CN103027924B - Application of trichoderma pseudokoningii exopolysaccharide as medicine for treating gastric cancer - Google Patents

Abstract

The invention relates to the application of Trichoderma pseudokonii exopolysaccharide as a drug for treating gastric cancer, and belongs to the technical field of medicine and health food. The Trichoderma pseudoconii exopolysaccharide with anti-tumor activity in the present invention can significantly inhibit the growth of human gastric cancer cells BGC-823, MKN-45, and MGC-803 by in vitro administration; cause the outbreak of active oxygen in BGC-823 cells and the decrease of mitochondrial membrane potential, break the nucleic acid in the tumor cells, produce DNA fragments, and make the BGC-823 cell cycle arrest in the S phase, and finally lead to the apoptosis of cancer cells, thereby producing anti-tumor effects; in vivo administration can inhibit BGC-823 tumor-bearing mice, MKN-45 tumor-bearing mice, and MGC-803 tumor-bearing mice grew tumors, reduced tumor weight, and had significant anti-tumor effects.

Description

Application of exopolysaccharide from Trichoderma pseudoconii as a drug for gastric cancer

technical field

The invention belongs to the technical field of medicine, and belongs to the application of Trichoderma pseudoconii exopolysaccharide as a drug for treating gastric cancer.

Background technique

Tumor is a common and frequently-occurring disease, among which malignant tumor is the most serious disease that endangers human health. According to statistics from the World Health Organization in 2008, about 12.7 million people in the world were diagnosed with tumors, and nearly 7.6 million people died of tumors. According to reports, more than 21 million people will suffer from tumors in 2030. Thirteen million people will die from cancer. According to the data survey, the cases of death due to tumor account for about 13% of all deaths in the world. The most serious and harmful tumors in my country include gastric cancer, lung cancer, nasopharyngeal cancer, esophageal cancer, colorectal cancer, liver cancer, breast cancer, cervical cancer, leukemia and lymphoma, and about 1.8 million people die of cancer every year.

Early diagnosis of gastric cancer is difficult, and most patients have already reached the advanced stage when they are diagnosed. Therefore, comprehensive treatment based on chemotherapy has become the main treatment for advanced gastric cancer. Worldwide, the death rate of gastric cancer accounts for the second place in the death rate of all kinds of tumors, about 23.2%. In my country, gastric cancer ranks first in the morbidity and mortality of malignant tumors.

Surgery, radiotherapy, and chemotherapy are the three traditional methods for treating tumors. Chemotherapy is one of the main methods used in the systemic treatment of malignant tumors. However, most anti-tumor chemical drugs also seriously damage normal cells while killing tumor cells. , so it is of great significance to find anti-tumor drugs with significant anti-tumor activity and little side effects on human body.

Polysaccharides are polyhydroxy polymers containing aldehyde or ketone groups and their derivatives formed by connecting more than 10 monosaccharides through glycosidic bonds. Polysaccharides have a wide range of biological functions. They can not only serve as energy-supplying substances in the body and basic components of cells, but also participate in processes such as cell recognition, immune function regulation, intercellular material transport, cell transformation, and cell apoptosis.

Trichoderma pseudokoningii fungus with strain preservation number CGMCC No.1443 is isolated from corn stalks, has a strong growth advantage, and secretes a large amount of exopolysaccharide during liquid fermentation culture. Chinese patent document CN101220101A (application number 200810014047.8) discloses a Trichoderma pseudoconii exopolysaccharide, which is characterized in that: (1) detected by high performance gel filtration chromatography, it has a single symmetrical peak with a molecular weight of 18325 ; (2) Through the improved sulfuric acid-phenol method and m-phenylphenol method, the neutral polysaccharide content is 65.2%, and the uronic acid content is 32.6%; (3) Through the GC analysis of sugar alcohol acetate, its single The sugar composition is rhamnose, glucose and galactose, and the molar ratio is RHA:GLC:GAL=5.6:2.7:1.0. After the polysaccharide is completely reduced, the molar ratio of rhamnose, glucose and galactose is RHA:GLC:GAL=14.5:9.3:1.0, and it contains glucuronic acid with a molar content of 26.6%. The preparation method of the exopolysaccharide of Trichoderma pseudokonii comprises the preparation of crude polysaccharide and the purification of polysaccharide.

At present, the functional research on the exopolysaccharide of Trichoderma pseudokonii has become a hotspot in the research of Trichoderma pseudokonii, but the research on the effect of T. pseudokonii exopolysaccharide on the growth of human gastric cancer cells has not been reported.

Contents of the invention

The content of the present invention is the application of Trichoderma pseudokonii exopolysaccharide as a drug for treating gastric cancer.

The Trichoderma pseudokoningii is CGMCC No.1443 Trichoderma pseudokoningii.

The exopolysaccharide of Trichoderma pseudoconii is the polysaccharide prepared by CN101220101A.

The mechanism by which the Trichoderma pseudokonii exopolysaccharide inhibits the growth of human gastric cancer cells is to induce apoptosis of cancer cells through the mitochondrial pathway: after the Trichoderma pseudokonii exopolysaccharide acts on human gastric cancer cells, the mitochondrial membrane potential of cancer cells Decrease, cytochrome C is released and binds to apoptosis-related factors, leading to apoptosis of cancer cells; at the same time, the concentration of reactive oxygen species ROS in cancer cells increases, and excessive ROS aggravates the decline of mitochondrial membrane potential and the release of cytochrome C.

Compared with the prior art in the present invention, the exopolysaccharide of Trichoderma pseudoconii can reduce the cell viability of human gastric cancer cells BGC-823, MKN-45, and MGC-803 in vitro; it can block the cell cycle of BGC-823 at In the S phase, the intracellular active oxygen concentration is increased, and the mitochondrial membrane potential in the cell is decreased, which causes human gastric cancer cell BGC-823 cell membrane phosphatidylserine to evert, and induces BGC-823 cell nucleus to shrink and fragment to produce apoptotic bodies , Apoptosis occurs so as to kill tumor cells and inhibit tumor growth; Trichoderma pseudoconii exopolysaccharide can inhibit BGC-823 tumor-bearing mice, MKN-45 tumor-bearing mice, MGC-803 tumor-bearing mice in vivo The growth of tumors in mice is significantly reduced, and it has anti-tumor effects.

Description of drawings

Figure 1 Inhibition rate of different concentrations of exopolysaccharide (EPS) from Trichoderma pseudoconii in vitro on the proliferation activity of human gastric cancer cell BGC-823 in vitro;

Among them: * indicates that after t test, there is a significant difference compared with the control group (P<0.05)

**Indicates that after the t test, there is a very significant difference compared with the control group (P<0.01)

Figure 2 Inhibition rate of different concentrations of exopolysaccharide from Trichoderma pseudoconii in vitro on the proliferation activity of human gastric cancer cell MKN-45 in vitro.

Among them: * indicates that after t test, there is a significant difference compared with the control group (P<0.05)

**Indicates that after the t test, there is a very significant difference compared with the control group (P<0.01)

Figure 3 Inhibition rate of different concentrations of exopolysaccharide from Trichoderma pseudoconii in vitro on the proliferation activity of human gastric cancer cell MBGC-803 in vitro.

Among them: * indicates that after t test, there is a significant difference compared with the control group (P<0.05)

**Indicates that after the t test, there is a very significant difference compared with the control group (P<0.01)

Figure 4 Effects of different concentrations of exopolysaccharide from Trichoderma pseudoconii in vitro on the morphology of human gastric cancer cell BGC-823;

Figure 5 Effects of Trichoderma pseudoconii exopolysaccharide administered in vitro on the DNA of human gastric cancer cell BGC-823

Fig. 6 Different concentrations of Trichoderma pseudoconii exopolysaccharides in vitro induce apoptosis of human gastric cancer cell BGC-823;

Fig. 7 Effects of Trichoderma pseudoconii exopolysaccharide administered in vitro on reactive oxygen species in human gastric cancer cell BGC-823.

Fig. 8 Effects of different concentrations of Trichoderma pseudoconii exopolysaccharide administered in vitro on the mitochondrial membrane potential of human gastric cancer cell BGC-823.

Fig. 9 Effect of in vivo administration of exopolysaccharide from Trichoderma pseudokonii on tumor growth of human gastric cancer cell BGC-823 mice.

Detailed ways

The present invention will be further described below in conjunction with the examples, but the protection scope of the present invention is not limited thereto.

In the examples, Trichoderma pseudokoningii (Trichoderma pseudokoningii) was purchased from China General Microorganism Culture Collection and Management Center, and the strain preservation number is CGMCC No.1443;

Trichoderma pseudoconii exopolysaccharide is a polysaccharide prepared by CN101220101A

SRB was purchased from Sigma-Aldrich;

Hoechst 33258 staining solution, Annexin V-FITC cell apoptosis detection kit, reactive oxygen species detection kit, DNA Ladder extraction kit, Rhodamine 123, PI were purchased from Biyuntian Biotechnology Co., Ltd.;

Human gastric cancer cells BGC-823, MKN-45, and MGC-803 were purchased from the Cell Resource Center, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences;

Trypan blue was purchased from Shanghai Huyu Biological Co., Ltd.;

RPMI-1640 is commercially available.

Other reagents are commercially available reagents commonly used in this field, analytically pure,

BALB/C-nu/nu nude mice were purchased from Shanghai Experimental Animal Center, Chinese Academy of Sciences.

PBS: a buffer solution prepared from distilled water, containing per liter: potassium dihydrogen phosphate 0.27g, disodium hydrogen phosphate 1.42g, sodium chloride 8g, potassium chloride 0.2g, pH 7.4.

Example 1

Effects of Trichoderma pseudoconii exopolysaccharide administered in vitro on the proliferation of human gastric cancer cells BGC-823, MKN-45, MGC-803

SRB is a protein-binding dye that can combine with the basic amino acids of cellular proteins fixed by trichloroacetic acid to present a bright pink color, and its color change is proportional to the protein in living cells. ELISA The absorbance is measured by the detector to obtain an accurate value of cell protein content, which can be used to calculate the number of cells and reflect the proliferation activity of cells.

experimental method

Human gastric cancer cells BGC-823, MKN-45, and MGC-803 in the logarithmic growth phase were taken to make a cell suspension with a density of 6× ¹⁰ cells/ml, and inoculated in a 96-well plate, 150 μL/well. After culturing for 24 hours, 50 μL of exopolysaccharide solution of Trichoderma pseudoconii at different concentrations was added to each well (final concentrations were 0.2, 0.4, 0.6, 0.8, 1 mg/ml), the total volume of each well was 200 μL, and each drug concentration was set 6 duplicate wells, and set blank control (no cells, only add RPMI-1640 culture solution) and normal control wells (no drug, add the same amount of RPMI-1640 culture solution), 37°C, 5% CO ₂ , saturated humidity (The following cell culture conditions are the same) Culture for 24, 48, 72 hours. After the incubation, 50 μL of 50 wt% trichloroacetic acid (TCA) solution at 4°C was added to each well to make the final concentration of TCA 10 wt%, and they were left to stand for 5 min and fixed in a refrigerator at 4°C for 1 h. Wash 5 times with deionized water, add 100 μL 0.4wt% SRB to each well after drying, let stand at room temperature for 30 min, discard the liquid in each well, wash 5 times with 1wt% acetic acid, add 150uL pH 10.5 SRB to each well after air drying 10mmol/L Tris base (trishydroxymethylaminomethane) solution, shake on a plate shaker for 5min until the dye is completely dissolved, measure the OD value at 490 nm with an enzyme-linked immunosorbent detector and record it.

Inhibition rate = (OD value of control group - OD value of treatment group) / OD value of control well × 100%

Trichoderma pseudokonii exopolysaccharide has inhibitory effect on the proliferation of human gastric cancer cells in vitro, and the experimental results are shown in Figures 1, 2, and 3. Trichoderma pseudoconii exopolysaccharide significantly inhibited the proliferation of human gastric cancer cells in vitro in a time- and dose-dependent manner. Different concentrations of exopolysaccharides from Trichoderma pseudoconii were treated for 24h, 48h and 72h, all of which inhibited the growth of human gastric cancer cells, with the highest inhibition rate of 42.57% for BGC-823 and 26.54% for MKN-823 , the highest inhibitory rate to MGC-803 was 36.34%.

Example 2

Effects of exopolysaccharide from Trichoderma pseudoconii in vitro on the chromatin morphology of human gastric cancer cell BGC-823

When cell apoptosis occurs, the permeability of the cell membrane changes, and the chromatin in the nucleus condenses and produces apoptotic bodies. Hoechst33258 dye can penetrate the cell membrane and bind to DNA, showing blue fluorescence. The nucleus or cytoplasm can be seen under a fluorescent microscope Densely dyed and dense granular block-like fluorescence appeared inside, and the morphological changes of cell nuclei were visually observed.

experimental method

Place sterile coverslips in a six-well plate, seed human gastric cancer cell BGC-823 and incubate for 24 hours. The next day, different concentrations of Trichoderma pseudoconii exopolysaccharide solutions were added to stimulate the cells for 48 hours, the culture medium was removed, 0.5ml of fixative solution (4% paraformaldehyde) was added, and the cells were fixed for 10 minutes. Blot off the fixative, add 0.5ml Hoechst 33258 staining solution after washing with PBS, stain for 5 minutes, wash off the staining solution with PBS. Observe and take pictures under a fluorescence microscope, the excitation wavelength is about 350nm, and the emission wavelength is about 460nm.

In vitro administration of exopolysaccharide from Trichoderma pseudoconii led to changes in the chromatin morphology of human gastric cancer cell BGC-823 cells, as shown in Figure 4. Hoechst 33238 staining solution staining results showed that the cells in the control group were uniform in size, regular in shape, and lightly stained in the nucleus. apoptotic body. The generation of apoptotic bodies is a significant sign of cell apoptosis, indicating that Trichoderma pseudoconii exopolysaccharide can induce apoptosis of human gastric cancer cell BGC-823.

Example 3

Effects of Trichoderma pseudoconii exopolysaccharide administered in vitro on the DNA of human gastric cancer cell BGC-823

A notable feature of apoptosis is the DNA degradation of cell chromosomes, resulting in 180-200 bp double DNA fragments, which are shown in agarose gel electrophoresis as a specific ladder ladder pattern, while necrosis presents a diffuse continuous pattern .

experimental method

Human gastric cancer cells BGC-823 in the logarithmic growth phase were planted in a six-well plate and incubated for 24 hours. The next day, different concentrations of Trichoderma pseudoconii exopolysaccharide solutions were added, and the cells were harvested after 48 hours of treatment. DNA was extracted separately, and then 1% agarose gel was prepared, and the samples were loaded separately at a voltage of 80v for electrophoresis.

In vitro administration of exopolysaccharide from Trichoderma pseudoconii can induce the apoptosis of human gastric cancer cell BGC-823, and produce apoptotic DNA fragments. Agarose gel electrophoresis presents a specific ladder-like Ladder pattern. The results are shown in Figure 5. The bands are from From left to right is Mark, the electrophoretic pattern of BGC-823 treated with Trichoderma pseudoconii exopolysaccharide at concentrations of 0, 0.2, 0.6, and 1.0 mg/mL.

Example 4

In vitro administration of exopolysaccharide from Trichoderma pseudokonii induces apoptosis of human gastric cancer cell BGC-823

The eversion of phosphatidylserine on the cell membrane is one of the important signs of early cell apoptosis. Annexin-V is a calcium ion-dependent phospholipid-binding protein with high affinity for phosphatidylserine. Annexin V-FITC is used with The green fluorescent fluorescent probe FITC-labeled Annexin V is used to detect phosphatidylserine that appears on the surface of the cell membrane for the detection of apoptosis.

experimental method

Take human gastric cancer cell BGC-823 in the logarithmic growth phase, adjust the concentration to 5×10 ⁵ cells/ml, plant in a six-well plate, and incubate for 24 hours. The next day, different concentrations of Trichoderma pseudoconii exopolysaccharide solutions were added, and the cells were harvested after 48 hours of treatment. Take 50,000-100,000 cells, add 195 μL Annexin V-FITC binding solution, then add 5 μL Annexin V-FITC, incubate at room temperature (20-25 ℃) in the dark for 10 minutes, centrifuge at 1000 g for 5 minutes, discard the supernatant, and add 190 μL Annexin V Gently resuspend the cells in -FITC binding solution, add 10 μL of propidium iodide staining solution, mix gently, place in an ice bath in the dark, and then test on the machine, the excitation light of Annexin V-FITC is green fluorescence, and the excitation light of propidium iodide Fluorescent red.

In vitro administration of Trichoderma pseudokonii exopolysaccharide induced apoptosis of human gastric cancer cell BGC-823, the results are shown in Figure 6. After human gastric cancer cell BGC-823 was treated with Trichoderma pseudoconii exopolysaccharide for 48 hours, compared with the control group, the proportion of early apoptotic cells (lower right quadrant) of human gastric cancer cell BGC-823 in the treatment group increased from 0.71% to 29.2% , late apoptotic and necrotic cells (upper right quadrant) increased from 0.74% to 14.9%, with the concentration increasing, the number of apoptotic cells gradually increased in a dose-dependent manner, indicating that Trichoderma pseudoconii exopolysaccharide can induce human gastric cancer cell BGC -823 undergoes apoptosis.

Example 5

Effects of exopolysaccharide from Trichoderma pseudoconii in vitro on the cell cycle of human gastric cancer cell BGC-823.

When cell apoptosis occurs, the activation of endogenous endonuclease causes DNA to be widely degraded and broken, and the DNA structure undergoes drastic changes. Small molecule DNA can escape from the cell through the cell membrane, and apoptotic bodies can also take away part of the DNA. As a result, the total amount of DNA in the cells decreases, and a group of cells with low DNA staining appears before the normal G ₀ /G ₁ phase cells, which is called the apoptotic peak. The percentage of apoptotic cells can be obtained by analyzing the percentage of the peak. Flow cytometry can analyze DNA changes in apoptotic cells.

experimental method

Human gastric cancer cell BGC-823 was treated with exopolysaccharide of Trichoderma pseudokonii at different concentrations for 48 hours, the cells were collected, centrifuged, the supernatant was discarded, fixed in 75% ethanol pre-cooled at 4 ℃, centrifuged after fixing for 24 hours, the fixative solution was discarded, and added PI staining at 100 μg/mL, incubated at 4°C in the dark for 30 min, and detected by flow cytometry.

In vitro administration of Trichoderma pseudokonii exopolysaccharide can arrest the cycle of human gastric cancer cell BGC-823 in the S phase. The results are shown in Table 1. Among Fig. 7, A, B, C, D are the cycle distribution figure after the Trichoderma pseudoconii exopolysaccharide acting on human gastric cancer cell BGC-823 48h with a concentration of 0, 0.2, 0.6, 1.0 mg/mL respectively, and it can be observed As the concentration increased, the number of hypodiploid cells increased significantly. When the concentration reached 1 mg/mL, the peak of hypodiploid apoptosis increased significantly, and the proportion of apoptotic cells reached 16.8%. It can be seen from Table 1 that with the increase of the concentration of the treatment group, the G ₂ /M phase cells decreased significantly, from 22.78% in the control group to 7.30%; the S phase cells increased from 13.01% in the control group to 32.95%, and the dose Dependence, the results showed that after Trichoderma pseudoconii exopolysaccharides treated cells, the tumor cells were mainly arrested in the S phase, and the cells accumulated in large quantities in the S phase, unable to undergo cell division, and eventually led to rapid apoptosis of tumor cells.

Table 1

Example 6

Effects of Trichoderma pseudoconii exopolysaccharide administration in vitro on reactive oxygen species in human gastric cancer cell BGC-823 cells.

Excess reactive oxygen species ROS play an important role in many apoptotic signaling pathways. The fluorescent probe DCFH-DA can be used to detect ROS produced in cells. DCFH-DA itself has no fluorescence and can freely pass through the cell membrane and be hydrolyzed by intracellular esterases. When DCFH is generated, ROS in the cell can oxidize non-fluorescent DCFH to fluorescent DCF, and the level of reactive oxygen species in the cell can be known by detecting the fluorescence of DCF.

experimental method

Human gastric cancer cell BGC-823 was treated with different concentrations of Trichoderma pseudoconii exopolysaccharide (0, 0.2, 0.6, 1.0 mg/mL). After 24 hours of treatment, the cells were stained with DCFH-DA fluorescence, and then confocal laser Scanning inverted microscope for detection.

In vitro administration of exopolysaccharide from Trichoderma pseudoconii can increase the reactive oxygen species in human gastric cancer cell BGC-823 cells. The results are shown in FIG. 7 . A, B, C, and D in Figure 7 are human gastric cancer cell BGC-823 cells treated with Trichoderma pseudoconii exopolysaccharide at a concentration of 0, 0.2, 0.6, and 1.0 mg/mL, respectively, and scanned under a laser confocal scanning inverted microscope The results of the pictures taken showed that the overall fluorescence intensity of the cells in the control group was weak, while the fluorescence intensity of the group added with Trichoderma pseudoconii exopolysaccharide gradually increased with the increase of the concentration, indicating that the concentration of ROS in the cells continued to increase.

Example 7

Effects of exopolysaccharide from Trichoderma pseudoconii in vitro on the mitochondrial membrane potential of human gastric cancer cell BGC-823 cells.

The decline of mitochondrial membrane potential is another landmark event in the early stage of apoptosis. Mitochondria are the main place for ROS. When mitochondria are damaged, excess ROS is released from mitochondria, and excessive ROS will further damage the membrane potential of mitochondria. , leading to a decrease in membrane potential and ultimately cell apoptosis. Rhodamine123 is a lipophilic cationic fluorescent dye that can penetrate the cell membrane and selectively stain the mitochondria of living cells. Its high fluorescence intensity indicates strong electronegativity and high membrane potential in the mitochondria. It is widely used as a fluorescent probe for detecting mitochondrial membrane potential.

experimental method

Human gastric cancer cells BGC-823 in the logarithmic growth phase were inoculated into six-well plates, cultured for 24 hours, and treated with different concentrations of Trichoderma pseudoconii exopolysaccharide (0, 0.2, 0.6, 1.0 mg/mL) the next day Act on human gastric cancer cell BGC-823, after 48 hours of treatment, wash the six-well plate with preheated PBS, add 1 mL of Rhodamine123 dye solution with a concentration of 10 μg/mL, incubate for 1 hour at 37 °C, in the dark, absorb the dye solution, and wash with PBS Clean and detect under confocal scanning inverted microscope.

In vitro administration of Trichoderma pseudokonii exopolysaccharide can reduce the mitochondrial membrane potential of human gastric cancer cell BGC-823, the results are shown in Figure 8. It can be seen from the figure that the fluorescence intensity in the treatment group was significantly lower than that in the control group, indicating that the mitochondrial membrane potential decreased after the exopolysaccharide of Trichoderma pseudoconii treated human gastric cancer cell BGC-823.

Example 8

Effects of in vivo administration of exopolysaccharide from Trichoderma pseudokonii on the body weight, tumor volume and tumor weight of tumor-bearing mice.

Human gastric cancer cells in the logarithmic growth phase were collected, and the viable cell content was checked by trypan blue staining to be above 95%. The cell density was adjusted to 1×10 ⁸ cells/ml with PBS buffer, and BALB/C-nu was inoculated subcutaneously in the right scapula. 50 nude mice of /nu strain, 0.2ml/only. One week later, tumor-forming mice were selected and randomly divided into 5 groups, control group, Trichoderma pseudoconii exopolysaccharide 20mg/kg.day group, 50mg/kg.day group, 100mg/kg.day group, cyclophosphamide 20mg /kg.day group, 10 rats in each group, and the difference in average body weight between groups is not more than 1 gram. Trichoderma pseudokonii exopolysaccharide normal saline solution with different concentrations was intragastrically administered at a fixed time every day, and the control group was given the same amount of normal saline for 20 consecutive days. The growth of the mice in each group was observed, the body weight of the mice was weighed every 5 days, and the tumor volume was measured. 24 hours after the last administration, the mice were sacrificed by bloodletting, and the subcutaneous solid tumor mass was removed, fat and connective tissue were removed, and weighed.

In vivo administration of Trichoderma pseudoconii exopolysaccharide can inhibit the growth of xenograft tumors and promote the weight gain of tumor-bearing mice. The results are shown in Table 2, Table 3, Table 4, Table 5, and Figure 9. Table 2 shows the effect of oral administration of Trichoderma pseudokonii exopolysaccharide on the body weight of tumor-bearing mice. The body weight of the mice increased significantly, and the average body weight of the mice in the 100mg/kg.day group was 16.33% higher than that of the control group after 20 days. Table 3, Table 4, and Table 5 show the effect of Trichoderma pseudoconii exopolysaccharide gavage administration on the tumor tumor weight of tumor-bearing mice. With the increase of the dosage, the tumor weight of tumor-bearing mice decreased, and tumor inhibition The rate was increased, which had a dose-dependent effect, and the tumor inhibition rate in the 100mg/kg.day group was as high as 48.88%. As shown in Figure 9, the tumor volume of BGC-823 tumor-bearing mice changed with the course of administration, and the increase rate of the tumor volume of mice in the treatment group was slower than that in the control group. The above results indicated that administration of exopolysaccharide from Trichoderma pseudoconii in vivo inhibited tumor growth and reduced tumor weight in tumor-bearing mice.

Table 2 Effect of in vivo administration of exopolysaccharide from Trichoderma pseudokonii on the body weight (grams) of BGC-823 tumor-bearing mice (n=10, )

Table 3 Effect of in vivo administration of exopolysaccharide from Trichoderma pseudoconii on tumor weight in BGC-823 tumor-bearing mice.

Trichoderma pseudoconii exopolysaccharide concentration Tumor weight (g) Tumor inhibition rate (%) 0 (control group) 1.78 - 20mg/kg.day 1.54 13.48 50mg/kg.day 1.21 32.02 100mg/kg.day 0.91 48.88

Table 4 Effect of in vivo administration of exopolysaccharide from Trichoderma pseudoconii on tumor weight in MKN-45 tumor-bearing mice.

Trichoderma pseudoconii exopolysaccharide concentration Tumor weight (g) Tumor inhibition rate (%) 0 (control group) 1.58 - 20mg/kg.day 1.41 10.76 50mg/kg.day 1.29 18.35 100mg/kg.day 1.05 33.54

Table 5 Effect of in vivo administration of exopolysaccharide from Trichoderma pseudokonii on tumor weight in MGC-803 tumor-bearing mice.

Trichoderma pseudoconii exopolysaccharide concentration Tumor weight (g) Tumor inhibition rate (%) 0 (control group) 1.69 - 20mg/kg.day 1.53 9.47 50mg/kg.day 1.32 21.89 100mg/kg.day 1.12 33.73

Claims (2)

1. trichoderma pseudokoningii exocellular polysaccharide is as the application of preparation treatment gastric cancer medicament, and the cancerous cell of described gastric cancer is BGC-823, MKN-45, MGC-803; Described trichoderma pseudokoningii exocellular polysaccharide suppresses the mechanism of gastric carcinoma cells growth to be by mitochondria pathway inducing cancer cell generation apoptosis: after trichoderma pseudokoningii exocellular polysaccharide acts on gastric carcinoma cells, the mitochondrial membrane potential of cancerous cell declines, cell plain color C discharges, and combine in Apoptosis-Related Factors, make cancerous cell generation apoptosis; In cancerous cell, active oxygen ROS concentration increases simultaneously, and excessive ROS aggravates the decline of mitochondrial membrane potential and the release of cytochrome C.

2. trichoderma pseudokoningii exocellular polysaccharide according to claim 1 is as the application of preparation treatment gastric cancer medicament, it is characterized in that: described trichoderma pseudokiningii is CGMCC No.1443.