CN103638322B - A kind of preparation method of KidneyHarmonizing Pill and application - Google Patents
A kind of preparation method of KidneyHarmonizing Pill and application Download PDFInfo
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Abstract
The invention provides a kind of preparation method of KidneyHarmonizing Pill, by liquorice Radix Morindae Officinalis 96g, wine Herba Cistanches (processed) 96g, salt Fructus Psoraleae (processed) 96g, Radix Glycyrrhizae Flos Lonicerae processs Radix Aconiti 48g, Cortex Cinnamomi 48g, Rhizoma Atractylodis Macrocephalae (parched with bran) 96g, Rhizoma Dioscoreae 96g, Poria 96g, salt processs Fructus Tribuli 96g, Rhizoma Dioscoreae Hypoglaucae 96g, Herba Dendrobii 96g, Semen Persicae 96g make as crude drug, employing supercritical extraction is prepared from, content is improved a lot, dose reduces, and present invention also offers the application of KidneyHarmonizing Pill in preparation suppression National People's Congress cell lung cancer NCI-H460 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 KidneyHarmonizing Pill.
Background technology
KidneyHarmonizing Pill is recorded in Ministry of Public Health standard WS3-B-0275-90, prescription is liquorice Radix Morindae Officinalis 96g, wine Herba Cistanches (processed) 96g, salt Fructus Psoraleae (processed) 96g, Radix Glycyrrhizae Flos Lonicerae process Radix Aconiti 48g, Cortex Cinnamomi 48g, Rhizoma Atractylodis Macrocephalae (parched with bran) 96g, Rhizoma Dioscoreae 96g, Poria 96g, salt process Fructus Tribuli 96g, Rhizoma Dioscoreae Hypoglaucae 96g, Herba Dendrobii 96g, Semen Persicae 96g, above 12 tastes, be ground into fine powder, sieve, mixing, with water pill, drying, to obtain final product.Can the kidney invigorating cold expelling.For failure of the kidney in receiving QI, the emission that raw invasion and attack cause, the scrotum is clammy, and lose and drench nebulousurine, umbilicus abdomen is had a pain, tired mind, forgetful insomnia, waist and leg ache, dizziness and tinnitus difficulty in urination and defecation.
In prior art, not yet have KidneyHarmonizing Pill 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.
In prior art, by the amount of this prescription, KidneyHarmonizing Pill can obtain 1000g, every 100 6g, each 6g, 3 times on the one, the KidneyHarmonizing Pill 500g adopting formula preparation of the present invention to become also is every 100 6g, but the medical material amount that every ball contains is original 2 times, therefore each 3g, 3 times on the one, greatly reduces dose under the condition with more active component.
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 KidneyHarmonizing Pill.
Another object of the present invention is to provide a kind of KidneyHarmonizing Pill to suppress the application in National People's Congress cell lung cancer NCI-H460 cell proliferation in preparation.
Technical scheme: the object of the invention is by following scheme realize:
A kind of preparation method of KidneyHarmonizing Pill, by liquorice Radix Morindae Officinalis 96g, wine Herba Cistanches (processed) 96g, salt Fructus Psoraleae (processed) 96g, Radix Glycyrrhizae Flos Lonicerae processs Radix Aconiti 48g, Cortex Cinnamomi 48g, Rhizoma Atractylodis Macrocephalae (parched with bran) 96g, Rhizoma Dioscoreae 96g, Poria 96g, Fructus Tribuli 96g processed by salt, Rhizoma Dioscoreae Hypoglaucae 96g, Herba Dendrobii 96g, Semen Persicae 96g makes as crude drug, described method is made up of the following step: extracting liquorice Radix Morindae Officinalis (processed) 96g, wine Herba Cistanches (processed) 96g, salt Fructus Psoraleae (processed) 96g, Radix Glycyrrhizae Flos Lonicerae processs Radix Aconiti 48g, Cortex Cinnamomi 48g, Rhizoma Atractylodis Macrocephalae (parched with bran) 96g, Rhizoma Dioscoreae 96g, Poria 96g, Fructus Tribuli 96g processed by salt, Rhizoma Dioscoreae Hypoglaucae 96g, Herba Dendrobii 96g, Semen Persicae 96g, 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, sieves, mixing, with water pill, dry, obtains 500g, 100 6g.
The preparation method of above-mentioned a kind of KidneyHarmonizing Pill, 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 KidneyHarmonizing Pill, 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 KidneyHarmonizing Pill suppresses the application in National People's Congress cell lung cancer NCI-H460 cell proliferation in preparation, KidneyHarmonizing Pill is by liquorice Radix Morindae Officinalis 96g, wine Herba Cistanches (processed) 96g, salt Fructus Psoraleae (processed) 96g, Radix Glycyrrhizae Flos Lonicerae processs Radix Aconiti 48g, Cortex Cinnamomi 48g, Rhizoma Atractylodis Macrocephalae (parched with bran) 96g, Rhizoma Dioscoreae 96g, Poria 96g, Fructus Tribuli 96g processed by salt, Rhizoma Dioscoreae Hypoglaucae 96g, Herba Dendrobii 96g, Semen Persicae 96g makes as crude drug, preparation method is made up of the following step: extracting liquorice Radix Morindae Officinalis (processed) 96g, wine Herba Cistanches (processed) 96g, salt Fructus Psoraleae (processed) 96g, Radix Glycyrrhizae Flos Lonicerae processs Radix Aconiti 48g, Cortex Cinnamomi 48g, Rhizoma Atractylodis Macrocephalae (parched with bran) 96g, Rhizoma Dioscoreae 96g, Poria 96g, Fructus Tribuli 96g processed by salt, Rhizoma Dioscoreae Hypoglaucae 96g, Herba Dendrobii 96g, Semen Persicae 96g, 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, sieves, mixing, with water pill, dry, obtains 500g, 100 6g.
Above-mentioned KidneyHarmonizing Pill suppresses the application in National People's Congress cell lung cancer NCI-H460 cell proliferation in preparation, and the percent by volume that the entrainer of CO2 supercritical extraction described in KidneyHarmonizing Pill preparation method accounts for total extractant is 5%.
Above-mentioned KidneyHarmonizing Pill suppresses the application in National People's Congress cell lung cancer NCI-H460 cell proliferation in preparation, the extracting pressure 20MPa of CO2 supercritical extraction described in KidneyHarmonizing Pill preparation method, temperature 40 DEG C, CO2 flow 2ml/g crude drug min, extraction time 160min.
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
Extracting liquorice Radix Morindae Officinalis (processed) 96g, wine Herba Cistanches (processed) 96g, salt Fructus Psoraleae (processed) 96g, Radix Glycyrrhizae Flos Lonicerae processs Radix Aconiti 48g, Cortex Cinnamomi 48g, Rhizoma Atractylodis Macrocephalae (parched with bran) 96g, Rhizoma Dioscoreae 96g, Poria 96g, Fructus Tribuli 96g processed by salt, Rhizoma Dioscoreae Hypoglaucae 96g, Herba Dendrobii 96g, Semen Persicae 96g joins in CO2 supercritical extraction device, ethanol is as entrainer, the percent by volume that entrainer accounts for total extractant is 4%, extracting pressure 15MPa, temperature 30 DEG C, CO2 flow 1m1/g crude drug min, extraction time 150min, obtain supercritical extract, be ground into fine powder, sieve, mixing, with water pill, dry, obtain 500g, 100 6g.
Embodiment 2
Extracting liquorice Radix Morindae Officinalis (processed) 96g, wine Herba Cistanches (processed) 96g, salt Fructus Psoraleae (processed) 96g, Radix Glycyrrhizae Flos Lonicerae process Radix Aconiti 48g, Cortex Cinnamomi 48g, Rhizoma Atractylodis Macrocephalae (parched with bran) 96g, Rhizoma Dioscoreae 96g, Poria 96g, salt processs Fructus Tribuli 96g, Rhizoma Dioscoreae Hypoglaucae 96g, Herba Dendrobii 96g, Semen Persicae 96g 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, sieves, mixing, with water pill, dry, obtains 500g, 100 6g.
Embodiment 3
Extracting liquorice Radix Morindae Officinalis (processed) 96g, wine Herba Cistanches (processed) 96g, salt Fructus Psoraleae (processed) 96g, Radix Glycyrrhizae Flos Lonicerae process Radix Aconiti 48g, Cortex Cinnamomi 48g, Rhizoma Atractylodis Macrocephalae (parched with bran) 96g, Rhizoma Dioscoreae 96g, Poria 96g, salt processs Fructus Tribuli 96g, Rhizoma Dioscoreae Hypoglaucae 96g, Herba Dendrobii 96g, Semen Persicae 96g 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, sieves, mixing, with water pill, dry, obtains 500g, 100 6g.Embodiment 4: KidneyHarmonizing Pill suppresses the experimentation data of National People's Congress cell lung cancer NCI-H460 cell proliferation
1 experiment material
1.1 experiment cell strains
National People's Congress cell lung cancer NCI-H460 cell, Nanjing Medical University's laboratory cell storehouse, DMEM+10%FBS cellar culture.
1.2 Experimental agents
Drugs: KidneyHarmonizing Pill of the present invention: prepare by embodiment 3 method.
Medicinal liquid liquid storage: take 100mg KidneyHarmonizing Pill, 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) National People's Congress's cell lung cancer NCI-H460 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 × 10
4individual/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 KidneyHarmonizing Pill 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 National People's Congress's cell lung cancer NCI-H460 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 KidneyHarmonizing Pill increases National People's Congress cell lung cancer NCI-H460 cell
influence research processed (X ± SD)
Note: compare with matched group,
*p<0.01;
*p<0.001
5 experiment conclusion
KidneyHarmonizing Pill can suppress National People's Congress cell lung cancer NCI-H460 cell proliferation, and reduce the Growth of Cells number of National People's Congress cell lung cancer NCI-H460 cell, this effect is dose dependent.
Claims (3)
1. the application of KidneyHarmonizing Pill in preparation suppression National People's Congress cell lung cancer NCI-H460 cell proliferation, it is characterized in that KidneyHarmonizing Pill is by liquorice Radix Morindae Officinalis 96g, wine Herba Cistanches (processed) 96g, salt Fructus Psoraleae (processed) 96g, Radix Glycyrrhizae Flos Lonicerae processs Radix Aconiti 48g, Cortex Cinnamomi 48g, Rhizoma Atractylodis Macrocephalae (parched with bran) 96g, Rhizoma Dioscoreae 96g, Poria 96g, Fructus Tribuli 96g processed by salt, Rhizoma Dioscoreae Hypoglaucae 96g, Herba Dendrobii 96g, Semen Persicae 96g makes as crude drug, preparation method is made up of the following step: extracting liquorice Radix Morindae Officinalis (processed) 96g, wine Herba Cistanches (processed) 96g, salt Fructus Psoraleae (processed) 96g, Radix Glycyrrhizae Flos Lonicerae processs Radix Aconiti 48g, Cortex Cinnamomi 48g, Rhizoma Atractylodis Macrocephalae (parched with bran) 96g, Rhizoma Dioscoreae 96g, Poria 96g, Fructus Tribuli 96g processed by salt, Rhizoma Dioscoreae Hypoglaucae 96g, Herba Dendrobii 96g, Semen Persicae 96g, 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, sieves, mixing, with water pill, dry, obtains 500g, 100 6g.
2. a kind of KidneyHarmonizing Pill suppresses the application in National People's Congress cell lung cancer NCI-H460 cell proliferation in preparation according to claim 1, it is characterized in that CO described in KidneyHarmonizing Pill preparation method
2the percent by volume that supercritical extraction entrainer accounts for total extractant is 5%.
3. a kind of KidneyHarmonizing Pill suppresses the application in National People's Congress cell lung cancer NCI-H460 cell proliferation in preparation according to claim 1, it is characterized in that CO described in KidneyHarmonizing Pill 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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| CN103028005B (en) * | 2012-10-08 | 2014-06-11 | 毛杰 | Preparation method of anti-lumbago tablet and application thereof |
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