CN101028311B - Use of selaginella tamariscina - Google Patents
Use of selaginella tamariscina Download PDFInfo
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- CN101028311B CN101028311B CN200610017494XA CN200610017494A CN101028311B CN 101028311 B CN101028311 B CN 101028311B CN 200610017494X A CN200610017494X A CN 200610017494XA CN 200610017494 A CN200610017494 A CN 200610017494A CN 101028311 B CN101028311 B CN 101028311B
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Abstract
An application of tamariskoid spikemoss herb, its liquid extract, and the water component and n-butanol component contained in said liquid extraction in preparing estrin medicines is disclosed.
Description
Technical field
The present invention relates to the new purposes of Chinese medicine Herba Selaginellae, i.e. the application of Herba Selaginellae in the preparation estrogenic.
Background technology
Herba Selaginellae is a Pteridophyta Selaginaceae Rock lily plant, and the successive dynasties famous expert all has it and discusses and research, thinks that its property is hot flat, and the removing blood stasis dissipating blood stasis is arranged, and promoting blood circulation and hemostasis cures mainly stomachache, amenorrhea, mass in the abdomen drenches knot, and spit blood, have blood in stool, the function of disease such as hematuria.Modern pharmacology and clinical research show that Herba Selaginellae has effects such as cancer preventing and treating, treatment gynaecopathia, antiinflammatory antiviral, analgesia, blood pressure lowering, blood sugar lowering, enhancing immune function of human body.Aspect treatment amenorrhea, metrorrhagia, induced labor, salpingemphraxis, blood stasis dispelling regulating menstruation, the hemostasis by removing blood stasis effect is being arranged also, it has brought into play important effect in the clinical difficult diseases such as tire, removing damp-heat detoxifcation, retention of dead fetus under removing blood stasis contracts the palace.
Summary of the invention
The object of the present invention is to provide the new purposes of Herba Selaginellae at the preparation estrogenic.
The present invention seeks to be achieved through the following technical solutions: the application of Herba Selaginellae in the preparation estrogenic.
The application of the total extract of Herba Selaginellae in the preparation estrogenic.
The application in the preparation estrogenic of water position in the Herba Selaginellae and n-butanol portion.
The inventor has proved the new purposes of medicine of the present invention in estrogen by experiment:
One, adopt zoopery in conjunction with the reporter gene technology, five kinds of Chinese medicines carried out the active experiment of phytoestrogen:
1, medicine
Select Chinese medicine for use: Herba Selaginellae (Selaginella tamariscina), Folium Pini (Pinus Massoniana Lamb), Herba Corallodisci Flabellati (Corallodiscus flabellata), Rhizoma Osmundae (Dryopteris sublaeta Ching et Hsu), Herba Leonuri (Leonurusheterophyllus Sweet).Each medical material boils with 10 times of water gagings respectively and carries twice, filters, and merging filtrate and concentrating under reduced pressure, vacuum drying are each medical material water extract.
Each medical material water extract dissolves with distilled water, and its dosage sees Table 1.Positive drug is a Nilestriol Tablets, and Hualian Pharmaceutical Co., Ltd., Shanghai produces, and makes the distilled water suspension, presses 1.7mgkg
-1D
-1Administration.
The animals administer amount of five kinds of Chinese medicines of table 1
2, laboratory animal and cell strain and plasmid
Kunming mouse, female, be born 21 days (ablactation just), body weight 9-12g purchases the Experimental Animal Center in Henan Province.(β-galactosidase, β-gal) control plasmid p β gal-Control, reorganization reporter gene carrier pERE-TAL-luc are so kind as to give by the dense doctor of leaf chess of BIO ENGINEERING INST MILITARY for HEK293 cell strain and beta galactosidase.Recombined human ER α (humanER α, hER α) expression vector pCXN2-hER α and recombined human ER β (humanER β, hER β) expression vector pCXN2-hER β are so kind as to give by Satoshi doctor Inoue of Tokyo University department of medial science.
3, main agents
RPMI1640 culture medium, no phenol red RPMI1640 culture medium and liposome Lipofectamine
TM2000Reagent is available from Gibco Invitrogen company; The luciferase fluorescence detection reagent kit is available from Promega company; (O-Nitrophenyl-β-D-galactopyranoside ONPG) is so kind as to give by the dense doctor of leaf chess of BIO ENGINEERING INST MILITARY neighbour-Nitrobenzol-β-D-gala pyranoside; 17 beta estradiols (17 β-estradiol, E
2) available from Sigma company as the positive control in the reporter gene detection method; (fetal bovine serum is FBS) available from Beijing ancient cooking vessel state Bioisystech Co., Ltd for hyclone; The no estrogen hyclone is a HyClong company product; All the other reagent are homemade analytical pure.
4, used key instrument
Electronical reading analytical balance (Japanese Shimadzu company make); The general surgery apparatus; CO2 gas incubator (Japanese SANYO company); Water bath chader (Dong Lian of Harbin City Electronic Technology Development Co.); Inverted microscope (Japanese Olympus); Desk type high speed freezing centrifuge (Fourth Ring, Beijing scientific instrument factory); UV-7504PC type ultraviolet-visible spectrophotometer (foreign scientific and technological Instr Ltd. is good in Foochow); 1250 fluorescence detectors (Finland Wallac company).
5, experimental technique
Mouse Uterus weightening finish method
Mice is balanced by body weight and principle is at random divided into groups.Administration continues 7 days.Blank group distilled water every day 0.3ml10g
-1Irritate stomach; Positive controls nilestriol every day suspension 0.3ml10g
-1Irritate stomach; Chinese medicine is respectively organized 0.3ml10g every day
-1Irritate stomach.24h after the last administration takes off neck and puts to death mice, cuts the uterus immediately and weighs, and calculates uterus coefficient (uterus weight in wet base (body weight)
-1100%).The result compares with the blank group, and calculating the investigation of P value has not statistically significant.
6, the reporter gene transient expression of ERE regulation and control detects
With containing the FBS of 10% no estrogen, no gentamycin and phenol red RPMI1640 culture medium inoculated in 12 orifice plates, cell density is 8 * 10 to 24h with the HEK293 cell before the transfection
4Cellsml
-1The hole
-1Every block of plate all is provided with the Chinese medicine hole of variable concentrations, simultaneously positive drug control wells, solvent control wells is set.The same day was used sterilization PBS rinsing cell once in transfection, add 800 μ l and contain the FBS of 10% no estrogen, no gentamycin and phenol red RPMI1640 culture medium, with pCXN2-hER α or pCXN2-hER β, pERE-TAL-luc and p β gal-Control cotransfection HEK293 cell strain, use E behind the 6h
2(4 * 10
-6Moll
-1), Chinese medicine total extract (saturated solution) intervenes, behind the 24h, collecting cell supernatant and cell pyrolysis liquid detect the activity of luciferase and β gal respectively, the normalized uciferase activity, thereby filter out ER α or the effective Chinese medicine of ER β.
7, statistical disposition
Experimental data is carried out statistical disposition with SPSS software.
8, experimental result
The result shows that heavy dose of and low dose of Herba Selaginellae group Mouse Uterus coefficient is compared with the blank group all has special significant difference (P＜0.01).All the other are respectively organized the uterus coefficient and compare not statistically significant with the blank group.See Table 2, Fig. 1.
Table 2 Herba Selaginellae, Herba Leonuri, Rhizoma Osmundae, Herba Corallodisci Flabellati, Folium Pini are to the influence of sex immature Mouse Uterus weight in wet base
Annotate: compare with the blank group, ☆ represents P＜0.05; ★ represents P＜0.01; Compare ▲ expression P＜0.01 with positive controls
9, Herba Selaginellae is to the inducing action of reporter gene carrier pTAL-ERE-luc transient expression
Result's demonstration, though beta mediated by ER α or ER, E
2Uciferase activity after the standardization of hole all is higher than solvent control hole (P＜0.01), and the uciferase activity after the standardization of Herba Selaginellae hole also all is higher than solvent control hole (P＜0.01), but still is lower than E
2Hole (P＜0.01).See Table 3, Fig. 2.
Table 3 Herba Selaginellae is to the inducing action (n=10) of reporter gene carrier pTAL-ERE-luc transient expression
Annotate: compare with the blank group, ☆ represents P＜0.05; ★ represents P＜0.01; Compare ▲ expression P＜0.01 with positive controls
Two, the water position that is filtered out in the employing zoopery proof Herba Selaginellae and the estrogen action of n-butanol portion
Select ether position, ethyl acetate position, n-butanol portion and the water position of from Herba Selaginellae, screening for use, adopt the experiment of Mouse Uterus weightening finish method again.
The result shows, heavy dose of and low dose of Herba Selaginellae water position group Mouse Uterus coefficient is compared with the blank group all has special significant difference (P＜0.01), heavy dose of and low dose of Herba Selaginellae n-butanol portion group Mouse Uterus coefficient is compared with the blank group also all significant difference (P＜0.05), but still well below positive controls (P＜0.01).All the other each groups are compared not statistically significant with the blank group.See Table 4, Fig. 3.
Table 4 Herba Selaginellae different parts is to the influence of sex immature Mouse Uterus weight in wet base
Annotate: compare with the blank group, ☆ represents P＜0.05; ★ represents P＜0.01; Compare ▲ expression P＜0.01 with positive controls
Three, conclusion
1, uterus weight experiment is the classical way of the detection estrogen activity set up the earliest, when estimating the estrogen activity of material, uterus experiment considered the absorption, metabolism of material, combine with plasma protein and many-sided factor such as pharmacokinetics, directly reflection is subjected to the overall biological effect of reagent thing behind internal metabolism, by above-mentioned experiment, the proof Herba Selaginellae has the phytoestrogen effect, and water position that filters out from Herba Selaginellae and n-butanol portion are effective site.
2, the reporter gene technology is the reporter gene carrier that makes up according to intravital estrogen signal transduction pathway, the complexity of the medicaments sifting model of cellular level on the 26S Proteasome Structure and Function that possesses the living cells that whole animal has, be easier to operation, be easy to infer experimental result, high specificity than whole animal experimentally simultaneously.In the inducing action experimentation of Herba Selaginellae, can infer roughly the active component of Herba Selaginellae with which kind of estrogen receptor combines, and act on the expression that estrogen response element (ERE) comes controlling gene reporter gene carrier pTAL-ERE-luc transient expression.This experimental result shows the affinity of Herba Selaginellae and ER β than big with the affinity of ER α, illustrates that the Herba Selaginellae activated gene transcribes mainly beta mediated by ER; Proved the estrogen action of Herba Selaginellae.
Also four groups of patients that suffer from the syndrome in menopause have been carried out clinical observation in addition: every group of patient 30 people; This 30 people all has hectic fever, dizziness, tinnitus, night sweat symptom at night, wherein also has 16 people also to suffer from the arthralgia symptom simultaneously.
First group of patient takes the total extracting solution of Herba Selaginellae; Dose and method are: get Herba Selaginellae 15 weight portions, add the water of 150 weight portions, boiled 20-30 minute, filter; The water that adds 150 weight portions in medicinal residues again boiled 15 minutes, filtered, and merging filtrate is also preserved under the situation of sterile vacuum, obeys 200 milliliters of merging filtrates at every turn, sooner or later respectively once.Take continuously hectic fever symptom that 18 people were arranged after 12 weeks and night the night sweat transference cure, tinnitus and dizzy symptom reduce, and have 8 person joint's pain phenomenons to reduce.
Second group of patient takes the water position of extracting from Herba Selaginellae, each dose is equivalent to restrain the amount of extracting the fresh Herba Selaginellae from 10, take night and morning every day, take continuously hectic fever symptom that 23 people were arranged after 12 weeks and night the night sweat transference cure, tinnitus and dizzy symptom reduce, and have 12 person joint's pain phenomenons to reduce.
The 3rd group of patient takes the n-butanol portion that extracts from Herba Selaginellae, each dose is equivalent to restrain the amount of extracting the fresh Herba Selaginellae from 10, take night and morning every day, take continuously hectic fever symptom that 20 people were arranged after 12 weeks and night the night sweat transference cure, tinnitus and dizzy symptom reduce, and have 11 person joint's pain phenomenons to reduce.
The 4th group of patient takes: by Herba Selaginellae 15 weight portions, the Rhizoma Anemarrhenae 12 weight portions, Poria 30 weight portions, Flos Chrysanthemi 10 weight portions, Rhizoma Dioscoreae 30 weight portions, the Radix Paeoniae Alba 12 weight portions, Fructus Corni 30 weight portions, Rhizoma Anemones Altaicae 12 weight portions, Herba Albiziae 12 weight portions, Semen Ziziphi Spinosae (parched) 12 weight portions, Radix Dipsaci 15 weight portions, the water that adds 6 times of total medicine weight, boiled 30 minutes, and filtered; Boil 20 minutes that add 4 times of total medicine weight in the medicinal residues again, filter, merging filtrate, 250 milliliters of the above-mentioned merging filtrates of each clothes, take night and morning every day, take continuously hectic fever symptom that 20 people were arranged after 12 weeks and night the night sweat transference cure, tinnitus and dizzy symptom reduce, and have 13 person joint's pain phenomenons to reduce.
From clinical observation, also proved the estrogen action of Herba Selaginellae to above-mentioned patient.
Description of drawings
Fig. 1 is Herba Selaginellae, Herba Leonuri, Rhizoma Osmundae, Herba Corallodisci Flabellati, the Folium Pini diagrammatic sketch to the influence of sex immature Mouse Uterus weight in wet base;
Fig. 2 is the diagrammatic sketch of Herba Selaginellae to the inducing action of reporter gene carrier pTAL-ERE-luc transient expression;
Fig. 3 is the diagrammatic sketch of Herba Selaginellae different parts to the influence of sex immature Mouse Uterus weight in wet base.
The specific embodiment
Embodiment 1: get Herba Selaginellae 15 weight portions, add the water of 150 weight portions, boiled 20-30 minute, filter; The water that adds 150 weight portions in medicinal residues again boiled 15 minutes, filtered, and merging filtrate is the total extract of Herba Selaginellae, and it is preserved under the situation of sterile vacuum, obeys 200 milliliters of merging filtrates at every turn.
Embodiment 2: take by weighing by Herba Selaginellae 15 weight portions, the Rhizoma Anemarrhenae 12 weight portions, Poria 30 weight portions, Flos Chrysanthemi 10 weight portions, Rhizoma Dioscoreae 30 weight portions, the Radix Paeoniae Alba 12 weight portions, Fructus Corni 30 weight portions, Rhizoma Anemones Altaicae 12 weight portions, Herba Albiziae 12 weight portions, Semen Ziziphi Spinosae (parched) 12 weight portions, Radix Dipsaci 15 weight portions, the water that adds 6 times of total medicine weight, boiled 30 minutes, and filtered; The water that adds 4 times of amounts of total medicine weight in the medicinal residues again boiled 20 minutes, filtered, and merging filtrate is also preserved it under the situation of sterile vacuum, obey 200 milliliters of merging filtrates at every turn.
Embodiment 3: take by weighing Herba Selaginellae 50 weight portions, add the water that medicine weighs 10 times of amounts, boiled 30 minutes, filter, add the water that medical material weighs 8 times of amounts in the medicinal residues again, boiled 25 minutes, filter merging filtrate, in temperature is 10-30 ℃, and pressure is to concentrate under the condition of 0.5～0.7MPa, concentrated solution is standby.
Embodiment 4: take by weighing fresh Herba Selaginellae 20 grams, after the aqueous acetone historrhexis extraction with 65-70%, put in 200 these aqueous acetone liquid of gram and soaked three days, filtering, get filtrate, is 45 ℃ in temperature, pressure is to be concentrated into paste under the 0.6MPa condition, in thick paste, add suitable quantity of water, use ether, ethyl acetate, n-butanol extraction successively, get ether, ethyl acetate, n-butyl alcohol and four positions of water.
N-butanol portion or water position are carried out column chromatography and preparation thin layer chromatographies such as Diaion HP-20, ToyoPearl HW-40, MCI Gel CHP-20, Sephadex LH-20, Silica Gel again, the separable monomeric compound that gets.
Above gained water position or n-butanol portion can or directly be taken for pharmaceutical purpose, and take at twice at each position.
The water position of gained and n-butanol portion also can use with the other medicines compatibility in the present embodiment, are used for making the estrogenic medicine of increase.
Embodiment 5: get Herba Selaginellae 100 parts by weight of crushed, cross the 120-140 mesh sieve, the powder after sieving adds refined honey 80-90 weight portion, makes the honeyed pill of every 3 weight portion, takes the 2-3 grain, every day 3 times at every turn.
Present embodiment is that Herba Selaginellae is directly as estrogenic.
Embodiment 6: get Herba Selaginellae 35 weight portions, add the water that medical material weighs 10 times of amounts, boiled 30 minutes, filter, add the water that medical material weighs 8 times of amounts in the medicinal residues again, boiled 25 minutes, filter merging filtrate, at pressure is 0.75～0.95MPa, and temperature is to concentrate under 35-60 ℃ the condition, concentrated solution is standby; Other gets Herba Selaginellae 65 parts by weight of crushed, crosses the 120-140 mesh sieve, and ball is made in powder after sieving and above-mentioned concentrated liquid flooding, takes the 3-4 weight portion at every turn, every day 2 times.
Claims (3)
1. the application of Herba Selaginellae in the preparation estrogenic.
2. purposes according to claim 1 is characterized in that: the application of the total extract of Herba Selaginellae in the preparation estrogenic.
3. purposes according to claim 1 is characterized in that: the application in the preparation estrogenic of water position in the Herba Selaginellae and n-butanol portion.
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| CN200610017494XA CN101028311B (en) | 2006-03-03 | 2006-03-03 | Use of selaginella tamariscina |
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|---|---|---|---|
| CN200610017494XA CN101028311B (en) | 2006-03-03 | 2006-03-03 | Use of selaginella tamariscina |
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| CN101028311B true CN101028311B (en) | 2010-04-21 |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102258570A (en) * | 2011-07-18 | 2011-11-30 | 贵州省中国科学院天然产物化学重点实验室 | Composition for inhibiting activity of alpha-glycuronide and preparation method of composition |
| CN102266359A (en) * | 2010-10-08 | 2011-12-07 | 河南中医学院 | Method for continuously extracting hypoglycemic active ingredient and estrogen active ingredient from tamariskoid spikemoss herbs |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1695643A (en) * | 2004-05-11 | 2005-11-16 | 上海先行医药开发有限公司 | Chinese medica preparation treating endometriosis and preparation method |
-
2006
- 2006-03-03 CN CN200610017494XA patent/CN101028311B/en active Active
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1695643A (en) * | 2004-05-11 | 2005-11-16 | 上海先行医药开发有限公司 | Chinese medica preparation treating endometriosis and preparation method |
Non-Patent Citations (2)
| Title |
|---|
| 周蜻 等.易修珍用卷柏治疗妇科病经验.中国民族民间医药杂志 2，总第37期.1999,(2，总第37期),第90页第2栏18-25行. |
| 周蜻等.易修珍用卷柏治疗妇科病经验.中国民族民间医药杂志 2，总第37期.1999,(2，总第37期),第90页第2栏18-25行. * |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102266359A (en) * | 2010-10-08 | 2011-12-07 | 河南中医学院 | Method for continuously extracting hypoglycemic active ingredient and estrogen active ingredient from tamariskoid spikemoss herbs |
| CN102266359B (en) * | 2010-10-08 | 2012-08-01 | 河南中医学院 | Method for continuously extracting hypoglycemic active ingredient and estrogen active ingredient from tamariskoid spikemoss herbs |
| CN102258570A (en) * | 2011-07-18 | 2011-11-30 | 贵州省中国科学院天然产物化学重点实验室 | Composition for inhibiting activity of alpha-glycuronide and preparation method of composition |
| CN102258570B (en) * | 2011-07-18 | 2012-10-03 | 贵州省中国科学院天然产物化学重点实验室 | Composition for inhibiting activity of alpha-glycuronide and preparation method of composition |
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|---|---|
| CN101028311A (en) | 2007-09-05 |
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