CN102357123B - Application of herba dicranostigmae alcohol extract in preparing medicines for resisting melanoma transfer - Google Patents
Application of herba dicranostigmae alcohol extract in preparing medicines for resisting melanoma transfer Download PDFInfo
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Abstract
The invention discloses a new application of a herba dicranostigmae alcohol extract in preparing medicines for resisting melanoma transfer. The extract has the antitumor transferance, particularly inhibiting melanoma spontaneous tumor transfer and experimental tumor transfer, and can be used for preparing medicines for resisting melanoma transfer.
Description
Technical field
The present invention relates to the new purposes of Chinese medicine extract, be specifically related to the application of Herba dicranostigmae alcohol extract in preparation melanoma diversion medicaments.
Background technology
According to World Health Organization (WHO) (WHO) statistics, the eighties in 20th century, the annual new cancered patient in the whole world approximately 7,000,000, died from approximately 5,000,000 people of cancer person every year.At the beginning of the fifties, China's cancer mortality accounts for the 9th or the 10th of whole causes of the death, and the seventies, China's cancer mortality leapt to the 3rd.Statistics in 1991, some big and medium-sized cities residents' of China cancer mortality have occupied the 1st of whole causes of the death.The annual new cancer stricken of China approximately 1,600,000 people, die from approximately 1,300,000 people of cancer person.Above-mentioned data shows: the cancer morbidity of the whole world and China is obviously increasing year by year.In addition, when China's cancer patient is medical, end-stage patients still occupy the majority, and have the operation of having lost and wait radical cure chance, the relatively poor characteristics of therapeutic effect.Numerous medical personnels strengthen the research of oncotherapy, grasp and improve the therapeutic effect of existing various treatment meanss, to reducing cancer mortality, reduce relapse rate, and the quality of life of improving cancer patient is significant.By the level of present China cancer mortality, the whole nation just had 2 people to die from cancer in every 1 minute, will have every day people more than 2800 to be seized life by cancer, will be equivalent to one and have the city of a population of one million to be destroyed in 1 year.As seen, cancer is commonly encountered diseases, frequently-occurring disease, to people's life and health person of having serious threat, is also huge on the impact of the development of the national economy.In the U.S., 83,000,000,000 dollars of annual treatment of cancer expenses account for 14% of national medical expense.Almost everyone experiences the threat of cancer.Because whole world every year up to 6,300,000 people, and China every year has reached 1,300,000 people because of the number of cancer mortality because of the number of cancer mortality, and cancer patient every year with 3% speed increase, being equivalent to just has 1 family to be subject to the threat of cancer in every 200 families.
Chemotherapy is an important means for the treatment of at present cancer, and also there is toxicity in chemotherapeutics to organism normal cell in kill cancer cell.In recent years, along with continuing to bring out of PTS, the rational Application of Combination chemotherapy, the curative effect of chemotherapy is enhanced, but these medicines normal tissue organ simultaneously have infringement or toxic action, thereby reduce body's immunity, a series of toxicities such as bone marrow depression occur, affect the curative effect of chemotherapy, even force chemotherapy to stop.Clinical research shows, when chemotherapy according to the toxicity symptom of patient's body constitution, cancer staging and appearance, be aided with treatment by Chinese herbs, can effectively strengthen the effect of chemotherapy, can avoid to a certain extent or alleviate the toxicity of chemicotherapy again, extend patient's life cycle and improve patient's life quality.And Chinese medicine is as the advantageous resource of China, and some medicine has obvious antitumor action, therefore screens high-efficiency low-toxicity from the traditional natural drug of China, antitumor drug tool that targeting is strong is of great significance.
Wang Guangfeng etc. adopt the kinds of tumors model, the antitumor action of the chromocor compound Nobiletin that extracts in the research Pericarpium Citri tangerinae.Result shows, the tumour inhibiting rate of Nobiletin 8～32mg/kg mouse melanoma B16 is 42.24%～65.95%, is 38.84%～59.09% to the tumour inhibiting rate of Mice Bearing Lewis Lung Cancer; Tumour inhibiting rate to mouse S 180 sarcoma is 36.02%～45.98%.When Nobiletin and low dose of paclitaxel, mitomycin, 52 fluorouracil, cisplatin and amycin coupling, show obvious cooperative effect.Nobiletin is 33.63%～38.94% to the suppression ratio that Mice Bearing Lewis Lung Cancer axil subcutaneous vaccination lung shifts; The suppression ratio that mice Colon26 colon cancer intraperitoneal inoculation peritoneum is shifted is 37.74%～43.40%.(Wang Guangfeng, Wang Xiaochen, Xiao's Lin, etc. the antitumor action research of. tangeritin Nobiletin, Chinese herbal medicine, 2007,38 (11): 1694-1697), but above research all can only play a role to certain tumor cell, studies deep not enough.
The Herba dicranostigmae main chemical compositions is multiple isoquinoline alkaloid.Herb contains: ICD, protopine, corydaline, isocorydine, magnoline, Sanguinarine, allocryptopine, boldine dimethyl ether.Root contains: protopine, chelerythrine, Sanguinarine, allocryptopine, cryptocavine, but do not report the application of Herba dicranostigmae alcohol extract in preparation melanoma diversion medicaments.
Summary of the invention
Goal of the invention: in order to address the above problem, the object of the present invention is to provide the application of a kind of Herba dicranostigmae alcohol extract in preparation melanoma diversion medicaments.
Technical scheme: the objective of the invention is to realize by following scheme:
The application of Herba dicranostigmae ethanol extract in preparation melanoma diversion medicaments.
The application of above-mentioned Herba dicranostigmae ethanol extract in preparation melanoma diversion medicaments, described Herba dicranostigmae ethanol extract is prepared from as follows: get the Herba dicranostigmae medical material, the volumetric concentration that adds 6-10 times of medical material weight is that 60%-75% ethanol carries out reflux, backflow 0.2-0.8 hour, repeat once, merge ethanol extract, when being concentrated into 95 ℃, relative density is 1.20～1.25 extractum, drying is ground into dry extract.
The application of above-mentioned Herba dicranostigmae ethanol extract in preparation melanoma diversion medicaments, described Herba dicranostigmae ethanol extract is prepared from as follows: get the Herba dicranostigmae medical material, the volumetric concentration that adds 8 times of medical material weight is that 70% ethanol carries out reflux, refluxed 0.5 hour, repeat once, merge ethanol extract, when being concentrated into 95 ℃, relative density is 1.20 extractum, drying is ground into dry extract.
Herba dicranostigmae is Papaveraceae Herba Dicranostigmatis Leptopodi platymiscium Herba Dicranostigmatis Leptopodi Dicranostigma leptopodum (Maxim.) Fedde, with all herbal medicine.
Beneficial effect:
In order to understand better essence of the present invention, the below illustrates its new purposes in pharmaceutical field with pharmacological testing and the experimental result of Herba dicranostigmae alcohol extract.Be below the research of Herba dicranostigmae alcohol extract inhibiting effect on tumor metastasis, experimental technique and result are as follows:
One, the preparation of Herba dicranostigmae alcohol extract:
Get 50 gram Herba dicranostigmae medical materials, the volumetric concentration that adds 8 times of medical material weight is that 70% ethanol carries out reflux, refluxes 0.5 hour, repeats once, merges ethanol extract, and when being concentrated into 95 ℃, relative density is 1.20 extractum, and drying is ground into dry extract.
Two, the impact of Herba dicranostigmae alcohol extract on spontaneous B16F10 melanoma metastasis model
1. experiment material
1.1 laboratory animal and cell
A cleaning level C57BL/6 mice (female, 6-8 age in week, 16-20g), Shanghai Slac Experimental Animal Co., Ltd., credit number: SCXK (Shanghai) 2007-0005; The B16F10 melanoma cell strain.
1.2 Experimental agents
Herba dicranostigmae alcohol extract: be prepared from according to the method described above.
1.3 experiment reagent
DMEM (U.S. GIBCO company lot number: 1340114), PBS (laboratory self-control), calf serum (Tianjin ocean biological product Science and Technology Ltd. lot number: QWTC0701), Trypsin (AMRESCO company lot number: 2009/08) EDTA (AMRESCO company lot number: 0105), formaldehyde (Shanghai hundred million chemical reagent company limited lot numbers of a specified duration: 20070610), normal saline (Shanghai Hua Yuanchang rich pharmaceutcal corporation, Ltd lot number: 070405045), NaHCO3 (Nanjing Chemistry Reagent Co., Ltd.'s lot number: 050480039), liquid paraffin (Shanghai hundred million chemical reagent company limited lot numbers of a specified duration: 20040701).
1.4 experiment equipment
Lycra inverted fluorescence microscope (German Lycra company model: DM1L), accurate pipettor (French Gilson Inc model: P2), full-automatic high-pressure autoclave (Japanese SANYO company model: MLS-3020), (safe and sound company of Su Jing group makes model to superclean bench: SW-CJ-ZFD), ultra cold storage freezer (U.S. Niu Aier company model: Nu-6511), CO2 incubator (FORMA model: 3111), pure water instrument (U.S. Spring company model: S/N020579), electronic balance (German Sai Duolisi company limited model: BT323S), table electrothermal air dry oven (Shanghai accurate experimental facilities company model: DHG9123A), refrigerator (Siemens Company's model: KG18V21TI), liquid nitrogen container (CBS model: 2001), centrifuge (Anting Scientific Instrument Factory, Shanghai's model: KA-1000), PH meter (HANNA company model: HI-221), syringe.
2. experimental technique
2.1 preparation B16F10 cell suspension
Cellar culture B16F10 tumor cell, front 4-5 days of experiment in Tissue Culture Flask, occurs not exclusively fusion by 1: 10 passage in order to avoid Growth of Cells is excessive.Each culture bottle approximately contains 6-8 * 106 cells, and the cell of the trophophase of taking the logarithm discards culture fluid, cleans with PBS, adds 1mL 0.25% pancreatin+0.02%EDTA Digestive system, puts into cell culture incubator, after 1-3min, raps culture bottle and makes cell detachment.Add 10mL DMEM, blow and beat cell with pipet, obtain single cell suspension, change the 50mL polypropylene centrifuge tube over to.Centrifuge cell (1200rpm * 10min), then wash twice with PBS.Expect blue counting cells with tongue, the adjustment cell concentration is 1 * 107/mL.
2.3 copy B16F10 spontaneous tumor metastasis model
Every C57BL/6 mice left hind foot lift hemostasis contains 5 * 10
5The suspension 0.05mL of individual B16F10 cell, these mices claimed a body weight in every five days.The 8th day large young pathbreaker mice according to the upper primary tumor of left foot pad is divided into four groups at random after inoculation: dosage group (4mg/kg Mouse Weight), Herba dicranostigmae alcohol extract high dose group (6g/kg Mouse Weight) in model group matched group (normal saline group), Herba dicranostigmae alcohol extract low dose group (2g/kg Mouse Weight), Herba dicranostigmae alcohol extract, every treated animal number is 10.Respectively organize from beginning in the 9th day of inoculation the medicine that gavage gives corresponding dosage, model group gives corresponding normal saline, once a day, until put to death animal, collect specimen.After inoculation the 23rd day, anesthetized mice, amputation lotus tumor lower limb, 10%PBS-formalin is the primary tumor tissue fixedly, the major axis (L1) and the longitudinal axis (L2) that take primary tumo(u)r weight and measure primary tumor with ruler are according to the volume of following formula calculating primary tumor: 0.5236 * L1 * (L2)
2Put to death mice in 22 days after the excision primary tumor, take out lung, avoid tweezers to touch the organ surface damaged tissue, clean tissue with PBS.10%PBS-formalin is lung tissue fixedly, takes lung heavy, takes pictures and calculate lung tumor tuberosity number under anatomic microscope.
3. experimental result
3.1 the impact that the Herba dicranostigmae alcohol extract shifts the spontaneous metastasis model lung of B16F10 melanoma
Experimental result adopts the Excel student t of statistical software check to test, can be found out by experimental data, the lung Nodules number average that each dosage group of Herba dicranostigmae alcohol extract and matched group compare the spontaneous metastasis model of melanoma has inhibitory action, and presents dosage correlation: Herba dicranostigmae alcohol extract 2g/kg dosage group (P＜0.05), Herba dicranostigmae alcohol extract 4g/kg dosage group (P＜0.01), Herba dicranostigmae alcohol extract 6g/kg dosage group (P＜0.01).For the heavy impact of lung, the lung weight of Herba dicranostigmae alcohol extract 6g/kg dosage group is starkly lower than the lung heavy (P＜0.01) of matched group, and other two groups without obvious effect.
3.2 the impact of Herba dicranostigmae alcohol extract on former position gross tumor volume of B16F10 melanoma and weight
Compare with the normal saline group, former position tumor of the mouse insole section of Herba dicranostigmae alcohol extract after for inoculation B16F10 melanoma cell is that volume and weight all do not have obvious influence.
4. conclusion
The Herba dicranostigmae alcohol extract can suppress B16F10 melanoma lung to be shifted, and the growth of inhibition metastasis, but on the not impact of primary tumo(u)r cell proliferation, change and to find out from Mouse Weight, the toxic and side effects of Herba dicranostigmae alcohol extract is very little, for safe medication in body from now on provides the basis.
Three, the Herba dicranostigmae alcohol extract affects B16F10 melanoma cell experimental lung metastasis
1. experiment material is the same
2. experimental technique
2.1 preparation cell suspension is the same
2.2 copy B16F10 experimental tumor metastasis model
Get the C57BL/6 mice and be placed in the injected in mice holder, tail vein injection 0.2mL cell suspension (contains 5 * 10
5Tumor cell).The mice that inoculated tumor cell is divided into 4 groups at random: dosage group (4mg/kg Mouse Weight), Herba dicranostigmae alcohol extract high dose group (6g/kg Mouse Weight) in model group matched group (normal saline group), Herba dicranostigmae alcohol extract low dose group (2g/kg Mouse Weight), Herba dicranostigmae alcohol extract, every treated animal number is 10, begin respectively to organize the medicine that gavage gives corresponding dosage after inoculation, model group gives corresponding normal saline, once a day, administration 23 days, put to death mice, get lungs.Avoid tweezers to touch organ surface, prevent damaged tissue, clean tissue with PBS.10%PBS-formalin is lung tissue fixedly, takes lung heavy, takes pictures, and estimates the impact that the Herba dicranostigmae alcohol extract shifts B16F10 tumor blood lung.
3. experimental result
Experimental result adopts the Excel student t of statistical software check, can find out from the lung tissue that each group is taken out, the lung tissue metastatic tumour tuberosity number of model group is significantly more than the administration group, because the lung tuberosity is too many, can't counting statistics, the statistical analysis heavy to lung show, in Herba dicranostigmae alcohol extract high dose group (6g/kg) and Herba dicranostigmae alcohol extract, dosage group (4g/kg) all can suppress mouse lung tissue weight, and the P value is respectively P＜0.01 and P＜0.05.Before and after the administration of Herba dicranostigmae alcohol extract, Mouse Weight difference is little, and compares with matched group, and Mouse Weight finds no significantly and reduces, the mortality rate demonstration of each group, and Herba dicranostigmae alcohol extract medicine is less for the toxicity in vivo of mice.
4. conclusion
Mortality rate and the matched group of Herba dicranostigmae alcohol extract administration group mice are suitable, and significantly do not reduce before body weight and administration yet, the toxic and side effects that the Herba dicranostigmae alcohol extract is described is very little, the Herba dicranostigmae alcohol extract can well suppress experimental B16F10 melanoma lung to be shifted, directly into the blood process after blood, sticking process and passing the blood vessel process has certain inhibitory action to the Herba dicranostigmae alcohol extract for the B16F10 cell.
The specific embodiment
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 only limits to following example, all technology that realizes based on foregoing of the present invention all belong to scope of the present invention.
Preparation Example 1:
Get 50 gram Herba dicranostigmae medical materials, the volumetric concentration that adds 8 times of medical material weight is that 70% ethanol carries out reflux, refluxes 0.5 hour, repeats once, merges ethanol extract, and when being concentrated into 95 ℃, relative density is 1.20 extractum, and drying is ground into dry extract.
Preparation Example 2:
Get 50 gram Herba dicranostigmae medical materials, the volumetric concentration that adds 6 times of medical material weight is that 70% ethanol carries out reflux, refluxes 0.5 hour, repeats once, merges ethanol extract, and when being concentrated into 95 ℃, relative density is 1.25 extractum, and drying is ground into dry extract.
Preparation Example 3:
Get 50 gram Herba dicranostigmae medical materials, the volumetric concentration that adds 10 times of medical material weight is that 70% ethanol carries out reflux, refluxes 0.6 hour, repeats once, merges ethanol extract, and when being concentrated into 90 ℃, relative density is 1.20 extractum, and drying is ground into dry extract.
Preparation Example 4:
Get 50 gram Herba dicranostigmae medical materials, the volumetric concentration that adds 8 times of medical material weight is that 60% ethanol carries out reflux, refluxes 0.5 hour, repeats once, merges ethanol extract, and when being concentrated into 95 ℃, relative density is 1.20 extractum, and drying is ground into dry extract.
Preparation Example 5:
Get 50 gram Herba dicranostigmae medical materials, the volumetric concentration that adds 6 times of medical material weight is that 75% ethanol carries out reflux, refluxes 0.5 hour, repeats once, merges ethanol extract, and when being concentrated into 95 ℃, relative density is 1.25 extractum, and drying is ground into dry extract.
Preparation Example 6:
Get 50 gram Herba dicranostigmae medical materials, the volumetric concentration that adds 10 times of medical material weight is that 60% ethanol carries out reflux, refluxes 0.6 hour, repeats once, merges ethanol extract, and when being concentrated into 90 ℃, relative density is 1.20 extractum, and drying is ground into dry extract.
Claims (2)
1. the application of Herba dicranostigmae ethanol extract in preparation melanoma diversion medicaments, it is characterized in that: described Herba dicranostigmae ethanol extract is prepared from as follows: get the Herba dicranostigmae medical material, the volumetric concentration that adds 6-10 times of medical material weight is that 60%-75% ethanol carries out reflux, backflow 0.2-0.8 hour, repeat once, merge ethanol extract, when being concentrated into 95 ℃, relative density is 1.20 ~ 1.25 extractum, drying is ground into dry extract.
2. the application of Herba dicranostigmae ethanol extract in preparation melanoma diversion medicaments as claimed in claim 2, it is characterized in that: described Herba dicranostigmae ethanol extract is prepared from as follows: get the Herba dicranostigmae medical material, the volumetric concentration that adds 8 times of medical material weight is that 70% ethanol carries out reflux, refluxed 0.5 hour, repeat once, merge ethanol extract, when being concentrated into 95 ℃, relative density is 1.20 extractum, drying is ground into dry extract.
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| CN107007591A (en) * | 2017-03-03 | 2017-08-04 | 黄冈师范学院 | A kind of application of polymethoxyflavone on the medicine for preparing treatment melanoma |
| CN110882305A (en) * | 2019-11-19 | 2020-03-17 | 天水师范学院 | Method for purifying total alkaloids of dicranostigma leptopodum by using acidic cation resin |
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