CN104815002A - Acquisition method and application of tinospora sinensis merr extract - Google Patents
Acquisition method and application of tinospora sinensis merr extract Download PDFInfo
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- CN104815002A CN104815002A CN201510238817.7A CN201510238817A CN104815002A CN 104815002 A CN104815002 A CN 104815002A CN 201510238817 A CN201510238817 A CN 201510238817A CN 104815002 A CN104815002 A CN 104815002A
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Abstract
The invention discloses an acquisition method and application of a tinospora sinensis merr extract, and relates to the field of Tibetan medicines. The tinospora sinensis merr extract can play a role in inhibiting influenza virus H3N2 in vitro. The acquisition method comprises the following steps: performing percolation extraction on tinospora sinensis merr dry powder for three times by using 95% ethanol at room temperature, maintaining for 2 hours in each percolation extraction process, and filtering and concentrating extracting solutions obtained from the three times of percolation extraction to obtain a first part of extract; then performing heating reflux extraction on residues which are obtained from previous extraction for three times by using 70% ethanol, maintaining for 2 hours in each heating reflux extraction process, and filtering and concentrating the extracting solutions obtained from the three times of heating reflux extraction to obtain a second part of extract; and combining the first part of extract and the second part of extract to acquire the tinospora sinensis merr extract.
Description
Technical field
The present invention relates to Tibetan medicine field, particularly relate to a kind of preparation method and application thereof of Herba Lycopodii extract.
Background technology
Herba Lycopodii (Tinospora sinensis (Lour.) Merr) is the stem of menispermaceous plants Herba Lycopodii.Tibetan medicine name " Le Zhe " is medicinal herbs most in use in Tibetan medicine, and micro-hardship is cool.Hardship, returns liver, spleen channel, is usually used in treating expelling wind and removing dampness, relaxing muscles and tendons and activating QI and blood in the collateral, the diseases such as wind-expelling pain-stopping.For this reason, a kind of method studying effective extraction Herba Lycopodii seems particularly important.
Summary of the invention
The object of this invention is to provide a kind of preparation method and application thereof of Herba Lycopodii extract, can vitro inhibition influenza virus H3N2.
For achieving the above object, the following technical scheme of employing of the present invention:
A preparation method for Herba Lycopodii extract, comprises the following steps:
1) Herba Lycopodii dry powder is at room temperature used the ethanol percolate extraction three times of 95%, each 2 hours, three gained extracting liquid filterings are concentrated, obtain Part I extractum;
2) said extracted residue is extracted three times with the alcohol heating reflux of 70% again, each 2 hours, three gained extracting liquid filterings are concentrated, obtain Part II extractum;
3) merge described Part I extractum and described Part II extractum, obtain Herba Lycopodii extract.
4) application of the extract of above-mentioned Herba Lycopodii, is applied to and suppresses In Vitro Anti influenza virus H3N2.
The preparation method of a kind of Herba Lycopodii extract that technique scheme provides and application thereof, by obtaining above-mentioned Herba Lycopodii extract with ethanol extraction, this Herba Lycopodii extract infected by influenza H3N2 has In-vitro Inhibitory Effect.
Accompanying drawing explanation
Fig. 1 is the schematic flow sheet of the preparation method of a kind of Herba Lycopodii extract provided by the invention.
Detailed description of the invention
Below in conjunction with the accompanying drawing in the embodiment of the present invention, technical scheme of the present invention is clearly and completely described.
The invention provides a kind of preparation method of Herba Lycopodii extract, described method comprises:
101: by the ethanol percolate extraction three times with 95% under Herba Lycopodii dry powder room temperature, each 2 hours, three gained extracting solution are merged filtering and concentrating, obtains Part I extractum.
102: the residue after above method being extracted, the alcohol heating reflux with 70% extracts three times, each 2 hours, three gained extracting solution is merged filtering and concentrating, obtains Part II extractum.
103: merge described Part I extractum and described Part II extractum, obtain Herba Lycopodii extract.
Herba Lycopodii extract provided by the invention, may be used for suppressing influenza virus H3N2.Apply concrete test below to illustrate, the Herba Lycopodii extract that the embodiment of the present invention provides may be used for suppressing influenza virus H3N2.Embodiment tests the influenza A virus H3N2 (section/30/95, A3/ capital) used, and by Shanghai City, health and epidemic prevention station provides.Cell model is Madin-Darby canine kidney(cell line) mdck cell (being provided by Chinese Academy of Sciences's cell).
Embodiment
1) Herba Lycopodii extract embodiment of the present invention provided carries out concentration configuration: first Herba Lycopodii extract is mixed with 20mg/ml with dimethyl sulfoxine.
2) then use mdck cell maintenance medium (cell culture fluid of namely not asking containing blood) respectively dilution for concentration be the different medicinal liquids of 200 μ g/ml, 100 μ g/ml, 50 μ g/ml, 25 μ g/ml, 12.5 μ g/ml.
3) toxotest is carried out to the medicinal liquid of variable concentrations: mdck cell 96 well culture plate monolayer culture 37 DEG C, 5%CO
2cultivate 24 hours, inhale and abandon supernatant, adding concentration is respectively 200 μ g/ml, 100 μ g/ml, 50 μ g/ml, 25 μ g/ml, 12.5 μ g/ml medicinal liquids, parallel 4 holes of each medicinal liquid, every hole dosing 100 μ l.Toxicity is observed through 72 hours.
4) cell phenotype conversion, experiment establishes cell controls group, virus control group, positive controls (ribavirin) and medicine group.Mdck cell 96 well culture plate monolayer culture 37 DEG C, 5%CO
2cultivate 24 hours, inhale and abandon supernatant, cell, through washing, adds 30TCID
50virus liquid, 37 DEG C, 5%CO
2adsorb 2 hours, suck virus liquid, add medicinal liquid under maximal non-toxic concentration, each sample parallel 4 hole, every hole adds medicinal liquid 100 μ l.Through 37 DEG C, 5%CO
2cultivate 72 hours, observe CPE (pathological changes), experiment establishes cell controls group, virus control group, and positive controls (ribavirin) and medicine group, observe CPE simultaneously, is greater than 50%, then calculates IC if suppress
50(half effective inhibition concentration) and therapeutic index TI value.Result of the test is as shown in table 1, TC in table
50for half toxic concentration.
Table 1
Obviously, described embodiment is only the present invention's part embodiment, instead of whole embodiments.Based on the embodiment in the present invention, those of ordinary skill in the art, not making the every other embodiment obtained under creative work prerequisite, belong to the scope of protection of the invention.
Claims (2)
1. the preparation method of Herba Lycopodii extract and an application thereof, is characterized in that, comprising:
1) by the ethanol percolate extraction three times with 95% under Herba Lycopodii dry powder room temperature, each 2 hours, three gained extracting solution are merged filtering and concentrating, obtains Part I extractum;
2) by 1) in extract after residue extract three times with the alcohol heating reflux of 70% again, each 2 hours, three gained extracting solution are merged filtering and concentrating, acquisition Part II extractum;
3) merge described Part I extractum and described Part II extractum, obtain Herba Lycopodii extract.
2. the application of Herba Lycopodii extract according to claim 1, is characterized in that, described Herba Lycopodii extract is applied to and suppresses influenza virus H3N2.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201510238817.7A CN104815002A (en) | 2015-05-12 | 2015-05-12 | Acquisition method and application of tinospora sinensis merr extract |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
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| CN201510238817.7A CN104815002A (en) | 2015-05-12 | 2015-05-12 | Acquisition method and application of tinospora sinensis merr extract |
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| CN104815002A true CN104815002A (en) | 2015-08-05 |
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| CN201510238817.7A Pending CN104815002A (en) | 2015-05-12 | 2015-05-12 | Acquisition method and application of tinospora sinensis merr extract |
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105851082A (en) * | 2016-04-04 | 2016-08-17 | 孙爱华 | Medicament for sterilization of anaesthesia operating room |
| CN106236842A (en) * | 2016-08-30 | 2016-12-21 | 青海民族大学 | Herba Lycopodii suppresses HDAC1 enzyme effective site and preparation method and application |
| CN106265930A (en) * | 2016-08-30 | 2017-01-04 | 青海民族大学 | Herba Lycopodii AntiHIV1 RT activity 1 virus effective site and preparation method and application |
| CN107897924A (en) * | 2017-12-04 | 2018-04-13 | 德兴市宋氏葛业有限公司 | A kind of kudzuvine root starch electuary for liver-protecting sobering up and preparation method thereof |
-
2015
- 2015-05-12 CN CN201510238817.7A patent/CN104815002A/en active Pending
Non-Patent Citations (3)
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| 代正福等: "《海南中药资源名录》", 31 August 2009, 中国农业出版社 * |
| 刘友平等: "《基本理化机能训练》", 31 August 2014, 中国医药科技出版社 * |
| 张海霞等: "GS-MS分析宽筋藤中不同萃取物成分比较", 《生物技术世界》 * |
Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105851082A (en) * | 2016-04-04 | 2016-08-17 | 孙爱华 | Medicament for sterilization of anaesthesia operating room |
| CN106236842A (en) * | 2016-08-30 | 2016-12-21 | 青海民族大学 | Herba Lycopodii suppresses HDAC1 enzyme effective site and preparation method and application |
| CN106265930A (en) * | 2016-08-30 | 2017-01-04 | 青海民族大学 | Herba Lycopodii AntiHIV1 RT activity 1 virus effective site and preparation method and application |
| CN106265930B (en) * | 2016-08-30 | 2019-11-12 | 青海民族大学 | Herba Lycopodii anti-HIV-1 virus active component and preparation method and application |
| CN106236842B (en) * | 2016-08-30 | 2019-12-10 | 青海民族大学 | Effective part of tinospora sinensis for inhibiting HDAC1 enzyme, preparation method and application |
| CN107897924A (en) * | 2017-12-04 | 2018-04-13 | 德兴市宋氏葛业有限公司 | A kind of kudzuvine root starch electuary for liver-protecting sobering up and preparation method thereof |
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Application publication date: 20150805 |
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