CN104784321A - Application of radix bupleuri and herba houttuyniae in preparation of medicines for prevention and treatment of ionising radiation damage - Google Patents

Abstract

The present invention relates to the field of medicines, is named as application of radix bupleuri and herba houttuyniae in preparation of medicines for prevention and treatment of ionising radiation damage, and discloses application a combination of two pure traditional Chinese medicine preparations of bupleurum chinense or bupleurum scorzonerifolium total extract and herba houttuyniae extract in preparation of medicines for prevention and treatment of ionising radiation damage. The invention discloses that the radix bupleuri extract and the herba houttuyniae extract can play a radiation protective effect in cell and living animal levels, and has a protective effect in a wide dose range comprising lethal dose, sub-lethal dose and low dose radiation, the combination of the two active extracts is better than single use of the extracts, the combination is low in cost and wide in sources, and can be used for preparation of the medicines for prevention and treatment of ionising radiation-caused relative damages and diseases.

Description

Radix Bupleuri and Herba Houttuyniae are preparing the application in prevention and therapy ionization radiation injury medicine

Relate to field

The present invention relates to field of medicaments, being specifically the effective ingredient come from extracting from Radix Bupleuri or Radix Bupeuri Scorzonerfolii. and the application of effective ingredient in the associated injury treatment caused for the preparation of prevention and therapy ionizing radiation and disease treatment come by extracting in plant Herba Houttuyniae.

Background technology

Flourish along with global core cause, nuclear technology is in every field extensive uses such as nuclear power station, aerospace industry, national defence and biological medicines, human contact's ionizing radiation also causes the chance damaged to increase, the nuclear war simultaneously brought along with world's nuclear safety situation is nervous and nuclear terrorism secret worry, the protection of the body injury (referred to as radiation damage) that ionizing radiation causes is being subject to increasing attention with treatment.

On the other hand, the sickness rate of malignant tumor and patient populations are in recent years always in the trend continuing to rise, radiotherapy is as one of essential therapeutic arsenals, play a part indispensable, but high dose radiation irradiates the acute radiation injury of tumor surrounding normal tissue and organ and even the whole body that can cause unavoidably, the side reaction that radiation damage brings seriously limits the extensive use of radiotherapy in oncotherapy, also obviously have impact on the quality of life after tumor patient Radiotherapy and treatment.

The radiation damage treatment related drugs of putting on market at present mainly contains: sulfur-containing compound, hormones, cytokine class and Chinese herbal medicine etc., there is respective inherent shortcoming in them: the general side effect of such as sulfur-containing compound larger respectively, amifostine (having another name called Amifostine) is the best compound of the protection effect of generally acknowledging at present as the representative of this compounds, it is international managing body pass through first selectively wide spectrum cytoprotective, but half-life extremely short (7 minutes) and expensive (domestic medical market price 400-500 unit /) limit its application, and hormone medicine to the control of radiation damage mainly to bone marrow nucleated cell, hematopoietic stem cell and CFU-GM, the impact of this type of medicine on sexual organ and reproductive system limits widely using of it, the hemopoietic function of bone marrow system injury that radiation causes can be alleviated and give treatment to cytokine class medicine such as interleukin class, colony stimulating factor class medicine, but its Study On The Radioprotective and administration time closely related (in the medical practice for preventing and give treatment to, this type of medicine requires high to medical care detection level and degree of concern), there is obvious proinflammatory effect, and expensive, be difficult to room temperature and preserve, Chinese herbal medicine class radioprotective composition mainly contains phenols, polysaccharide, natural flavonoid, has the features such as active component is indefinite, low toxicity, studies through for many years, so far without listing or similar drugs to be gone on the market.

The present invention relates to two kinds of natural plants: the effective ingredient extracted in Radix Bupleuri and Herba Houttuyniae is preparing the application in the treatment of prevention and therapy ionization radiation injury and tumour radiotherapy ancillary drug.

Extract provided by the invention is divided into the compositions of two kinds of medicines: a kind of is Radix Bupleuri extract, for the dry root total extract of umbelliferae bupleurum (Bupleurumchinense DC) or Radix Bupeuri Scorzonerfolii. (Bupleurum scorzonerifolium Wild), composition comprises saikoside, volatile oil, sterol (bupleurumol, eugenol etc.), flavonoid, lignanoids, coumarin and polysaccharide etc., clinical injection of Radix Bupleuri used is Radix Bupleuri total extract, water extraction method, has antipyretic-antalgic, antiinflammatory, antiviral, Nnti-Bacterial endotoxin effect.Another kind is plant Herba Houttuyniae extract, has heat-clearing and toxic substances removing, eliminating carbuncle evacuation of pus effect.

Up to now, Radix Bupleuri extract, Herba Houttuyniae extract, and both compositionss for the preparation of prevention and therapy ionization radiation injury treatment and tumour radiotherapy ancillary drug in application there is not been reported.

Summary of the invention

The object of this invention is to provide the novelty teabag of Radix Bupleuri and Herba Houttuyniae composite reagent.

The novelty teabag of Radix Bupleuri provided by the present invention and Herba Houttuyniae is: for the preparation of the treatment of prevention and therapy ionization radiation injury and the application of tumour radiotherapy ancillary drug aspect.

Ionization radiation injury of the present invention comprises fatal dose irradiation, sublethal dose and low dose exposure damage, also comprises the radiation damage that tumour radiotherapy produces.

Radix Bupleuri provided by the invention and Herba Houttuyniae type of service, according to clinical needs, can add corresponding adjuvant, exists with dosage forms such as tablet, pill, capsule, suspension, solution, syrup, injection, solvent, cream, ointment, sprays.

The using dosage of Radix Bupleuri provided by the present invention and Herba Houttuyniae novelty teabag 1-10g/Kg body weight (people), but is not limited thereto scope, and both preferred proportions are 1: 2 ~ 1: 5, and preferred formulation form is oral+injection.

Objects and advantages of the present invention are by for illustration and explanation by the non-limitative illustration of following preferred embodiments, and these embodiments provide as an example with reference to accompanying drawing.

Accompanying drawing explanation

Fig. 1 Radix Bupleuri and Herba Houttuyniae significant prolongation C57 Mus fatal dose irradiate, and (wherein, χ roentgenization: Fig. 1 a, gamma-rays irradiates: natural law of Fig. 1 b) surviving afterwards.

Fig. 2 Radix Bupleuri and Herba Houttuyniae significantly improve C57 Mus sublethal dose (9Gy gamma-rays) and irradiate latter 30 days survival rates.

Fig. 3 Radix Bupleuri and Herba Houttuyniae irradiate the Hemoprotection of the radiation damage caused to low dosage (5Gy gamma-rays).

Fig. 4 Radix Bupleuri is to the radiate protective action of MCF-7 Human Breast Cancer Cells (cytological map, 1. matched group under microscope; 2. irradiation group; 3. Radix Bupleuri 1mg/ml; 4. irradiation+Radix Bupleuri 1mg/ml).

Fig. 5 Herba Houttuyniae is to the radiate protective action of MCF-7 Human Breast Cancer Cells (cytological map, 1. matched group under microscope; 2. irradiation group; 3. fish raw meat injection 1 μ l/ml culture fluid; 4. irradiation+fish raw meat injection 1 μ l/ml culture fluid)

Detailed description of the invention

Embodiment one

Single fatal dose (13Gy χ ray/gamma-rays) irradiates the radiation damage treatment caused

Radix Bupleuri extract, can prepare voluntarily with reference to published patent of invention description or pertinent literature, also can buy in medical market, administering mode is lumbar injection, and dosage is 200 ~ 400mg/Kg body weight (Mus).Herba Houttuyniae injectio, Hebei Shineway Pharmaceutical Co., Ltd, the accurate word Z13020888 of traditional Chinese medicines, lot number 13041222, administering mode is lumbar injection, and dosage is 100 ~ 200 μ l/ (Mus).

χ beam exposure apparatus X-Rad RS-2000 irradiation instrument, voltage 160KV, band radiation energy is 50KeV, close rate 1.01Gy/min.

Gamma-rays irradiation unit ¹³⁷cs irradiation instrument, close rate 0.8Gy/min.

C57BL/6 mice, male and female half and half, body weight 18-20g, quality certification SCXK (capital) 2012-0001, grouping situation is: do not irradiate group, irradiation and to normal saline group, irradiation and to Radix Bupleuri group, irradiation and to Herba Houttuyniae group, irradiation and give Radix Bupleuri+Herba Houttuyniae group, often organize 10.

Adopt the total irradiation of single fatal dose, individually dosed group, dosage regimen is pre-irradiation 30 minutes intraperitoneal administrations, supplement immediately after irradiation and be administered once, within every 24 hours afterwards, be administered once, continuous 14 days, Radix Bupleuri extract dosage is 200mg/Kg, and Herba Houttuyniae injectio dosage is 200 μ l/.Composite reagent, dosage regimen is pre-irradiation 1 hour intraperitoneal administration Radix Bupleuri 200mg/Kg, irradiate intraperitoneal administration Herba Houttuyniae injectio 100 μ l/ in latter 30 minutes and only, then within every 24 hours, only continue administration, continuous 14 days by Radix Bupleuri 200mg/Kg+ Herba Houttuyniae injectio 200 μ l/.

Testing index: survival natural law.

Experimental result is shown in Fig. 1, and Radix Bupleuri and Herba Houttuyniae combination medicine-feeding can the postradiation survival natural law of significant prolongation C57 Mus fatal dose, are better than both individually administrations.

Embodiment two

Radix Bupleuri extract and Herba Houttuyniae extract oral administration irradiate the radiation damage treatment caused to single sublethal dose (9Gy gamma-rays)

Herba Houttuyniae ethanol extraction process: after the Herba Houttuyniae herb 10g of drying is ground, be placed in 250ml flask, the alcoholic solution of concentration 70% is added according to solid-liquid ratio 1: 20 (g/ml), reflux lixiviate in 80 DEG C of water-baths, each 2 hours, reflux 2 times, then by centrifugal for extracting solution 4000r/min 20min, supernatant Rotary Evaporators is concentrated.

Radix Bupleuri extract, can with reference to published patent of invention description or pertinent literature preparation, and dosage is 1g/Kg body weight (mice).Herba Houttuyniae ethanol extraction, dosage is 1.575g/Kg body weight (mice), and above dosage is crude drug weight.Animal information, irradiation devices are identical with embodiment one.

Exposure dose is 9Gy.

Testing index: 30 days survival rates.

Individually dosed group, dosage regimen: pre-irradiation continuous oral administration three days, supplements immediately after irradiation and is administered once, within every 24 hours afterwards, be administered once, continuous 30 days.Composite reagent group, dosage regimen: Radix Bupleuri and Herba Houttuyniae extract mixing, according to Radix Bupleuri: the administration of Herba Houttuyniae (1g/Kg:1.575g/Kg) continuous oral, irradiate after three days, irradiate in latter 30 minutes and supplement administration, Radix Bupleuri: Herba Houttuyniae (1g/Kg:2.5g/Kg), then (1: 2.5) administration in every 24 hours, continuous 30 days.

Experimental result is shown in Fig. 2, and Radix Bupleuri and Herba Houttuyniae combination medicine-feeding can significantly improve the survival rate after C57 Mus Sublethal Doses, is better than both individually administrations.

Embodiment three

Low dosage (5Gy gamma-rays) irradiates the Hemoprotection of the radiation damage caused

Medicine information, animal information, irradiation devices, administering mode are identical with embodiment one.

Exposure dose is 5Gy.

Successive administration 7 days after irradiating, drug withdrawal is observed and is put to death animal two days later, and sampling detects.

Testing index: body weight, peripheral blood counting, one-sided femur counting, CFU-GM, index and spleen index, thymus index.

Experimental result is shown in Fig. 3, and Radix Bupleuri and Herba Houttuyniae combination medicine-feeding significantly can show the hemopoietic system damage alleviating the animal that low dose exposure causes.

Embodiment four

Radix Bupleuri, Herba Houttuyniae injectio are to kinds of tumor cells/Normocellular radiate protective action

Medicine information, irradiation devices are identical with embodiment one with parameter.

Cell line adopts MCF-7 Breast Cancer Cell, breast carcinoma MDA-MB-231 cell, hepatoma Hep G 2 cells, lung cancer A549 cell, Human Embryonic Kidney HEK 293 cell, people's microvascular endothelial HMEC-1 cell.

Experimentation:

1. cell line and cultivation

Adopt human hepatoma HepG2 cell, typeⅡ pneumocyte, respectively with containing 10%FBS (Gibico, REF10099-141), containing 100 μ g/ml streptomycins and l00U/ml penicillin, DMEM in high glucose (Hyclone, SH30243.01B) and PRMI1640 (Hyclone, SH30809.01B) cultivate, be placed in 37 DEG C, 5%CO ₂constant incubator in routine passage cultivate.

2. drug treating and irradiation

Cell culture, to exponential phase, changes the culture fluid containing variable concentrations medicine, continues cultivation 24 hours, accepts various dose gamma-rays and irradiates.

3. colony formation

After cell irradiation, inoculate different cell quantity by different exposure doses and be inoculated into 60mm culture dish.Cell continuous culture was fixed with methanol after 3 weeks, and add Ji's nurse Sa application liquid and dye 30 minutes, flowing water is cleaned, clone's number of counting cells number >=50.Cloning efficiency and surviving fraction is calculated by cloning efficiency PE=(matched group clone number/experimental group cell number) × 100% and surviving fraction=clone's number/(experimental group cell number × PE).And click models fitting drafting cell survival curve by many targets.

Testing index: IC50 (killing the radiological dose of 50% cell)

Experimental result is shown in Fig. 4 ~ 5 and table 1 ~ 2, and Radix Bupleuri and Herba Houttuyniae can improve kinds of tumor cells or Normocellular radiation tolerance dose respectively, have certain radiate protective action.

Table 1 Radix Bupleuri irradiates IC to kinds of tumor cells/normal cell gamma-rays irradiating gamma-ray ₅₀impact (Gy)

Table 2 Herba Houttuyniae injectio irradiates IC to kinds of tumor cells/normal cell gamma-rays ₅₀impact (Gy)

Comprehensive above embodiment result, explanation the invention has the advantages that:

Radix Bupleuri and Herba Houttuyniae is individually dosed and combination can both play radiate protective action in cellular level and living animal level to medication; and under radiation dose range widely; all there is protective effect; wherein combination medicine-feeding is better than individually administration; therefore, Radix Bupleuri and Herba Houttuyniae can be used for the treatment of prevention and therapy ionization radiation injury and tumour radiotherapy ancillary drug.

Claims (6)

1. Radix Bupleuri and Herba Houttuyniae are preparing the application in prevention and therapy ionization radiation injury treatment medicine.

2. application according to claim 1, wherein said ionization radiation injury treatment includes but not limited to that single fatal dose irradiates the acute ionization radiation injury treatment caused.

3. application according to claim 1, wherein said ionization radiation injury treatment includes but not limited to that the hemopoietic system damage that single/repeatedly low dose exposure causes is given treatment to.

4. application according to claim 1, wherein the use preferred proportion of Radix Bupleuri and Herba Houttuyniae is 1: 2 ~ 1: 5.

5. Radix Bupleuri and Herba Houttuyniae are preparing the application in tumour radiotherapy ancillary drug.

6. application according to claim 4, wherein said tumour radiotherapy includes but not limited to breast carcinoma.