CN102824614A - Application of Wujin tablets for preparing medicines for suppressing PC-3 cell proliferation - Google Patents

Abstract

The invention discloses an application of Wujin tablets for preparing medicines for suppressing PC-3 cell proliferation. The formula of the Wujin tablets comprises the components of pollen typhae, plant soot, leonurus, prepared rehmannia root, charred folium artemisiae argyi, rhizoma sparganii, rhizoma corydalis, rhizoma cyperi, white paeony root, fructus psoraleae, fructus evodiae, rhizoma chuanxiong, curcuma zedoary, elecampane, angelica sinensis and fennel oil. The Wujin tablets have the effects of regulating menstruation and removing stasis and are used for qi depression hysteresis, chest and hypochondrium pain, postpartum blood stasis, lower abdomen pain, dysphoria in chestpalms-soles and emaciation with sallow complexion, the specification is 0.6g per tablet, the tablets are taken orally, four tablets are taken once in the mode of two times per day. The tablets can suppress the PC-3 cell proliferation.

Description

The coal sheet suppresses the application in the PC-3 cell proliferation medicine in preparation

Technical field

The present invention relates to a kind of new purposes of Chinese patent medicine, relate in particular to the coal sheet and suppress the application in the PC-3 cell proliferation medicine in preparation.

Background technology

Carcinoma of prostate is exactly the malignant tumor that betides in the male prostate tissue, is the result of the unusual unordered growth of prostate acinous cell.The sickness rate of carcinoma of prostate has tangible geography and race difference.In developed country and areas such as America and Europes, it is the modal malignant tumor of male, and its mortality rate occupies second of various cancers; In the Asia, its sickness rate is lower than western countries, but is rapid ascendant trend in recent years.

The coal sheet is the exclusive product of Yunnan Prov YongZiTang Pharmaceutical Co., Ltd, does not also have it to suppress the bibliographical information of carcinoma of prostate (PC-3) cell proliferation at present.

Summary of the invention

Goal of the invention: the object of the present invention is to provide the coal sheet to suppress the application in the PC-3 cell proliferation medicine in preparation.

Technical scheme: the coal sheet suppresses the application in the PC-3 cell proliferation medicine in preparation.

The coal tablet recipe is Pollen Typhae, Pulvis Fumi Carbonisatus, Herba Leonuri, Radix Rehmanniae Preparata, Folium Artemisiae Argyi (charcoal), triangular, Rhizoma Corydalis, Rhizoma Cyperi, the Radix Paeoniae Alba, Fructus Psoraleae, Fructus Evodiae, Rhizoma Chuanxiong, Rhizoma Curcumae, the Radix Aucklandiae, Radix Angelicae Sinensis, fennel oil, and regulating menstruation and absorbing clots is used for stagnation of QI knot and stagnates, the twinge of the breast side of body; Postnatal blood stasis, lower abdomen pain, dysphoria with feverish sensation in the chest palms and soles; Emaciation and sallow complexion, specification is every 0.6g, and is oral; One time 4,2 times on the one, find that at present it can suppress the PC-3 cell proliferation.

Beneficial effect: the coal sheet suppresses the PC-3 cell proliferation, and is respond well.

The specific embodiment

The coal sheet suppresses the experimentation data of PC-3 cell proliferation

1 experiment material

1.1 cell strain is used in experiment

Carcinoma of prostate (PC-3), from our company's laboratory cell bank, DMEM+10%FBS is conventional to be cultivated.

1.2 experiment medicine

Research medicine: coal sheet, Yunnan Prov YongZiTang Pharmaceutical Co., Ltd, lot number: 20110423

The medicinal liquid liquid storage: take by weighing 100mg coal sheet and pulverize, be dissolved in the 5ml dehydrated alcohol, 0.2 μ m filter filters, and 500 μ l doff manage packing ,-20 ℃ of storages, and 0.2 μ m filter filters the usefulness of dehydrated alcohol in order to matched group simultaneously.

1.3 experiment reagent

DMEM (the Cat.No.12100-061 Lot.No.758137 of GIBCO company); Hyclone (Hangzhoupro, sky, Zhejiang bio tech ltd 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); Penicillin G Sodium Salt (AMRESCO company lot number: 2010242); Streptomycin Sulfate (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); It is thus clear that-ultraviolet light microwell plate detector (U.S. MD company model: SPECTRAMAX 190); CO2 incubator (FORMA model: 3111); Super-clean bench (the safe and sound manufactured model of Su Jing group: SW-CJ-ZFD); Pure water appearance (U.S. Spring company model: S/N 020579); 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); Desk-top electric drying oven with forced convection (the accurate experimental facilities in Shanghai company 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 μ m 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) the PC-3 cell carries out routine with DMEM+10%FBS in 37 ℃, 5%CO2 and cultivates (10cm culture dish), when cell grows to logarithmic (log) phase, and collecting cell; Discard culture fluid, PBS fine laundering 3 times adds 3ml 0.25% trypsin-0.04%EDTA; Behind 37 ℃ of digestion 2min,, behind the piping and druming cell it is changed in the centrifuge tube to wherein adding 5ml complete medium neutralization reaction; The centrifugal 5min of 1000rpm, 3 * 104/ml of adjustment concentration of cell suspension.

2) the cell kind is gone in 96 well culture plates, every hole adds cell suspension 180 μ l, and culture plate is put into cell culture incubator, and (37 ℃, 5%CO2) routine is cultivated.

3) according to the cell growing state, generally grow to 50%-70%, add coal sheet solution, continue to cultivate 24h.

4) add 20 μ l MTT solution (5mg/ml, i.e. 0.5%MTT) behind the 24h, continue to cultivate 4h.

5) the buckle method is removed supernatant behind the 4h, claps driedly gently with absorbent paper, and every hole adds 200 μ l dimethyl sulfoxide, puts low-speed oscillation 10min on the shaking table, and crystal is fully dissolved.Measure the light absorption value in each hole at enzyme-linked immunosorbent assay instrument 490nm place.

6) background (do not add cell, only add culture fluid) is set simultaneously, control wells (the medicine dissolution medium of cell, same concentrations, culture fluid, MTT, dimethyl sulfoxide), 6 multiple holes of every settings.

7) result representes with the suppression ratio of medicine pair cell:

Cell increment suppression ratio (%)=(control wells OD value-dosing holes OD value)/control wells OD value * 100%.Experiment repetition 3 times.

3 statistical dispositions

Adopt correlation analysis and Student t check in Microsoft Excel 2003 softwares, data are represented with mean ± S.D..

4 experimental results

Mtt assay experiment back statistical result showed; Compare with matched group; When dosage reaches 5mg/ml; To PC-3 cell inhibitory effect variant (P < 0.05), dosage this difference when 10mg/>ml has significance (P < 0.01), and utmost point significant difference (P < 0.001) is arranged when dosage reaches 15-20mg/>ml.

Table 1 coal sheet is to PC-3 cell inhibitory effect influence research

Annotate: compare * P < 0.01 with matched group; * P < 0.001

5 experiment conclusion

The coal sheet can suppress the PC-3 cell proliferation, reduces the cell growing number of PC-3 cell, and this effect is time and dose dependent.

Claims (1)

1. the coal sheet suppresses the application in the PC-3 cell proliferation medicine in preparation.