KR101800534B1 - Pharmaceutical compositions for prevention or treatment of oral mucositis induced by 5-fluorouracil comprising an extract of Salviae Miltiorrhizae Radix - Google Patents

Abstract

The present invention relates to a pharmaceutical composition for the prevention or treatment of oral mucositis comprising as an active ingredient an extract of Panax ginseng, and more particularly, to a pharmaceutical composition for preventing or treating oral mucositis comprising DPSH radical scavenging activity, protection against cytotoxicity, ROS speicies), inhibition of apoptosis, and inhibition of cytokine production, thereby effectively treating oral mucositis, which is one of the side effects caused by chemotherapy and radiation therapy in cancer patients.

Description

FIELD OF THE INVENTION The present invention relates to a pharmaceutical composition for preventing or treating oral mucositis induced by 5-fluorouracil containing an extract of Panax ginseng as an active ingredient. The pharmaceutical composition for prevention or treatment of oral mucositis induced by 5-fluorouracil comprises an extract of Salviae Miltiorrhizae Radix

The present invention relates to a pharmaceutical composition for the prevention or treatment of oral mucositis comprising an extract of Panax ginseng as an active ingredient.

Danshen (Salvia miltiorrhiza Bunge) is a perennial plant belonging to the family Lamiaceae (Labiatae). It has an efficacy that makes it active in swallowing, Postpartum abdominal pain and hemorrhoids, heartburn, treatment of bruises, insomnia, and skin rash.

The main chemical components of danshans are diterpene compounds including tanshinone I, IIA and IIB, salvianic acid A, protocatechuic aldehyde, (Vitamin E) and tannin (tannin), which is a phenolic compound, including salicylic acid B, salicylic acid B, and baicaline, β-sitosterol, ursolic acid, vitamin E and tannin ) Are known.

Antimicrobial, antioxidant, antitumor, antimutagenic and antimutagenic effects of magnesium salvianolate (magnesium lithospermate) on hepatitis are known as the physiological activities of Dansam. Antithrombotic efficacy of warfarin wafarin), but the mechanism of the mechanism is insufficient.

Oral mucositis (oral mucositis) is one of the common side effects after chemotherapy and radiation therapy. Oral mucositis causes severe pain in many patients and lowers quality of life. In some patients, oral mucositis may not be able to provide adequate dose of chemotherapy or prevent planned chemotherapy.

Accordingly, the inventors of the present invention completed the present invention by studying functional pharmaceutical materials derived from natural products, confirming that the extract of Dansamp extract can effectively treat oral mucositis induced by chemotherapy.

Accordingly, an object of the present invention is to provide a pharmaceutical composition for preventing or treating oral mucositis comprising an extract of Panax ginseng as an active ingredient.

Another object of the present invention is to provide an oral hygiene composition containing the dried persimmon extract as an active ingredient.

Another object of the present invention is to provide a health functional food for preventing or ameliorating oral mucositis comprising the extract of Panax ginseng as an active ingredient.

In order to achieve the above object, the present invention provides a pharmaceutical composition for preventing or treating oral mucositis comprising an extract of Panax ginseng as an active ingredient.

In one embodiment of the present invention, the oral mucositis may be one caused by chemotherapy.

In one embodiment of the present invention, the extract of Panax notoginseng can be obtained by extracting Panax ginseng with distilled water and concentration under reduced pressure.

In one embodiment of the present invention, the extract may be one having the effects of DPPH radical scavenging activity, protection against cytotoxicity, inhibition of ROS (reative oxygen speicies) generation, cell death and cytokine production inhibition.

In addition, the present invention provides an oral care composition comprising an extract of Panax ginseng as an active ingredient.

In one embodiment of the present invention, the oral care composition may have at least one formulation selected from the group consisting of a toothpaste, a mouthwash, an oral spray, and an oral ointment.

The present invention also provides a health functional food for preventing or ameliorating oral mucositis comprising the extract of Panax ginseng as an active ingredient.

The extract of Dansamp extract of the present invention has an effect on PPH radical scavenging activity, protection against cytotoxicity, inhibition of ROS (reative oxygen speicies) generation, cell death and restoration of cytokine production, It can be used to effectively treat oral mucositis, one of the side effects.

FIG. 1 is a graph showing DPPH radical scavenging activity according to the concentration of the extract of Radix Salviae Radix.
FIG. 2 is a graph showing the results of confirming the effect of the extract on the cell viability of human pharyngeal cell of the present invention.
FIG. 3 is a graph showing the results of examining the effect of the extract of Rawalpital on the production of intracellular ROS.
FIG. 4 is a graph showing the results of confirming the effect of the extract of the extract of the present invention on the expression of apoptosis protein in the humanpharyngeal cell line.
FIG. 5 is a graph showing the results of examining the effect of the extract of Dansamp ginseng according to the present invention on the tissue of cheek pouches of a mucositis-induced hamster.
FIG. 6 is a graph showing the results of observation of apoptosis in the tissues of cheek pouches of oral mucositis-induced hamster of the extract of R. giltii of the present invention.
FIG. 7 is a graph showing the results of examining the effect of the extract of the present invention on the expression of NF-kB, IL-1β, TNF-α and Caspase-3 proteins in the tissues of cheek pouches of oral mucositis-induced hamsters.

Hereinafter, the present invention will be described in detail.

The present invention is characterized in that it can be used to treat oral mucositis, which is one of the common side effects occurring after chemotherapy and radiation therapy. Specifically, it is characterized in that oral ginseng extract The present invention provides a pharmaceutical composition for preventing or treating mucositis.

Oral mucositis is a common side effect after chemotherapy and radiation therapy. The most important side effect of chemotherapy in the past was reduction of immunity due to vomiting and bone marrow suppression. However, On the other hand, mucositis is recently recognized as the most serious side effect of cancer patients.

Oral mucositis is a clinically important disease as it causes serious pain in many patients and not only lowers the quality of life but is a high risk factor for sepsis in cancer patients with hypopnea. Furthermore, some patients may not have adequate chemotherapy due to mucositis, or they may cancel or delay the planned chemotherapy.

The epithelial cells of the oral mucosa are the most rapidly growing tissues in the body. Normally, the mucosal cells of the oral and gastrointestinal tracts are replaced by the 7-14 day cycle. Anticancer drugs interfere with recurrence of mucosal cells, and malnutrition in patients makes them worse. Typically, mucositis occurs 5 to 7 days after the administration of anticancer drugs. In the absence of leukopenia, the mucositis usually heals between 2 and 3 weeks. It usually affects the lips, cheeks, and research dogs and shows various lesions ranging from simple eruptions to ulcers.

General stomatitis is known to occur due to the involvement of various factors. Stress, trauma, and genetic problems are caused by complex interactions of exacerbation factors such as bacteria, and defense factors such as saliva and mucosal keratin that protect the oral cavity. On the other hand, oral mucositis induced by anticancer drugs has a different pattern. The toxicity of anticancer drugs damages the basal epithelium layer, which decreases the regenerative capacity of the epithelium, resulting in clonogenic cell death, atrophy, and ulceration. This is a phenomenon confined to epithelial tissues that appear as nonspecific damage to the dividing epithelial cells. Resulting in epithelial changes.

In one embodiment, it is the role of cytokines in the development of oral mucositis. The levels of IL-1β and TNF-α in mucosa were correlated with the degree of side effects after chemotherapy and the concentration of TNF-α, interleukin 1 (IL-1), and IL-6 in peripheral blood of patients. The degree of expression is associated with the development of mucositis. In addition, it has been confirmed in laboratory and clinical studies that drug therapy to reduce the concentration of these cytokines can prevent the development of mucositis.

The first occurrence of oral mucositis is the development of oxidative stress and reactive oxygen species (ROS) due to anticancer drugs. It is known that ROS plays a major role in most of the pathways leading to mucositis. ROS is a series of drugs that directly damage various cells, tissues, and blood vessels regardless of their cause (chemotherapy, radiation) Activates transcription factors that cause development. Several studies have shown that agents that effectively inhibit or eliminate oxygen-free radicals can successfully delay mucosal damage.

Therefore, the inventors of the present invention confirmed that the extract of Dansamp extract is active in oral mucositis. According to one embodiment of the present invention, DPPH radical scavenging activity of human pharyngeal cells, anti-cancer agent 5 (5-FU) induced cytotoxicity and 5-FU-induced inhibition of reactive oxygen species (ROS) production by 5-FU, recovery of histopathological inflammation of cheek pouches, recovery of apoptosis, expression of anti-inflammatory proteins and significant changes of caspase 3 were confirmed.

The extract of Dansamp ginseng according to the present invention is characterized in that the extract is extracted with distilled water and concentrated under reduced pressure.

The extract of Dansamp according to the present invention can be obtained by extracting and separating from nature using an extraction and separation method known in the art, and the 'extract' defined in the present invention can be extracted from Danshen will be.

All.

As an appropriate solvent for extracting the extract from the raw ginseng, any pharmaceutically acceptable organic solvent may be used, and water or an organic solvent may be used, and for example, purified water, methanol acetone, ether, benzene, chloroform (such as methanol, ethanol, propanol, isopropanol, and butanol) chloroform, ethyl acetate, methylene chloride, hexane, and cyclohexane may be used alone or in combination. Preferably, distilled water is used.

As the extraction method, any one of the methods such as hot water extraction method, cold extraction method, reflux cooling extraction method, solvent extraction method, steam distillation method, ultrasonic extraction method, elution method and compression method can be selected and used. In addition, the desired extract may be further subjected to a conventional fractionation process or may be purified using a conventional purification method. There is no limitation on the production method of the extract of Radix Salviae of the present invention, and any known method can be used.

For example, the extract of Dansampula contained in the composition of the present invention can be prepared into a powdery state by an additional process such as vacuum distillation, freeze drying, spray drying, or the like, with the primary extract extracted by the hot water extraction or solvent extraction method described above .

Further, the primary extract can be further purified by using various chromatographies such as silica gel column chromatography, thin layer chromatography, high performance liquid chromatography and the like, You can get it.

Therefore, in the present invention, the ginseng extract is a concept including all the extract, fraction and purified product obtained in each step of extraction, fractionation or purification, a diluted solution thereof, a concentrate or a dried product.

The present invention provides an oral hygiene composition comprising an extract of Panax ginseng as an active ingredient.

The oral care composition may be prepared with a toothpaste, a mouthwash, an oral spray, and an oral ointment.

The present invention provides a health functional food for preventing or ameliorating oral mucositis comprising an extract of Panax ginseng as an active ingredient.

The health functional food of the present invention can be manufactured and processed in the form of tablets, capsules, powders, granules, liquids, and circles for the purpose of improving oral mucositis diseases.

In the present invention, the term " health functional food " refers to foods manufactured and processed using raw materials or ingredients having useful functions in accordance with Law No. 6727 on Health Functional Foods. Or to obtain a beneficial effect in health use such as physiological action.

The health functional foods of the present invention may contain conventional food additives and, unless otherwise specified, whether or not they are suitable as food additives are classified according to the General Rules for Food Additives approved by the Food and Drug Administration, Standards and standards.

Examples of the items listed in the above-mentioned "food additives" include chemical compounds such as ketones, glycine, calcium citrate, nicotinic acid, and cinnamic acid; Natural additives such as persimmon extract, licorice extract, crystalline cellulose, high color pigment and guar gum; L-glutamic acid sodium preparations, noodle-added alkalis, preservative preparations, tar coloring preparations and the like.

For example, the health functional food in the form of tablets may be prepared by granulating a mixture of the extract of Dansamp, an effective ingredient of the present invention, with an excipient, a binder, a disintegrant, and other additives in a usual manner and then adding a lubricant, Alternatively, the mixture can be directly compression molded. In addition, the health functional food of the tablet form may contain a mating agent or the like if necessary.

The hard capsule of the capsule type health functional food may be prepared by filling a normal hard capsule with a mixture of the extract of Dansamp, an effective ingredient of the present invention, with an additive such as an excipient, and the soft capsule may contain additives such as excipients And filling the mixture with a capsule base such as gelatin. The soft capsule may contain a plasticizer such as glycerin or sorbitol, a coloring agent, a preservative and the like, if necessary.

The ring-shaped health functional food can be prepared by molding a mixture of the extract of Dansamp, the active ingredient of the present invention, excipient, binder, disintegrant and the like by a conventionally known method, and if necessary, Or the surface may be coated with a material such as starch, talc.

As shown in the following examples, the health functional food containing the extract of Dansamp extract of the present invention has DPPH radical scavenging activity, protective effect against 5-fluorouracil (5-FU) induced cytotoxicity, 5- FU-induced inhibition of reactive oxygen species (ROS) production, and 5-FU-induced stomatitis animal models were used to evaluate the recovery of histological inflammation of hamster's cheek pouches, the effect of restoration of apoptosis, caspase 3, and thus can be effectively used for prevention and treatment of oral mucositis.

Hereinafter, the present invention will be described in more detail with reference to Examples. These examples are for further illustrating the present invention, and the scope of the present invention is not limited to these examples.

< Example >

Hereinafter, the present invention will be described in more detail with reference to Examples. It is to be understood by those skilled in the art that these examples are for illustrative purposes only and that the scope of the present invention is not construed as being limited by these examples.

Materials and methods

1. Materials and reagents

For the present invention, Fetal bovine serum (FBS; GIBCO BRL, USA), dulbecco's modified eagle medium (DMEM; GIBCO BRL, USA), trypsin-EDTA (GIBCO BRL, USA), 2,2-diphenyl-1-picrylhydrazyl ; Sigma, USA), ascorbic acid (Sigma, USA), ethanol 99.9% (Duksan, Korea), tetrazolium salt 3, [4,5-dimethylthiazol-2-yl] -2,5- diphenyl tetrazolium bromide , USA), dimethyl sulfoxide (DMSO; Sigma, USA) and 5-fluorouracil (Sigma, USA) were used.

(Bio-Rad, USA) and a 30% acrylamide mix (Bio-Rad, USA) for SDS-polyacrylamide gel electrophoresis (SDS-PAGE) and Western blotting assays USA), anti-ß-tubulin (Santa Cruz Biotechnology, USA), SeePico CBB stain kit (Benebiosis, Korea), anti-NF-κB, anti-caspase 3, anti- (Sigma Chemical Co., USA), ammonium persulfate (Sigma Chemical Co., USA), N, N, N, N ' (Sigma Chemical Co., USA), Tween 20 (Sigma Chemical Co., USA), glycine (Sigma Chemical Co., USA), dithiothreitol (DTT, Sigma Chemical Co., USA), bromophenol blue (SDS, Sigma Chemical Co., USA), skim milk (Becton Dickison, USA), glycerol (Showa, Japan), nitrocellulose membranes (Schleicher & Schuell, Germany), sodium dodecyl sulfate Sigma Chemical Co., USA), and gel blott ing paper (Schleicher & Schuell, Germany).

2. Preparation of sample

300 g of Salviae Miltiorrhizae Radix was accurately weighed and placed in 3,000 ml of primary distilled water. The temperature was elevated to near 100 ° C., and the mixture was heated for 120 minutes from the boiling point of the bath liquid. The filtrate was filtered with a filter paper, and the filtrate was concentrated using a rotary vacuum evaporator. This concentrate was dried using a freeze dryer and used as a sample. The yield of freeze dried extract of Danshang was 69.1g and the yield was 23.03%.

3. Cell line and cell culture

The human pharyngeal cell line (Detroit 562, ATCC CCL138) was purchased from the American Type Culture Collection (Manassas, VA, USA). The Detroit 562 cell line was cultured in a modified Eagle's Medium (MEM) (Gibco-BRL, Grand Island, NY, USA) supplemented with 10% fetal bovine serum (FBS; Gibco-BRL), 100 U / mL penicillin and 100 μg / mL streptomycin. using the medium was cultured at 37 ℃, 5% CO ₂ condition.

4. Dansh DPPH  Measurement of radical scavenging activity

To investigate DPPH radical scavenging activity, lyophilized DPPH radicals were dissolved in distilled water and the solutions were prepared at concentrations of 1, 5, 10, 50, 100 and 500 μg / ml. As a positive control, ascorbic acid was used at the same concentration as the sample. To the 96-well microplate (Corning, USA), 0.1 mM DPPH dissolved in ethanol and the same amount of each test solution were added to each well, shaken well, left at room temperature for 30 minutes, and absorbance was measured at 517 nm using a microplate spectrophotometer. Radical scavenging activity was calculated using the following equation (1).

DPPH radical scavenging activity (%) = [(AB-AT) / AB] x 100
AB: absorbance of blank sample
AT: absorbance of tested extract solution

5. For Detroit 562 cells Dansh  Cytotoxicity measurement

In order to investigate the protective effect of 5-fluorouracil-induced cytotoxicity, the MTT assay was applied. 100 μl of 1 × 10 5 cells / ml was added to each well of a 96-well plate and incubated for 24 h at 37 ° C in a 5% CO 2 incubator. The medium was discarded and the surface of the cells was washed with PBS. 10 μM 5-fluorouracil dissolved in PBS (1, 5, 10, 50, 100 μg / ml) and FBS free DMEM was added to each well and cultured for 48 hours. After the incubation, the medium was discarded and washed with PBS. Then, 20 μl of 5 mg / ml MTT dissolved in PBS and 200 μl of FBS free DMEM were treated in each well, shaded with aluminum foil, and incubated for 4 hours under the same conditions. After removing 200 μl of DMSO and incubating at 37 ° C for 2 hours, absorbance was measured at 570 nm using a microplate spectrophotometer.

6. ROS  (reactive oxygen species) production measurement

In order to investigate the inhibitory effect of 5-fluorouacil on the production of ROS, we used the manual of ROS detection kit (Cell biolabs, USA). (100 μg / ml), 10 μM of 5-fluorouracil, and 100 μl (100 μM) of DCF-DA were added to the cells and cultured at 37 ° C for 1 hour. Media was removed and washed twice with 1X PBS. 100 μl of fresh serum free media and 100 μl of 2X Cell Lysis buffer were added and incubated at room temperature for 5 minutes. 150 μl of the mixture was transferred to a 96-well plate and measured at 485 nm excitation and 530 nm emission wavelength using a fluorescence plate reader (Fluoroskan Ascent 6-96 Well Plate Reader, ThermoFisher Scientific, USA).

7. Animal test method

7-1. Experimental animal

Experimental animals were fed with 24 male Golden Syrian hamsters (8 weeks, 100 ~ 110 gm) from the central experimental animals. Experimental animals were temperature 22 ± 2 ℃, humidity 45-55%, illumination 300lux (50~90cm single-phase ²⁾ and light-dark cycle was kept in a 12-hour environmental hamster solid feed for (Purina, bio-PIA) and drinks are available in unlimited Respectively. The experimental animals were divided into 6 groups (normal, negative, positive control, drug treatment group (100, 500, 1000 mg / kg)) after 1 week of purification.

7-2. 5- On Fluorouracil  by Oral mucositis  cause

Hamsters were randomly divided into 6 groups as follows: normal (N: vehicle-treated, n = 6), negative control (C: 5-fluorouracil treated, n = 6), positive control (P: 0.15% benzydimine HCL, op only treated, n = 6 treatment with 5'-flu + 5'-flu 80 mg / kg, op, n = 6 / group)

To induce oral mucositis, 80 mg / kg 5-Fluorouracil (5'-flu) was injected intraperitoneally to the control group and the drug group except for the normal group on days 0 and 2 of the experiment. A scratching wound was applied under an operating microscope using an 18-gauge needle to provide the same environment as the damage to the oral mucosa that could be caused by normal chewing motion in the human body. The larynx was stimulated three times in a linear manner daily for 2 days until the change of erythema occurred in the eversion of the buccal mucosa. The positive control group, benzydimine HCL (0.15%), and 100, 500 and 1000 mg / kg of ginseng were administered orally for 2 weeks after oral mucositis induction.

7-3. Hematoxylin -eosin (HE) staining

The cheek pouches were cut into 5 μm sections using a paraffin cutter and fixed in 4% PFA fixative (4% paraformaldehyde, 0.1 M PB, pH = 7.0) for 30 minutes. The sections were washed for 10 minutes in flowing water and stained for 1 minute in hematoxylin solution. Washed with water for 1 minute and then stained with 70% hydrochloric acid in ethanol. When the nuclei were stained, they were immersed in 80% ethanol, immersed in eosin for 1 minute, washed again, dehydrated and sealed with Canadian balm.

7-4. For cell death confirmation TUNEL  dyeing

ApoTag Peroxidase In Situ Apoptosis Detection Kit (Chemicon, USA) was used to observe the programmed cell death (apoptosis). (TUN) -ditiated biotin nick end-labeling (TUN) method using a 4% PFA fixative. The cells were washed 3 times with PBS, blocked with 0.3% hydrogen peroxide for 15 minutes, and rinsed 3 times for 10 minutes with PBS. After incubation with Eilibrium buffer for 10 minutes, terminal deoxynucleotidyl transferase (TdT) was reacted in a 2: 1 ratio for 1 hour at 37 ℃ incubator. Stop-Wash buffer for 10 minutes, washed three times with PBS, and reacted with anti-digoxigenin for 30 minutes. The cells were washed 3 times with PBS, developed with DAB for 3 minutes, stained with hematoxylin, stained with mounting media, and observed under an optical microscope.

7-5. Western blot analysis

25 μg of protein was electrophoresed on a 6 to 15% SDS-PAGE gel and electrophoretically transferred to a polyvinylidene fluoride (PVDF) membrane at 100 v. Blocking buffer (TPBS containing 5% skimmed milk powder) was added to the electrophoresis membrane and allowed to stand at room temperature for 1 hour. The primary antibody diluted 1: 1000 in blocking buffer was added and reacted at 4 ° C for one day. (Sata Cruz Biotechnology, Inc., USA), TNF-? (Sata Cruz Biotechnology, Inc., USA), NF-? B,? -Tubulin . After washing with PBS (Washing buffer, 0.05% Tween 20), horseradish peroxidase-conjugated anti-goat IgG or anti-rabbit IgG (Cell Signalin Technology Inc., Danvers, 1000 and reacted at room temperature for 2 hours. (Agfa Healthcare, Greenville, SC, USA) after treatment with ECL (Thermo Fisher Scientific Inc, Waltham, Mass., USA)

8. Statistical Analysis

All the results obtained in the present invention were calculated by using the statistical program SPSS (ver 11.0, Chicago, USA) for Window, and the descriptive statistics (mean ± SD) for each variable were calculated. (Independent test). The hypothesis was set at α = 0.05.

Example  One: Dansh DPPH  Confirming radical scavenging activity

The DPPH radical scavenging activities of ascorbic acid and danshen were increased in a concentration - dependent manner. Dansamp showed DPPH radical scavenging activities of 20.3, 24.21, 24.95, 58.65 and 61.96% at the concentrations of 1, 5, 10, 50 and 100 μg / ml, respectively.

In the positive control group, ascorbic acid (AC) showed 20.35, 54.08, 53.39, 59.95 and 64.23% scavenging activity (see FIG. 1). The concentration (IC50) of 50% DPPH radical scavenging activity calculated from the dose-response curve is 4.6 μg / ml for ascorbic acid and 42.6 μg / ml for sclerotin.

Example 2: Dansam  Identification of the effects of human pharyngeal cells on cell survival

MTT assay was performed to determine the effect of Dansam on the growth and proliferation of human pharyngeal cells. The human pharyngeal cells, Detroit 562 cells, were maintained in growth-arrested condition for 24 h in MEM medium without FBS, treated with 10 μM 5'flu and 1, 5, 10, 50, 100 μg / Respectively. 1, 5, 10, 50, and 100 μg / mL, respectively. 120.15, p <0.01; 118.90, p <0.01; 111.86, p <0.05; 128.97%, p < 0.01, and the cell survival rate was higher than that of the normal group (see Fig. 2A). (5, 10, 50, and 100 μg / m) treated group was significantly higher than the control group (p <0.001) , p <0.01; 78.43, p <0.001; 81.33, p <0.001; 102.1, p <0.01; 95.97, p &lt; 0.01 and showed a significant protective effect on cytotoxicity induced by 5'flu (see Fig. 2B).

Example  3: Dansam  Intracellular ROS  Identify the impact on production

We examined the effect of Dansamp on ROS production in human pharyngeal cell line and examined the effect of Dansamp on ROS production and protection using ROS detection kit. In the 5'-flu-treated control group, ROS production was significantly increased compared to the normal group. However, it was found that the production of ROS was inhibited in the group treated with 10 μM 5'-flu and treated with 1 μg, 10 μg and 100 μg / ml of ROS (see FIG. 3).

Example  4: Dansam apoptosis  Confirm the effect on protein expression

 The expression of NF-κB and Cleaved caspase-3 in the nucleus was increased in the negative control group treated with 10 μM 5'-flu compared to the normal group (127.6% and 126.7%, respectively) in the human pharyngeal cell line The expression of NF-κB was significantly decreased (103.2%, p <0.01 and 101.95%, p <0.01) in the group treated with 10 μM 5'- p &lt; 0.05).

In addition, the expression of cleaved caspase-3 was 105.1 and 101.6%, respectively (p <0.05) at a concentration of 100 ug / ml of dansamp (p <0.05)

Example  5: Dansam  Determine the impact of cheek pouches on your organization

Hematoxylin and Eosin staining was performed to investigate the histological changes of cheek pouches. No lesions were observed in the normal cheek pouches tissues and no abnormalities were found in the granules and basal layer. However, in the mucositis model group induced by 5-fluorouracil (5'-flu: 80 mg / kg, ip), deformation of the stratum corneum was observed and thickening of the epithelium of the mucosal membrane and destruction of the boundary between granule and basal layer was observed 4B). However, the positive control group, benzydine HCL treated group showed recovery of the epithelial layer (see FIG. 4C) and 500 and 1000 mg / kg treated group (FIGS. 4E and 4F).

Example  6: Oral mucositis  Observation of cell death in induced hamster cheek pouches

The induction of apoptosis and recovery of medicinal products in cheek pouches tissues of 5'flu induced oral mucositis were studied by TUNEL staining. In the TUNEL test, apoptosis was observed in the mucositis-induced cheek pouches tissues as compared with the normal group (see arrows in FIG. 5B), whereas in the positive control group and in the 100, 500 and 1000 mg / 5E and FIG. 5F), it was confirmed that there is a recovery effect in the granular layer.

Example  7: Dansam  Identification of the effects on anti-inflammatory protein expression

Western blotting was performed to examine the expression levels of pro-inflammatory cytokines (NF-κB, IL-1β, TNF-α, Caspase-3) related to inflammation in 5'-flu-treated induced oral mucositis model. β-tubulin was used for internal control.

As a result, the expression of NF-κB in the nucleus was significantly increased in the negative control group (136.01%, p <0.01) at the concentration of 1000 mg / kg of the benzydimine HCL treated group and the positive control group, 94.7% &Lt; 0.01 and 108.7%, p < 0.05). Protein expression of TNF-α and IL-1β was increased in the mucosal-induced model, negative control (141.5%, p <0.05; 147.0, p <0.01, respectively). However, in the positive control, protein expression of TNF-α and IL-1β was 110.1%, p <0.01; 129.9%, p <0.01; compared to the negative control group. In addition, the expression level of TNF-α and IL-1β decreased (118.19%, p <0.05 and 115.3%, p <0.05, respectively, Fig. 6) in the simultaneous treatment with 1000 mg / kg of ginseng and 5'flu. Expression of Cleaved caspase-3 was also increased in the negative control group (136.62%, p <0.05) and decreased in the positive control group and 500 and 1000 mg / kg (108.3%, p <0.05, 103.7%, p <0.05; and 96.1%, p &lt; 0.01, respectively) (see Fig. 6).

The present invention has been described with reference to the preferred embodiments. It will be understood by those skilled in the art that various changes in form and details may be made therein without departing from the spirit and scope of the invention as defined by the appended claims. Therefore, the disclosed embodiments should be considered in an illustrative rather than a restrictive sense. The scope of the present invention is defined by the appended claims rather than by the foregoing description, and all differences within the scope of equivalents thereof should be construed as being included in the present invention.

Claims (7)

A pharmaceutical composition for preventing or treating oral mucositis comprising Salviae Miltiorrhizae Radix extract,
The oral mucositis is characterized by being generated by chemotherapy,
The oral mucositis is characterized by being induced by 5-fluorouracil (5-FU)
The dried ginseng extract is characterized in that the ginseng extract is extracted with distilled water and concentrated under reduced pressure.
The extract of Panax notoginseng is characterized by exhibiting scavenging activity of DPPH radical induced by 5-fluorouracil (5-FU)
The extract is characterized by exhibiting a protective effect against 5-fluorouracil (5-FU) -induced cytotoxicity,
The extract of Rawan ginseng is characterized by exhibiting an inhibitory effect of 5-fluorouracil (5-FU) on the production of ROS (reative oxygen species)
The pharmaceutical composition for preventing or treating oral mucositis according to claim 1, wherein the extract has an effect of inhibiting 5-fluorouracil (5-FU) induced apoptosis and cytokine production. delete delete delete delete delete delete

Images