CN104435318B - A kind of extracting method for treating hemorrhagic stroke Chinese medical extract components group and application - Google Patents

A kind of extracting method for treating hemorrhagic stroke Chinese medical extract components group and application Download PDF

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CN104435318B
CN104435318B CN201410734891.3A CN201410734891A CN104435318B CN 104435318 B CN104435318 B CN 104435318B CN 201410734891 A CN201410734891 A CN 201410734891A CN 104435318 B CN104435318 B CN 104435318B
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medical extract
hemorrhagic stroke
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CN104435318A (en
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尹莲
李国春
黄西
吴勉华
程海波
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Nanjing University of Chinese Medicine
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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
    • A61K36/18—Magnoliophyta (angiosperms)
    • A61K36/185—Magnoliopsida (dicotyledons)
    • A61K36/70—Polygonaceae (Buckwheat family), e.g. spineflower or dock
    • A61K36/708—Rheum (rhubarb)
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
    • A61K36/18—Magnoliophyta (angiosperms)
    • A61K36/185—Magnoliopsida (dicotyledons)
    • A61K36/25—Araliaceae (Ginseng family), e.g. ivy, aralia, schefflera or tetrapanax
    • A61K36/258—Panax (ginseng)
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
    • A61K36/18—Magnoliophyta (angiosperms)
    • A61K36/185—Magnoliopsida (dicotyledons)
    • A61K36/71—Ranunculaceae (Buttercup family), e.g. larkspur, hepatica, hydrastis, columbine or goldenseal
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
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    • A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
    • A61K36/18—Magnoliophyta (angiosperms)
    • A61K36/185—Magnoliopsida (dicotyledons)
    • A61K36/80—Scrophulariaceae (Figwort family)
    • A61K36/804—Rehmannia
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
    • A61K2236/10—Preparation or pretreatment of starting material
    • A61K2236/15—Preparation or pretreatment of starting material involving mechanical treatment, e.g. chopping up, cutting or grinding
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
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    • A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
    • A61K2236/30—Extraction of the material
    • A61K2236/33—Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones
    • A61K2236/331—Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones using water, e.g. cold water, infusion, tea, steam distillation, decoction
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
    • A61K2236/30—Extraction of the material
    • A61K2236/39—Complex extraction schemes, e.g. fractionation or repeated extraction steps
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
    • A61K2236/50—Methods involving additional extraction steps
    • A61K2236/55—Liquid-liquid separation; Phase separation

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Abstract

The invention discloses a kind of extracting method for treating hemorrhagic stroke Chinese medical extract components group and its application.Rheum officinale, the root bark of tree peony, the radix paeoniae rubrathe, the dried rhizome of rehmannia, pseudo-ginseng add water to cook extraction according to a certain weight ratio, after crushing, and with after dichloromethane extraction, water layer uses extracting n-butyl alcohol to extract again, and it is Chinese medical extract components group that butanol extraction liquid is reclaimed after doing.Chinese medical extract HPLC MS are analyzed by Chinese herbaceous peony glycoside, tannin class, Paeonol glycoside, Anthraquinones, stilbene class, saponins and iridoid glycosides 33 into being grouped into, male spontaneously hypertensive rat neuronspecific enolase, 100 β albumen of S, IL 1, TNF α, plasma fibrinogen level can be significantly reduced, can be used to prepare the purposes for the treatment of hemorrhagic stroke medicine.

Description

A kind of extracting method for treating hemorrhagic stroke Chinese medical extract components group and application
First, technical field
The present invention relates to technical field of Chinese medicine, specifically one kind treat hemorrhagic stroke Chinese medical extract components group Extracting method and its application.
2nd, background technology
Hemorrhagic stroke (cerebral hemorrhage) is disease clinically common both at home and abroad, and the disease morbidity is hurried, and progress is rapid, dies of illness Rate is high, and patient's prognosis situation directly influences life quality after being ill.Neuronspecific enolase (SNE) and S-100 β eggs It is the important indicator for judging the cerebral lesion order of severity in vain, wherein, SNE is a mark of neuron and neuroendocrine cell Enzyme, when brain tissue occurs hypoxic-ischemic, in cerebrospinal fluid, SNE levels are raised.S-100 β albumen is brain specific proteins, in serum S-100 β protein levels are higher, and degree of cerebral damage is heavier.The method of current western medical treatment cerebral hemorrhage is that operative treatment and internal medicine are comprehensive Two big class for the treatment of are closed, is had certain risk and curative effect is not notable enough.
The traditional Chinese medical science thinks that hemorrhagic stroke comes from stagnated heat in xuefen, and fight is not understood, impatency brain key, and makes heat symptoms caused by an exopathgen more vehement, Jin Erhua The raw phlegm humorously of fire, causes stagnant heat resistance key, therefore based on being expelled the heat-evil with cool blood dissipates the stasis of blood, logical internal organs during treatment.Chinese herb rhubarb clearing heat-fire, the root bark of tree peony Cool blood dissipates the stasis of blood, radix paeoniae rubrathe cooling and activating blood, Sanqi Tongmai stagnation resolvation, dried rhizome of rehmannia nourishing Yin and clearing heat.Therefore, the present invention is carried with these Chinese medicine compositions Take and prepare Chinese medical extract, evaluated its anti-inflammatory, invigorate blood circulation and prevented and treated brain damage effect with spontaneous hypertensive rat, for treating out The research and development of courageous and upright apoplexy medicine.
3rd, the content of the invention
For the demand of clinical Hemorrhage apoplexia, it is an object of the invention to provide a kind of have treatment hemorrhagic stroke The extracting method of Chinese medical extract components group and its application.
The purpose of the present invention is realizing by following technical scheme：
1st, a kind of extracting method for treating hemorrhagic stroke Chinese medical extract components group, extraction step are as follows：
(1) bulk drug：Rheum officinale 1-5 parts, root bark of tree peony 1-5 parts, radix paeoniae rubrathe 1-5 parts, dried rhizome of rehmannia 1-8 parts, pseudo-ginseng 1-6 parts；
(2) above-mentioned raw materials medicinal powder is broken to into 20-40 mesh, plus 5-15 times is measured water and extracted twice, merging filtrate is concentrated into 0.5- 1.5g crude drugs/ml, measure dichloromethane with 1-3 times and extract 3-6 time, abandon dichloromethane layer, and water layer measures extracting n-butyl alcohol with 1-3 times again 3-6 time, merge butanol extraction liquid recovery and do as Chinese medical extract components group；
(3) Chinese medical extract constituent analysis：The chemical composition of Chinese medical extract is analyzed with HPLC-MS, mainly by Paeoniflorin Class, tannin class, Paeonol glycoside, Anthraquinones, stilbene class, saponins and iridoid glycosides 33 are into being grouped into.
2nd, application of a kind of Chinese medical extract components group in treatment hemorrhagic stroke medicine is prepared.
Beneficial effect：
The Chinese medical extract components group of the present invention, mainly by Chinese herbaceous peony glycoside, tannin class, Paeonol glycoside, Anthraquinones, stilbene Class, saponins and iridoid glycosides 33 can significantly reduce spontaneous hypertensive rat neuronal specificity alkene into being grouped into Enolase, S-100 β albumen, IL-1, TNF-α, plasma fibrinogen level, can be used to prepare treatment hemorrhagic stroke medicine Purposes.
4th, the pharmacological testing of Chinese medical extract components group
1. modeling and administration
Male spontaneously hypertensive rat is randomly divided into 4 groups, 8 per group, with Wistar male rats as blank control group (physiological saline group), Hypertensive Rats are respectively model group (physiological saline group), and positive controls (aspirin group), Chinese medicine are carried Take thing high and low dose group.Each group is pressed into 10mLkg-1Daily gastric infusion 1 time, continuous 15 days.After 15th day gastric infusion 1h Arteria carotis takes blood 5ml in added with 0.038gml-1In the centrifuge tube of sodium citrate, with 3000rmin-1Rotating speed is centrifuged 10min, Take upper plasma, Elisa kit measurement rat S-100B albumen, NSE, IL-1, TNF-α, plasma fibrinogen.
Statistics carries out the inspection of double sample t and variance analysis using SPSS13.0 statistical softwares to data, experimental data withRepresent.
2. the impact of couple S-100B and NSE
Experimental result (table 1) shows：Model group S-100B and NSE significantly raise (P ＜ 0.01) compared with blank group, with model group Relatively, positive controls and Chinese medical extract various dose group can significantly reduce S-100B and NSE levels (P ＜ 0.01).
Impact of 1 Chinese medical extract of table to S-100B and NSE
Note：With blank group than ##P ﹤ 0.01；With model group than * * P ﹤ 0.01
3rd, the impact to inflammatory factor
Experimental result (table 2) shows：Spontaneously hypertensive rat model group IL-1, TNF-α level are significantly risen compared with blank group High (P ＜ 0.01).Compare with model group, positive drug control group and Chinese medical extract high and low dose group can significantly reduce spontaneity Hypertensive Rats IL-1, TNF-α level (P ＜ 0.01).
Impact of 2 Chinese medical extract of table to inflammatory factor
Note：Compare * * P ﹤ 0.01 with model group；Compare ##P ﹤ 0.01 with blank group
4th, the impact to plasma fibrinogen
Experimental result (table 3) shows：Male spontaneously hypertensive rat fibrinogen level is significantly raised compared with blank group (P ＜ 0.01).Compare with model group, positive controls and Chinese medical extract high and low dose group can significantly reduce spontaneous high blood Pressure male rat plasma fibrinogen level (P ＜ 0.01).
3 Chinese medical extract of table is to rat annulus proteinogen measurement result
Note：Compare * * P ﹤ 0.01 with model group；Compare ##P ﹤ 0.01 with blank group
5th, traditional Chinese medicine extraction chemical constituent HPLC-MS analyses
Chromatographic column (Thermo ODS-2HYPERSIL 250mm × 4.6mm)；10 μ L of sample size；Column temperature：35℃；Flow velocity is 1.0mL.min-1；Mobile phase：- 0.5% formic acid (B) of methyl alcohol (A), elution program is：0～10min, 22%～30%A；10～ 25min, 30%～40%；25～30min, 40%～50%A；30～50min, 50%～62%A；Sample introduction 10ul；Detection ripple It is long：254nm.Ion gun：Electron spray；Scanning of the mass spectrum mode：Anion；Atomization gas pressure：55psi；Auxiliary heating atmospheric pressure： 55psi；Air curtain pressure：40psi；Ionization temperature：550℃；Collision energy：10V；Capillary voltage：5500V；Remove cluster electricity Pressure：-60V；Sweep limits：M/z50～1000.
Chinese medical extract Jing HPLC-MS are analyzed, and total ion current figure is shown in Fig. 1.By each chromatogram molecular ion peak and main broken The parsing at piece peak reference literature, identify the Chinese medical extract mainly altogether by Chinese herbaceous peony glycoside, tannin class, Paeonol glycoside, anthracene Quinones, stilbene class, saponins and iridoid glycosides 33 are into being grouped into (table 4).
4 n-butanol portion main chemical compositions of table
Result of study shows, by the Chinese medical extract of present invention preparation, mainly by Chinese herbaceous peony glycoside, tannin class, root bark of tree peony phenolic glycoside Class, Anthraquinones, stilbene class, saponins and iridoid glycoside constituents etc. 33 can significantly reduce SH into being grouped into Male rat neuronspecific enolase, S-100 β albumen, IL-1, TNF-α, plasma fibrinogen level, can be used for Prepare the purposes for the treatment of hemorrhagic stroke medicine.
6th, illustrate
Fig. 1 Chinese medical extract HPLC-MS ion flow graphs.
Seven. specific embodiment
Embodiment 1：The extracting method of the Chinese medical extract components group for the treatment of hemorrhagic stroke, extraction step are as follows：
(1) bulk drug：1 part of rheum officinale, 1 part of the root bark of tree peony, 1 part of the radix paeoniae rubrathe, 1 part of the dried rhizome of rehmannia, 1 part of pseudo-ginseng；
(2) above-mentioned raw materials medicinal powder is broken to into 20 mesh, plus 5 times of amount water is extracted twice, merging filtrate, be concentrated into 0.5g crude drugs/ Ml, is extracted 3 times with 1 times of amount dichloromethane, abandons dichloromethane layer, and water layer is again with 1 times of amount extracting n-butyl alcohol 3 times, merging n-butanol Extract is reclaimed and is done as Chinese medical extract components group；
(3) Chinese medical extract constituent analysis：The chemical composition of Chinese medical extract is analyzed with HPLC-MS, mainly by Paeoniflorin Class, tannin class, Paeonol glycoside, Anthraquinones, stilbene class, saponins and iridoid glycosides 33 are into being grouped into.
Embodiment 2：The extracting method of the Chinese medical extract components group for the treatment of hemorrhagic stroke, extraction step are as follows：
(1) bulk drug：2 parts of rheum officinale, 3 parts of the root bark of tree peony, 3 parts of the radix paeoniae rubrathe, 5 parts of the dried rhizome of rehmannia, 3 parts of pseudo-ginseng；
(2) above-mentioned raw materials medicinal powder is broken to into 30 mesh, plus 10 times of amount water is extracted twice, merging filtrate, be concentrated into 1.0g crude drugs/ Ml, is extracted 4 times with 2 times of amount dichloromethane, abandons dichloromethane layer, and water layer is again with 2 times of amount extracting n-butyl alcohols 4 times, merging n-butanol Extract is reclaimed and is done as Chinese medical extract components group；
(3) Chinese medical extract constituent analysis：The chemical composition of Chinese medical extract is analyzed with HPLC-MS, mainly by Paeoniflorin Class, tannin class, Paeonol glycoside, Anthraquinones, stilbene class, saponins and iridoid glycosides 33 are into being grouped into.
Embodiment 3：The extracting method of the Chinese medical extract components group for the treatment of hemorrhagic stroke, extraction step are as follows：
(1) bulk drug：5 parts of rheum officinale, 5 parts of the root bark of tree peony, 5 parts of the radix paeoniae rubrathe, 8 parts of the dried rhizome of rehmannia, 6 parts of pseudo-ginseng；
(2) above-mentioned raw materials medicinal powder is broken to into 40 mesh, plus 15 times of amount water is extracted twice, merging filtrate, be concentrated into 1.5g crude drugs/ Ml, is extracted 6 times with 3 times of amount dichloromethane, abandons dichloromethane layer, and water layer is again with 3 times of amount extracting n-butyl alcohols 6 times, merging n-butanol Extract is reclaimed and is done as Chinese medical extract components group；
(3) Chinese medical extract constituent analysis：The chemical composition of Chinese medical extract is analyzed with HPLC-MS, mainly by Paeoniflorin Class, tannin class, Paeonol glycoside, Anthraquinones, stilbene class, saponins and iridoid glycosides 33 are into being grouped into.

Claims (2)

1. a kind of extracting method for treating hemorrhagic stroke Chinese medical extract components group, extraction step are as follows：
（1）Bulk drug：Rheum officinale 1-5 parts, root bark of tree peony 1-5 parts, radix paeoniae rubrathe 1-5 parts, dried rhizome of rehmannia 1-8 parts, pseudo-ginseng 1-6 parts；
（2）Above-mentioned raw materials medicinal powder is broken to into 20-40 mesh, plus 5-15 times is measured water and extracted twice, merging filtrate is concentrated into 0.5-1.5g Crude drug/ml, measures dichloromethane with 1-3 times and extracts 3-6 time, abandon dichloromethane layer, and water layer measures extracting n-butyl alcohol 3- with 1-3 times again 6 times, merge butanol extraction liquid recovery and do as Chinese medical extract components group；
（3）Chinese medical extract constituent analysis：The chemical composition of Chinese medical extract is analyzed with HPLC-MS, mainly by Chinese herbaceous peony glycoside, tan Matter class, Paeonol glycoside, Anthraquinones, stilbene class, saponins and iridoid glycosides 33 are into being grouped into.
2. application of the Chinese medical extract components group as claimed in claim 1 in treatment hemorrhagic stroke medicine is prepared.
CN201410734891.3A 2014-12-04 2014-12-04 A kind of extracting method for treating hemorrhagic stroke Chinese medical extract components group and application Expired - Fee Related CN104435318B (en)

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