CN105168301A - Application of semen lepidii water extract in preparation of estrogen medicines - Google Patents
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Abstract
The invention discloses an application of a semen lepidii water extract in preparation of estrogen medicines. Firstly, the semen lepidii water extract is screened through a mice uterus growth test and has estrogenic activity; secondly, the condition that the semen lepidii water extract has estrogenic activity is proved again in vitro by an MCF-7 cell proliferation assay; thirdly, the condition that semen lepidii plays a role through a classic estrogen signal transduction pathway is shown through an MCF-7 cell proliferation reversal test; and finally, reporter gene transient expression on ERE regulation and control is detected by a recombinant DNA technology, so as to primarily investigate the mechanism of the semen lepidii for playing an estrogen-like function. The semen lepidii water extract is simple in preparation method, and good in repeatability; through multiple tests, the same or similar results are obtained; medicinal materials are expanded for research of phytoestrogen; and a new medicinal application of the semen lepidii is developed.
Description
Technical field
The present invention relates to estrogens medicine field, be specifically related to a kind of Semen Lepidii (Semen Descurainiae) water extract and preparing the application in estrogens medicine.
Background technology
Semen Lepidii (Semen Descurainiae) is the dry mature seed of crucifer descurainia sophia (l.) webb ex prantl Descurainiasophia (L) Webb.exPrantl. (DSW) or Lepidium sativum L. LepidiumapetalumWilld. (LAW).Tap plant during fruit maturation in summer, dry, rub seed, removing impurity.The former practises title Semen Descurainiae, and the latter practises title lepidii,semen.Because the medicine source of Semen Descurainiae is comparatively wide, therefore Semen Descurainiae research is selected in this experiment.Semen Lepidii (Semen Descurainiae) begins to be loaded in Shennong's Herbal, is classified as low-grades, acrid in the mouth, hardship; Property Great Cold; Return lung, urinary bladder channel.The traditional Chinese medical science thinks that it can eliminating pathogen from the lung for relieving asthma, inducing diuresis to remove edema; Stop up lung for sputum, cough with asthma abundant expectoration, fullness and distention in the chest and hypochondrium, must not put down sleeping, and ascites pleural fluid swells, dysuria etc.Modern pharmacological research shows, Semen Lepidii (Semen Descurainiae) water extract obviously can improve the retention of sodium and water of Heart Failure Wistar Rats, and has significant diuresis.Meanwhile, Semen Lepidii (Semen Descurainiae) has antioxidation to have experiment to show.Semen Lepidii (Semen Descurainiae) can also regulate the blood lipid level of hyperlipidemia rat.But there is not yet the research of Semen Lepidii (Semen Descurainiae) estrogenic activity aspect.
Goal of the invention
The object of the invention is research Semen Lepidii (Semen Descurainiae) water extract and prepare the application in estrogens medicine.
Technical scheme of the present invention is: described Semen Lepidii (Semen Descurainiae) water extract is preparing the application in estrogens medicine.
The application of described Semen Lepidii (Semen Descurainiae) water extract in the Uterine coefficient increasing sex immature female mice.
Described Semen Lepidii (Semen Descurainiae) water extract is to the application of MCF-7 cell proliferation.
Described Semen Lepidii (Semen Descurainiae) water extract in vivo, externally have estrogenic activity, play estrogenic activity by activating estrogen receptor ER β.
The preparation method of described Semen Lepidii (Semen Descurainiae) water extract is: Semen Lepidii (Semen Descurainiae) is with the soak by water 3 times of 10 times amount, and each 1h, when first time decocts, first invades bubble 30 minutes, merges 3 decocting liquid, and concentrating under reduced pressure drying, obtains Semen Lepidii (Semen Descurainiae) water extract.
The invention has the beneficial effects as follows: the present invention uses the estrogenic activity of Mouse Uterus weightening finish experiment and external MCF-7 cell proliferation experiment screening Semen Lepidii (Semen Descurainiae) water extract in body; Adopt the mechanism of action of the reporter gene Luc transient expression detection technique determination Semen Lepidii (Semen Descurainiae) water extract estrogenic activity of MCF-7 cell proliferation antagonistic experiment and ERE regulation and control.Semen Lepidii (Semen Descurainiae) water extract significantly can increase the Uterine coefficient of sex immature female mice, has a significant difference (P<0.01) compared with normal group; MCF-7 cell proliferation experiment shows, multiple concentration of Semen Lepidii (Semen Descurainiae) water extract have obvious proliferation (P<0.01), and this proliferation can by estrogen receptor blocker ICI182, and 780 suppress.The reporter gene transient expression result display of ERE regulation and control, time alpha mediated by ER, the uciferase activity after each concentration standard of Semen Lepidii (Semen Descurainiae) water extract does not have significant difference compared with normal group; Time beta mediated by ER, the uciferase activity after the Semen Lepidii (Semen Descurainiae) water extract standardization of 0.05mg/ml is significantly higher than normal group (P<0.01).Semen Lepidii (Semen Descurainiae) water extract in vivo, externally have obvious estrogenic activity, and play estrogenic activity mainly through activating estrogen receptor ER β.
First the present invention selects Semen Lepidii (Semen Descurainiae) water extract by Mouse Uterus weightening finish testing sieve and has estrogenic activity, then again prove that Semen Lepidii (Semen Descurainiae) water extract has estrogenic activity by MCF-7 cell proliferation experiment from external, then find that Semen Lepidii (Semen Descurainiae) is worked by the estrogen receptor transduction pathway of classics by MCF-7 cell proliferation antagonistic experiment, finally use recombinant DNA technology, the reporter gene transient expression of ERE regulation and control is detected, thus Primary Study Semen Lepidii (Semen Descurainiae) plays the mechanism of estrogen-like effects.
Semen Lepidii (Semen Descurainiae) water extract preparation method of the present invention is simple, and favorable reproducibility, through test of many times, all achieves identical or akin result, for the research of phytoestrogen expands medicine resource, opens the medicinal novelty teabag of Semen Lepidii (Semen Descurainiae).
Accompanying drawing explanation
Fig. 1 is the impact of Semen Lepidii (Semen Descurainiae) water extract on sex immature Mouse Uterus coefficient;
Fig. 2 is the impact of Semen Lepidii (Semen Descurainiae) water extract on MCF-7 cell proliferation;
Fig. 3 is the impact of Semen Lepidii (Semen Descurainiae) water extract on MCF-7 cell proliferation antagonism;
Fig. 4 be Semen Lepidii (Semen Descurainiae) water extract alpha mediated by ER time to ERE regulation and control reporter gene transient expression testing result;
Fig. 5 be Semen Lepidii (Semen Descurainiae) water extract beta mediated by ER time to ERE regulation and control the discussion of reporter gene transient expression testing result.
Detailed description of the invention
1 experiment material
1.1 medicine
Semen Descurainiae (being purchased from Zhengzhou City's Chinese Medicinal Materials Markets, through being accredited as the dry seed of crucifer descurainia sophia (l.) webb ex prantl Descurainiasophia (L.) Webb.exPrantl.) awards qualification by Henan College Of Traditional Chinese Medicine Dong Cheng penetrating judgment.
Semen Lepidii (Semen Descurainiae) is with the soak by water 3 times of 10 times amount, and each 1h, when first time decocts, first invades bubble 30 minutes, merges 3 decocting liquid, and concentrating under reduced pressure drying, obtains Semen Lepidii (Semen Descurainiae) water extract.
1.2 animal
SPF level Kunming mouse, female, body weight 9-12g, is born 21 days (just weaning), is purchased from Henan Province's zoopery animal center (credit number: SCXK (Henan) 2010-0002).
1.3 cell strains and plasmid
HEK293 cell strain is purchased from China typical culture collection center.Human breast cancer cell (MCF-7) is purchased from Institute of Basic Medical Sciences, Chinese Academy of Medical Sciences's cellular resources center.Restructuring reporter gene pERE-TAL-luc, beta galactosidase (β-galactosidase, β-gal) control plasmid p β gal-Control are so kind as to give by the BIO ENGINEERING INST MILITARY dense doctor of leaf chess.Recombined human ER β (humanER β, hER β) expression vector pCXN2-hER β and recombined human ER α (humanER α, hER α) expression vector pCXN2-hER α is so kind as to give by department of medial science of Tokyo University doctor SatoshiInoue.
1.4 reagent
Cationic-liposome LipofectamineTM2000Reagent and DMEM high glucose medium are purchased from GibcoInvitrogen company; Hyclone is purchased from Guangzhou Ilex purpurea Hassk.[I.chinensis Sims company; Glucosan-active carbon removes hormone hyclone, without phenol red DMEM high glucose medium purchased from Hyclone company; Ampicillin (Amp), 17 beta estradiols (17 β-estrogen, E
2), Tris Jian Jun available from Sigma; O-Nitrobenzol-β-D-synthesis (O-Nitrophenyl-β-D-galactopyranoside, ONPG), MTT, EDTA and DMSO are Amresco Products; Estradiol valerate tablet (Bayer medicine); Glittering lysis buffer (GlolysisBuffer), Steady-Glo stablize Luciferase Assay System test kit (Steady-GloLuciferaseAssaySysterm) purchased from Promega company; Tryptone (OXOID company); Agarose (Biowest company); Yeast powder (OXOID 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.
1.5 instrument
RD-7080 full-automatic novel air dry oven (Shanghai ZHICHENG Anaiytical Instrument Manufacturing Co., Ltd.); SB-1000 type rotary evaporator (Shanghai Ai Lang Instrument Ltd.); Z micro sample adding appliance (NichipetEXPLUS); AB204-N electronical reading analytical balance (Mettler-Toledo Instrument's product); Microplate reader (BIO-RAD680); SK6200H type ultrasonic cleaner (Shanghai High Kudos Science Instrument Co., Ltd.); 90-3 magnetic stirring apparatus (Shanghai Zhen Jie experimental facilities company limited); Inverted microscope (NIKONECLIPSETS100); CO2 gas incubator (STIK); SORVALLST16RCentrifuge high-speed low temperature refrigerated centrifuger (ThermoFisherScientific company); Pure water instrument (Sartorius611VF); 10cm culture dish, 96 well culture plates, cryopreservation tube are Corning company and produce; Veritas
tMmicroplate luminometer (TurnerBioSystems company); Superclean bench (Jiangsu Su Jing group); BioMate3S ultraviolet spectrophotometer (ThermoFisherScientific 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
The female mice of sex immature, according to the grouping of Stochastic sum body weight homeostatic principle, continues medication 7 days.Blank group gavage distilled water, dosage 0.02mlg
-1d
-1; Positive controls estradiol valerate every day (0.33mgkg
-1d
-1) gavage; Semen Lepidii (Semen Descurainiae) low dosage (1.667gkg
-1d
-1): the 10 times amount × extraction ratio of clinical application; Dosage (3.333gkg in Semen Lepidii (Semen Descurainiae)
-1d
-1): the 20 times amount × extraction ratio of clinical application; Semen Lepidii (Semen Descurainiae) high dose (6.667gkg
-1d
-1): the 40 times amount × extraction ratio of clinical application, after Semen Lepidii extract (Semen Descurainiae extract) is made into the suspension of corresponding dosage, by 0.02mlg
-1d
-1gavage; To get before result fasting 12 hours, can't help water; Last administration, after 24 hours, puts to death mice, takes out uterus at once 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
Human breast carcinoma cell lines MCF-7 cell is through containing 5% go the DMEM culture medium culturing of estrogen serum after 1 week without phenol red, and choose exponential phase cell, PBS washes twice, after 0.05% trypsinization, adds 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 every hole cumulative volume is 200 μ l.Cultivate 24h after cell attachment, add test medicine and E
2intervene.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.Measure each hole absorbance (A) under 490nm, calculate average A-value and the rate of increase (ProliferationRate, PR).Test in triplicate parallel.
PR%=(experimental group A value-blank group A value) × blank group A value
-1× 100%
2.3MCF-7 cell proliferation antagonistic experiment
MCF-7 cell goes the DMEM culture medium culturing of hormone serum after one week with containing 5%, will be in the cell of exponential phase with 6 × 10
4the concentration of individual/ml is inoculated in 96 orifice plates.Use instead after adherent pastille without phenol red DMEM culture medium and estrogen receptor blocker ICI182,780 (10
-8mol/L).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 (ProliferationRate, PR).Test in triplicate parallel.
PR%=(experimental group A value-blank group A value) × blank group A value
-1× 100%
2.4 reporter gene technology
2.4.1ERE the reporter gene transient expression regulated and controled detects
1. the HEK293 cell (HEKC) being in exponential phase is inoculated in 24 orifice plates, density is 1.6 × 10
5individual/0.5ml/ hole, with 10% remove estrogen serum do not cultivate 24 hours without phenol red DMEM containing antibiotic.
2. by pERE-TAL-luc plasmid (400ng/ μ l), p β gal-Control plasmid (200ng/ μ l), pCXN2-hER α plasmid (80ng/ μ l) or pCXN2-hER β plasmid (80ng/ μ l) cotransfection in HEK293 cell strain, cultivate 6 hours.
3. add pharmaceutical intervention, normal group, positive group (17 beta estradiol) and medicine group to be measured are set respectively, 37 DEG C, 5%CO
2cultivate 24 hours.
4. cultivate after 24 hours, take out from incubator, equilibrium at room temperature 20min, sucks culture fluid.Luciferase Assay System test kit (Steady-GloLuciferaseAssaySysterm) and the operation of glittering lysis buffer (GlolysisBuffer) 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.
2.4.2 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 40mlZ-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 lONPG, reversing mixing;
(4) 37 DEG C of incubators are placed, to displaing yellow;
(5) Na is added
2cO
31ml cessation reaction;
(6) the OD value at 420nm place is surveyed;
(7) uciferase activity of normalized;
2.5 statistical disposition
Experimental data with
represent, SPSS18.0 carries out one factor analysis of variance statistical disposition.
3 experimental results
3.1 Semen Lepidii (Semen Descurainiae) water extracts are on the impact of sex immature Mouse Uterus
Compared with blank group, the basic, normal, high dosage of Semen Lepidii (Semen Descurainiae) significantly can increase the Uterine coefficient (p<0.01) of sex immature female mice, and prompting Semen Lepidii (Semen Descurainiae) water extract has estrogenic activity in vivo.The results are shown in Table 1, Fig. 1.
Table 1 Semen Lepidii (Semen Descurainiae) water extract is on the impact of sex immature Mouse Uterus coefficient
Note: compared with blank group, * represents P<0.05, * * represents P<0.01
3.2 Semen Lepidii (Semen Descurainiae) water extracts are on the impact of MCF-7 cell proliferation
Compared with normal group, Semen Lepidii (Semen Descurainiae) water extract 0.0001mg/ml, 0.00002mg/ml, 0.000004mg/ml, 0.0000008mg/ml have significant proliferation (P<0.01) to MCF-7 cell, show that Semen Lepidii (Semen Descurainiae) water extract also has significant estrogen-like effects in vitro.The results are shown in Table 2, Fig. 2.
Table 2 Semen Lepidii (Semen Descurainiae) water extract is on the impact of MCF-7 cell proliferation
Note: compared with normal group, * represents P<0.05, and * * represents P<0.01
3.3 Semen Lepidii (Semen Descurainiae) water extracts are on the impact of MCF-7 cell proliferation antagonism
17 beta estradiols can promote the propagation (P<0.01) of MCF-7 cell extremely significantly compared with normal group, but add estrogen receptor antagon ICI182, after 780, the proliferation of 17 beta estradiols disappears, no difference of science of statistics.
Semen Lepidii (Semen Descurainiae) water extract 0.0001mgml
-1, 0.00001mgml
-1, 0.000001mgml
-1compared with normal group, the propagation (P<0.01) of MCF-7 cell can be promoted significantly, and add estrogen receptor antagon ICI182,780, proliferation equal no difference of science of statistics compared with normal group of Semen Lepidii (Semen Descurainiae) water extract.Prompting Semen Lepidii (Semen Descurainiae) water extract promotes MCF-7 cell proliferation by the approach of Mediated by Estrogen Receptor.The results are shown in Table 3, Fig. 3.
Table 3 Semen Lepidii (Semen Descurainiae) water extract is on the impact of MCF-7 cell proliferation antagonism
Note: compared with normal group, * represents P<0.05, and * * represents P<0.01
3.4 Semen Lepidii (Semen Descurainiae) water extracts are to the inducing action of the reporter gene transient expression that ERE regulates and controls
Time alpha mediated by estrogen receptor ER, 17 beta estradiols (10
-8mol/l), after interacting with estrogen receptor, the uciferase activity after standardization is significantly higher than Normal group (P<0.01).But for Semen Lepidii (Semen Descurainiae) water extract when concentration is 0.05mg/ml, 0.005mg/ml, 0.0005mg/ml, uciferase activity time alpha mediated by estrogen receptor ER after standardization does not have significant difference compared with Normal group.Prompting Semen Lepidii (Semen Descurainiae) water extract does not play estrogen-like effects by estrogen receptor ER α.The results are shown in Table 4, Fig. 4.
Time beta mediated by estrogen receptor ER, 17 beta estradiols (10
-8mol/l), after interacting with estrogen receptor, the uciferase activity after standardization is significantly higher than Normal group (P<0.01).When Semen Lepidii (Semen Descurainiae) water extract concentration is 0.05mg/ml, uciferase activity time beta mediated by estrogen receptor ER after standardization also has significant significant difference (P<0.01) compared with Normal group.Result prompting Semen Lepidii (Semen Descurainiae) water extract can play estrogen-like effects by estrogen receptor ER β.In table 5, Fig. 5.
To the reporter gene transient expression testing result of ERE regulation and control when table 4 Semen Lepidii (Semen Descurainiae) water extract is alpha mediated by ER
Note: compared with normal group, * represents P<0.05, and * * represents P<0.01
To the reporter gene transient expression testing result of ERE regulation and control when table 5 Semen Lepidii (Semen Descurainiae) water extract is beta mediated by ER
Note: compared with normal group, * represents P<0.05, and * * represents P<0.01
Mouse Uterus weightening finish method is the classical way of detection of drugs estrogenic activity.This method economy, easy, save time, and highly sensitive, the material that some need metabolism in vivo, activation just has estrogen activity and intermediate product can detect by this method.The principle of Mouse Uterus weightening finish experiment is that a large amount of estrogen receptor is contained in uterus, so estrogen and there is the compound of estrogenic activity by being combined with this receptor, thin intramicellar reaction can be brought out, the estrogen-induced protein content in uterus is increased, shows as uterine cancer cell hypertrophy.Have an impact to experimental result for reducing endogenous estrogen secretion in this experiment, the animal selected is the female mice of firm ablactation, sex immature as far as possible, eventually through the ratio of Uterine coefficient and uterine wet weight and body weight as the index evaluating estrogen activity.Experimental result shows that the basic, normal, high dosage of Semen Lepidii (Semen Descurainiae) is compared with blank group, and Uterine coefficient has remarkable rising, thus proves that Semen Lepidii (Semen Descurainiae) water extract has estrogenic activity in vivo.
Human breast cancer cell (MCF-7 cell strain) is estrogen receptor positive cell line, estrogen or estrogen-like activity material all can induce the propagation of this cell line, and the proliferation function of this estrogen-like is very responsive, therefore MCF-7 cell has been widely used in the activity of screening and evaluation phytoestrogen.Next we select mtt assay to detect Semen Lepidii (Semen Descurainiae) water extract to the impact of MCF-7 cel l proliferation.Result shows, and Semen Lepidii (Semen Descurainiae) water extract 0.0001mg/ml, 0.00002mg/ml, 0.000004mg/ml, 0.0000008mg/ml all have significant proliferation to MCF-7 cell.This represents that Semen Lepidii (Semen Descurainiae) water extract also has significant estrogen-like effects in vitro.
Above-mentioned test is carried out repeatedly repeatedly, all achieves identical or close result, prompting, Semen Lepidii (Semen Descurainiae) water extract in vivo, external all there is certain estrogen-like effects.
MCF-7 cell proliferation antagonistic experiment, adds estrogen receptor antagon ICI182,780.It can weaken significantly or suppress to be combined excited by part with estrogen receptor, and by the estrogen receptor transduction pathway that estrogen response element (ERE) regulates and controls.Experimental result shows, after adding estrogen receptor antagon, positive drug 17 beta estradiol and Semen Lepidii (Semen Descurainiae) water extract the proliferation of MCF-7 cell is disappeared and compared with Normal group not statistically significant.This result prompting Semen Lepidii (Semen Descurainiae) water extract has played estrogen-like effects by classical estrogen receptor pathway.
Reporter gene (reportergene) refers to that its expression product is easily detected, and is easy to the gene that same endogenous background proteins distinguishes.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.Carry in the inducing action experiment of pERE-TAL-luc transient expression at Semen Lepidii (Semen Descurainiae) water extract to reporter gene, by the expression of visual report gene, namely the height of fluorescence intensity can detect whether tested material can 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 Lepidii (Semen Descurainiae) water extract estrogen-like effects and itself and ER α or ER β affinity.Result shows, time alpha mediated by estrogen receptor ER, and 17 beta estradiols (10
-8mol/l) uciferase activity after standardization is significantly higher than Normal group.Uciferase activity after the Semen Lepidii (Semen Descurainiae) water extract standardization of each concentration does not have significant difference compared with Normal group.This shows that Semen Lepidii (Semen Descurainiae) water extract does not play estrogen-like effects by estrogen receptor ER α.Time beta mediated by estrogen receptor ER, 17 beta estradiols (10
-8mol/l) uciferase activity after standardization is significantly higher than Normal group.When Semen Lepidii (Semen Descurainiae) water extract concentration is 0.05mg/ml, the uciferase activity after standardization also has significant significant difference compared with Normal group.This result represents that Semen Lepidii (Semen Descurainiae) water extract plays estrogen-like effects by estrogen receptor ER β.
Traditionally, Semen Lepidii (Semen Descurainiae) is applied to clinical as a relieving cough and asthma, edema-alleviating diuretic; The Late Cambrian Semen Lepidii (Semen Descurainiae) water extracts such as the Reporter Gene Experiments that this research is regulated and controled by Mouse Uterus weightening finish experiment, external MCF-7 cell proliferation antagonistic experiment, ERE have estrogen-like effects, and are played a role by estrogen receptor ER β.This shows not only the popularity of Chinese medicine effect, also be that the various diseases (as osteoporosis, climacteric syndrome) solved because body inner estrogen hyposecretion causes provide better medication reference clinically, thus improve prescription curative effect when drug matching.
Claims (5)
1. Semen Lepidii (Semen Descurainiae) water extract is preparing the application in estrogens medicine.
2. Semen Lepidii (Semen Descurainiae) water extract according to claim 1 is preparing the application in estrogens medicine, it is characterized in that: the application of described Semen Lepidii (Semen Descurainiae) water extract in the Uterine coefficient increasing sex immature female mice.
3. Semen Lepidii (Semen Descurainiae) water extract according to claim 1 is preparing the application in estrogens medicine, it is characterized in that: described Semen Lepidii (Semen Descurainiae) water extract is to the application of MCF-7 cell proliferation.
4. Semen Lepidii (Semen Descurainiae) water extract according to claim 1 is preparing the application in estrogens medicine, it is characterized in that: described Semen Lepidii (Semen Descurainiae) water extract in vivo, externally have estrogenic activity, play estrogenic activity by activating estrogen receptor ER β.
5. the Semen Lepidii (Semen Descurainiae) water extract according to claim 1 or 2 or 3 is preparing the application in estrogens medicine, it is characterized in that: the preparation method of described Semen Lepidii (Semen Descurainiae) water extract is: Semen Lepidii (Semen Descurainiae) is with the soak by water 3 times of 10 times amount, each 1h, first time is when decocting, first invade bubble 30 minutes, merge 3 decocting liquid, concentrating under reduced pressure is dry, obtains Semen Lepidii (Semen Descurainiae) water extract.
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| CN107056644A (en) * | 2017-05-26 | 2017-08-18 | 河南中医药大学 | A kind of method and its application that 2 phenyl acetamide compounds are extracted from lepidii,semen |
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| CN103006741A (en) * | 2012-11-19 | 2013-04-03 | 江苏大学 | Semen lepidii anti-tumor effective component extract, as well as preparation method and application thereof |
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