CN104586924A - Red-backed Christmas bush root extract and preparation method as well as application of red-backed Christmas bush root extract in preparation of medicine for treating hepatic fibrosis - Google Patents
Red-backed Christmas bush root extract and preparation method as well as application of red-backed Christmas bush root extract in preparation of medicine for treating hepatic fibrosis Download PDFInfo
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Abstract
The invention discloses a red-backed Christmas bush root extract as well as a preparation method and application of the red-backed Christmas bush root extract in preparation of a medicine for treating hepatic fibrosis. The method comprises the following steps: performing alcohol extraction, centrifugal condensation, macroreticular resin column, ethanol elution and reduced pressure drying to obtain an effective part extract of the red-backed Christmas bush root. The extraction process of the red-backed Christmas bush root extract is simple and reasonable, and the obtained effective part extract of the red-backed Christmas bush root can reduce ALT and AST, as well as HA, LN and III procollagens and TGFbeta1 and TIMP-1 in serums of model animals so as to prove that the red-backed Christmas bush root extract has an ideal anti-hepatic fibrosis function, is a promising candidate medicine and can be applied to preparation of anti-hepatic fibrosis medicines.
Description
Technical field
The invention belongs to the field of Chinese medicines, particularly a kind of effective part extract of redback christmashush root, also relate to preparation method and the purposes of this extract.
Background technology
Hepatic fibrosis is one of difficult disease of serious harm human physical and mental health, still finds no the prevention and treatment method of effect so far.Hepatic fibrosis is the early stage process of chronic hepatitis to cirrhosis progress, vital effect is played in the factors determining patients with chronic liver prognosis, if before developing into liver cirrhosis, block or reverse generation and the development of hepatic fibrosis by certain means, at chronic hepatopathy therapeutically by generation important breakthrough.Modern medicine aspect, colchicine is comparatively early applied to clinical anti-hepatic fibrosis medicines, but toxic and side effects is comparatively strong, and clinical practice is restricted.Glucocorticoid and Beracilline etc. are not widely adopted greatly and due to side effect.How at the experimental stage proline-4-hydroxylase mortifier, prostaglandin E similar medicine, gamma interferon, precollagen peptide, hepatocyte growth factor, IL-10 etc. are at present, and its preparation and source all exist shortcomings, Western medicine aspect there is no effectively ripe and that side effect is little anti-hepatic fibrosis medicines at present.Chinese medicine also achieves encouraging progress in recent years about the research work of anti-hepatic fibrosis.Many prescriptions prove to have certain effect of anti hepatic fibrosis through experiment or clinical research, concentrate on the research of the anti-hepatic fibrosis of the Chinese medicines such as dispersing liver and promoting blood circulation blood stasis dispelling, vital energy benefiting and the kidney invigorating, Ruan Jian Xiao Disorder, supporting vital QI and dispersing blood stasis at present, as strong liver softening the hard mass soup, Fuzheng Huayu 319 Recipe, Semen Persicae extract, artificial cordyceps mycelia, compound recipe 861 mixture etc.But mostly rest on experimentation or clinical observation on the therapeutic effect, there is no the medicine listing with significant curative effect at present.
Redback christmashush root is the root of dicotyledon medicine euphorbia plant red back of the body Cortex Alchorneae Davidii Alchornea trewioides (Benth.) Muell. – Arg..Call red back of the body ma, Folium Malloti barbati (" Guangxi Chinese herbal medicine "), the red skirt of thin silk (Guangzhou air force " conventional Chinese herbal medicine handbook ").Distribution Central China and the southeast, south China, be usually used as medicine with root, leaf.Its root sweet in the mouth, property is put down.Effect heat-clearing and toxic substances removing, expelling wind and removing dampness, dissipating blood stasis stops blooding, and relievings asthma, killing parasites for relieving itching.Tradition is applied to controls enteritis, diarrhea, dysentery, icterohepatitis, urinary tract infection and calculus, nephritis, dysuria, hematuria, metrorrhagia, leucorrhea, eczema, urticaria, carbuncle pyogenic infections from tumour or sore, and skin infection such as not to close up for a long time the disease.Modern medicine research shows, has cough-relieving, phlegm-dispelling functions in redback christmashush root, and anti-acetylcholine effect suppresses golden yellow and Staphylococcus albus effect with slight.
Redback christmashush root water extract and water extract-alcohol precipitation gained extractum have hepatitis virus resisting and stop the effect of hepatic fibrosis, can clinically in order to treat chronic hepatitis and liver cirrhosis.Result of study shows that redback christmashush root significantly can suppress two important factors (TGF-β 1, TIMP-1) of the liver fibrosis due in carbon tetrachloride (CCL4) complex factors and alcoholic fibrosis rat blood serum, and animal pattern liver fiber index (HA, III procollagen type), liver function indexes (ALT, AST) has remarkable reduction, and have protecting the liver, fall enzyme, the effect of anti-hepatic fibrosis.Therefore, further the screening of anti-hepatic fibrosis effective site is carried out to redback christmashush root and purposes research significant.
Summary of the invention
Technical problem to be solved by this invention is, provides a kind of redback christmashush root extract and its production and use.
The technical solution adopted for the present invention to solve the technical problems is as follows: a kind of preparation method of redback christmashush root extract, is characterized in that comprising the following steps:
(1) redback christmashush root medical material 6-12 times amount 60%-90% ethanol extraction 1-2 hour, extracts 1-3 time, merges ethanol extract, concentrating under reduced pressure after filtering;
(2) by centrifugal for step (1) gained concentrated solution, supernatant concentrating under reduced pressure is for subsequent use;
(3) by the concentrated solution that step (2) obtains, with 1.3-1.7 column volume/hour flow velocity, upper macroporous adsorptive resins;
(4) with after deionized water eluting removing impurity, then use ethanol elution, elution flow rate be 2.3-2.7 column volume/hour;
(5) collect ethanol elution drying under reduced pressure, after pulverizing, obtain redback christmashush root effective part extract.
In described step (1), concentration of alcohol is 60%-90%.
In described step (1), concentration of alcohol is 70%-80%.
In described step (4), concentration of alcohol is 20%-60%.
In described step (4), concentration of alcohol is 30%-50%.
The extract that the redback christmashush root effective part extract that described step (5) obtains is is main active with alkaloids and flavonoid.
The redback christmashush root extract obtained by above-mentioned preparation method.
Above-mentioned redback christmashush root extract is preparing the application in anti-hepatic fibrosis medicines.
Beneficial effect of the present invention is as follows:
1. extraction process of the present invention is rationally easy, can obtain redback christmashush root effective part extract.
2. redback christmashush root extract of the present invention, in liver function index context of detection, medicine of the present invention 3 dosage groups significantly can reduce ALT and AST (p<0.01) in animal pattern serum, and colchicine is also reduction trend (p<0.05) to a certain degree to ALT and AST in serum.Other extract groups also can reduce ALT and AST (p<0.05) in animal pattern serum, but action intensity extract comparatively of the present invention is poor.In the context of detection of hepatic fibrosis index (in serum HA, LN, III precollagen), medicine of the present invention 3 dosage groups significantly can reduce HA, LN in animal pattern serum, III precollagen (p<0.01), colchicine to the HA in serum, LN, III precollagen, type Ⅳ collagen also in reduction trend (p<0.05) to a certain degree.Other extract groups also can reduce ALT and AST in animal pattern serum to a certain extent, but action intensity extract comparatively of the present invention is poor.The context of detection of TGF β 1 and TIMP-1 in serum, medicine of the present invention 3 dosage groups significantly can reduce TGF β 1 in animal pattern serum and TIMP-1, colchicine to the TGF β 1 in serum and TIMP-1 also in reduction trend (see table 1).Other extract groups also can reduce TGF β 1 in animal pattern serum and TIMP-1 to a certain extent, but action intensity extract comparatively of the present invention is poor.The above results illustrates that extract of the present invention has comparatively ideal effect of anti hepatic fibrosis, is more promising drug candidate.
Detailed description of the invention
The present invention is a kind of preparation method of redback christmashush root extract, and it comprises the following steps:
(1) redback christmashush root medical material 6-12 times amount ethanol extraction 1-2 hour, extracts 1-3 time, merges ethanol extract, concentrating under reduced pressure after filtering.Preferably, in step (1), concentration of alcohol is 60%-90%, and preferred, concentration of alcohol is 70%-80%.Redback christmashush root is the root of dicotyledon medicine euphorbia plant red back of the body Cortex Alchorneae Davidii Alchorneatrewioides (Benth.) Muell. – Arg..
(2) by centrifugal for step (1) gained concentrated solution, supernatant concentrating under reduced pressure is for subsequent use.
(3) by the concentrated solution that step (2) obtains, with 1.3-1.7 column volume/hour flow velocity, upper middle polarity or nonpolar macroporous adsorptive resins.
(4) with after deionized water eluting removing impurity, then use ethanol elution, elution flow rate be 2.3-2.7 column volume/hour.Preferably, in step (4), concentration of alcohol is 20%-60%, and preferred, concentration of alcohol is 30%-50%.
(5) collect ethanol elution drying under reduced pressure, namely obtain redback christmashush root effective part extract after pulverizing, the extract that this redback christmashush root effective part extract is is main active with alkaloids and flavonoid.
Above-mentioned obtained Radix seu Folium Alchorneae trewioidis effective site root extract is preparing the application in anti-hepatic fibrosis medicines.
Describe the present invention below in conjunction with embodiment.But the present invention is not limited to these given embodiments.Most preferred embodiment 1.
Embodiment 1
By redback christmashush root medical material, with alcohol reflux 2 times, concentration of alcohol 75%, alcohol adding amount is 10 times of medical material weight, extraction time 90min.Merge extractive liquid, filters and concentrating under reduced pressure, and concentrated solution is centrifugal, gets supernatant concentrating under reduced pressure for subsequent use.
Get middle polarity macroporous adsorbent resin, post is filled in the ratio with crude drug weight ratio 1:2, then by above-mentioned concentrated solution for subsequent use with 1.5 column volumes/hour flow velocity by this macroporous resin column, after passing through completely, first use deionized water eluting, carry out gradient elution with 30-50% ethanol again, elution flow rate be 2.5 column volumes/hour.Collect ethanol elution, drying under reduced pressure, pulverizing, obtain redback christmashush root extract.Yield is 0.38%.
Embodiment 2
By redback christmashush root medical material, with alcohol reflux 3 times, concentration of alcohol 90%, alcohol adding amount is 12 times of medical material weight, extraction time 120min.Merge extractive liquid, filters and concentrating under reduced pressure, and concentrated solution is centrifugal, gets supernatant concentrating under reduced pressure for subsequent use.
Get middle polarity macroporous adsorbent resin, post is filled in the ratio with crude drug weight ratio 1:2, then by above-mentioned concentrated solution for subsequent use with 1.5 column volumes/hour flow velocity by this macroporous resin column, after passing through completely, first use deionized water eluting, carry out gradient elution with 30-60% ethanol again, elution flow rate be 2.5 column volumes/hour.Collect ethanol elution, drying under reduced pressure, pulverizing, obtain redback christmashush root extract.Yield is 0.29%.
Embodiment 3
By redback christmashush root medical material, with alcohol reflux 1 time, concentration of alcohol 60%, alcohol adding amount is 6 times of medical material weight, extraction time 60min.Merge extractive liquid, filters and concentrating under reduced pressure, and concentrated solution is centrifugal, gets supernatant concentrating under reduced pressure for subsequent use.
Get middle polarity macroporous adsorbent resin, post is filled in the ratio with crude drug weight ratio 1:2, then by above-mentioned concentrated solution for subsequent use with 1.5 column volumes/hour flow velocity by this macroporous resin column, after passing through completely, first use deionized water eluting, carry out gradient elution with 20-40% ethanol again, elution flow rate be 2.5 column volumes/hour.Collect ethanol elution, drying under reduced pressure, pulverizing, obtain redback christmashush root extract.Yield is 0.31%.
Embodiment 4
By redback christmashush root medical material, with alcohol reflux 3 times, concentration of alcohol 70%, alcohol adding amount is 7 times of medical material weight, extraction time 80min.Merge extractive liquid, filters and concentrating under reduced pressure, and concentrated solution is centrifugal, gets supernatant concentrating under reduced pressure for subsequent use.
Get middle polarity macroporous adsorbent resin, post is filled in the ratio with crude drug weight ratio 1:2, then by above-mentioned concentrated solution for subsequent use with 1.5 column volumes/hour flow velocity by this macroporous resin column, after passing through completely, first use deionized water eluting, carry out gradient elution with 40-50% ethanol again, elution flow rate be 2.5 column volumes/hour.Collect ethanol elution, drying under reduced pressure, pulverizing, obtain redback christmashush root extract.Yield is 0.35%.
Embodiment 5
By redback christmashush root medical material, with alcohol reflux 1 time, concentration of alcohol 120%, alcohol adding amount is 10 times of medical material weight, extraction time 60min.Merge extractive liquid, filters and concentrating under reduced pressure, and concentrated solution is centrifugal, gets supernatant concentrating under reduced pressure for subsequent use.
Get middle polarity macroporous adsorbent resin, post is filled in the ratio with crude drug weight ratio 1:2, then by above-mentioned concentrated solution for subsequent use with 1.5 column volumes/hour flow velocity by this macroporous resin column, after passing through completely, first use deionized water eluting, carry out gradient elution with 40-60% ethanol again, elution flow rate be 2.5 column volumes/hour.Collect ethanol elution, drying under reduced pressure, pulverizing, obtain redback christmashush root extract.Yield is 0.27%.
Embodiment 6 pharmacological experimental example
For the anti-hepatic fibrosis effect evaluating redback christmashush root extract carries out the experimentation of its anti-hepatic fibrosis.
1 experiment material
1.1 laboratory animal regular grade SD rats, male and female half and half, weight is 220 ± 10g about.
1.2 medicine redback christmashush root extracts are extract (HE) prepared by experimental example 1; Colchicines tablets, Xishuangbanna pharmaceutcal corporation, Ltd; Prepare redback christmashush root 3 kinds of different extracts according to a conventional method: ligroin extraction (PE), ethyl acetate extract (EE), n-butanol extract (BE).
1.2 reagent TGF β 1 test kits, Senxiong Science & Technology Industry Co., Ltd., Shanghai; TIMP-1 test kit, Senxiong Science & Technology Industry Co., Ltd., Shanghai.Analytical pure CCL4, Shantou brilliance chemical reagent work; Oleum Arachidis hypogaeae semen (100%), Luhua Group Co., Ltd., Shandong; Cholesterol, Chinese Hui Guang biochemical reagents company limited.
2. experimental technique
2.1 modeling Liver Fibrosis Model groups and each administration group rat subcutaneous injection in the morning 40% carbon tetrachloride Oleum Arachidis hypogaeae semen (the 1st 0.5mL/100g weight, after every 3 days subcutaneous 0.3mL/l00g weight), afternoon, drug group gave relative medicine, model group gives normal saline gavage (every day 1 time, each lmL/100g weight).Model and administration group all give high fat diet (79.5% Semen Maydis powder+20% Adeps Sus domestica+0.5% cholesterol), and normal group gives normal diet, and experiment carries out 6 weeks altogether.
Animal is divided into 15 groups at random by 2.2 animal groupings from modeling: Normal group, Liver Fibrosis Model group, colchicine group (gastric infusion, 0.9 × 10
-5g/kg) and the different extract group of redback christmashush root 4 kinds: extract of the present invention (HE), ligroin extraction (PE), ethyl acetate extract (EE), n-butanol extract (BE) divide equally height (high), in (middle), low dosage (low) group (write a Chinese character in simplified form h, m, l), totally 12 groups.The high, medium and low dosage group of redback christmashush root 4 kinds of extracts then presses 12g respectively
.kg
-1, 6g
.kg
-1, 3g
.kg
-1crude drug amount and distilled water are mixed and made into aqueous solution, once a day property gavage.Model control group is with equal-volume normal saline gavage.Repetition measurement body weight once weekly, adjustment dose gavage, gavage l4d altogether.
After 2.3 Indexs measure experiments expire, water 12h is can't help in animal fasting, and with 10% chloral hydrate intraperitoneal anesthesia, ventral aorta is taken a blood sample, and collect with vacuum negative pressure blood-taking pipe, room temperature leaves standstill 1.5h, and centrifugal 15min 2500r/min, gets upper serum, is sub-packed in centrifuge tube; Serum ALT, AST is measured with AU5400 automatic clinical chemistry analyzer; Serum HA, LN, PCIII is measured with putting method of exempting from; Serum TG F β 1 and TIMP-1 (all being undertaken by reagent description) is measured by ELISA method.
The above-mentioned indices of 2.4 statistical method all calculates mean and standard deviation, carries out variance analysis and non parametric tests with statistic software SPSS 11.5.
3. experimental result the results are shown in Table 1
3.1 the change of liver function (serum alt, AST) and compare medicine of the present invention 3 dosage groups and significantly can reduce ALT and AST (p<0.01) in animal pattern serum, colchicine to ALT and AST in serum also in reduction trend (p<0.05) to a certain degree.Other extract groups also can reduce ALT and AST (p<0.05) in animal pattern serum, but action intensity extract comparatively of the present invention is poor.
2.2.2 hepatic fibrosis index (in serum HA, LN, III precollagen) change and compare medicine of the present invention 3 dosage groups and significantly can reduce HA, LN in animal pattern serum, III precollagen (p<0.01), colchicine to the HA in serum, LN, III precollagen, type Ⅳ collagen also in reduction trend (p<0.05) to a certain degree.Other extract groups also can reduce ALT and AST in animal pattern serum to a certain extent, but action intensity extract comparatively of the present invention is poor.
2.2.3 in serum TGF β 1 and TIMP-1 change and compare medicine of the present invention 3 dosage groups and significantly can reduce TGF β 1 in animal pattern serum and TIMP-1, colchicine to the TGF β 1 in serum and TIMP-1 also in reduction trend (see table 1).Other extract groups also can reduce TGF β 1 in animal pattern serum and TIMP-1 to a certain extent, but action intensity extract comparatively of the present invention is poor.
Claims (8)
1. a preparation method for redback christmashush root extract, is characterized in that comprising the following steps:
(1) redback christmashush root medical material 6-12 times amount ethanol extraction 1-2 hour, extracts 1-3 time, merges ethanol extract, concentrating under reduced pressure after filtering;
(2) by centrifugal for step (1) gained concentrated solution, supernatant concentrating under reduced pressure is for subsequent use;
(3) by the concentrated solution that step (2) obtains, with 1.3-1.7 column volume/hour flow velocity, upper macroporous adsorptive resins;
(4) with after deionized water eluting removing impurity, then use ethanol elution, elution flow rate be 2.3-2.7 column volume/hour;
(5) collect ethanol elution drying under reduced pressure, after pulverizing, obtain redback christmashush root effective part extract.
2. the preparation method of redback christmashush root extract according to claim 1, is characterized in that: in described step (1), concentration of alcohol is 60%-90%.
3. the preparation method of redback christmashush root extract according to claim 2, is characterized in that: in described step (1), concentration of alcohol is 70%-80%.
4. the preparation method of redback christmashush root extract according to claim 1, is characterized in that: in described step (4), concentration of alcohol is 20%-60%.
5. the preparation method of redback christmashush root extract according to claim 4, is characterized in that: in described step (4), concentration of alcohol is 30%-50%.
6. the preparation method of redback christmashush root extract according to claim 1, is characterized in that: the extract that the redback christmashush root effective part extract that described step (5) obtains is is main active with alkaloids and flavonoid.
7. by the redback christmashush root extract that preparation method described in claim 1-6 is obtained.
8. redback christmashush root extract described in claim 7 is preparing the application in anti-hepatic fibrosis medicines.
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| CN104586925A (en) * | 2015-01-29 | 2015-05-06 | 南方医科大学 | Redback christmashush extract, and preparation method and application thereof in preparation of drugs for treating hepatitis B |
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| CN104586925B (en) * | 2015-01-29 | 2018-12-04 | 南方医科大学 | Redback christmashush root extract and preparation method and preparation treatment hepatitis B medicine application |
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