CN104288239A - Application of semen plantaginis water extract in preparation of estrogen drugs - Google Patents
Application of semen plantaginis water extract in preparation of estrogen drugs Download PDFInfo
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- CN104288239A CN104288239A CN201410481701.1A CN201410481701A CN104288239A CN 104288239 A CN104288239 A CN 104288239A CN 201410481701 A CN201410481701 A CN 201410481701A CN 104288239 A CN104288239 A CN 104288239A
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Classifications
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/18—Magnoliophyta (angiosperms)
- A61K36/185—Magnoliopsida (dicotyledons)
- A61K36/68—Plantaginaceae (Plantain Family)
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
- A61K2236/30—Extraction of the material
- A61K2236/33—Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones
- A61K2236/331—Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones using water, e.g. cold water, infusion, tea, steam distillation or decoction
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
- A61K2236/30—Extraction of the material
- A61K2236/39—Complex extraction schemes, e.g. fractionation or repeated extraction steps
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- Health & Medical Sciences (AREA)
- Natural Medicines & Medicinal Plants (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- Alternative & Traditional Medicine (AREA)
- Biotechnology (AREA)
- Botany (AREA)
- Medical Informatics (AREA)
- Medicinal Chemistry (AREA)
- Microbiology (AREA)
- Mycology (AREA)
- Pharmacology & Pharmacy (AREA)
- Epidemiology (AREA)
- Animal Behavior & Ethology (AREA)
- General Health & Medical Sciences (AREA)
- Public Health (AREA)
- Veterinary Medicine (AREA)
- Medicines Containing Plant Substances (AREA)
Abstract
The invention relates to application of a semen plantaginis water extract in preparation of estrogen drugs. The medication for treating deficiency of estrogens can be effectively solved, so that the problem that adverse reactions are brought by artificially synthesized estrogens in long-term clinical application is solved. According to the technical scheme, the invention discloses application of the semen plantaginis water extract in preparation of estrogen drugs. The semen plantaginis water extract is prepared by the following steps: adding water in an amount of 10 times that of the weight of semen plantaginis for decocting for three times and an hour each time, soaking for 30 minutes during decocting at the first time, merging the water decoction three times, performing vacuum concentration, and drying, thereby obtaining the semen plantaginis water extract. The semen plantaginis water extract has a certain estrogen-like effects in vivo and in vitro; through repeated tests, the same or similar result is achieved, and the water extract is safe and effective, does not have any adverse reaction and is effectively used for preparing the estrogen drugs; novel medical application of semen plantaginis is developed, and the application refers to an innovation of application of semen plantaginis.
Description
Technical field
The present invention relates to field of medicaments, particularly a kind of Semen Plantaginis water extract is preparing the application in estrogenic.
Background technology
Perimenopausal syndrome is due to women's ovary deterioration, estrogen secretion level obviously declines, the nerve caused, cardiovascular, hormonal system dysfunction, cause the symptoms such as dysfunctional uterine hemorrhage, paroxysmal hectic fever and perspiration, osteoporosis, pudendum and vaginal atrophy and occur spirit and neuro symptom.In climacteric women, its sickness rate is higher, directly has influence on their quality of life.Controversies in hormone replacement in the elderly to alleviating climacteric women menopausal symptom, prevent and treat osteoporosis and have good effect.But prolonged application synthetic estrogen can make the risk of carcinoma of endometrium increase.Apply with progestin combinations, although can this risk be reduced, the sickness rate of breast carcinoma and cardiovascular and cerebrovascular disease can be made to increase.The untoward reaction that synthetic estrogen brings in clinical prolonged application annoyings people always.
Phytoestrogen derives from plant, has the polyol of analog structure with mammiferous steroid hormone-17 beta estradiol.The extensive concern of people is caused because of the side effect caused by phytoestrogen prosthetic synthetic estrogen alternative medicine.The estrogen-like effects of phytoestrogen is dual: on the one hand, have estrogenic activity when estrogen level is lower in vivo; On the other hand, antiestrogenic can be played when body inner estrogen level is higher.Therefore, phytoestrogen is otherwise known as " selective estrogen receptor modulators " (seleetive estrogen receptor modulators, SERMs) in recent years, and its clinical effect is just receiving the concern of numerous scholar.
Semen Plantaginis is the dry mature seed of Plantaginaceae plant Herba Plantaginis Plantagoasiatica L. or Plantago depressa Willd PlantagodepressaWilld., and fruit ear of gathering when summer, autumn two season seed maturity, dries, rub seed, removing impurity.Its main component has phenethyl alcohol glycoside, iridoids, flavone, polysaccharide etc.Semen Plantaginis sweet in the mouth cold in nature, returns liver, kidney, lung, small intestine meridian.According to pharmacopeia record, there is clearing away heat and promoting diuresis treating stranguria, eliminating dampness by diuresis antidiarrheal, improving eyesight, the effect of eliminating the phlegm.Modern pharmacology and clinical research show that Semen Plantaginis also has antioxidation, immunomodulating, antiinflammatory, and antiviral protects the liver, and promote wound healing, the effect of antitumor and blood pressure lowering.But not yet there is the open report in Semen Plantaginis estrogenic activity.
Summary of the invention
For above-mentioned situation, for solving the defect of prior art, the object of the present invention is just to provide a kind of Semen Plantaginis water extract and is preparing the application in estrogenic, effectively can solve the medication for the treatment of oestrogen deficiencies, to overcome the problem of the untoward reaction that synthetic estrogen brings in clinical prolonged application.
The technical scheme that the present invention solves is, Semen Plantaginis water extract is preparing the application in estrogenic, this Semen Plantaginis water extract is, add the soak by water 3 times of Semen Plantaginis 10 times of weight, each 1h, when first time decocts at every turn, first soak 30 minutes, merge 3 decocting liquid, concentrating under reduced pressure is dry, obtains Semen Plantaginis water extract; First by Mouse Uterus weightening finish experiment, the screening of estrogenic activity has been carried out to Semen Plantaginis water extract, then MCF-7 cell proliferation experiment is utilized to carry out the research of external estrogenic activity to Semen Plantaginis water extract, finally use recombinant DNA technology, the reporter gene transient expression of ERE regulation and control is detected, have studied the mechanism of action that Semen Plantaginis plays estrogenic activity.Prove that Semen Plantaginis water extract has estrogen-like effects, it effectively can solve the medication problem of the relevant disease (as climacteric syndrome etc.) caused due to estrogen hyposecretion in body.
Semen Plantaginis water extract of the present invention in vivo, external all there is certain estrogen-like effects, and through repeated tests, all achieve identical or close result, safe and effective, have no adverse reaction, being effective to prepare estrogens medicine, opening the medicinal novelty teabag of Semen Plantaginis, is that in Semen Plantaginis purposes innovates greatly.
Accompanying drawing explanation
Fig. 1 be Semen Plantaginis water extract of the present invention on the impact of sex immature female mice Uterine coefficient (
n=10) schematic diagram.
Fig. 2 be Semen Plantaginis water extract of the present invention MCF-7 is bred impact (
n=6) schematic diagram.
Fig. 3 be Semen Plantaginis water extract of the present invention by the alpha mediated reporter gene transient expression testing result to ERE regulation and control of ER (
n=3) schematic diagram.
Fig. 4 be Semen Plantaginis water extract of the present invention by the beta mediated reporter gene transient expression testing result to ERE regulation and control of ER (
n=3) schematic diagram.
Detailed description of the invention
Below in conjunction with drawings and Examples, the specific embodiment of the present invention is described in further detail.
Embodiment 1
Semen Plantaginis water extract is preparing the application in estrogenic, this Semen Plantaginis water extract is, Semen Plantaginis 100g is added at every turn the soak by water 3 times of 1000ml, each 1h, when first time decocts, first soak 30 minutes, merge 3 decocting liquid, concentrating under reduced pressure is dry, obtains Semen Plantaginis water extract, three times repeatedly continuously, average yield is 12.14%.
Embodiment 2
Semen Plantaginis water extract is preparing the application in estrogenic, this Semen Plantaginis water extract is, Semen Plantaginis 500g is added at every turn the soak by water 3 times of 5000ml, each 1h, when first time decocts, first soak 30 minutes, merge 3 decocting liquid, concentrating under reduced pressure is dry, obtains Semen Plantaginis water extract, three times repeatedly continuously, average yield is 12.91%.
Embodiment 3
Semen Plantaginis water extract is preparing the application in estrogenic, this Semen Plantaginis water extract is, Semen Plantaginis 1000g is added at every turn the soak by water 3 times of 10000ml, each 1h, when first time decocts, first soak 30 minutes, merge 3 decocting liquid, concentrating under reduced pressure is dry, obtains Semen Plantaginis water extract, three times repeatedly continuously, average yield is 13.42%.
The Semen Plantaginis water extract of any amount can be obtained by suitability for industrialized production in the ratio of Semen Plantaginis and water according to said method, for the preparation of estrogens medicine, effectively can solve the medication problem for the treatment of the disease caused due to estrogen hyposecretion in body, and obtain sufficient proof through test, correlation test data is as follows:
1 experiment material
1.1 Experimental agents
Herba Plantaginis is the dry mature seed of Plantaginaceae plant Herba Plantaginis Plantagoasiatica L. or Plantago depressa Willd PlantagodepressaWilld., the summer, and fruit ear of gathering during autumn two season seed maturity, dries, rub seed, removing impurity.Semen Plantaginis adds the soak by water 3 times of 10 times of weight, each 1h, when first time decocts, first soaks 30 minutes, merges 3 decocting liquid, and concentrating under reduced pressure drying, obtains Semen Plantaginis water extract.
1.2 laboratory animals and cell strain and plasmid
Kunming mouse, female, be born 21 days (just weaning), body weight 9 ~ 12g, is purchased from Henan Province's Experimental Animal Center.Human breast cancer cell (MCF-7) is purchased from Institute of Basic Medical Sciences, Chinese Academy of Medical Sciences's cellular resources center.HEK293 cell strain is purchased from China typical culture collection center.Beta galactosidase (β-galactosidase, β-gal) control plasmid p β gal-Control, restructuring reporter gene pERE-TAL-luc are so kind as to give by the BIO ENGINEERING INST MILITARY dense doctor of leaf chess.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 is so kind as to give by department of medial science of Tokyo University Satoshi doctor Inoue.
1.3 main agents
DMEM high glucose medium and cationic-liposome LipofectamineTM2000Reagent are purchased from Gibco Invitrogen company; Hyclone is purchased from Guangzhou Ilex purpurea Hassk.[I.chinensis Sims company; Go hormone hyclone purchased from Hyclone company without phenol red DMEM high glucose medium, glucosan-active carbon; 17 beta estradiols (17 β-estrogen, E2), ampicillin (Amp), Tris Jian Jun available from Sigma; Estradiol valerate tablet (Bayer medicine); O-Nitrobenzol-β-D-synthesis (O-Nitrophenyl-β-D-galactopyranoside, ONPG), EDTA, MTT and DMSO are Amresco Products; Steady-Glo stablizes Luciferase Assay System test kit (Steady-Glo Luciferase Assay Systerm), glittering lysis buffer (Glolysis Buffer) purchased from Promega company; Yeast powder (OXOID company); Tryptone (OXOID company); Agarose (Biowest company); Competence bacillus coli DH 5 alpha (TransGen company); The little extraction reagent kit of plasmid (Tian Gen company); All the other reagent are domestic analytical pure.
1.4 key instrument
General surgery apparatus; CO2 gas incubator (STIK); Inverted microscope (NIKON ECLIPSE TS100); SORVALL ST16R Centrifuge high-speed low temperature refrigerated centrifuger (Thermo Fisher Scientific company); Microplate reader (BIO-RAD680); Pure water instrument (Sartorius611VF); 90-3 magnetic stirring apparatus (Shanghai Zhen Jie experimental facilities company limited); ZRD-7080 full-automatic novel air dry oven (Shanghai ZHICHENG Anaiytical Instrument Manufacturing Co., Ltd.); Micro sample adding appliance (Nichipet EX PLUS); AB204-N electronical reading analytical balance (Mettler-Toledo Instrument's product); 10cm culture dish, 96 well culture plates, cryopreservation tube are Corning company and produce; Superclean bench (Jiangsu Su Jing group); Bio Mate3S ultraviolet spectrophotometer (Thermo Fisher Scientific company); Veritas
tMmicroplate luminometer (TurnerBioSystems company); SK6200H type ultrasonic cleaner (Shanghai High Kudos Science Instrument Co., Ltd.); LRH array of biochemical incubator (the permanent Science and Technology Ltd. in Shanghai one).
2 experimental techniques
2.1 Mouse Uterus weightening finish experiment
Mice is divided into groups by the principle that body weight is balanced and random.Administration continues 7 days.Blank group distilled water every day 0.2ml10g
-1gavage; Positive controls estradiol valerate every day (0.33mgkg
-1d
-1) gavage; Semen Plantaginis water extract is low dosage by the 20 times amount × extraction ratio of clinical application, and the 40 times amount × extraction ratio of clinical application is that high dose calculates, and be made into the suspension of respective concentration, every day presses 0.2ml10g
-1gavage.After last administration 24h, de-neck puts to death mice, wins uterus immediately and weighs, and calculates Uterine coefficient (uterine wet weight × (body weight)
-1× 100%).Test in triplicate parallel.Result, compared with blank group, calculates P value, investigates and has not statistically significant.
2.2MCF--7 cell proliferation experiment
MCF-7 cell (human breast cancer cell) is through containing 5% go the DMEM culture medium culturing of estrogen serum after 1 week without phenol red, choose exponential phase cell, PBS washes twice, after 0.05% trypsinization, add without phenol red DMEM culture medium piping and druming evenly, with 6 × 10
4the concentration of individual/ml is inoculated in 96 orifice plates, and it is 200 μ l that cumulative volume is cultivated in every hole.Cultivate 24h after cell attachment, be changed to pastille culture fluid and continue to cultivate.37 DEG C, 5%CO
2after cultivating 48h, every hole adds MTT solution (5mgml
-1) 20 μ l, 37 DEG C are continued to cultivate 4h, and carefully exhaust culture fluid, and every hole adds 150 μ lDMSO, and concussion 5-10min, makes purple crystal dissolve completely.Under 490nm, measure each hole absorbance (A) by microplate reader, calculate average A-value and the rate of increase (Proliferation Rate, PR).Test in triplicate parallel.
PR%=(experimental group A value-blank group A value) × blank group A value
-1× 100%
The reporter gene transient expression of 2.3ERE regulation and control detects
2.3.1 cell inoculation
Before transfection, the HEK293 cell (HEKC) being in exponential phase is inoculated in 24 orifice plates by 24h, and density is 16.0 × 10
4individual/0.5ml/ hole, and culture fluid is go not containing of estrogen serum antibiotic without phenol red DMEM culture fluid containing 10%, arranges blank control wells, estradiol hole and medicine hole to be measured respectively.
2.3.2 the transfection of plasmid
(1) when cell cloth ware 80%-90%, the former culture medium of careful sucking-off, adds 420 μ l and goes not containing of estrogen serum antibiotic without phenol red DMEM culture medium containing 10%.
(2) get 3 EP pipes, add 490 μ l in ER alphatrons without phenol red DMEM culture medium, 9 μ lpCXN2-hER α plasmid (80ng μ l
-1), 9 μ lpERE-TAL-luc plasmid (400ng μ l
-1), 9 μ lp β gal-Control plasmid (200ng μ l
-1); 490 μ l are added without phenol red DMEM culture medium, 9 μ lpCXN2-hER β plasmid (80ng μ l in ER β pipe
-1), 9 μ l pERE-TAL-Iuc plasmid (400ng μ l
-1), 9 μ l β gal-Control plasmid (200ng μ l
-1); 1000 μ l are added without phenol red DMEM culture medium and 36 μ l cationic-liposomes in liposome pipe.
(3) reversing mixing liposome pipe, room temperature places 5min, gets 510 μ l and adds ER α and ER β respectively and manage, mix gently, room temperature placement 20min;
(4) get liposome/DNA complex in 80 μ lER alphatrons and add 24 orifice plate ER α experimental ports respectively, get liposome/DNA complex in 80 μ lER β pipes and add 24 orifice plate ER β experimental ports respectively.Front and back shake up, and put into incubator.
(5) control drug and detection of drugs is added after 4-6h.
2.3.3ERE the reporter gene transient expression regulated and controled detects
After dosing, the cell of 24h, takes out from incubator, and equilibrium at room temperature 20min, sucks culture fluid.Luciferase Assay System test kit (Steady-Glo Luciferase Assay Systerm) and the operation of glittering lysis buffer (Glolysis Buffer) evolutionary operation step is stablized according to Steady-Glo, collecting cell cracking supernatant, add luciferase reaction substrate (fluorescein), be placed in dark place, use Veritas
tMmicroplate luminometer fluorescence intensity.
2.3.4 β-gal Activity determination
In order to investigate transfection efficiency, internal reference β-gal activity is detected.
(1) 54 μ l beta-mercaptoethanol adds in 40ml Z-buffer, mixing, and get EP pipe, often pipe adds 1800 μ l;
(2) often pipe adds 20 μ l cracking supernatant;
(3) often pipe adds 400 μ l ONPG, reversing mixing;
(4) 37 DEG C of incubators are placed, to displaing yellow;
(5) Na is added
2cO
31ml cessation reaction;
(6) OD420 value is surveyed;
(7) uciferase activity of normalized;
3 statistical procedures
Experimental data with
represent, SPSS18.0 carries out one factor analysis of variance statistical disposition.
4 experimental results
4.1 Semen Plantaginis water extracts are on the impact of sex immature female mice Uterine coefficient
Result shows, Semen Plantaginis water extract low dosage can the increase (P<0.01) of extremely remarkable inductivity immaturity female mice Uterine coefficient compared with blank group, and Semen Plantaginis high dose group can the increase (P<0.05) of remarkable inductivity immaturity female mice Uterine coefficient compared with blank group.Prompting Semen Plantaginis water extract has estrogenic activity in vivo.In table 1, Fig. 1.
Table 1 Semen Plantaginis water extract on the impact of sex immature female mice Uterine coefficient (
n=10)
Note: compare with blank group; # represents P<0.05, and ## represents P<0.01; Compare with estradiol valerate group, * represents P<0.05, and * * represents P<0.01.
4.2 administrations after 72 hours Semen Plantaginis water extract on the impact of MCF--7 cell proliferation
Result shows, and Semen Plantaginis water extract is at 0.0001mgml
-1, 0.001mgml
-1, 0.1mgml
-1, 1mgml
-1time with blank group ratio, all have extremely significant proliferation (P<0.01) to MCF-7 cell.0.01mgml
-1time with blank group ratio, have significant proliferation (P<0.05) to MCF-7 cell.Prompting Semen Plantaginis water extract has estrogenic activity in vitro.In table 2, Fig. 2.
Table 2 Semen Plantaginis water extract MCF-7 is bred impact (
n=6)
Note: compare with blank group, # represents P<0.05, and ## represents P<0.01; With E
2group compares, and * represents P<0.05, and * * represents P<0.01.
4.3 Semen Plantaginis water extracts are to the inducing action of the reporter gene transient expression that ERE regulates and controls
Reporter gene technology is utilized to detect Semen Plantaginis water extract and estrogen receptor (Estrogen Receptors, ERs) interactional impact.
Positive controls i.e. 17 beta estradiol (E
2) final concentration that adds is 10
-8molL
-1, Semen Plantaginis water extract adding consistency is respectively 1mgml
-1, 0.1mgml
-1, 0.01mgml
-1, 0.001mgml
-1.Result shows, no matter be beta mediated by ER α or ER, and E
2uciferase activity after standardization is all significantly higher than blank well (P<0.01).Time alpha mediated by ER, Semen Plantaginis water extract 1mgml
-1, 0.1mgml
-1, 0.01mgml
-1uciferase activity after standardization is significantly higher than normal hole (P<0.01), table 3, Fig. 3.Time beta mediated by ER, Semen Plantaginis water extract 1mgml
-1and 0.01mgml
-1uciferase activity after two standard sets is also all significantly higher than blank well (P<0.01), table 4, Fig. 4.Prompting Semen Plantaginis water extract can be passed through the beta mediated activated gene of estrogen receptor ER α and ER and transcribes performance estrogen-like effects.
Table 3 Semen Plantaginis water extract by ER alpha mediated to ERE regulation and control reporter gene transient expression testing result (
n=3)
Note: compare with blank group, # represents P<0.05, and ## represents P<0.01; With E
2group compares, and * represents P<0.05, and * * represents P<0.01.
Table 4 Semen Plantaginis water extract by ER beta mediated to ERE regulation and control reporter gene transient expression testing result (
n=3)
Note: compare with blank group, # represents P<0.05, and ## represents P<0.01; With E
2group compares, and * represents P<0.05, and * * represents P<0.01.
The display of Mouse Uterus weightening finish experimental result, Semen Plantaginis water extract significantly can increase sex immature female mice Uterine coefficient, and prompting Semen Plantaginis water extract has estrogenic activity in vivo.
MCF-7 cell proliferation experiment result shows, and Semen Plantaginis water extract is 1mgml in concentration
-1, 0.1mgml
-1, 0.01mgml
-1, 0.001mgml
-1and 0.0001mgml
-1time to MCF-7 cell, all there is significant proliferation, prompting Semen Plantaginis water extract has estrogenic activity in vitro.
The reporter gene transient expression testing result display of ERE regulation and control, the estrogen-like effects of Semen Plantaginis water extract mediates performance jointly by ER α and ER β.
In sum, animal and the display of cell experiment result, Semen Plantaginis water extract in vivo, external all there is certain estrogen-like effects, and be jointly mediate by estrogen receptor ER α and ER β and play estrogen-like effects.
5 conclusions
Uterus growth test is the classical way of the detection estrogen activity set up the earliest, when evaluating estrogen activity, it considers the absorption of material, metabolism, to be combined with plasma protein and the many factors such as pharmacokinetics, directly reflects the total Biology effect of test medicine after internal metabolism.Therefore, this experiment employing sex immature female mice uterine wet weight and the ratio (Uterine coefficient) of body weight are as the index evaluating estrogenic activity.By above-mentioned experiment, prove that Semen Plantaginis water extract significantly can increase sex immature Mouse Uterus coefficient, prompting Semen Plantaginis water extract has estrogen-like effects in vivo.
MCF-7 cell is the Breast cancer lines of estrogen receptor positive, and can regulate by estrogen or estrogenic activity material and breed specifically, be the cell strain of the most frequently used detection estrogenic activity.MCF-7 cell estrogen-like proliferation test is very sensitive, can distinguish estrogen agonist and antagonist, is thus widely used in external a large amount of, rapid screening and evaluation environmental estrogens and phytoestrogen.Urge in the experiment of MCF-7 cell proliferation at Semen Plantaginis water extract, Semen Plantaginis water extract is 1mgml in concentration
-1, 0.1mgml
-1, 0.01mgml
-1, 0.001mgml
-1and 0.0001mgml
-1time have significant proliferation to MCF-7 cell, prompting Semen Plantaginis water extract has estrogenic activity in vitro.
Above-mentioned test is carried out repeatedly repeatedly, all achieves identical or close result, prompting, Semen Plantaginis water extract in vivo, external all there is certain estrogen-like effects.
Reporter gene (reporter gene) is the gene of a kind of coding protein that can be detected or enzyme, and namely its expression product of a class is very easy to identified, and with endogenous background proteins more easily distinguish gene.Reporter gene technology is connected with reporter gene by response element, by examining report activity of gene expression, known signal transduction pathway and on genes within cells express regulation and control and impact.Reporter gene technology is highly sensitive, selectivity is strong and simple to operate.In the inducing action experiment of Semen Plantaginis water extract to Reporter gene vector pERE-TAL-luc transient expression, by the expression of visual report gene, namely whether the height of fluorescence intensity can detect tested material and be combined with estrogen receptor, and act on the expression that estrogen response element (ERE) carrys out controlling gene, thus understand the size of the power of Semen Plantaginis water extract estrogen-like effects and itself and ER α or ER β affinity.Experimental result shows, time alpha mediated by ER, and Semen Plantaginis water extract 1mgml
-1, 0.1mgml
-1, 0.01mgml
-1uciferase activity after standardization is significantly higher than normal hole (P<0.01).Time beta mediated by ER, Semen Plantaginis water extract 1mgml
-1and 0.01mgml
-1uciferase activity after two standard sets is also all significantly higher than blank well (P<0.01).Prompting, Semen Plantaginis water extract can be passed through Mediated by Estrogen Receptor activated gene and transcribes.
Organism is a complicated system, and vivo and vitro result exists certain deviation.In this experimental applications In vitro cell experiment, body, Integral animal experiment and reporter gene technology combine and jointly verify the estrogenic activity of tested material, ensure that the accuracy of result.
By above-mentioned experiment, prove that Semen Plantaginis water extract can promote the propagation of MCF-7 cell and increase the weight in wet base of sex immature Mouse Uterus, Semen Plantaginis water extract can also activate by the beta mediated genetic transcription of ER α and ER and play estrogenic activity simultaneously.From above experimental result, Semen Plantaginis water extract has estrogen-like effects, it effectively can solve the medication problem of the relevant disease (as climacteric syndrome etc.) caused due to estrogen hyposecretion in body, and provide reference to clinician, consider when being intended to drug matching that this factor improves prescription curative effect, clinical meaning is huge.
Claims (4)
1. Semen Plantaginis water extract is preparing the application in estrogenic, and this Semen Plantaginis water extract is, adds the soak by water 3 times of Semen Plantaginis 10 times of weight at every turn, each 1h, when first time decocts, first soaks 30 minutes, merge 3 decocting liquid, concentrating under reduced pressure is dry, obtains Semen Plantaginis water extract.
2. Semen Plantaginis water extract according to claim 1 is preparing the application in estrogenic, it is characterized in that, described Semen Plantaginis water extract is, Semen Plantaginis 100g adds the soak by water 3 times of 1000ml at every turn, each 1h, first time, when decocting, first soaks 30 minutes, merges 3 decocting liquid, concentrating under reduced pressure is dry, Semen Plantaginis water extract, repeatedly three times continuously, average yield is 12.14%.
3. Semen Plantaginis water extract according to claim 1 is preparing the application in estrogenic, it is characterized in that, described Semen Plantaginis water extract is, Semen Plantaginis 500g adds the soak by water 3 times of 5000ml at every turn, each 1h, first time, when decocting, first soaks 30 minutes, merges 3 decocting liquid, concentrating under reduced pressure is dry, Semen Plantaginis water extract, repeatedly three times continuously, average yield is 12.91%.
4. Semen Plantaginis water extract according to claim 1 is preparing the application in estrogenic, it is characterized in that, described Semen Plantaginis water extract is, Semen Plantaginis 1000g adds the soak by water 3 times of 10000ml at every turn, each 1h, first time, when decocting, first soaks 30 minutes, merges 3 decocting liquid, concentrating under reduced pressure is dry, Semen Plantaginis water extract, repeatedly three times continuously, average yield is 13.42%.
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| U.SEBNEM HARPUT: "Cytotoxic and antioxidative activities of Plantago lagopus L. and characterization of its bioactive compounds", 《FOOD AND CHEMICAL TOXICOLOGY》 * |
| 郝庆秀: "熟地等4味中药的植物雌激素作用的实验研究", 《中国中药杂志》 * |
| 陈民勤: "清热利湿助孕汤治疗不孕症30例", 《湖北中医杂志》 * |
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