solution 50 microlitres and add 200 microlitre 0.4mol/L sodium hydroxide solutions, leave standstill 3～4 minutes after, measure shading value with microplate reader in 450nm.
Drug effect all data is as a result represented with the mean value variance.Adopt monolateral variance analysis and T check carrying out statistical analysis.Compare with model group, P＜0.05 is thought significantly effectively.Drug effect the results are shown in Table 2.According to endotheliocyte lactic acid dehydrogenase testing result, moutan bark effective constituent has the highly significant effect to reducing LDH release.
Table 2
| Component | Mean | Std | P |
| The moutan bark effective constituent model group | 0.104 0.122 | 0.016 0.006 | 0.049 |
Need not further to elaborate, believe and adopt the disclosed content in front, those skilled in the art can use to greatest extent.Therefore, the preferred specific embodiments of front should be understood that only to illustrate, but not limits the scope of the invention by any way.
Claims (10)
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| 丹皮的化学成分研究 吴少华，马云保，罗晓东，郝小红，吴大刚，中草药，第33卷第8期 2002 * |
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