CN104586924B - Redback christmashush root extract and preparation method and preparation treatment hepatic fibrosis medicines application - Google Patents
Redback christmashush root extract and preparation method and preparation treatment hepatic fibrosis medicines application Download PDFInfo
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- CN104586924B CN104586924B CN201510047470.8A CN201510047470A CN104586924B CN 104586924 B CN104586924 B CN 104586924B CN 201510047470 A CN201510047470 A CN 201510047470A CN 104586924 B CN104586924 B CN 104586924B
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Abstract
The invention discloses a kind of redback christmashush root extract and preparation method and in the application of preparation treatment hepatic fibrosis medicines comprising alcohol extracting, upper large pore resin absorption column, ethanol elution, is dried under reduced pressure centrifugal concentrating, obtains redback christmashush root effective part extract.Extraction process of the invention is easy rationally, obtained redback christmashush root effective part extract can reduce ALT and AST in animal pattern serum, and HA, LN, III precollagen and TGF β 1, TIMP-1, illustrate that there is comparatively ideal effect of anti hepatic fibrosis, it is more promising drug candidate, can be applied to prepare anti-hepatic fibrosis medicines.
Description
Technical field
The invention belongs to the field of Chinese medicines, in particular to a kind of effective part extract of redback christmashush root further relates to the extraction
The preparation method and purposes of object.
Background technique
Liver fibrosis is to seriously endanger one of the difficult disease of human physical and mental health, and discovery effectively preventing is not done yet so far
Method.Liver fibrosis is early period process of the chronic hepatitis to cirrhosis progress, in the factors for determining patients with chronic liver prognosis
In play a crucial role, as can blocking or reversing liver fibrosis by certain means before developing into cirrhosis
Occur and develop, will therapeutically generate an important breakthrough in chronic liver disease.In terms of modern medicine, colchicin is relatively early answers
For clinical anti-hepatic fibrosis medicines, but toxic side effect is stronger, and clinical application is restricted.Glucocorticoid and Beracilline
It is not widely adopted Deng big due to side effect.Proline-4-hydroxylation enzyme inhibitor, prostaglandin E similar medicine, γ-interference
How at the experimental stage element, precollagen peptide, hepatocyte growth factor, IL-10 etc. be at present, and its preparation and source exist
Shortcomings, Western medicine aspect there is no mature effective and Small side effects anti-hepatic fibrosis medicines at present.Traditional Chinese medicine is fine in relation to anti-liver
The research work of dimensionization also achieves encouraging progress in recent years.Much prescriptions prove there is certain resist through experiment or clinical research
Fibrosis effect, the anti-liver for concentrating on the Chinese medicines such as dispersing liver and promoting blood circulation stagnation resolvation, invigorating Qi and tonifying kidney, softening hard tissue, supporting vital QI and dispersing blood stasis at present are fine
The research of dimensionization, such as strong liver softening hard masses soup, Fuzheng Huayu 319 Recipe, Semen Persicae extract, artificial cordyceps mycelia, 861 mixture of compound.
But experimental study or clinical observation on the therapeutic effect are rested on mostly, the drug with significant curative effect there is no to list at present.
Redback christmashush root is dicotyledon medicine euphorbia plant red back mountain hemp rod Alchornea trewioides
(Benth.) root of Muell.-Arg..The red back ma of alias, red crown (" Guangxi Chinese herbal medicine "), (Guangzhou air force is " common for the red skirt of thin silk
Chinese herbal medicine handbook ").It is distributed Central China and the southeast, south China, is usually used as medicine with root, leaf.Its root is sweet in flavor, mild-natured.Effect heat-clearing solution
Poison, dispelling wind and eliminating dampness, dissipating stasis and stanching bleeding are relievingd asthma, and desinsection is antipruritic.Tradition is applied to control enteritis, diarrhea, dysentery, icteric hepatitis, urinary tract
Infection and calculus, ephritis, difficult urination, blood urine, metrorrhagia, leukorrhea, eczema, nettle rash, ulcerative carbuncle pyogenic infections, sore do not close up long
Disease.Studies have shown that having cough-relieving, phlegm-dispelling functions in redback christmashush root, anti-acetylcholine acts on and slightly inhibits golden yellow modern medicine
Color and staphylococcus albus act on.
Medicinal extract obtained by redback christmashush root water extract and water extract-alcohol precipitation has the function of hepatitis virus resisting and prevention liver fibrosis, can
Clinically to treat chronic hepatitis and cirrhosis.Result of study shows that redback christmashush root can significantly inhibit carbon tetrachloride (CCL4)
Two important factors (TGF-β 1, TIMP-1) of complex factors and the liver fibrosis due in alcoholic fibrosis rat blood serum,
And animal pattern liver fiber index (HA, III procollagen type), liver function indexes (ALT, AST) have significant decrease, and have liver protection, drop
The effect of enzyme, anti-hepatic fibrosis.Therefore, the screening of anti-hepatic fibrosis active component further is carried out to redback christmashush root and purposes is ground
Study carefully and is of great significance.
Summary of the invention
The technical problem to be solved by the invention is to provide a kind of redback christmashush root extracts and its preparation method and application.
The technical solution adopted by the present invention to solve the technical problems is as follows: a kind of preparation side of redback christmashush root extract
Method, it is characterised in that the following steps are included:
(1) redback christmashush root medicinal material is measured 60%-90% ethyl alcohol with 6-12 times and is extracted 1-2 hours, extracts 1-3 times, merges ethyl alcohol
Extracting solution is concentrated under reduced pressure after filtering；
(2) concentrate obtained by step (1) is centrifuged, supernatant is concentrated under reduced pressure spare；
(3) concentrate for obtaining step (2), with 1.3-1.7 column volume/hour flow velocity, upper large pore resin absorption column；
(4) after removing decontamination is washed with deionized water, then ethanol elution is used, elution flow rate is 2.3-2.7 column volume/small
When；
(5) it collects ethanol eluate to be dried under reduced pressure, redback christmashush root effective part extract is obtained after crushing.
Concentration of alcohol is 60%-90% in the step (1).
Concentration of alcohol is 70%-80% in the step (1).
Concentration of alcohol is 20%-60% in the step (4).
Concentration of alcohol is 30%-50% in the step (4).
The redback christmashush root effective part extract that the step (5) obtains is using alkaloids and flavonoids as chief active
The extract of ingredient.
The redback christmashush root extract as made from above-mentioned preparation method.
Above-mentioned redback christmashush root extract is preparing the application in anti-hepatic fibrosis medicines.
Beneficial effects of the present invention are as follows:
1. extraction process of the invention is easy rationally, available redback christmashush root effective part extract.
2. redback christmashush root extract of the invention, in liver function index context of detection, 3 dosage groups of drug of the present invention are significant
Can reduce the ALT and AST (p < 0.01) in animal pattern serum, colchicin in serum ALT and AST be also in a certain degree
Decreasing trend (p < 0.05).Other extract groups can also reduce ALT and AST (p < 0.05) in animal pattern serum, but make
It is poor with intensity extract more of the present invention.In the context of detection of hepatic fibrosis index (HA, LN, III precollagen in serum), the present invention
3 dosage groups of drug can significantly reduce HA, LN, III precollagen (p < 0.01) in animal pattern serum, and colchicin is to serum
In HA, LN, III precollagen, type Ⅳ collagen be also in a degree of decreasing trend (p < 0.05).Other extract groups can also be
The ALT and AST in animal pattern serum are reduced to a certain extent, but action intensity extract more of the present invention is poor.The TGF in serum
The context of detection of β 1 and TIMP-1,3 dosage groups of drug of the present invention can significantly reduce by 1 He of TGF β in animal pattern serum
TIMP-1, colchicin in serum TGF β 1 and TIMP-1 be also in decreasing trend (being shown in Table 1).Other extract groups can also be
The TGF β 1 and TIMP-1 in animal pattern serum are reduced to a certain extent, but action intensity extract more of the present invention is poor.Above-mentioned knot
Fruit illustrates that extract of the present invention has comparatively ideal effect of anti hepatic fibrosis, is more promising drug candidate.
Specific embodiment
The present invention is a kind of preparation method of redback christmashush root extract comprising following steps:
(1) redback christmashush root medicinal material is measured ethyl alcohol with 6-12 times and is extracted 1-2 hours, extracts 1-3 times, merges ethanol extract, mistake
It is concentrated under reduced pressure after filter.Preferably, concentration of alcohol is 60%-90% in step (1), it is furthermore preferred that concentration of alcohol is 70%-
80%.Redback christmashush root is dicotyledon medicine euphorbia plant red back mountain hemp rod Alchornea trewioides (Benth.)
Muell. the root of-Arg..
(2) concentrate obtained by step (1) is centrifuged, supernatant is concentrated under reduced pressure spare.
(3) concentrate for obtaining step (2), with 1.3-1.7 column volume/hour flow velocity, upper middle polarity or non-pole
The large pore resin absorption column of property.
(4) after removing decontamination is washed with deionized water, then ethanol elution is used, elution flow rate is 2.3-2.7 column volume/small
When.Preferably, concentration of alcohol is 20%-60% in step (4), it is furthermore preferred that concentration of alcohol is 30%-50%.
(5) it collects ethanol eluate to be dried under reduced pressure, obtains redback christmashush root effective part extract after crushing, the redback chrismashush root and leaf
Root effective part extract is using alkaloids and flavonoids as the extract of main active.
Above-mentioned obtained redback chrismashush root and leaf active component root extract is preparing the application in anti-hepatic fibrosis medicines.
Below with reference to embodiment, the present invention will be described in detail.But the present invention is not limited to these given realities
Apply example.Most preferred embodiment 1.
Embodiment 1
By redback christmashush root medicinal material, being extracted 2 times with alcohol reflux, concentration of alcohol 75%, alcohol adding amount is 10 times of medicinal material weight,
Extraction time 90min.Combined extract is filtered and is concentrated under reduced pressure, concentrate is centrifuged, and takes supernatant to be concentrated under reduced pressure spare.
Middle polarity macroporous absorbent resin is taken, fills column in the ratio of crude drug weight ratio 1:2, it then will be above-mentioned spare dense
Contracting liquid by this macroporous resin column, after passing through completely, is first washed with deionized water de-, then used with 1.5 column volumes/hour flow velocity
30-50% ethyl alcohol carries out gradient elution, and elution flow rate is 2.5 column volumes/hour.Ethanol eluate is collected, is dried under reduced pressure, powder
It is broken, obtain redback christmashush root extract.Yield is 0.38%.
Embodiment 2
By redback christmashush root medicinal material, being extracted 3 times with alcohol reflux, concentration of alcohol 90%, alcohol adding amount is 12 times of medicinal material weight,
Extraction time 120min.Combined extract is filtered and is concentrated under reduced pressure, concentrate is centrifuged, and takes supernatant to be concentrated under reduced pressure spare.
Middle polarity macroporous absorbent resin is taken, fills column in the ratio of crude drug weight ratio 1:2, it then will be above-mentioned spare dense
Contracting liquid by this macroporous resin column, after passing through completely, is first washed with deionized water de-, then used with 1.5 column volumes/hour flow velocity
30-60% ethyl alcohol carries out gradient elution, and elution flow rate is 2.5 column volumes/hour.Ethanol eluate is collected, is dried under reduced pressure, powder
It is broken, obtain redback christmashush root extract.Yield is 0.29%.
Embodiment 3
By redback christmashush root medicinal material, being extracted 1 time with alcohol reflux, concentration of alcohol 60%, alcohol adding amount is 6 times of medicinal material weight,
Extraction time 60min.Combined extract is filtered and is concentrated under reduced pressure, concentrate is centrifuged, and takes supernatant to be concentrated under reduced pressure spare.
Middle polarity macroporous absorbent resin is taken, fills column in the ratio of crude drug weight ratio 1:2, it then will be above-mentioned spare dense
Contracting liquid by this macroporous resin column, after passing through completely, is first washed with deionized water de-, then used with 1.5 column volumes/hour flow velocity
20-40% ethyl alcohol carries out gradient elution, and elution flow rate is 2.5 column volumes/hour.Ethanol eluate is collected, is dried under reduced pressure, powder
It is broken, obtain redback christmashush root extract.Yield is 0.31%.
Embodiment 4
By redback christmashush root medicinal material, being extracted 3 times with alcohol reflux, concentration of alcohol 70%, alcohol adding amount is 7 times of medicinal material weight,
Extraction time 80min.Combined extract is filtered and is concentrated under reduced pressure, concentrate is centrifuged, and takes supernatant to be concentrated under reduced pressure spare.
Middle polarity macroporous absorbent resin is taken, fills column in the ratio of crude drug weight ratio 1:2, it then will be above-mentioned spare dense
Contracting liquid by this macroporous resin column, after passing through completely, is first washed with deionized water de-, then used with 1.5 column volumes/hour flow velocity
40-50% ethyl alcohol carries out gradient elution, and elution flow rate is 2.5 column volumes/hour.Ethanol eluate is collected, is dried under reduced pressure, powder
It is broken, obtain redback christmashush root extract.Yield is 0.35%.
Embodiment 5
It by redback christmashush root medicinal material, is extracted 1 time with alcohol reflux, concentration of alcohol 120%, alcohol adding amount is the 10 of medicinal material weight
Times, extraction time 60min.Combined extract is filtered and is concentrated under reduced pressure, concentrate is centrifuged, and takes supernatant to be concentrated under reduced pressure spare.
Middle polarity macroporous absorbent resin is taken, fills column in the ratio of crude drug weight ratio 1:2, it then will be above-mentioned spare dense
Contracting liquid by this macroporous resin column, after passing through completely, is first washed with deionized water de-, then used with 1.5 column volumes/hour flow velocity
40-60% ethyl alcohol carries out gradient elution, and elution flow rate is 2.5 column volumes/hour.Ethanol eluate is collected, is dried under reduced pressure, powder
It is broken, obtain redback christmashush root extract.Yield is 0.27%.
6 pharmacological experimental example of embodiment
To evaluate the experimental study that the anti-hepatic fibrosis effect of redback christmashush root extract carries out its anti-hepatic fibrosis.
1 experimental material
1.1 experimental animal regular grade SD rats, half male and half female, weight about 220 ± 10g.
1.2 drug redback christmashush root extracts are extract (HE) prepared by experimental example 1；Colchicines tablets, Xishuangbanna medicine
Industry Co., Ltd；Prepare 3 kinds of redback christmashush root different extracts according to a conventional method: ligroin extraction (PE), ethyl acetate extract
Object (EE), n-butanol extract (BE).
1.2 reagent TGF β, 1 kit, Senxiong Science & Technology Industry Co., Ltd., Shanghai；TIMP-1 kit, Shanghai Sen Xiong section
Skill Industrial Co., Ltd..Analyze pure CCL4, Shantou brilliance chemical reagent work；Peanut oil (100%), Luhua Group Co., Ltd., Shandong；
Cholesterol, Chinese Hui Guang biochemical reagents Co., Ltd.
2. experimental method
40% carbon tetrachloride peanut oil of 2.1 modeling Liver Fibrosis Model groups and the subcutaneous injection of each administration group rat morning (the
1 0.5mL/100g weight, after every 3 days subcutaneous 0.3mL/l00g weight), give relative medicine, mould to medicine group in the afternoon
Type group gives physiological saline stomach-filling (one time a day, each lmL/100g weight).Model and administration group give high fat diet
(+0.5% cholesterol of+20% lard of 79.5% corn flour), normal group is given normal diet, and experiment carries out 6 weeks altogether.
Animal is randomly divided into 15 groups from the modeling by 2.2 animal packets: Normal group, Liver Fibrosis Model group,
Colchicin group (gastric infusion, 0.9 × 10-5) and the different extract groups of 4 kinds of redback christmashush root g/kg: extract (HE) of the present invention,
Ligroin extraction (PE), ethyl acetate extract (EE), n-butanol extract (BE) divide equally high (high), in (middle),
Low dosage (low) group (writing a Chinese character in simplified form h, m, l), totally 12 groups.4 kinds of high, medium and low dosage groups of extract of redback christmashush root then press 12g respectively·
kg-1、6g·kg-1、3g·kg-1Crude drug amount and distilled water are mixed and made into aqueous solution, property stomach-filling once a day.Model control group with etc.
ML normal saline stomach-filling.Repetition measurement weight is primary weekly, adjusts dose stomach-filling, altogether stomach-filling l4d.
After the experiment of 2.3 Indexs measures expires, animal is deprived of food but not water 12h, with 10% chloraldurate intraperitoneal anesthesia, abdomen master
Arterial blood drawing is collected with vacuum negative pressure blood-taking pipe, is stored at room temperature 1.5h, is centrifuged 15min 2500r/min, is taken upper serum, point
Loaded in centrifuge tube；Serum ALT, AST are measured with AU5400 automatic clinical chemistry analyzer；With radioimmunoassay measurement serum HA, LN,
PCIII；Serum TG F β 1 and TIMP-1 (carrying out by reagent specification) is measured with ELISA method.
The above-mentioned indices of 2.4 statistical methods calculate mean and standard deviation, with 11.5 side of progress of statistic software SPSS
Difference analysis and non-parametric test.
3. experimental result the results are shown in Table 1
It is dynamic that 3 dosage groups of variation and drug more of the present invention of 3.1 liver functions (serum alt, AST) can significantly reduce model
ALT and AST (p < 0.01) in object serum, colchicin in serum ALT and AST be also in a degree of decreasing trend (p
<0.05).Other extract groups can also reduce ALT and AST (p < 0.05) in animal pattern serum, but action intensity relatively this hair
Bright extract is poor.
2.2.2 3 dosage of variation and drug more of the present invention of hepatic fibrosis index (HA, LN, III precollagen in serum)
Group can significantly reduce HA, LN, III precollagen (p < 0.01) in animal pattern serum, colchicin in serum HA, LN, III
Precollagen, type Ⅳ collagen are also in a degree of decreasing trend (p < 0.05).Other extract groups can also drop to a certain extent
ALT and AST in low animal pattern serum, but action intensity extract more of the present invention is poor.
2.2.3 3 dosage groups of the variation of TGF β 1 and TIMP-1 and drug more of the present invention can significantly reduce model in serum
TGF β 1 and TIMP-1 in animal blood serum, colchicin in serum TGF β 1 and TIMP-1 be also in decreasing trend (being shown in Table 1).
Other extract groups can also reduce TGF β 1 and TIMP-1 in animal pattern serum to a certain extent, but action intensity is relatively originally
Invention extract is poor.
Claims (4)
1. redback christmashush root extract is preparing the application in anti-hepatic fibrosis medicines, it is characterised in that: the redback christmashush root extracts
The preparation method of object the following steps are included:
(1) redback christmashush root medicinal material is measured ethyl alcohol with 6-12 times and is extracted 1-2 hours, extracts 1-3 times, merges ethanol extract, after filtering
It is concentrated under reduced pressure, wherein concentration of alcohol is 60%-90%；
(2) concentrate obtained by step (1) is centrifuged, supernatant is concentrated under reduced pressure spare；
(3) concentrate for obtaining step (2), with 1.3-1.7 column volume/hour flow velocity, upper large pore resin absorption column；
(4) be washed with deionized water removing decontamination after, then use ethanol elution, elution flow rate be 2.3-2.7 column volume/hour,
Middle concentration of alcohol is 20%-60%；
(5) it collects ethanol eluate to be dried under reduced pressure, redback christmashush root effective part extract is obtained after crushing.
2. redback christmashush root extract according to claim 1 exists preparing the application in anti-hepatic fibrosis medicines, feature
In: concentration of alcohol is 70%-80% in the step (1).
3. redback christmashush root extract according to claim 1 exists preparing the application in anti-hepatic fibrosis medicines, feature
In: concentration of alcohol is 30%-50% in the step (4).
4. redback christmashush root extract according to claim 1 exists preparing the application in anti-hepatic fibrosis medicines, feature
In: the redback christmashush root effective part extract that the step (5) obtains is using alkaloids and flavonoids as main active
Extract.
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Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1939383A (en) * | 2006-09-25 | 2007-04-04 | 南方医科大学 | Use of redback christmashush in preparing medicine for hepatitis B |
| CN103585206A (en) * | 2013-10-31 | 2014-02-19 | 济南星懿医药技术有限公司 | Preparation method of Chrysosplenium extract and its application |
| 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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Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1939383A (en) * | 2006-09-25 | 2007-04-04 | 南方医科大学 | Use of redback christmashush in preparing medicine for hepatitis B |
| CN103585206A (en) * | 2013-10-31 | 2014-02-19 | 济南星懿医药技术有限公司 | Preparation method of Chrysosplenium extract and its application |
| 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 |
Non-Patent Citations (1)
| Title |
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| "红背叶根不同提取物体外抑制HBsAg和HBeAg的实验研究";沈海容等;《中药新药与临床药理》;20140131;第25卷(第1期);第43页左栏第1段 * |
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