CN102824380B - Application of Lianzhi anti-inflammation dropping pills in preparing medicine for suppressing HT1080 cell proliferation - Google Patents

Abstract

The invention discloses application of Lianzhi anti-inflammation dropping pills in preparing a medicine for suppressing HT1080 cell proliferation. The prescription of Lianzhi anti-inflammation dropping pills is common andrographis herb dry extract and screwtree root extract with specifications of 30 pills/bag and 6 bags/box; the dropping pills are taken orally, 30 pills once and 3 times per day; the dropping pills can realize the effects of clearing heat, removing toxin, cooling blood and diminishing swelling, and are used for treating the cough and asthma, mumps, acute tonsillitis and throat impediment and sore throat caused by heat and fire toxin in the lung and stomach; and the dropping pills can suppress HT1080 cell proliferation according to the current discovery.

Description

The application of lotus sesame anti-inflammatory dripping pills in preparation inhibition HT1080 cell proliferation medicine

Technical field

The present invention relates to a kind of new purposes of Chinese patent medicine, relate in particular to the application of lotus sesame anti-inflammatory dripping pills in preparation inhibition HT1080 cell proliferation medicine.

Background technology

Fibrosarcoma shows as deeply at the hard fixed joint of single-shot limitation, and the surface is nervous, and light is rubescent, is difficult for ulceration, and surface skin is normal usually, can move, but fixing can not the movement when invading adjacent tissue can infiltrate to subcutaneous fat, muscle, fascia etc.When originating from corium or invading corium afterwards as pathological changes, atrophy, pigmentation and ulceration can occur in surface skin, show as once in a while gill fungus sample lump, multiple infringement can occur near local excision's cicatrix.After repeatedly excising, still common recurrence.After multiple relapse, systemic symptom can appear.Metastasis is found in lung, and occasionally in liver, metastases in local lymph node is rarely found.More pernicious person's tumor is larger, and more soft, makes progress more rapid.Tumor also is found in the baby, has when even being born.Present treatment means effect is all not so good.Lotus sesame anti-inflammatory dripping pills is the exclusive product of Hainan Yi Shun pharmaceutcal corporation, Ltd, does not also have at present it to suppress the bibliographical information of fibrosarcoma (HT1080) cell proliferation.

Summary of the invention

Goal of the invention: the object of the present invention is to provide the application of lotus sesame anti-inflammatory dripping pills in preparation inhibition HT1080 cell proliferation medicine.

Technical scheme: the application of lotus sesame anti-inflammatory dripping pills in preparation inhibition HT1080 cell proliferation medicine.

Lotus sesame anti-inflammatory dripping pills prescription is Herba Andrographis dry extractum (with total andrographolides juice), Radix Helicteris extractum, specification is 30 ball/bags, 6 bags/box, oral, 30 balls, 3 times on the one, the energy heat-clearing and toxic substances removing, the removing heat from blood detumescence, being used for the lung stomach has cough and asthma, mumps, tonsillitis, the sore throat pharyngalgia that in heat, fire-toxin, Sheng causes, finds that now it can suppress the HT1080 cell proliferation.

Beneficial effect: lotus sesame anti-inflammatory dripping pills suppresses the HT1080 cell proliferation, and is respond well.

The specific embodiment

Lotus sesame anti-inflammatory dripping pills suppresses the experimentation data of HT1080 cell proliferation

1 experiment material

1.1 experiment cell strain

Fibrosarcoma (HT1080), from our company's laboratory cell bank, DMEM+10% FBS cellar culture.

1.2 Experimental agents

Drugs: lotus sesame anti-inflammatory dripping pills, Hainan Yi Shun pharmaceutcal corporation, Ltd, lot number: 20110219

The medicinal liquid liquid storage: take 100mg lotus sesame anti-inflammatory dripping pills content, be dissolved in the 5ml dehydrated alcohol, 0.2 μ m filter filters, 500 μ l doff pipe packing, and-20 ℃ of storages, 0.2 μ m filter filters dehydrated alcohol in order to the use of matched group simultaneously.

1.3 experiment reagent

The Cat.No.12100-061 Lot.No.758137 of DMEM(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); The autogamy of PBS(laboratory);

1.4 experiment equipment

Lycra inverted microscope (German Leica model: DM1L); As seen-ultraviolet light microwell plate detector (U.S. MD company model: SPECTRA MAX 190); CO2 incubator (FORMA model: 3111); (safe and sound company of Su Jing group makes model to super-clean bench: SW-CJ-ZFD); Pure water instrument (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); Table electrothermal air dry oven (Shanghai accurate experimental facilities 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 HT1080 cell carries out cellar culture (10cm culture dish) with DMEM+10% FBS in 37 ℃, 5% CO2, when Growth of Cells during to logarithmic (log) phase, collecting cell discards culture fluid, PBS fine laundering 3 times, add 3ml 0.25% trypsin-0.04%EDTA, after 37 ℃ of digestion 2min, add wherein 5ml complete medium neutralization reaction, after the piping and druming cell, it is changed in centrifuge tube, the centrifugal 5min of 1000rpm adjusts concentration of cell suspension 3 * 10 ⁴Individual/ml.

2) the cell kind is entered in 96 well culture plates, every hole adds cell suspension 180 μ l, culture plate put into cell culture incubator (37 ℃, 5% CO ₂) cellar culture.

3) according to the Growth of Cells situation, generally grow to 50%-70%, add lotus sesame anti-inflammatory dripping pills solution, continue to cultivate 24h.

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

5) after 4h, the buckle method is removed supernatant, pats dry gently with absorbent paper, and every hole adds 200 μ l dimethyl sulfoxide, puts low-speed oscillation 10min on shaking table, and crystal is fully dissolved.Extinction OD value in each hole of measurement, 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) is set 6 multiple holes for every group.

7) result represents the value-added suppression ratio of cell with medicine:

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

3 statistical dispositions

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

4 experimental results

Statistical result showed after the mtt assay experiment, compare with matched group, when dosage reaches 5mg/ml, to HT1080 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 lotus sesame anti-inflammatory dripping pills is on HT1080 cell inhibitory effect impact research

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

5 experiment conclusion

Lotus sesame anti-inflammatory dripping pills can suppress the HT1080 cell proliferation, reduces the Growth of Cells number of HT1080 cell, and this effect is dose dependent.

Claims (1)

1. the application of lotus sesame anti-inflammatory dripping pills in preparation inhibition HT1080 cell proliferation medicine, lotus sesame anti-inflammatory dripping pills is produced by Hainan Yi Shun pharmaceutcal corporation, Ltd, prescription is Herba Andrographis dry extractum, Radix Helicteris extractum, and specification is 30 ball/bags, 6 bags/box, oral, 30 balls, 3 times on the one, the energy heat-clearing and toxic substances removing, the removing heat from blood detumescence, being used for the lung stomach has cough and asthma, mumps, tonsillitis, the sore throat pharyngalgia that in heat, fire-toxin, Sheng causes.