CN103656016B - A kind of preparation method of antiasthmatic pill of perilla fruit and application - Google Patents
A kind of preparation method of antiasthmatic pill of perilla fruit and application Download PDFInfo
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- CN103656016B CN103656016B CN201310712487.1A CN201310712487A CN103656016B CN 103656016 B CN103656016 B CN 103656016B CN 201310712487 A CN201310712487 A CN 201310712487A CN 103656016 B CN103656016 B CN 103656016B
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Abstract
The invention provides a kind of preparation method of antiasthmatic pill of perilla fruit, by Fructus Perillae (parched) 160g, Cortex Magnoliae Officinalis 160g, Radix Peucedani 160g, Radix Glycyrrhizae 160g, Rhizoma Pinelliae (processed with Rhizoma Zingiberis Recens) 160g, Pericarpium Citri Reticulatae 160g, Lignum Aquilariae Resinatum 112g, Radix Angelicae Sinensis 112g as crude drug, employing supercritical extraction is prepared from, content is improved a lot, dose reduces, and present invention also offers the application of antiasthmatic pill of perilla fruit in preparation suppression human tissue cell lymphoma U937 cell proliferation.
Description
Technical field
The present invention relates to technical field of traditional Chinese medicine preparation, be specifically related to a kind of preparation method and application of antiasthmatic pill of perilla fruit.
Background technology
Antiasthmatic pill of perilla fruit is recorded in Ministry of Public Health standard WS3-B-0278-90, prescription is Fructus Perillae (parched) 160g, Cortex Magnoliae Officinalis 160g, Radix Peucedani 160g, Radix Glycyrrhizae 160g, Rhizoma Pinelliae (processed with Rhizoma Zingiberis Recens) 160g, Pericarpium Citri Reticulatae 160g, Lignum Aquilariae Resinatum 112g, Radix Angelicae Sinensis 112g, above eight tastes, except Fructus Perillae, all the other wait seven tastes to be ground into fine powder, grind with Folium Perillae substring again, sieve, mixing; The general ball of juice is decocted with Rhizoma Zingiberis Recens 40g, Fructus Jujubae 80g; Cold drying obtains 1100g, to obtain final product.Energy lowering the adverse-rising QI to resolve phlegm, the kidney warming improving inspiration by invigorating kidney-QI.Stop up for QI rising in reverse order expectorant, cough is panted, septum pectorale painful abdominal mass plug.
In prior art, not yet have antiasthmatic pill of perilla fruit extracting the report adopting supercritical technology in preparation, and adopt the method for beating powder, technique is coarse, backward, and impurity is many, causes patient's consumption excessive, is inconvenient to take, and has had a strong impact on this product and has applied clinically.
Summary of the invention
Goal of the invention: in order to solve the problem, the object of the present invention is to provide a kind of preparation method of antiasthmatic pill of perilla fruit.
Another object of the present invention is to provide a kind of antiasthmatic pill of perilla fruit to suppress the application in human tissue cell lymphoma U937 cell proliferation in preparation.
Technical scheme: the object of the invention is by following scheme realize:
A kind of preparation method of antiasthmatic pill of perilla fruit, by Fructus Perillae (parched) 160g, Cortex Magnoliae Officinalis 160g, Radix Peucedani 160g, Radix Glycyrrhizae 160g, Rhizoma Pinelliae (processed with Rhizoma Zingiberis Recens) 160g, Pericarpium Citri Reticulatae 160g, Lignum Aquilariae Resinatum 112g, Radix Angelicae Sinensis 112g makes as crude drug, described method is made up of the following step: get Fructus Perillae (parched) 160g, Cortex Magnoliae Officinalis 160g, Radix Peucedani 160g, Radix Glycyrrhizae 160g, Rhizoma Pinelliae (processed with Rhizoma Zingiberis Recens) 160g, Pericarpium Citri Reticulatae 160g, Lignum Aquilariae Resinatum 112g, Radix Angelicae Sinensis 112g, join in CO2 supercritical extraction device, ethanol is as entrainer, the percent by volume that entrainer accounts for total extractant is 4-6%, extracting pressure 15-30MPa, temperature 30-50 DEG C, CO
2flow 1-3m1/g crude drug min, extraction time 150-180min, obtains supercritical extract, is ground into fine powder, and water pill obtains 550g.
The preparation method of above-mentioned a kind of antiasthmatic pill of perilla fruit, described CO
2the percent by volume that supercritical extraction entrainer accounts for total extractant is 5%.
The preparation method of above-mentioned a kind of antiasthmatic pill of perilla fruit, described CO
2the extracting pressure 20MPa of supercritical extraction, temperature 40 DEG C, CO
2flow 2ml/g crude drug min, extraction time 160min.
Above-mentioned a kind of antiasthmatic pill of perilla fruit suppresses the application in human tissue cell lymphoma U937 cell proliferation in preparation, antiasthmatic pill of perilla fruit is made up as crude drug of Fructus Perillae (parched) 160g, Cortex Magnoliae Officinalis 160g, Radix Peucedani 160g, Radix Glycyrrhizae 160g, Rhizoma Pinelliae (processed with Rhizoma Zingiberis Recens) 160g, Pericarpium Citri Reticulatae 160g, Lignum Aquilariae Resinatum 112g, Radix Angelicae Sinensis 112g, preparation method is made up of the following step: get Fructus Perillae (parched) 160g, Cortex Magnoliae Officinalis 160g, Radix Peucedani 160g, Radix Glycyrrhizae 160g, Rhizoma Pinelliae (processed with Rhizoma Zingiberis Recens) 160g, Pericarpium Citri Reticulatae 160g, Lignum Aquilariae Resinatum 112g, Radix Angelicae Sinensis 112g, join CO
2in supercritical extraction device, ethanol is as entrainer, and the percent by volume that entrainer accounts for total extractant is 4-6%, extracting pressure 15-30MPa, temperature 30-50 DEG C, CO
2flow 1-3m1/g crude drug min, extraction time 150-180min, obtains supercritical extract, is ground into fine powder, and water pill obtains 550g.
Above-mentioned antiasthmatic pill of perilla fruit suppresses the application in human tissue cell lymphoma U937 cell proliferation in preparation, CO described in antiasthmatic pill of perilla fruit preparation method
2the percent by volume that supercritical extraction entrainer accounts for total extractant is 5%.
Above-mentioned antiasthmatic pill of perilla fruit suppresses the application in human tissue cell lymphoma U937 cell proliferation in preparation, CO described in antiasthmatic pill of perilla fruit preparation method
2the extracting pressure 20MPa of supercritical extraction, temperature 40 DEG C, CO
2flow 2ml/g crude drug min, extraction time 160min.
In prior art, by former explained hereafter, above-mentioned recipe quantity medical material obtains antiasthmatic pill of perilla fruit 1100g, every 13 heavy 1g, each 6g, 1-2 time on the one, adopt the antiasthmatic pill of perilla fruit 550g that the present invention is prepared into, every 13 heavy 1g, but the medical material amount contained is original 2 times, therefore each 3g, 1-2 time on the one, greatly reduces dose under the condition with more active component.
Detailed description of the invention
Form by the following examples, foregoing of the present invention is described in further detail again, but this should be interpreted as that the scope of the above-mentioned theme of the present invention is only limitted to following example, all technology realized based on foregoing of the present invention all belong to scope of the present invention.
Embodiment 1
Get Fructus Perillae (parched) 160g, Cortex Magnoliae Officinalis 160g, Radix Peucedani 160g, Radix Glycyrrhizae 160g, Rhizoma Pinelliae (processed with Rhizoma Zingiberis Recens) 160g, Pericarpium Citri Reticulatae 160g, Lignum Aquilariae Resinatum 112g, Radix Angelicae Sinensis 112g join CO
2in supercritical extraction device, ethanol is as entrainer, and the percent by volume that entrainer accounts for total extractant is 4%, extracting pressure 15MPa, temperature 30 DEG C, CO
2flow 1m1/g crude drug min, extraction time 150min, obtains supercritical extract, is ground into fine powder, and water pill obtains 550g.
Embodiment 2
Get Fructus Perillae (parched) 160g, Cortex Magnoliae Officinalis 160g, Radix Peucedani 160g, Radix Glycyrrhizae 160g, Rhizoma Pinelliae (processed with Rhizoma Zingiberis Recens) 160g, Pericarpium Citri Reticulatae 160g, Lignum Aquilariae Resinatum 112g, Radix Angelicae Sinensis 112g join CO
2in supercritical extraction device, ethanol is as entrainer, and the percent by volume that entrainer accounts for total extractant is 6%, extracting pressure 30MPa, temperature 50 C, CO
2flow 3m1/g crude drug min, extraction time 180min, obtains supercritical extract, is ground into fine powder, and water pill obtains 550g.
Embodiment 3
Get Fructus Perillae (parched) 160g, Cortex Magnoliae Officinalis 160g, Radix Peucedani 160g, Radix Glycyrrhizae 160g, Rhizoma Pinelliae (processed with Rhizoma Zingiberis Recens) 160g, Pericarpium Citri Reticulatae 160g, Lignum Aquilariae Resinatum 112g, Radix Angelicae Sinensis 112g join CO
2in supercritical extraction device, ethanol is as entrainer, and the percent by volume that entrainer accounts for total extractant is 5%, extracting pressure 20MPa, temperature 40 DEG C, CO
2flow 2m1/g crude drug min, extraction time 160min, obtains supercritical extract, is ground into fine powder, and water pill obtains 550g.
Embodiment 4: antiasthmatic pill of perilla fruit suppresses the experimentation data of human tissue cell lymphoma U937 cell proliferation
1 experiment material
1.1 experiment cell strains
Human tissue cell lymphoma U937 cell, Nanjing Medical University's laboratory cell storehouse, DMEM+10%FBS cellar culture.
1.2 Experimental agents
Drugs: antiasthmatic pill of perilla fruit of the present invention: prepare by embodiment 3 method.
Medicinal liquid liquid storage: take 100mg antiasthmatic pill of perilla fruit, is dissolved in 5ml dehydrated alcohol, 0.2 μm of frit, 500 μ ldoff pipe subpackages ,-20 DEG C of storages, and simultaneously 0.2 μm of frit dehydrated alcohol is in order to the use of matched group.
1.3 experiment reagent
DMEM(GIBCO company Cat.No.12100-061Lot.No.758137); Hyclone (Tian Hang bio tech ltd, Zhejiang Lot.No.100419); NaHCO3(Shanghai hundred million chemical reagent company limited Cat.No.11810-033Lot.No.1088387 of a specified duration); Trypsin(AMRESCO company lot number: 2010/04); EDTA(AMRESCO company lot number: 2009/10); PenicillinGSodiumSalt(AMRESCO company lot number: 2010242); StreptomycinSulfate(AMRESCO company lot number: 2010382); Dehydrated alcohol (Nanjing Chemistry Reagent Co., Ltd.'s lot number: 080310182); MTT (Biosharp lot number: 0793); PBS(laboratory autogamy); 1.4 experiment equipment
Lycra inverted microscope (German Leica model: DM1L); Visible-ultraviolet light microwell plate detector (MD company of U.S. model: SPECTRAMAX190); CO2 incubator (FORMA model: 3111); Super-clean bench (safe and sound Inc. of Su Jing group moulding number: SW-CJ-ZFD); Pure water instrument (Spring company of U.S. model: S/N020579); Accurate pipettor (French Gilson Inc model: P2); Electronic balance (German Sai Duolisi company limited model: BT323S); Full-automatic high-pressure autoclave (Japanese SANYO company model: MLS-3020); Table electrothermal air dry oven (the accurate experimental facilities company in Shanghai model: DHG9123A); Refrigerator (Siemens Company's model: KG18V21TI); Liquid nitrogen container (CBS model: 2001); Low speed centrifuge (Anting Scientific Instrument Factory, Shanghai's model: KA-1000); 0.2 μm of filter (MILLIPORE model: SLGP033RB); 10cm culture dish (NEST company), 96 well culture plates (NEST company); Cell counting count board; Centrifuge tube, pipet, Tips are some.
2 experimental techniques
1) human tissue cell lymphoma U937 cell DMEM+10%FBS in 37 DEG C, 5%CO2 carries out cellar culture (10cm culture dish), when Growth of Cells is to logarithmic (log) phase, collecting cell, discards culture fluid, PBS fine laundering 3 times, add 3ml0.25% trypsin-0.04%EDTA, after 37 DEG C of digestion 2min, add 5ml complete medium neutralization reaction wherein, proceeded in centrifuge tube after piping and druming cell, the centrifugal 5min of 1000rpm, adjustment concentration of cell suspension 3 × 104/ml.
2) enter in 96 well culture plates by cell kind, every hole adds cell suspension 180 μ l, and culture plate puts into cell culture incubator (37 DEG C, 5%CO2) cellar culture.
3) according to cell growth status, generally grow to 50%-70%, add antiasthmatic pill of perilla fruit solution, continue to cultivate 24h.
4) add 20 μ lMTT solution (5mg/ml, i.e. 0.5%MTT) after 24h, continue to cultivate 4h.
5) after 4h, buckle method removes supernatant, pats dry gently with absorbent paper, and every hole adds 200 μ l dimethyl sulfoxide, puts low-speed oscillation 10min on shaking table, crystal is fully dissolved.The light absorption value in each hole is measured at enzyme-linked immunosorbent assay instrument 490nm place.
6) background (do not add cell, only add culture fluid) is set, control wells (the medicine dissolution medium of cell, same concentrations, culture fluid, MTT, dimethyl sulfoxide) simultaneously, often organizes the multiple hole of setting 6.
7) result represents with the suppression ratio of medicine to cell:
Cell proliferation suppression ratio (%)=(control wells OD value-dosing holes OD value)/control wells OD value × 100%.Experiment repetition 3 times.
3 statistical dispositions
Adopt the correlation analysis in MicrosoftExcel2003 software and Studentt inspection, data represent with mean ± S.D..
4 experimental results
Statistical result showed after mtt assay experiment, compare with matched group, when dosage reaches 5mg/ml, to human tissue cell's lymphoma U937 cell inhibitory effect variant (P<0.05), dosage this difference when 10mg/ml has significance (P<0.01), has pole significant difference (P<0.001) when dosage reaches 15-20mg/ml.
Table 1 antiasthmatic pill of perilla fruit is to human tissue cell lymphoma U937 cell inhibitory effect influence research (X ± SD)
Note: compare with matched group, * P<0.01; * P<0.001
5 experiment conclusion
Antiasthmatic pill of perilla fruit can suppress human tissue cell lymphoma U937 cell proliferation, and reduce the Growth of Cells number of human tissue cell lymphoma U937 cell, this effect is dose dependent.
Claims (3)
1. the application of antiasthmatic pill of perilla fruit in preparation suppression human tissue cell lymphoma U937 cell proliferation, it is characterized in that antiasthmatic pill of perilla fruit is made up as crude drug of Fructus Perillae (parched) 160g, Cortex Magnoliae Officinalis 160g, Radix Peucedani 160g, Radix Glycyrrhizae 160g, Rhizoma Pinelliae (processed with Rhizoma Zingiberis Recens) 160g, Pericarpium Citri Reticulatae 160g, Lignum Aquilariae Resinatum 112g, Radix Angelicae Sinensis 112g, it is characterized in that described method is made up of the following step: get Fructus Perillae (parched) 160g, Cortex Magnoliae Officinalis 160g, Radix Peucedani 160g, Radix Glycyrrhizae 160g, Rhizoma Pinelliae (processed with Rhizoma Zingiberis Recens) 160g, Pericarpium Citri Reticulatae 160g, Lignum Aquilariae Resinatum 112g, Radix Angelicae Sinensis 112g, join CO
2in supercritical extraction device, ethanol is as entrainer, and the percent by volume that entrainer accounts for total extractant is 4-6%, extracting pressure 15-30MPa, temperature 30-50 DEG C, CO
2flow 1-3m1/g crude drug min, extraction time 150-180min, obtains supercritical extract, is ground into fine powder, and water pill obtains 550g.
2. a kind of antiasthmatic pill of perilla fruit suppresses the application in human tissue cell lymphoma U937 cell proliferation in preparation according to claim 1, it is characterized in that CO described in antiasthmatic pill of perilla fruit preparation method
2the percent by volume that supercritical extraction entrainer accounts for total extractant is 5%.
3. a kind of antiasthmatic pill of perilla fruit suppresses the application in human tissue cell lymphoma U937 cell proliferation in preparation according to claim 1, it is characterized in that CO described in antiasthmatic pill of perilla fruit preparation method
2the extracting pressure 20MPa of supercritical extraction, temperature 40 DEG C, CO
2flow 2ml/g crude drug min, extraction time 160min.
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Inventor after: Li Mei Inventor before: Li Qiang |
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Effective date of registration: 20160128 Address after: 273300 No. 33 Hing Shui street, Pingyi County, Linyi, Shandong Applicant after: Li Mei Address before: 266000 Shandong city of Qingdao province Zhongshan Road City, No. 10 room 2106 Applicant before: QINGDAO QINCHENG MEDICINE TECHNOLOGY CO., LTD. |
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