CN107837311A - Application of the red dragon analgesia piece in neurocyte protection preparation is prepared - Google Patents
Application of the red dragon analgesia piece in neurocyte protection preparation is prepared Download PDFInfo
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Abstract
The invention discloses application of the red dragon analgesia piece in neurocyte protection preparation is prepared; including forming application of the extract prepared in conventional manner in neurocyte protection preparation is prepared according to red dragon analgesia slice prescription, its mechanism of action is removing free radical, reduces NO or NF κ B inflammation.Instant invention overcomes technology prejudice; it is proposed that red dragon analgesia piece and its extract have neuroprotection first; damage by nerve cell caused by removing free radical, reducing a large amount of NO etc.; improve Neuronal Survival rate; and the expression of NF κ B in ischemical reperfusion injury nerve cell can be significantly inhibited, so as to suppress damaging action of the inflammatory reaction to nerve cell.
Description
Technical field
The present invention relates to application of the red dragon analgesia piece in neurocyte protection preparation is prepared, belong to pharmaceutical technology field.
Background technology
Red dragon analgesia piece is Tibetan medicine's Gu proved recipe, is recorded in Tibetan medicine ministry standard, standard number：WS-10659(ZD-0659)-
2002-2011Z, by keel 112.0g, Indian Herba Swertiae bimaculatae 67.0g, safflower 67.0g, Chrysosplenium nudicaule Bunge 14.0g, west szechwan pyrethrum
67.0g, Aconitum Szechenyianum Gay 14.0g, Tangut aconite 67.0g, bear gall powder 6.6g, sinter 13.5g, Semen Herpetospermi 13.5g, RADIX PRZEWALSKIAE TANGUTICAE
13.5g, parmelia saxatilis 13.5g, Lagotis brachystachya Maxim 67.0g, styrax 13.0g, mother-of-pearl 35.0g compositions, have consciousness regaining, dredging collateral
The effect of analgesic, for the antimigraine caused by blood stagnant in cerebral venation syndrome, angioneurotic headache.
The clinical antimigraine caused by blood stagnant in cerebral venation syndrome of red dragon analgesia piece, angioneurotic headache effect is preferable, at present
Document without specific pharmacology study mechanism, analyze its formula speculate the main pharmacological mechanism of its analgesic activity for improve blood supply and
Anti-inflammatory and analgesic effect.Existing literature does not find that red dragon analgesia piece is directed to the research of neurocyte protection effect.
The content of the invention
It is an object of the invention to provide application of the red dragon analgesia piece in neurocyte protection preparation is prepared.
Technical scheme is as follows：
Application of the red dragon analgesia piece in neurocyte protection preparation is prepared.
Preferably, the red dragon analgesia piece is removing free radical, reduces NO to the mechanism of action of neurocyte protection preparation
Or NF- κ B inflammatory reaction mechanism.
Preferably, red dragon analgesia piece includes the extract prepared in conventional manner according to red dragon analgesia slice prescription composition.
Preferably, said extracted thing includes the one or more in ethyl acetate extract, ethanol extract, water extract.
Wherein, each extract is prepared as follows
Ethyl acetate extract：Medicinal material is taken in red dragon analgesia formula herbs and its ratio, in addition to bear gall powder, styrax,
The ethyl acetate that remaining medicinal material adds 6-10 times of w/v extracts 2 times, each 2h, and it is dense that acetic acid ethyl acetate extract merges decompression
Contract and continue drying, add bear gall powder, styrax and it is well mixed after, obtain ethyl acetate extract；
Ethanol extract：Medicinal material is taken in red dragon analgesia formula herbs and its ratio, in addition to bear gall powder, styrax, remaining
The ethanol that medicinal material adds the volumetric concentration 70% of 6-10 times of w/v extracts 2 times, first time 2h, second of 1h, merges two
Secondary ethanol extract, it is concentrated under reduced pressure after filtration and continues drying, obtain ethanol extract；
Water extract：Medicinal material is taken in red dragon analgesia formula herbs and its ratio, in addition to bear gall powder, styrax, remaining medicinal material
Add 6-10 times of water of w/v to extract 2 times, first time 2h, second of 1.5h, merge Aqueous extracts twice, depressurized after filtration dense
Contract and continue drying, obtain water extract.
The relation of parts by weight and parts by volume of the present invention is g/ml or kg/L.
The present invention compared with prior art, has the beneficial effect that：
1st, instant invention overcomes prior art prejudice, propose that red dragon analgesia piece and its extract have neuroprotection work first
With.
2nd, the neuroprotection of red dragon analgesia piece mainly passes through nerve cell caused by removing free radical, reducing a large amount of NO
Damage etc., Neuronal Survival rate is improved, and the expression of NF- κ B in ischemical reperfusion injury nerve cell can be significantly inhibited, from
And suppress damaging action of the inflammatory reaction to nerve cell.
Embodiment
Following experimental examples and embodiment are used to further illustrate but be not limited to the present invention.
Experimental example 1：Red dragon analgesia piece causes the protective effect after PC12 cell line chemical damages to glutamic acid
First, experiment material
1st, sample and its preparation
Cell line：PC12 cell lines (source：Rattus norvegicus Adrenal Pheochromocytomas, Nanjing Keygen Biotech
Development in science and technology Co., Ltd)；
Reagent：H-DMEM cell culture mediums (Hyclone, match silent winged generation that biochemistry product Beijing Co., Ltd)；Tire
Cow's serum (Israel Bioind)；Pancreatin cell dissociation buffer (the green skies), penicillin streptomycin mixed liquor is dual anti-, cisplatin for injection
(Qilu Pharmaceutical Co., Ltd.), citrate buffer (Beijing Bioisystech Co., Ltd of Zhong Shan Golden Bridge), paraformaldehyde
(solarbio), Pidolidone (Klonetech, Japan), compound danshen dripping pills (Tianjin Tasly Pharmaceutical Co., Ltd,
20131202)
Red dragon analgesia piece each sample referred to as control is as follows：
Red dragon analgesia piece (Qinghai gold scolds Tibetan medicine, lot number 20141022)
Red dragon analgesia piece ethyl acetate extract (ester is carried, and embodiment 1 is made by oneself)
Red dragon analgesia piece ethanol extract (alcohol extracting, embodiment 1 are made by oneself)
Red dragon analgesia piece water extract (water extraction, embodiment 1 are made by oneself)
The red mesh of dragon analgesia piece micro mist 300 (micro mist, embodiment 1 are made by oneself)
2nd, laboratory apparatus
Vertical pressure steam sterilization pan (LDZX-50FBS, Shanghai Shen peace)；Double one side clean work station (SW-CJ-1C,
Suzhou purifies)；CO2gas incubator (BB16 μ V/BB5060 μ V, HERAE US)；Desk centrifuge (H3, Sigma)；Enzyme mark
Instrument (Multiskan MK3, Thermo Scientific)；(101, Shanghai roc has electric heating constant-temperature blowing drying box along scientific instrument
Limit company)；Inverted microscope (XDS-1B, Chongqing optical instrument factory)；Water-bath constant temperature oscillator (SHZ-82, Jintan City's medical instrument
Device factory)；Pipettor (Gilson, Eppendorf)；Blake bottle (Corning)；96 orifice plates (Costar, USA).
2nd, experimental method
PC12 cytochemistry damage models are prepared with glutamic acid, absorption is determined at 570nm using tetrazolium bromide (MTT) method
Value, repair of the observation drug candidate to PC12 cellular damages.Each index is detected using SOD, NO, LDH, MDA kit to contain
Amount, illustrate the mechanism of action of each medicine.
Modeling：It is thin that 30mmol/l glutamic acid (this is sample-adding final concentration, is dissolved in incomplete culture medium) acts on PC12
Born of the same parents 24h.
Packet：Blank group, chemical damage model group, chemical damage+medicine group, chemical damage+positive control drug group.Because of 96
Orifice plate number limits, and cultivates detection simultaneously using two plates when sample size is larger, sets model group blank group per plate and the positive is right
According to group, to ensure the reliability of result and uniformity.
3rd, experiment content and step
1st, cell culture
The PC12 cell recoveries frozen are taken from ultra low temperature freezer in blake bottle, with the H-DMEM containing 10% hyclone
Medium culture.When PC12 cell growths to 80% fusion, with 0.25% pancreatin cell dissociation buffer (containing 0.02%EDTA)
Digested, be inoculated in 1 × 104 cell per well in 96 orifice plates, zeroing hole is not added with cell.
2nd, cellular damage
After the cell in 96 orifice plates covers with individual layer, culture medium is discarded, sterile PBS liquid cleans 2 times, except zeroing group, blank
Outside control group, final concentration of 30mmol/l glutamic acid (being dissolved in incomplete culture medium) is added per hole in being cultivated in incubator
24h, that is, cause PC12 cellular damage models.
3rd, dosing reparation
Culture medium is discarded, is cleaned 2 times with sterile PBS liquid, administration group is separately added into the final concentration of the medicine containing screening per hole successively
For 100 μ g/ml, 10 μ g/ml, 1 μ g/ml the μ l of DMEM culture mediums 100, the medicine of each concentration adds 6 holes.With not drug containing
DMEM culture mediums are blank control group, and positive controls add the plasma-free DMEM medium of the compound danshen dripping pills of various concentrations
100μl.Then cell inserted 37 DEG C, continue in 5%CO2 incubators to cultivate 24h.
4th, index determining
4.1MTT methods determine cell survival rate
100 μ l plasma-free DMEM mediums are added per hole after discarding culture medium, 5mg/ml MTT solution is added then at every hole
20 μ l react 4h in lucifuge in incubator.Discard culture medium and add 100 μ l DMSO ELIASAs in 570nm measure OD values, calculate
Cell survival rate：
Calculation formula：Cell survival rate (%)=(medication group or model group A570/ control group As 570) × 100%.
4.2 nitric oxides (NO), lactic dehydrogenase (LDH), superoxide dismutase (SOD), MDA (MDA) are containing measurement
It is fixed
Cell supernatant is collected with sterile tube within 24 hours after administration, centrifuge 20 minutes (2000r/min), carefully collect supernatant
Liquid, by kit specification time-and-motion study LDH, NO content.
24 hours supernatant discardings, add 2%Trion-100X after administration, and cell suspension is mixed after standing 12 hours in 4 DEG C, from
The heart 20 minutes (2000r/min), carefully collects supernatant, by kit specification time-and-motion study MDA, SOD content.
4th, experimental result
All data represent that one-way analysis of variance and the multigroup means of LSD compare the notable of group difference two-by-two with X ± SD
Property, P<0.05 is significant difference.
1st, influence of the red dragon analgesia piece to the PC12 cell survival rates of damage
The result (n=8) that the red dragon analgesia piece of table 1 influences on the PC12 cell survival rates of damage
Note：* P is represented<0.05
The result of table 1 shows, in final concentration of 1 μ g/ml drug candidate, alcohol extracting, and micro mist, water extraction, the cell survival that ester carries
Rate has significant difference compared with corresponding model group.In the μ g/ml of final concentration 10 drug candidate, ester carries, the cell of micro mist
Survival rate is compared with model group, there is significant difference；Alcohol extracting, the cell survival rate of water extraction compare no significant difference with model group.
Show, the protective effect of liposoluble constituent and red dragon analgesia piece to external PC12 nerve cells is stronger.
2nd, influence of the red dragon analgesia piece to LDH, NO, MDA, SOD content
Influence of the 2.1 red dragon analgesia pieces to the PC12 cell NO contents of in vitro culture
The NO assays result (n=3) of the red dragon each extract of analgesia piece of table 2 and Contained Serum to PC12 cells
Note：*P<0.05
The result of table 2 shows, the μ g/ml of final concentration 1 and 10 μ g/ml alcohol extracting and ester carries and 10 μ g/ml water extraction and mould
Type group has significant difference；It is administered in serum, blood medicine is compared with blank serum group, and there were significant differences.Show each extract and contain
Medicine serum has considerable influence to the NO contents of PC12 cells.
Influence of the 2.2 red dragon analgesia pieces to the PC12 cell LDH contents of in vitro culture
Each extract of the red dragon analgesia piece of table 3 and Contained Serum LDH assays result (n=3)
Note：*P<0.05
The result of table 3 shows that μ g/ml of micro mist 1,10 μ g/ml, μ g/ml of alcohol extracting 10, the μ g/ml of water extraction 10 and Contained Serum can show
Writing reduces the leakage for being damaged LDH in PC12 cell culture fluids.
Influence of the 2.3 red dragon analgesia pieces to the PC12 cell SOD contents of in vitro culture
Each extract of the red dragon analgesia piece of table 4 and Contained Serum SOD assays result (n=3)
Note：*P<0.05
The result of table 4 shows that ester carries 1 μ g/ml, μ g/ml of water extraction 1, ester and carries 10 μ g/ml, the μ g/ml of water extraction 10 and model group ratio
Compared with there is significant difference.Show it is red dragon analgesia piece ester extract and water extract can significantly improve damaged nerve cells SOD it is dense
Degree, there is provided its anti-oxidant left and right.
Influence of the 2.4 red dragon analgesia pieces to the PC12 cell MDA contents of in vitro culture
Each extract of the red dragon analgesia piece of table 5 and Contained Serum MDA assays result (n=3)
Note：*P<0.05
The result of table 5 shows that each group is compared with model group, difference that there are no significant.
3rd, red dragon analgesia piece in-vitro pharmacological experiments research conclusion
The red dragon analgesia each preparation result list of piece of table 6
+ represent that said preparation or extract compared with model group, there is significant statistical significance (P<0.05).
The influence of 3.1 pairs of cell survival rates
Mtt assay measure PC12 cell survival rate results show, in final concentration of 1 μ g/ml drug candidate, alcohol extracting, and micro mist,
Water extraction, the cell survival rate that ester carries have significant difference compared with corresponding model group.In the μ g/ml of final concentration 10 drug candidate
In, alcohol extracting, the cell survival rate of micro mist has significant difference compared with corresponding model group.Positive control drug compound danshen dripping pills
It is administered with 27mg/ml final concentrations, the cell survival rate highest in each group.Other each group unknown significance differences, or significantly
Less than model control group.
The influence of 3.2 pairs of SOD contents
Ester carries 1 μ g/ml, μ g/ml of water extraction 1, ester and carries 10 μ g/ml and the μ g/ml of water extraction 10 compared with model group, there is conspicuousness
Difference.
The influence of 3.3 pairs of MDA contents
Each sample unknown significance difference compared with model group.
The influence of 3.4 pairs of NO contents
The μ g/ml of final concentration 1 and 10 μ g/ml alcohol extractings, ester carry, 10 μ g/ml water extractions and model group have significant difference；Drug containing
Serum is compared with blank serum group, and there were significant differences.
The influence of 3.5 pairs of LDH contents
μ g/ml of micro mist 1,10 μ g/ml, μ g/ml of alcohol extracting 10, the μ g/ml of water extraction 10, and Contained Serum and model group are more equal
There is significant difference.
The synthesis conclusion of 3.6 red dragon analgesia pieces
Mtt assay cytoactive the selection result confirms：Compared with model group, red dragon analgesia piece alcohol extract, Ultramicro-powder, water extract
There is significant repair to PC12 cellular damages caused by glutamic acid under suitable concentration.Compared with model group, Hong Long towns
Pain piece ethyl acetate extract and water extract can significantly increase cell SOD activity.Above medicine is prompted to PC12 caused by glutamic acid
Caused free radical has certain scavenging action after cellular damage.
μ g/ml of micro mist 1,10 μ g/ml, the μ g/ml of water extraction 10, the sample of the μ g/ml concentration of alcohol extracting 10, and Contained Serum can drop
LDH leakage in low nutrient solution, prompt each screening medicine of the above to reduce PC12 damaged nerve cells and hinder degree, to glutamic acid
Caused neural cell injury has certain restoration and protection effect.
Red dragon analgesia piece ethyl acetate extract Contained Serum can substantially reduce NO contents in cell culture fluid, reduce a large amount of
The damage of nerve cell caused by NO.
The protective effect of experimental example 2, red dragon analgesia piece to nerve cell after ischemia-reperfusion
1 materials and methods
1.1 material
(1) animal:The pregnant mouse of pregnant two weeks Kunming kind two level is provided by Shandong University's animal center；
(2) main agents and instrument:Hyclone is provided by Hangzhou Chinese holly company, and DMEM culture mediums are by GIBCO companies
There is provided；Pancreatin is provided by magnificent company；Rabbit-anti mouse NF- κ Bp65 polyclonal antibodies (identification 1-286 amino acid sequences) are by the U.S.
Santa companies provide；Fluorescein isothiocynate (flourescernisothiocyanate, FITC) marks goat anti-rabbit igg by U.S.
Hua companies provide；'beta '-tubulin (β-tubulin) monoclonal antibody is provided by Progma companies；Red dragon analgesia piece scolds Tibetan by gold
Medicine joint-stock company is provided, and its ethanol extract and acetic acid ethyl acetate extract are prepared by embodiment 1；CO2gas incubator by
SANYO companies provide；Flow cytometer is U.S.'s CoulterEliteEPS types.
1.2 method
(1) after taking some sterilizations together of 25mL blake bottles, handled through poly-D-lysine coating；Artificial cerebrospinal fluid is prepared, its
Form and be: NaCl123mmol/L, KCl3.75mmol/L, KH2PO41.25mmol/L, NaHCO326mmol/L,
MgCl21mmol/L, CaCl22mmol/L, glucose 10mmol/L, pH:7.4.
(2) cell culture:The pregnant mouse of pregnant two weeks Kunming kind two level is taken, dislocation is put to death, the tincture of iodine, ethanol disinfection belly, successively cutd open
Skin of abdomen, muscle, peritonaeum are opened, takes out tire mouse, fetal membrane amnion is carefully peeled off, exposure tire mouse, stereoscopic micro- lower stripping meninx, divides
From the cortex of tire mouse brain brain vesicles outside portion, brain tissue is shredded, digests (37 DEG C/15min) through the pancreatin containing 125mg/L, centrifugation
1000r/min, 10min, supernatant is abandoned, the DMEM of 200mL/L hyclones terminates digestion, with complete medium (10mL/L tire oxen
Serum, penicillin and streptomycin 100U/mL) it is diluted to 7 × 10^5/mL density and is inoculated in the blake bottle treated through poly-D-lysine,
50mL/LCO2 incubator is sent into, after 37 DEG C are cultivated 48h, adds cytarabine (5 μ g/mL) to suppress the continuation of non-neuron
Growth, measured every 2d half and change liquid, after cultivating 10d, it is seen that typical nerve cell, it is glimmering through β-tubulin monoclonal antibody immunities
Light dyeing shows more than 80% cell for the positive, it is believed that is pure neuronal cell cultures, carries out subsequent experimental research.
(3) class ischemia model is established:Class ischemic mould is established according to OGD/R (missing of sugar and other nutriments) method
Type, cell culture medium are changed to the artificial cerebrospinal fluid without sugar, and temperature is maintained at (30 ± 2) DEG C, and mixed gas is changed to containing 80mL/
LCO2 nitrogen.Class ischemic preconditioning 10min, artificial cerebrospinal fluid is replaced by culture medium and puts back to incubator row reoxygenation again.
(4) flow cytometry count detection:The primary neural cell being inoculated in blake bottle (1 × 10^6 bottles) is divided at random
For 4 groups, one group is non-ischemia group, and another group of reoxygenation 1/2h again after class ischemic preconditioning, is Ischemia Reperfusion group, third and fourth group is
I group and II group for the treatment of are treated, red dragon analgesia piece ethanol extract (I group for the treatment of) and acetic acid second are separately added into after class ischemic preconditioning
Ester extract solution (II group for the treatment of) pretreatment 10min, two of which extract add coordinative solvent dilute with the extract of embodiment 1 respectively
It is interpreted into tablet containing 1mg/ml liquid.Every group of 4 parts of sample.4 groups are washed 5min with 0.01moL/LPBS respectively, totally 3 times, with 0.25%
Cell dissociation into single cell suspension, is added the μ L of 1: 200NF- κ Bp65 polyclonal antibodies 200 by trypsase after centrifuge washing,
37 DEG C are put, 45min, is washed with 0.01mol/LPBS, totally 3 times, supernatant is abandoned in centrifugation, adds the anti-rabbit IgG200 of 1: 50FITC mark
μ L, 4 DEG C are put, 45min, is washed with 0.01mol/LPBS.Another to set two groups of negative control groups, one group is not added with that I is anti-and II is anti-, for glimmering
Luminous intensity baseline correction；Another group of list adds II to resist, and as non-specific binding control, all samples are separately added into 400 μ L,
Machine testing on 0.01mol/LPBS liquid, every part of sample count more than 5000 cells, as a result take average.
1.3 statistical method
As a result represented with x ± s, data compares with variance analysis and t inspections between group.
2 results
Flow cytomery result NF- Κ b/P65 in non-ischemia group neuron basal expression for (1.38 ±
0.56) %, the 1/2h expression significantly rise (P ＜ 0.01) after ischemia-reperfusion.Through red dragon analgesia 2 kinds of extract pretreatments of piece
Afterwards, expression rate is remarkably decreased (P ＜ 0.05), and compared to there is significant difference (P between four groups<0.05).Wherein, Ischemia Reperfusion
Note group has pole significant difference (P compared with treating II group<0.01).Show ethyl acetate extract to ischemical reperfusion injury mould
The protective effect of type is stronger.
The comparison of NF- κ B expression rates between each group of table 1
Note：Compared with non-ischemia group, ##P ＜ 0.01；Compared with ischemia-reperfusion group, * P ＜ 0.05, * * P ＜ 0.01
3 brief summaries
NF- κ B are present in cerebrovascular endothelial cell, nerve cell and spongiocyte, and its expression is bright after cerebral ischemia re-pouring
It is aobvious to increase and closely related with the inflammation mechanism of cerebral ischemia.This experiment investigates red dragon analgesia piece intervention by index of NF- κ B
It is cerebral ischemia re-pouring injured to treat mouse, it is found that it significantly reduces effect in cerebral ischemia perfusion injury to NF- κ B expression, so as to
Reduce damaging action of the inflammation mechanism to ischemia-reperfusion to nerve cell.
Following examples are used to further illustrate the present invention.
The preparation of embodiment 1, red dragon analgesia piece each group extract
1) preparation of red dragon analgesia piece ethyl acetate extract：
Taken medicinal material in red dragon analgesia formula herbs and its ratio and amounted to 292g, crushed.In addition to bear gall powder, styrax, its
Remaining medicinal material extract 2 times, respectively plus 2250ml ethyl acetate extraction 2h, acetic acid ethyl acetate extract, which merges, is concentrated under reduced pressure and continues drying,
Add bear gall powder, styrax and it is well mixed after, obtain red dragon analgesia piece ethyl acetate extract 2.11g.
2) preparation of red dragon analgesia piece ethanol extract：
Taken medicinal material in red dragon analgesia formula herbs and its ratio and amounted to 292g, crushed.In addition to bear gall powder, styrax, its
Remaining medicinal material adds the ethanol of 2250ml volume ratios 70% and extracted 2 times, first time 2h, second of 1h, merges ethanol extract twice, filter
Later it is concentrated under reduced pressure and continues drying, obtains red dragon analgesia piece ethanol extract 11.78g.
3) preparation of red dragon analgesia piece water extract：
Taken medicinal material in red dragon analgesia formula herbs and its ratio and amounted to 292g, crushed.In addition to bear gall powder, styrax, its
Remaining medicinal material adds 2820ml water and extracted 2 times, first time 2h, second of 1.5h, merges Aqueous extracts twice, is concentrated under reduced pressure simultaneously after filtration
Continue drying, obtain red dragon analgesia piece water extract 17.84g.
4) red dragon analgesia piece micro mist：Red dragon analgesia piece is ground into fine powder.
5) preparation of Contained Serum：
Red dragon analgesia piece ethanol extract is taken, Cmax aqueous suspension, oral feeding mouse 5 is made, dosage is
0.0236g/g body weight, wins eyeball after 30min, and not anti-freezing takes blood, and it is standby that 3000rpm centrifuges 15 minutes separation serum, as sample
Product (blood medicine).
The application of embodiment 2, red dragon analgesia piece in neurocyte protection preparation is prepared.
The application of embodiment 3, red dragon analgesia piece in neurocyte protection preparation is prepared, mechanism of action are free to remove
Base, reduce NO or NF- κ B inflammatory reaction mechanism.
The application of embodiment 4, red dragon analgesia piece ethyl acetate extract in neurocyte protection preparation is prepared, its acetic acid
Ethyl ester method for preparing extractive is the same as embodiment 1.
The application of embodiment 5, red dragon analgesia piece ethanol extract in neurocyte protection preparation is prepared, the extraction of its ethanol
Thing preparation method is the same as embodiment 1.
The application of embodiment 6, red dragon analgesia piece water extract in neurocyte protection preparation is prepared, its water extract system
Preparation Method is the same as embodiment 1.
The application of embodiment 7, red dragon analgesia piece extractive composition in neurocyte protection preparation is prepared, extract group
Compound includes ethyl acetate extract and ethanol extract, and method for preparing extractive is the same as embodiment 1.
The application of embodiment 8, red dragon analgesia piece extractive composition in neurocyte protection preparation is prepared, extract group
Compound includes ethyl acetate extract and water extract, and method for preparing extractive is the same as embodiment 1.
The application of embodiment 9, red dragon analgesia piece extractive composition in neurocyte protection preparation is prepared, extract group
Compound includes water extract and ethanol extract, and method for preparing extractive is the same as embodiment 1.
The application of embodiment 10, red dragon analgesia piece extractive composition in neurocyte protection preparation is prepared, extract
Composition includes ethyl acetate extract, ethanol extract and water extract, and method for preparing extractive is the same as embodiment 1.
Medicine prepared by above-described embodiment has reached experimental example 1 and the effect described in experimental example 2.
Claims (7)
1. application of the red dragon analgesia piece in neurocyte protection preparation is prepared.
2. application of the red dragon analgesia piece according to claim 1 in neurocyte protection preparation is prepared, it is characterised in that
The mechanism of action of the neurocyte protection preparation is removing free radical, reduces NO or NF- κ B inflammatory reaction mechanism.
3. application of the red dragon analgesia piece according to claim 1 or 2 in neurocyte protection preparation is prepared, its feature exist
In described red dragon analgesia piece includes the extract prepared in conventional manner according to red dragon analgesia slice prescription composition.
4. application of the red dragon analgesia piece according to claim 3 in neurocyte protection preparation is prepared, it is characterised in that
Described extract includes the one or more in ethyl acetate extract, ethanol extract, water extract.
5. application of the red dragon analgesia piece according to claim 4 in neurocyte protection preparation is prepared, it is characterised in that
The preparation method of described ethyl acetate extract is as follows：
Medicinal material is taken in red dragon analgesia formula herbs and its ratio, in addition to bear gall powder, styrax, remaining medicinal material adds bulking value
Than g/ml 6-10 times ethyl acetate extract 2 times, each 2h, and acetic acid ethyl acetate extract, which merges, to be concentrated under reduced pressure and continue drying, adds
Bear gall powder, styrax and it is well mixed after, obtain ethyl acetate extract.
6. application of the red dragon analgesia piece according to claim 4 in neurocyte protection preparation is prepared, it is characterised in that
The preparation method of described ethanol extract is as follows：
Medicinal material is taken in red dragon analgesia formula herbs and its ratio, in addition to bear gall powder, styrax, remaining medicinal material adds bulking value
The ethanol of volumetric concentration 70% than g/ml 6-10 times extracts 2 times, first time 2h, second of 1h, merges ethanol extract twice,
It is concentrated under reduced pressure after filtration and continues drying, obtains ethanol extract.
7. application of the red dragon analgesia piece according to claim 4 in neurocyte protection preparation is prepared, it is characterised in that
The preparation method of described water extract is as follows：
Medicinal material is taken in red dragon analgesia formula herbs and its ratio, in addition to bear gall powder, styrax, remaining medicinal material adds bulking value
Than g/ml 6-10 times water extract 2 times, first time 2h, second of 1.5h, merge Aqueous extracts twice, be concentrated under reduced pressure after filtration and after
Continuous drying, obtains water extract.
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Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102631467A (en) * | 2012-05-14 | 2012-08-15 | 山东阿如拉药物研究开发有限公司 | Honglong antalgic medicinal composition, and preparation method and application thereof |
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2016
- 2016-09-20 CN CN201610834515.0A patent/CN107837311B/en active Active
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102631467A (en) * | 2012-05-14 | 2012-08-15 | 山东阿如拉药物研究开发有限公司 | Honglong antalgic medicinal composition, and preparation method and application thereof |
Non-Patent Citations (3)
| Title |
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| 李光华等: "龙骨对小鼠抗坐骨神经损伤及单核细胞吞噬廓清率的观察", 《宁夏医学院学报》 * |
| 柏蕙英等: "红花对鼠视网膜神经细胞损伤的保护作用", 《上海第二医科大学学报》 * |
| 王品等: "脑脉舒胶囊的药效学试验研究", 《齐鲁药事》 * |
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