CN104435095B - The total terpene active component of common rabdosia leaf and its extracting method, application, composition - Google Patents
The total terpene active component of common rabdosia leaf and its extracting method, application, composition Download PDFInfo
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Abstract
The invention discloses the total terpene active component of common rabdosia leaf and its extracting method, application, composition.The total terpene active component of the common rabdosia leaf contains oleanolic acid and ursolic acid, contains 0.02 0.5% oleanolic acid, 0.05 1% ursolic acid by raw material weight percentage composition.The main component production and processing of the present invention using medicine turns into suitable pharmaceutical preparation as the immune liver damage disease for the treatment of, can improve liver state, have no side effect, pharmacodynamic tests prove that its is evident in efficacy；Medicine material source in the present invention is easy to get, and is easy to industrialization, and various formulations can be made as needed, and the modern Chinese herbal medicine more convenient, more efficient, quality is more controllable is provided for clinic, more interests is brought for patient, so as to produce huge social benefit.Invention additionally discloses the extracting method of the total terpene active component of the common rabdosia leaf and the application in anti-immunity liver damage disease, the pharmaceutical composition with the total terpene active component of the common rabdosia leaf.
Description
Technical field
The present invention relates to Chinese medicine common rabdosia leaf to extract field, and in particular to a kind of common rabdosia leaf total terpene active component, the therefrom king of medicine
The method of the extraction total terpene active component of common rabdosia leaf and the total terpene active component of the common rabdosia leaf are in anti-immunity liver damage disease in Chinese date
Application in medicine, the pharmaceutical composition with the total terpene active component of the common rabdosia leaf.
Background technology
Virus hepatitis is a kind of whole world common disease, and China's category is popular serious regional, and all kinds virus hepatitis is tight
The health of the people is threaten again, and western countries are most with hepatitis C, China's Major Epidemic hepatitis B.In China, B-mode liver
Scorching virus carrier accounts for the 10% of population, accounts for more than the 1/3 of whole world carrier.Existing hepatitis B patient is about 30,000,000.This
The course of disease is delayed, and is easily changed into CAH, cirrhosis and liver cancer.China is changed into liver cancer because suffering from hepatitis B every year
And the dead is caused up to more than 160,000 people, if plus more if causing the dead because of acute liver failure and cirrhosis, therefore, Chronic Liver
Scorching treatment is become as a significant problem in the urgent need to address.
Types of drugs at present both at home and abroad for chronic hepatitis treatment is a lot of, is summed up, can substantially be divided into three classes：
(1) antiviral agent, mainly there are interferon and several ucleosides analogs, interferon is to B-mode and hepatitis C therapeutic effect
About 30-50%, comparison hepatitis B is slightly good to be seemed to the effect of hepatitis C, but most of disease in latter year of discontinuing medication
People is recurred, and expenses for medicine is fairly expensive, and one patient of import interferon needs the tens thousand of members of RMB, domestic one patient of interferon
For expenses for medicine also more than 10,000 yuan, interferon needs intramuscular injection, there is the side reaction of influenza-like symptom after some patients' medication.It is some
Ucleosides analog is not so good as interferon to the therapeutic effect of hepatitis, also exists and occurs rebound rebound phenomenon, medicine after discontinuing medication
Price is also fairly expensive, and some patients have side reaction.To improve curative effect, external someone tries out interferon and some ucleosides are disease-resistant
Toxic agent, which merges, to be used, and as a result curative effect has no raising.(2) improve liver function medicine, also known as anti-hepatocellular injury medicine, as DDB,
Fruit of Chinese magnoliavine preparation and some Chinese herbal medicines, glycyrrhizin are first succeeded in developing by Japan, and silymarin is German product.This kind of medicine pair
Serum transaminase is reduced, removing jaundice subcutaneous ulcer has certain effect, wherein the double cruel effects of the biphenyl voluntarily developed by China are same better than other
The medicine of type.But this kind of medicine, after stopping using, also there is knock-on and recurred in most patient, slow to hepatitis viruse
Property index is turned out cloudy relatively inexpensive like inefficacy, expenses for medicine, and side reaction is few.(3) immunomodulator, as transfer factor, interleukins-
2nd, thymic peptide etc., is mainly derived from non-specific immunomodulator, is used in the patient that can not apply interferon therapy mostly, or make
For drug combination, chronic hepatitis B is treated, HBeAg negative conversion rates are in 20-30% or so, the definite treatment of such drug therapy hepatitis
Imitate and need more to verify, price is also fairly expensive.From described above satisfied slow virus is controlled as can be seen that there is no at present
The medicine of property hepatitis.
Chronic viral hepatitis particularly hepatitis B, due to hepatitis viruse have can be fitted together to DNA in human liver cell,
The biological heredity material such as RNA, is replicated using the inhereditary material of host cell, above-mentioned molecular biosciences, is caused antiviral
The searching of agent is the problem that the world today faces.In addition, almost all of viral generation, depends not only on virus,
Additionally depend on the individual factors of people.Current research finds that the response of body itself epidemic disease has in the development mechanism of virus hepatitis
There is very important meaning, hepatitis type B virus is to liver cell wound mainly as caused by immune response, while hepatitis viruse sense
The hepatocellular apoptosis of induction or the downright bad damage phenomenon all degraded with fragmentation or diffusivity are contaminated, therefore in addition to antiviral, promote
Enter to damage the reparation of liver cell to look for the stronger immunoregulation medicament of specific aim, also will be the side of such disease therapeuticing medicine research
One of to.
The field that hepatic is a great prospect is researched and developed from Chinese herbal medicine.The existing successful precedent of foreign countries, such as day
The silymarin that this glycyrrhizin class medicine developed from radix glycyrrhizae and Europe are developed, oneself turns into international natural drug city
Important liver-protecting product on field.Clinically also there is the treatment that many natural medicinal formulations are used for hepatitis in China at present, but from facing
Bed curative effect, kind sales volume and added value see that Chinese herbal and crude drugs preparations is not dominant.From China in 1985《New drug approval method》Implement
Since, the national new Chinese medicine of approved about 1000, wherein the new Chinese medicine available for hepatitis treatment about 70, and greatly
The anti-hepatitis new Chinese medicine in part is three classes, four classes, is illustrated for refractory diseases such as chronic hepatitis, and traditional Chinese medicine has certain only
Special curative effect, while also illustrate, there is low-level repetition phenomenon in anti-hepatitis new drug development, due to bioactive components and work
Indefinite with mechanism, the modern anti-hepatitis Chinese medicine for not having high-efficiency low-toxicity so far emerges.
Labiatae Rabdosia contains 25 in the Rabdosia that various active material, Cheng Peiyuan etc. have been studied China
New diterpene-kind compound is summarized more than the 70 of kind, find wherein considerable part compound have cell toxicant, it is antitumor and
Antibacterial activity, and diterpene-kind compound antitumor activity gene in Rabdosia plant and structure-activity relationship are inquired into.In medicine
In terms of imitating, document report protection of the Amethystoidin A to carbon tetrachloride hepatic injury, to send out it have obvious liver protection effect.
Clinically document report Rabdosia plant Folium solani verbascifolii Rterifdia Haraa compound preparations treatment hepatitis B has obvious curative effects.
Liver is the major organs for producing cell factor, is one of first target organs of cell factor again, cell factor net
The not normal balance of network is hepatic diseases, the hepatocellular injury as caused by chronic viral hepatitis, the main disease of liver cell fibrosis
Reason mechanism.The most commonly used transcription regulatory factor NF- κ B of Recent study are regulation pro-inflammatory cytokines, cell surface receptor
With the most important transcription regulatory factor of adhesion factor expression.In terms of the inflammation damnification of liver, NF- κ B are pro-inflammatory by regulating and controlling
The transcription of cell factor (IL-6, TNF-α), chemotactic factor (CF) and CAF directly participates in the acute and chronic inflammation of liver
Disease.
Recent studies indicate that tumor necrosis factor-alpha (TNF-α) inflammation damage in play it is extremely important
Effect.The hour after ConA injections 2, TNF-α has been peak value, gives anti-tnf-alpha serum in advance, and serum TNF-cc level declines,
And protect liver damage to hinder completely, in view of the ability of macrophage synthesis TNF-α is more than T lymphocytes, can speculate that T drenches
Stimulating expression of macrophage release TNF-α, may play a significant role in this pathologic process after bar cell activation.
Common rabdosia leaf Rabdosia amethy stoides (Benth) Hara systems Labiatae Rabdosia plant, our province are main
The mountain area of the northern band in Suzhou is grown in, is Wild Medicinal.For a long time, the local masses take common rabdosia leaf soup to treat
The tradition of disease, and spread the saying of " being that sore is not sore, three bowls of common rabdosia leaf soup ".It is among the people to be used to treat acute infective liver
Inflammation, there is antibacterial, anti-inflammatory and antitumor action etc..Common rabdosia leaf contains many chemical compositions, such as common rabdosia leaf extract I G-1 and H-
1st, H-9, common rabdosia leaf IG-1 and A prime, common rabdosia leaf B prime, Diterpenes, triterpenes, total terpene, flavones and glucoside etc., wherein always
The immune hepatic injury that ter penoidses are induced concanavalin A (ConA) has significant protective effect.
In the pathogenesis of virus hepatitis, liver cell immunologic mjury caused by the activation of T lymphocytes is to cause liver
The direct factor of functional lesion.The experimental liver damaged (such as bacteria lipopolysaccharide, D amine-galactoses) applied for many years is situated between
The hepatic lesion led, this pathological characteristic can not be reflected well more, the immunomodulator or influence for not being suitable for having liver protection effect are exempted from
The evaluating drug effect of the hepatinica of epidemic disease system.Mouse specificity liver damage is induced from the successful application concanavalin A such as Tiegs in 1992
The pathomechanism of wound, its pathogenic characteristic and our human viral's hepatitis known today has the similarity of many.Typically recognize
For immune response is one of important mechanisms for causing viral hepatic injury in liver, such as in the pathogenic process of hepatitis B, is caused
Hepatocellular injury and the direct factor destroyed are the cell immune responses of T cell mediation.The hepatic injury induced with amynologic mechanism
The foundation of model, new approach is opened for the research of hepatic injury, the preventing and treating to research virus hepatitis is significant.Cause
This, we carry out the research of the total terpene liver protection effect of common rabdosia leaf from Immune liver injury.But the effective portion of the total terpene of common rabdosia leaf
Position is not easy to extract, and this application to the total terpene of common rabdosia leaf serves certain obstruction, and the medicine for being unfavorable for the total terpene of common rabdosia leaf pushes away
Extensively.
The content of the invention
It is effective that the total terpene of the common rabdosia leaf is extracted the invention discloses a kind of total terpene active component of common rabdosia leaf, therefrom in king of medicine Chinese date
The application in the medicine of anti-immunity hepatic injury of the method at position and the total terpene active component of the common rabdosia leaf, there is the total terpene of the common rabdosia leaf
The pharmaceutical composition of active component.The present invention have studied the preliminary molecular mechanisms of action of the total terpene liver protection effect of common rabdosia leaf, be king
The medicine of Chinese date total terpene exploitation into preventing and treating hepatic injury provides theoretical foundation.The present invention the total ter penoidses of common rabdosia leaf are extracted,
Isolation and purification, and based on the pathological characteristic of human viral's hepatitis immunologic derangement, the model of immunological liver injury is established,
Influence of the total terpene of common rabdosia leaf to immunological liver injury in mice serum enzyme and Histopathology is observed, the total terpene of common rabdosia leaf is to being immunized liver
The influence of critical inflammatory medium T NF- α and IFN-γ protein level is damaged, and molecule is recorded to TNF-α upstream signaling molecule consideration convey
NF- κ B regulatory mechanism research, during inquiring into immunological liver injury, the preliminary molecular action machine of the total terpene liver protection effect of common rabdosia leaf
Reason, to provide theoretical foundation the medicine of common rabdosia leaf total terpene exploitation into preventing and treating hepatic injury.The present invention is achieved in that a Seed King
The total terpene active component of Chinese date, it contains oleanolic acid and ursolic acid, contains 0.02% to 0.5% by raw material weight percentage composition
Oleanolic acid, 0.05% to 1% ursolic acid.By the percentage by weight of the total terpene active component of the common rabdosia leaf, oleanolic acid contains
Amount accounts for 0.8% to 8%, and ursolic acid content accounts for 1.0% to 12%.
Wherein, the total terpene active component of the common rabdosia leaf is extracted using following steps：
(1) take clean common rabdosia leaf to crush, add 15 to 35 times of weight, 70% to 95% ethanol, 50 to 80 DEG C of ultrasounds carry
40 to 100min are taken, extract solution filtering, filtrate adjusts pH value=1-3 with hydrochloric acid, and hydrolysis 20 to 40min is heated to reflux to filtrate, to
The water that 0.5-1.0 times of volume is added in hydrolyzate fully mixes, then, merges extraction with isometric chloroform extraction several times
Liquid；
(2) the common rabdosia leaf extract of gained is concentrated under reduced pressure, carries out post separation, with water and ethanol elution, collect respective streams
Part, freeze.
The present invention also provides a kind of method that total terpene active component is extracted in therefrom king of medicine Chinese date, and it comprises the following steps：
(1) take clean common rabdosia leaf to crush, add 15 to 35 times of weight, 70% to 95% ethanol, 50 to 80 DEG C of ultrasounds carry
40 to 100min are taken, extract solution filtering, filtrate adjusts pH value=1-3 with hydrochloric acid, and hydrolysis 20 to 40min is heated to reflux to filtrate, to
The water that 0.5-1.0 times of volume is added in hydrolyzate fully mixes, then, merges extraction with isometric chloroform extraction several times
Liquid；
(2) the common rabdosia leaf extract of gained is concentrated under reduced pressure, carries out post separation, with water and ethanol elution, collect respective streams
Part, freeze, produce the total terpene active component of above-mentioned common rabdosia leaf.
As the further improvement of such scheme, the splitter is macroporous resin column, the macroporous absorption of the macroporous resin column
Resin is at least one of D101, AB-8, HPD100A, HPD400, HPD600, HPD750.Preferably, adsorption time be 6 to
After 10 hours, eluted with 1 to 4BV/h flow velocitys, first with 3 to 6BV washing post remove water-solubility impurity, then with 3 to
6BV40% merges ethanol eluate to 60% ethanol elution, freezes, and produces the total terpene active component of common rabdosia leaf of purity >=50%.
As the further improvement of such scheme, the splitter is polyamide column, with water elution, the weight of extract solution and filler
It is 1 ︰ 25-40 to measure ratio, after eluting 1-3 column volume, starts collection 2-6 column volume of fraction, lyophilized, produces the king of purity >=50%
The total terpene active component of Chinese date.
As the further improvement of such scheme, the splitter is sephadex column, and the sephadex column is selected from G-
10、G-25、G-50、LH-20.Preferably, with water elution, the weight ratio of extract solution and filler is 1 ︰ 20-50, elutes 1-3 post
After volume, start to collect 2-6 column volume of fraction, freeze, produce purity >=effective portion of the total terpene of 50% common rabdosia leaf.
As the further improvement of such scheme, the splitter is reverse C18, with water elution, the weight of extract solution and filler
It is 1 ︰ 10-20 to measure ratio, after eluting 2-4 column volume, starts to collect 5-11 column volume of fraction, freezes, produce purity >=50%
The total terpene active component of common rabdosia leaf.
As the further improvement of such scheme, the splitter is C8 posts, with water elution, the weight ratio of extract solution and filler
For 1 ︰ 20-50, after eluting 1-3 column volume, start to collect 2-6 column volume of fraction, it is lyophilized, produce the common rabdosia leaf of purity >=50%
Total terpene active component.
The present invention also provides a kind of application of the total terpene active component of above-mentioned common rabdosia leaf in the medicine of anti-immunity hepatic injury.It is special
Be not treatment virus hepatitis, oneself immunity hepatitis, hepatonecrosis, fatty liver, cholestasia, liver fibrosis, hepatic sclerosis and
The application of liver cancer etc..
The present invention also provides a kind of pharmaceutical composition for treating immune liver damage disease, and one kind is included in the pharmaceutical composition
The total terpene active component of above-mentioned common rabdosia leaf as main component.
Beneficial effects of the present invention are as follows：
(1) the total terpene active component extracted in a kind of common rabdosia leaf from Chinese medicine is provided, produced with the main component of medicine
Suitable pharmaceutical preparation is processed into as the immune liver damage disease for the treatment of, liver state can be improved, had no side effect, through pharmacodynamics
Experiment proves that its is evident in efficacy；
(2) the medicine material source in the present invention is easy to get, and is easy to industrialization, various formulations can be made as needed, to face
The modern Chinese herbal medicine that bed offer is more convenient, more efficient, quality is more controllable, more interests are brought for patient, so as to produce
Huge social benefit；
(3) the immune liver damage disease that medicine of the invention is treated mainly includes：Virus hepatitis, LADA liver
Inflammation, hepatonecrosis, fatty liver, cholestasia, liver fibrosis, hepatic sclerosis and liver cancer etc..
Brief description of the drawings
Fig. 1 is the chromatogram that oleanolic acid and ursolic acid reference substance determine.
Fig. 2 is the oleanolic acid and ursolic acid chromatogram of common rabdosia leaf extract solution sample measure.
Fig. 3 oleanolic acid canonical plottings.
Fig. 4 A, Fig. 4 B, Fig. 4 C, Fig. 4 D, Fig. 4 E, Fig. 4 F are protective effect of the total terpene various dose of common rabdosia leaf to hepatic injury
HE colored graphs：Fig. 4 A Normal groups, Fig. 4 B Normal groups+common rabdosia leaf active component high dose group, Fig. 4 C model groups, Fig. 4 D
Common rabdosia leaf active component low dose group, Fig. 4 E common rabdosia leaf active component middle dose groups, Fig. 4 F common rabdosia leaf active component high dose groups.
Embodiment
In order to make the purpose , technical scheme and advantage of the present invention be clearer, it is right below in conjunction with drawings and Examples
The present invention is further elaborated.It should be appreciated that specific embodiment described herein is only to explain the present invention, not
For limiting the present invention.
The total terpene active component of common rabdosia leaf contains oleanolic acid and ursolic acid, contains 0.02% by raw material weight percentage composition
Oleanolic acid, 0.05% to 1% ursolic acid to 0.5%.The molecular formula of oleanolic acid and ursolic acid difference is as follows：
The method that total terpene active component is extracted in Chinese medicine common rabdosia leaf, is comprised the following steps that.
(1) take clean common rabdosia leaf to crush, be put into ultrasonic tank, add the ethanol of 15 to 35 times of weight 70% to 95%, 50
PH value=1-3 is adjusted with hydrochloric acid, filtrate is heated to reflux hydrolyzing to 80 DEG C of ultrasonic extractions 40 to 100min, extract solution filtering, filtrate
20 to 40min, to hydrolyzate in add the water of 0.5-1.0 times of volume and fully mix, then with isometric chloroform extraction 1-3
It is secondary, combining extraction liquid.
(2) the common rabdosia leaf extract of gained is concentrated under reduced pressure to proper volume, carries out post separation, splitter is selected from macropore tree
Fat post, polyamide column, sephadex column, reverse C18 and C8 posts, with water and the ethanol elution of various concentrations, collect respective streams
Part, freeze, produce the total terpene active component of common rabdosia leaf.
Wherein, 1, when being separated using large pore resin absorption column, preferable macroporous absorbent resin be D101, AB-8,
One or more kinds of macroporous absorbent resins in HPD100A, HPD400, HPD600, HPD750, maximum applied sample amount be 1 to
4BV resin demands, pH are 3 to 6, and resin blade diameter length ratio is the ︰ 15 of 1 ︰ 6 to 1, and adsorption rate is 1 to 4BV/h, and adsorption time is 6 to 10
After hour, eluted with 1 to 4BV/h flow velocitys, first remove water-solubility impurity with 3 to 6BV washing post, then with 3 to 6BV
40% to 60% ethanol elution, merge ethanol eluate, freeze, produce the total terpene active component of common rabdosia leaf of purity >=50%；
2nd, when being separated using polyamide column, with water elution, the weight ratio of extract solution and filler is 1 ︰ 25-40, elutes 1-3
After individual column volume, start to collect 2-6 column volume of fraction, it is lyophilized, produce the total terpene active component of common rabdosia leaf of purity >=50%；
3rd, when using sephadex post separation, preferable sephadex is selected from G-10, G-25, G-50, LH-20,
With water elution, the weight ratio of extract solution and filler is 1 ︰ 20-50, preferably 1 ︰ 25-40, after eluting 1-3 column volume, starts to receive
Collect 2-6 column volume of fraction, freeze, produce purity >=effective portion of the total terpene of 50% common rabdosia leaf；
4th, when using anti-phase C18 post separations, with water elution, the weight ratio of extract solution and filler is 1 ︰ 10-20, elutes 2-
After 4 column volumes, start to collect 5-11 column volume of fraction, it is lyophilized, produce the total terpene active component of common rabdosia leaf of purity >=50%；
5th, when using anti-phase C8 post separations, with water elution, the weight ratio of extract solution and filler is 1 ︰ 20-50, elutes 1-3
After individual column volume, preferable elution volume is 2 column volumes, starts to collect 2-6 column volume of fraction, lyophilized, produce purity >=
The total terpene active component of 50% common rabdosia leaf.
It is as follows to the determination method of oleanolic acid and ursolic acid in above-mentioned gained common rabdosia leaf extract solution.
Experimental agents：Oleanolic acid and ursolic acid reference substance, are purchased from Nat'l Pharmaceutical ＆ Biological Products Control Institute's chromatographic condition
With system suitability test：Shim-pack vp-ODS C18Chromatographic column (250mm × 4.6mm, 5 μm).Mobile phase methanol-water (90
∶10)；Detection wavelength：210nm；Flow velocity：0.8mL/min；Column temperature：25℃.Theoretical cam curve is calculated with oleanolic acid and ursolic acid
It is not less than 4000.
Reference substance solution：Precision weigh 105 DEG C of dryings to constant weight oleanolic acid and ursolic acid reference substance 5.8mg,
2.4mg, dissolved with methanol and be settled to 10mL, be made into the solution that concentration is 0.58mg/mL, 0.24mg/mL, cross 0.45 μm of filter
Film, as reference substance stock solution, it is standby.
Need testing solution：It is appropriate that precision pipettes common rabdosia leaf extract, is evaporated, residue adds methanol to dissolve and is settled to 10mL amounts
In bottle, shake up, 0.45 μm of miillpore filter filtration, produce need testing solution.
Sample size determines：It is accurate respectively to draw reference substance and each 20 μ l of need testing solution, liquid chromatograph is injected, is surveyed
It is fixed, as a result see Fig. 1-2, reference substance is consistent with the retention time of ursolic acid with oleanolic acid in sample chromatogram figure, and separating effect is good
It is good, reach the related request of separating degree.
As a result, content of oleanolic acid accounts for the 0.02% to 0.5% of common rabdosia leaf medicinal material in common rabdosia leaf extract solution, ursolic acid content
Account for the 0.05% to 1% of common rabdosia leaf medicinal material.
It is as follows to the determination method of ter penoidses in the total terpene active component of above-mentioned gained common rabdosia leaf.
Experimental agents：Oleanolic acid reference substance, it is purchased from Nat'l Pharmaceutical ＆ Biological Products Control Institute's reference substance solution：Precision weighs
105 DEG C of dryings are dissolved with methanol to the oleanolic acid reference substance 5.0mg of constant weight and are settled to 25mL, it is 0.20mg/ to be made into concentration
ML solution, as reference substance stock solution, it is standby.
Need testing solution：It is appropriate that precision pipettes the total terpene active component freeze-dried powder of common rabdosia leaf, adds methanol to dissolve and be settled to
In 10mL measuring bottles, shake up, produce need testing solution.
Standard curve is established：It is accurate respectively to draw oleanolic acid reference substance solution (0.20mg/mL) 0.2,0.4,0.6,
0.8,1.0mL in different tool plug test tubes and numbers, and water-bath volatilizes, and adds 5% vanillic aldehyde-glacial acetic acid solution of brand-new
0.4mL, perchloric acid 1.6mL, with methanol constant volume to 5mL, shaking is mixed after 70 DEG C of heating water baths, frozen water cooling.With examination of accompanying
Blank is done in agent, and spectral scan result is shown, there is maximum absorption band at 517nm, selects to determine absorption value at 517nm.With extinction
Angle value A is ordinate, and reference substance concentration C is that abscissa is returned, and obtains equation A=0.8125C+0.0043, r=0.9996
(see Fig. 3).Oleanolic acid linear relationship in the range of 0.008~0.04mg/mL is good.
Total terpene content in the total terpene active component of determined by ultraviolet spectrophotometry common rabdosia leaf：It is accurate respectively to draw appropriate control
Product and sample solution, heating water bath solution volatilize solution to be measured, then add the 5% vanillic aldehyde-glacial acetic acid solution just made
0.4mL, perchloric acid 1.6mL, and with methanol constant volume to 5mL, be put in 70 DEG C of water-baths and heat after solution is rocked uniformly, frozen water
Cooling.Retinue reagent is done into blank control, its absorption value is determined at 517nm.
As a result, total terpene content (being calculated with oleanolic acid) is 52%-70% in the total terpene active component freeze-dried powder of common rabdosia leaf.
Oleanolic acid and ursolic acid content in the total terpene active component of high effective liquid chromatography for measuring common rabdosia leaf：By above-mentioned efficient
Liquid phase chromatogram condition is measured to the total terpene active component freeze-dried powder of common rabdosia leaf of purifying, as a result in the total terpene active component of common rabdosia leaf
Content of oleanolic acid accounts for the 0.8% to 8% of freeze-dried powder, and ursolic acid content accounts for the 1.0% to 12% of freeze-dried powder.
The pharmacodynamics test of the total terpene active component anti-immunity hepatic injury of common rabdosia leaf proves that it is carried out in the steps below：
1 experiment material
1.1 experimental animal
Male ICR mouse, the scholar 2g of body weight 20, Yangzhou University's medical experiment animal center provide.
1.2 Experimental agents
Concanavalin A (ConcanavalinA, ConA)：Sigma Products；Pyrrole alkane dithiocar-bamate
(Pyrrolidinedithioearbamate, PDTC), common rabdosia leaf active component, by Pharmaceutical Biotechnology research institute of Suzhou University
There is provided.
2 experimental methods
2.1 ConA cause immune liver injury model preparation method
With reference to relevant document, ConA 18mg/kg mouse tail vein injections are prepared into liver injury model.And calculated according to body weight
ConA amount, is then diluted to 0.3mL volume injections with sterile PBS needed for going out, and plucks eyeball blood sampling, blood after injection after 8 hours
3000rpm/min centrifuges 10min, takes serum ultra low temperature freezer to freeze in case in next step needed for detection, determines ALT, AST work
Property, and mouse was put to death in the 8th hour, take liver to detect corresponding index.
2.2 animal packets design
After ICR mouse adaptability is fed three days, 6 groups, every group 10 are randomly divided into.(1) Normal group：Daily gavage is same
Isometric physiological saline, once a day, continuous 5 days.(2) liver injury model group is immunized：Daily gavage equal volume sterile physiological
Salt solution, once a day, continuous 5 days, damaged in the 5th day gavage half an hour after tail vein injection ConA 18mg/kg with causing to be immunized liver
Wound model.(3) PDTC treatment groups：Daily gavage equal volume physiological saline, and on 5th before ConA is by the injection of above-mentioned dosage
1 hour, PDTC120mg/kg intraperitoneal injections, ConA and PDTC are all equally diluted to 0.3mL volumes.(4) the effective portion of common rabdosia leaf
Daily high dose 10mg/kg, middle dosage 3mg/kg, low dosage 1mg/kg gavage mouse, continuous gavage 5 days, in the 5th are pressed in position administration
1 hour tail vein injection ConA 18mg/kg after its gavage.
2.3 Testing index
2.3.1 serum enzyme inspection
2.3.1.1 reagent：Glutamic-pyruvic transaminase detection kit (ALT, reitman-frankel method), it is public purchased from the glad biotechnology of Shanghai section
Department；Glutamic-oxalacetic transaminease detection kit (AST, reitman-frankel method), purchased from Shanghai Ke Xin biotech companies.
2.3.2 histopathologic examination
2.3.2.1 reagent：10% neutral formalin (is formulated) by 40% formaldehyde 10mL, pH=7.4 PBS；Soviet Union
Another name for (element)：1 gram of haematine, 8.8 grams, absolute ethyl alcohol 10mL, distilled water 200mL of aluminum sulfate.First sulphur is dissolved by heating with distilled water
Sour aluminium, then poured into the bush seminal fluid for being dissolved in absolute ethyl alcohol in the sulfuric acid aluminium liquid dissolved, boil 1 minute, slightly cool down, slowly
0.1 gram of sodium iodate is added, dye liquor is changed into aubergine, with gauze lid bottleneck, every 100mL acetic acid on the rocks after being filtered using preceding filter paper
5mL；Yihong liquid：After the grinding of 1 gram of Yihong is dissolved in into alcohol with glass bar, instilling acetic acid makes in the pasty state, addend milliliter distilled water, mistake
Filter, the sediment filtered out is dried in an oven, dissolved in 95% alcohol 200mL, you can.
2.3.3 influence of the total terpene active component of common rabdosia leaf to serum levels of inflammatory cytokines TNF-α, IFN-γ
Experiment packet is same as above with administration, and plucking eyeball after modeling administration 8h takes blood, and serum is left and taken in centrifugation.ELISA method determines
The expression of TNF-α, IFN-γ (pg/mL).
2.3.4 the total terpene active component of common rabdosia leaf to the influence experiment packet of hepatic tissue NF- κ B p65 expression with giving
Medicine is same as above, and is put to death mouse after modeling administration 8h, is cut open the belly and take liver.Western-blot methods measure murine liver tissue NF- κ B p65
Expression.
3 statistical dispositions
The data obtained is represented with x ± s, using SPSS11.0 statistical softwares analyze data, using one-way analysis of variance:
Used when variance is neat during LSD methods (least significant difference method) heterogeneity of variance and use Dunnett T3 methods.Variance analysis test of homogeneity
Horizontal α1=0.05, level of significance test α2=0.05.
4 testing results
4.1 ALT and AS are checked：After ConA is injected 8 hours, Serum ALT and AST concentration significantly raise, with Normal group
Compare, difference has conspicuousness (p<0.01), it was demonstrated that modeling success.PDTC can resist ConA pairs as NF- kB activity inhibitor
Damage caused by mouse liver, can substantially reduce Serum ALT and AST is horizontal, and the heavy dose of group of common rabdosia leaf active component can also show
Writing reduces the level of Serum ALT and AST, and both have significant difference (p compared with model control group<0.01).It is and low dose of
Group has certain conspicuousness (p compared with model group<0.05).It the results are shown in Table 1.
Influence (n=10) of the common rabdosia leaf active component of table 1 to immune hepatic injury mice serum ALT and AST
Note：Compared with Normal group,#p<0.05；##p<0.01；Compared with model control group,*p<0.05；**p<0.01
4.2 pathological examination results
4.2.1 visual results：There is the necrotic plaque to differ in size on model group liver enlargement, surface.The effective portion of common rabdosia leaf
Hyte, the change of positive drug control group liver are light compared with model group.Normal group liver is shown no obvious abnormalities.
4.2.2 light microscopy checking：This experiment is in the case where following the principle of hepatic injury pathological grade classification respectively from degeneration of liver cells
Improvement with the aspect of necrosis of liver cells two evaluation common rabdosia leaf active component to blood lipid.HE coloration results see Fig. 4 A,
Fig. 4 B, Fig. 4 C, Fig. 4 D, Fig. 4 E, Fig. 4 F, using 6 groups, Fig. 4 A are Normal group, and Fig. 4 B are that Normal group+common rabdosia leaf is effective
Position high dose group, Fig. 4 C are model group, and Fig. 4 D are common rabdosia leaf active component low dose group, and Fig. 4 E are that model group+common rabdosia leaf has
Position middle dose group is imitated, Fig. 4 F are model group+common rabdosia leaf active component high dose group, and their histological scores are shown in Table 2.
In terms of degeneration of liver cells, χ2=55.835, p=0.000；In terms of necrosis of liver cells, χ2=40.731, p=0.000；
Show that each group curative effect has significant difference.Compared with normal group, the pathological change such as the denaturation of model group liver and necrosis is special
Significantly (p ＜ 0.05/5), Animal Model success.In terms of degeneration of liver cells and necrosis of liver cells, each administration group improves liver
The significant effect of dirty denaturation, and have certain dose dependent.Microscopic observation the results are shown in Table 2.
Influence (n=10) of the common rabdosia leaf active component of table 2 to the liver morphology of each group mouse
Note：#P ＜ 0.05/5,**P ＜ 0.001, compared with model group.
4.3 each group peripheral blood TNF-αs, the level of IFN-γ
As shown in table 3, compared with normal group, model group mouse peripheral blood TNF-α, IFN-γ level significantly raise
(p<0.05).Compared with model group, control group, common rabdosia leaf active component various dose group can obviously reduce mouse peripheral blood blood
Clear TNF-α, IFN-γ level, there is significant difference (p<0.05).
The comparison horizontal to each group mice serum TNF-α, IFN-γ of the common rabdosia leaf active component of table 3
Note：Compared with normal group,##p<0.01,#p<0.05, compared with model**P ＜ 0.01,*P ＜ 0.05.
Influence of the total terpene active component of 4.4 common rabdosia leafs to hepatic tissue NF- κ B p65 expression
Western-blot methods measure murine liver tissue NF- κ B p65 expression.Statistical result is shown in Table 4, and normal
Group compares, and model group NF- κ B p65's is horizontal significantly raised, statistics there were significant differences (p<0.05)；It is right compared with model group
There is a significant change according to the level of group and common rabdosia leaf active component various dose group NF- κ B p65, statistics there were significant differences (p<
0.05).Common rabdosia leaf active component various dose group can significantly reduce immunological liver injury in mice hepatic tissue NF- κ B p65 expression
It is horizontal.
Influence of the common rabdosia leaf active component of table 4 to each group hepatic tissue NF- κ B p65 expression
Note：Compared with normal group,##p<0.01,#p<0.05, compared with model**P ＜ 0.01,*P ＜ 0.05.
The present invention further discloses the medicine that the total terpene active component of the common rabdosia leaf containing oleanolic acid and ursolic acid is formed
Application of the compositions in the medicine for treating immune liver damage disease is prepared, particularly in treatment virus hepatitis, itself exempts from
The application of epidemic disease hepatitis, hepatonecrosis, fatty liver, cholestasia, liver fibrosis, hepatic sclerosis and liver cancer etc..
The present invention carries out extracting and developing and purifying to the total ter penoidses of common rabdosia leaf, and is immunized based on human viral's hepatitis
Disorderly pathological characteristic, the model of immunological liver injury is established, the observation total terpene of common rabdosia leaf is to immunological liver injury in mice blood
The influence of clear zymetology and Histopathology, the total terpene of common rabdosia leaf are expressed immune hepatic injury critical inflammatory medium T NF- α and IFN-γ
Horizontal influence, and molecule NF- κ B regulatory mechanism research is recorded to TNF-α and IFN-γ upstream signaling molecule consideration convey, inquire into
During immunological liver injury, the preliminary molecular mechanisms of action of the total terpene liver protection effect of common rabdosia leaf, be the total terpene of common rabdosia leaf develop into
The medicine for preventing and treating hepatic injury provides theoretical foundation.
The effect of the present invention：
(1) the total terpene active component extracted in a kind of common rabdosia leaf from Chinese medicine is provided, produced with the main component of medicine
Suitable pharmaceutical preparation is processed into as the immune liver damage disease for the treatment of, liver state can be improved, had no side effect, through pharmacodynamics
Experiment proves that its is evident in efficacy；
(2) the medicine material source in the present invention is easy to get, and is easy to industrialization, various formulations can be made as needed, to face
The modern Chinese herbal medicine that bed offer is more convenient, more efficient, quality is more controllable, more interests are brought for patient, so as to produce
Huge social benefit；
(3) the immune liver damage disease that medicine of the invention is treated mainly includes：Virus hepatitis, LADA liver
Inflammation, hepatonecrosis, fatty liver, cholestasia, liver fibrosis, hepatic sclerosis and liver cancer etc..
The foregoing is merely illustrative of the preferred embodiments of the present invention, is not intended to limit the invention, all essences in the present invention
All any modification, equivalent and improvement made within refreshing and principle etc., should be included in the scope of the protection.
Claims (10)
- A kind of 1. total terpene active component of common rabdosia leaf, it is characterised in that：It contains oleanolic acid and ursolic acid, by raw material weight percentage Than forming the ursolic acid containing 0.02% to 0.5% oleanolic acid, 0.05% to 1%；Wherein, the effective portion of the total terpene of the common rabdosia leaf Extracted using following steps position：(1) take clean common rabdosia leaf to crush, add 15 to 35 times of weight, 70% to 95% ethanol, 50 to 80 DEG C of ultrasonic extractions 40 To 100min, extract solution filtering, filtrate adjusts pH value=1-3 with hydrochloric acid, hydrolysis 20 to 40min is heated to reflux to filtrate, to hydrolysis The water that 0.5-1.0 times of volume is added in liquid fully mixes, then, combining extraction liquid. with isometric chloroform extraction several times；(2) the common rabdosia leaf extract of gained is concentrated under reduced pressure, carries out post separation, with water and ethanol elution, collect corresponding fraction, frozen It is dry.
- A kind of 2. total terpene active component of common rabdosia leaf, it is characterised in that：It contains oleanolic acid and ursolic acid, by the total terpene of the common rabdosia leaf The percentage by weight of active component, content of oleanolic acid account for 0.8% to 8%, and ursolic acid content accounts for 1.0% to 12%；Wherein, should The total terpene active component of common rabdosia leaf is extracted using following steps：(1) take clean common rabdosia leaf to crush, add 15 to 35 times of weight, 70% to 95% ethanol, 50 to 80 DEG C of ultrasonic extractions 40 To 100min, extract solution filtering, filtrate adjusts pH value=1-3 with hydrochloric acid, hydrolysis 20 to 40min is heated to reflux to filtrate, to hydrolysis The water that 0.5-1.0 times of volume is added in liquid fully mixes, then, combining extraction liquid. with isometric chloroform extraction several times；(2) the common rabdosia leaf extract of gained is concentrated under reduced pressure, carries out post separation, with water and ethanol elution, collect corresponding fraction, frozen It is dry.
- 3. the method for total terpene active component is extracted in a kind of therefrom king of medicine Chinese date, it is characterised in that it comprises the following steps：(1) take clean common rabdosia leaf to crush, add 15 to 35 times of weight, 70% to 95% ethanol, 50 to 80 DEG C of ultrasonic extractions 40 To 100min, extract solution filtering, filtrate adjusts pH value=1-3 with hydrochloric acid, hydrolysis 20 to 40min is heated to reflux to filtrate, to hydrolysis The water that 0.5-1.0 times of volume is added in liquid fully mixes, then, combining extraction liquid. with isometric chloroform extraction several times；(2) the common rabdosia leaf extract of gained is concentrated under reduced pressure, carries out post separation, with water and ethanol elution, collect corresponding fraction, frozen It is dry, produce the total terpene active component of common rabdosia leaf as claimed in claim 1 or 2.
- 4. the method for total terpene active component is therefrom extracted in king of medicine Chinese date as claimed in claim 3, it is characterised in that the separation Post is macroporous resin column, the macroporous absorbent resin of the macroporous resin column is D101, AB-8, HPD100A, HPD400, HPD600, At least one of HPD750, adsorption time are after 6 to 10 hours, are eluted with 1 to 4BV/h flow velocitys, first with 3 to 6BV's Wash post and remove water-solubility impurity, then with 3 to 6BV40% to 60% ethanol elution, merge ethanol eluate, freeze, produce pure Spend the total terpene active component of >=50% common rabdosia leaf.
- 5. the method for total terpene active component is therefrom extracted in king of medicine Chinese date as claimed in claim 3, it is characterised in that the separation Post is polyamide column, and with water elution, the weight ratio of extract solution and filler is 1 ︰ 25-40, after eluting 1-3 column volume, starts to receive Collect 2-6 column volume of fraction, it is lyophilized, produce the total terpene active component of common rabdosia leaf of purity >=50%.
- 6. the method for total terpene active component is therefrom extracted in king of medicine Chinese date as claimed in claim 3, it is characterised in that the separation Post is sephadex column, and the sephadex column is selected from G-10, G-25, G-50, LH-20, with water elution, extract solution and filler Weight ratio be 1 ︰ 20-50, after eluting 1-3 column volume, start to collect 2-6 column volume of fraction, it is lyophilized, produce purity >= The effective portion of the total terpene of 50% common rabdosia leaf.
- 7. the method for total terpene active component is therefrom extracted in king of medicine Chinese date as claimed in claim 3, it is characterised in that the separation Post is anti-phase C18 posts, and with water elution, the weight ratio of extract solution and filler is 1 ︰ 10-20, after eluting 2-4 column volume, starts to receive Collect 5-11 column volume of fraction, it is lyophilized, produce the total terpene active component of common rabdosia leaf of purity >=50%.
- 8. the method for total terpene active component is therefrom extracted in king of medicine Chinese date as claimed in claim 3, it is characterised in that the separation Post is anti-phase C8 posts, and with water elution, the weight ratio of extract solution and filler is 1 ︰ 20-50, after eluting 1-3 column volume, starts to receive Collect 2-6 column volume of fraction, it is lyophilized, produce the total terpene active component of common rabdosia leaf of purity >=50%.
- 9. a kind of total terpene active component of common rabdosia leaf as claimed in claim 1 or 2 is in the medicine for preparing anti-immunity hepatic injury Using.
- A kind of 10. pharmaceutical composition for treating immune liver damage disease, it is characterised in that：Make in the pharmaceutical composition comprising a kind of For the total terpene active component of common rabdosia leaf of main component, the total terpene active component of the common rabdosia leaf is king's jujube as claimed in claim 1 or 2 Sub total terpene active component.
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| CN105585471B (en) * | 2016-01-18 | 2018-08-07 | 大连理工大学 | A kind of extracting method of common rabdosia leaf active constituent |
| CN106800579A (en) * | 2017-01-10 | 2017-06-06 | 大连理工大学 | A kind of method of separation and Extraction Rabdosia infedus glycosides A from common rabdosia leaf and application |
| CN106581146A (en) * | 2017-01-11 | 2017-04-26 | 安徽科技学院 | Isodon amethystoides heat-clearing bacteriostatic granule and production method thereof |
| CN108432816B (en) * | 2018-03-14 | 2020-07-28 | 淮北师范大学 | Extraction method of active ingredients of Wangzao date and application of active ingredients in agriculture |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1522723A (en) * | 2003-02-21 | 2004-08-25 | 宿州绿源中医药科技有限公司 | Novel use of traditional Chinese medicine rabdosia rubeseens and Isodon longitubus |
-
2014
- 2014-11-14 CN CN201410648925.7A patent/CN104435095B/en active Active
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
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Non-Patent Citations (3)
| Title |
|---|
| HPLC测定蓝萼香茶菜中齐墩果酸和熊果酸的含量;沈晓丹等;《中成药》;20101231;第32卷(第12期);2182-2183 * |
| 王枣子三萜成分的研究;崔佳等;《安徽中医学院学报》;20110630;第30卷(第3期);57-59 * |
| 齐墩果酸和熊果酸保肝药理作用的研究进展;张明发等;《抗感染药学》;20120331;第9卷(第1期);13-19 * |
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