CN113876840A

CN113876840A - Application of tartary buckwheat flavone in preparation of medicine for treating pancreatic cancer

Info

Publication number: CN113876840A
Application number: CN202110764134.0A
Authority: CN
Inventors: 廖志勇; 程文文; 蔡慈峰; 周美亮
Original assignee: Wenzhou University
Current assignee: Wenzhou University
Priority date: 2021-07-06
Filing date: 2021-07-06
Publication date: 2022-01-04

Abstract

The invention discloses application of buckwheat flavone in preparation of a medicine for treating pancreatic cancer, wherein the aspect of activity of buckwheat flavone on pancreatic cancer cells is explored, and the finding that buckwheat flavone can obviously promote apoptosis of pancreatic cancer cells, inhibit proliferation of pancreatic cancer cells, break DNA and have great potential in the aspect of exerting antitumor effect is found. The tartary buckwheat flavone is prepared by alcohol leaching of tartary buckwheat powder and resin purification, not only is the preparation method simple, but also when the tartary buckwheat flavone is used for preparing a pharmaceutical composition for preventing, treating or improving pancreatic tumors, the tartary buckwheat flavone can promote the expression of apoptosis proteins Caspase-3, Caspase-8, cleared-Caspase-3, cleared-Caspase 8, cleared-Caspase 9 and Bax, and inhibit the expression of anti-apoptosis protein Bcl-2, so that the tartary buckwheat flavone has the activity function of promoting pancreatic cancer cell apoptosis, and further plays the activity role of resisting pancreatic cancer.

Description

Application of tartary buckwheat flavone in preparation of medicine for treating pancreatic cancer

Technical Field

The invention belongs to the field of medical industry, relates to medicinal application of tartary buckwheat flavone, and particularly relates to application of tartary buckwheat flavone in preparation of a medicament for treating pancreatic cancer.

Background

Pancreatic cancer is a malignant tumor mainly derived from the epithelium of a pancreatic duct, is one of common malignant tumors of the digestive system, has the incidence rate accounting for 1% -2% of the common malignant tumors, and has the good incidence age of 50-60 years, and more males than females (1.7: 1). In recent years, the incidence of pancreatic cancer has increased worldwide, and there are about 27000 new cases of pancreatic cancer in the united states each year, and 26000 patients who die of pancreatic cancer in the same year, with their fatality rate being at the 5 th position of the united states' tumor. The incidence of pancreatic cancer is also increasing year by year in China.

Pancreatic cancer is one of the worst-prognosis malignancies due to its high malignancy, occult onset, late clinical symptoms, lack of effective early diagnostic tools, and low surgical resection rate. Therefore, intensive basic and clinical research on pancreatic cancer, improvement of the diagnosis, prevention and treatment level of pancreatic cancer, and improvement of patient prognosis are one of the hot spots in the current research on digestive tract tumors.

At present, chemotherapy and radiotherapy are mainly adopted for treating pancreatic cancer for conservative treatment, but the pancreatic cancer cannot be completely cured and has great side effect.

Therefore, it is one of the important directions for treating pancreatic cancer to explore the expression of pancreatic cancer-related proteins, analyze their correlation, investigate their roles in pancreatic cancer development and relationship with clinical pathological parameters of pancreatic cancer, and provide effective theoretical basis for diagnosis, malignancy determination, prognosis analysis and biological treatment of pancreatic cancer. In addition, the discovery of novel effective drugs for preventing and treating pancreatic cancer from natural products and the elucidation of the active mechanism of action thereof are also hot spots of current research.

Disclosure of Invention

In order to overcome the problems, the inventor of the invention makes intensive research on a pancreatic cancer treatment method and a pancreatic cancer treatment medicine, finds that the buckwheat flavonoids can obviously promote apoptosis of pancreatic cancer cells, inhibit proliferation of pancreatic cancer cells, break DNA and have great potential in the aspect of exerting antitumor effect, and further researches the application of the buckwheat flavonoids in preparing the pancreatic cancer treatment medicine. The tartary buckwheat flavone is prepared by carrying out alcohol leaching and resin purification on tartary buckwheat powder, the preparation method is simple, the content of total flavone in the prepared tartary buckwheat flavone is not lower than 80 wt%, the source is wide, and when the tartary buckwheat flavone is used for preparing a pharmaceutical composition for preventing, treating or improving pancreatic tumors, the expression of apoptosis proteins Caspase-3, Caspase-8, cleared-Caspase-3, cleared-Caspase 8, cleared-Caspase 9 and Bax can be promoted, the expression of anti-apoptosis protein Bcl-2 is inhibited, the tartary buckwheat flavone has the activity function of promoting pancreatic cancer cell apoptosis, and further the activity function of resisting pancreatic cancer is exerted, so that the invention is completed.

Specifically, the present invention aims to provide the following:

on the one hand, the pharmaceutical composition for preventing, treating or improving pancreatic tumors is provided, wherein the pharmaceutical composition contains the tartary buckwheat flavone, and the content of the total flavone in the tartary buckwheat flavone is more than 80 wt%.

On the other hand, provides the application of the tartary buckwheat flavone in preparing the medicine for preventing, treating or improving the pancreatic tumor.

The invention has the advantages that:

(1) the tartary buckwheat flavone provided by the invention has the effect of treating, improving or preventing pancreatic cancer.

(2) The pharmaceutical composition for preventing, treating or improving pancreatic tumors provided by the invention contains the buckwheat flavonoids, and the buckwheat flavonoids are not only widely available, but also extracted from natural products, are safe to use, have no toxic or side effect, have strong operability, have an obvious effect of inhibiting pancreatic cancer cell proliferation, and have the characteristics of low toxic or side effect after long-term application.

(3) The tartary buckwheat flavone in the pharmaceutical composition for preventing, treating or improving the pancreatic tumor provided by the invention has potential clinical value in the aspect of treating or improving the pancreatic tumor by using the tartary buckwheat flavone alone or combining with the traditional medicine.

(4) The tartary buckwheat flavone is applied to preparation of the medicine for treating pancreatic cancer, can obviously promote apoptosis of pancreatic cancer cells, inhibits proliferation of the pancreatic cancer cells, enables DNA to be broken, is low in cost, can reduce diagnosis and treatment cost of patients and medical institutions, and improves survival rate of the patients.

(5) Experiments prove that the tartary buckwheat flavone can promote the expression of apoptosis proteins Caspase-3, Caspase-8, cleared-Caspase-3, cleared-Caspase 8, cleared-Caspase 9 and Bax, inhibit the expression of anti-apoptosis protein Bcl-2, and provide a new development idea for treating pancreatic cancer and other tumors or cancers.

Drawings

FIG. 1 shows photographs of the expression results of different proteins of example 3;

FIG. 2 shows photographs of the TUNEL apoptosis assay of example 4;

FIG. 3 is a photograph showing the results of the cell proliferation assay of CCK-8 of example 5.

Detailed Description

The invention is explained in more detail below with reference to the figures and examples. The features and advantages of the present invention will become more apparent from the description.

The word "exemplary" is used exclusively herein to mean "serving as an example, embodiment, or illustration. Any embodiment described herein as "exemplary" is not necessarily to be construed as preferred or advantageous over other embodiments. While the various aspects of the embodiments are presented in drawings, the drawings are not necessarily drawn to scale unless specifically indicated.

In one aspect of the invention, a pharmaceutical composition for preventing, treating or improving pancreatic tumors is provided, wherein the pharmaceutical composition contains tartary buckwheat flavonoids.

Tartary buckwheat is a small coarse cereal crop which can be used as both medicine and food, is rich in a plurality of nutrient substances, and most importantly, is flavonoid. Tartary buckwheat has the activity of reducing high blood pressure, high blood sugar, high blood fat, high blood sugar, high blood fat, high blood sugar, high blood fat and high blood sugar. How to safely apply the tartary buckwheat to actual treatment and exploring the aspect of the tartary buckwheat having the effect in treatment are still difficult problems in the current medical treatment.

The inventor carefully researches the contents of the anti-tumor and anti-cancer aspects in recent years, and unexpectedly discovers that the flavone of the tartary buckwheat can inhibit the proliferation of pancreatic cancer cells and break DNA, so that the flavone of the tartary buckwheat can be used for preparing a pharmaceutical composition for preventing, treating or improving pancreatic tumors.

Further, the preparation of the pharmaceutical composition comprises tablets, capsules, pills, granules, oral liquid and injection.

In the invention, the tartary buckwheat flavone is used alone or used as an active component and auxiliary materials to be prepared into tablets, capsules, pills or granules, wherein the auxiliary materials comprise one or more of adhesives, fillers and lubricants.

Wherein, the adhesive is selected from one or more of starch slurry, carboxymethyl cellulose, hydroxypropyl cellulose and ethyl cellulose, the filler is selected from one or more of starch, dextrin and lactose, and the lubricant comprises one or more of aerosil, gaseous silica gel, talcum powder and magnesium stearate.

In the invention, the content of total flavonoids in the tartary buckwheat flavonoids reaches more than 80 wt%, and the total flavonoids mainly comprise rutin and quercetin, wherein the rutin accounts for more than 75 wt% of the total flavonoids, and the quercetin accounts for more than 1 wt% of the total flavonoids; further, the content of total flavonoids in the tartary buckwheat flavonoids is more than 83 wt%, rutin accounts for more than 79 wt% of the total flavonoids, and quercetin accounts for more than 2 wt% of the total flavonoids.

In the invention, the buckwheat flavone is not limited to buckwheat flavone produced by any manufacturer on the market, and certainly, the buckwheat flavone can be self-made, and is preferably prepared by alcohol leaching and resin purification of buckwheat powder.

Further, the method for preparing the tartary buckwheat flavone by carrying out alcohol extraction and resin purification on the tartary buckwheat powder specifically comprises the following steps: extracting tartary buckwheat powder in an organic solvent I, concentrating to obtain an extracting solution, dissolving the extracting solution by using an organic solvent II, and purifying by using macroporous adsorption resin.

In the invention, the organic solvent I is an organic solvent capable of entering macromolecules of the tartary buckwheat powder, preferably alcohols, such as methanol, ethanol, propanol and the like, and more preferably, safe and nontoxic ethanol.

Among them, the organic solvent II is preferably the same as the organic solvent I, for example, ethanol.

According to the invention, when the tartary buckwheat flavone is purified by using the tartary buckwheat powder, the concentration of the organic solvent has great influence on the extraction rate of the tartary buckwheat flavone, and the concentration of the organic solvent is adjusted to control the extracted target product.

Preferably, the concentration of the organic solvent I is 60-65%, such as 63%; the concentration of the organic solvent II is 25 to 35%, for example 28%.

In the present invention, the total flavone yield is gradually increased with the increase of the extraction temperature, preferably 60-75 ℃, for example 70 ℃.

According to the invention, the resin is preferably a styrene macroporous resin, such as AB-8 macroporous adsorption resin and DM-2 macroporous adsorption resin, which has a large specific surface area and uniform pore size distribution. The resin is more preferably AB-8 macroporous adsorption resin, and has the advantages of large adsorption capacity, easiness in elution, good adsorption kinetics, stable property and long service life, and can not adsorb inorganic compounds and micromolecule hydrophilic organic matters, so that the tartary buckwheat flavone to be purified can be separated from the substances.

Optionally, after the purification is finished, the buckwheat flavonoids are further eluted, wherein the polarity of the eluent is larger than that of the macroporous resin, preferably alcohol, such as ethanol with the concentration of 60%.

In another aspect of the invention, the application of the tartary buckwheat flavone in preparing the medicine for preventing, treating or improving the pancreatic tumor is provided.

Further, the pancreatic tumors comprise pancreatic tumors and pancreatic cancers, and preferably provides application of the tartary buckwheat flavone in preparation of medicines for preventing, treating or improving the pancreatic cancers.

The inventor cultures pancreatic cancer cells, incubates the pancreatic cancer cells by using a tartary buckwheat flavone culture solution, and finds that: the buckwheat flavonoids can obviously promote the apoptosis of pancreatic cancer cells and inhibit the proliferation of the pancreatic cancer cells, and has great potential in the aspect of exerting anti-tumor effect.

According to the invention, the pancreatic cancer comprises ductal cell carcinoma, acinar cell carcinoma, islet cell carcinoma and rare types of pancreatic cancer, including undifferentiated carcinoma, carcinosarcoma and pancreatic maternal cell carcinoma.

In medical research, apoptosis is an active and programmed death process of cells under certain physiological or pathological conditions and accurately regulated by various genes, and is an important self-stabilization regulation mechanism of organisms.

Research shows that the occurrence, development and prognosis of tumors are closely related to the disturbance of the apoptosis process of cells. In the process of apoptosis, Caspase family plays an indispensable role, and as a group of apoptosis-promoting cysteine-containing proteases, apoptosis signals are transduced through hydrolysis of substrates or directly participate in the apoptosis process as apoptosis effector molecules. Caspase-3, Caspase-8, Caspase-9 are partial members of the family, and are key apoptosis-executing proteases in the process of apoptosis. In addition, the Bcl2 gene is one of the most important regulatory genes in the apoptosis regulation process, and the Bax protein, as a Bcl-2 related gene, has the capability of antagonizing Bcl-2 gene to inhibit apoptosis and participates in the regulation of the apoptosis process.

The inventor further researches the expression of pancreatic cancer apoptosis related proteins such as Caspase-3, Caspase-8, cleared-Caspase-3, cleared-Caspase 8, cleared-Caspase 9, Bax and Bcl2, and finds that buckwheat flavonoids can up-regulate the expression of the apoptotic proteins Caspase-3, Caspase-8, cleared-Caspase-3, cleared-Caspase 8, cleared-Caspase 9 and Bax and down-regulate or inhibit the expression of the anti-apoptotic protein Bcl-2. Therefore, the tartary buckwheat flavone has the activity function of promoting the apoptosis of pancreatic cancer cells, and further plays the role of resisting the pancreatic cancer.

Therefore, the tartary buckwheat flavone can be used as a novel medicine for preventing, treating or improving pancreatic cancer, pancreatic tumor and other related diseases, and can also provide a basis for evaluating the treatment effect of the pancreatic cancer, pancreatic tumor and other related diseases.

According to the invention, in the application of the tartary buckwheat flavone in preparing the medicine for preventing, treating or improving pancreatic tumors, the effective using dose of the tartary buckwheat flavone is 1-100 mu g/mL, preferably 5-50 mu g/mL, such as 5 mu g/mL, 10nmol/L, 20nmol/L, 40nmol/L and 50 nmol/L.

The inventor researches the influence of the buckwheat flavonoids on the proliferation activity of pancreatic cancer cells by using the buckwheat flavonoids with different treatment doses, and finds that the inhibition proliferation effect of the buckwheat flavonoids with high treatment dose on the pancreatic cancer cells is more obvious, and the treatment effect is optimal when the effective use dose of the buckwheat flavonoids is 1-100 mug/mL.

Wherein, the content of total flavone in the tartary buckwheat flavone reaches more than 80 wt%, the total flavone mainly comprises rutin and quercetin, wherein the rutin accounts for more than 75 wt% of the total flavone, and the quercetin accounts for more than 1 wt% of the total flavone; further, the content of total flavonoids in the tartary buckwheat flavonoids is more than 83 wt%, rutin accounts for more than 79 wt% of the total flavonoids, and quercetin accounts for more than 2 wt% of the total flavonoids.

According to the invention, the buckwheat flavone is not limited to buckwheat flavone produced by any manufacturer on the market or prepared by self-making, and preferably the buckwheat flavone is prepared by alcohol extraction and resin purification of buckwheat powder.

Examples

The present invention is further described below by way of specific examples, which are merely exemplary and do not limit the scope of the present invention in any way.

Example 1

Preparation of buckwheat flavone

Extracting radix Et rhizoma Fagopyri Tatarici powder in 63% ethanol at 70 deg.C for 2 hr, filtering with circulating water multi-purpose vacuum pump, collecting supernatant, concentrating with rotary evaporator for 30min at 60 deg.C to obtain extractive solution, and dissolving the extractive solution with 28% ethanol;

washing AB-8 macroporous adsorbent resin with 98% ethanol at room temperature, loading onto column, injecting the ethanol-dissolved extractive solution into the column, standing for 2h, slowly eluting with water, discarding water-washing solution, eluting with 60% ethanol, collecting eluate, recovering ethanol under reduced pressure, concentrating and drying Tartary buckwheat flavone to obtain powder, which is extracted Tartary buckwheat flavone, and analyzing by HPLC to obtain extract containing 83 wt% of total flavone, 80.7 wt% of rutin, and 2.1 wt% of quercetin.

Example 2

Cell culture

The cell complete medium (DMEM medium, purchased from Gibco, USA) was taken 30min in advance and returned to room temperature, and 10% fetal bovine serum and 0.1% penicillin-streptomycin were inoculated into the DMEM medium to obtain DMEM medium containing 10% fetal bovine serum and 0.1% penicillin-streptomycin double antibody. Subsequently, PANC-1 cells (pancreatic cancer ductal cells, purchased from American type culture Collection (ATCC, Manassas, VA, U.S.) were seeded in DMEM medium containing 10% fetal bovine serum and 0.1% dual anti-S, and placed at 37 ℃ in 5% CO₂Culturing in a cell culture box. Replacing the culture medium every 24h, finishing the culture operation of the PANC-1 cells when the PANC-1 cells are in the logarithmic growth phase, and collecting the cultured PANC-1 cells for later use.

Example 3

Western Blot Western Blot detection

Dividing the PANC-1 cells obtained by culturing in the example 2 into 3 groups on average, digesting each group by 0.25% pancreatin, inoculating the cells into a 96-well plate, and inoculating 6 multiple wells in each group, and performing Western Blot Western Blot detection according to the following steps when the PANC-1 cells grow to the density of 50-60% respectively:

a first group: the PANC-1 cells after growth were not treated with any drugs, and total protein was extracted with RIPA cell protein lysate (from Biyuntian Biotech Co., Ltd.), and the protein concentration was measured with BCA assay kit (from Biyuntian Biotech Co., Ltd.). After the protein sample is quantified, the protein sample is mixed with Loading Buffer (5X) (purchased from Biyuntian biotechnology Co., Ltd.) and boiled at 99 ℃ for 10min for later use. SDS-PAGE electrophoresis, after electrophoresis, loading into a membrane transfer tank, sealing the periphery with ice, and transferring by electricity for 90min to 300 mA. The membrane was removed with forceps and added to phosphate buffer containing 5% skimmed milk powder (purchased from Biyuntian biotechnology Co., Ltd.) and blocked for 1.5 h. After the sealing is finished, the membrane is washed by phosphate buffer solution for 3 times and 5 min/time. Specific antibodies (Caspase-3, Caspase-8, cleared-Caspase-3, cleared-Caspase 8, cleared-Caspase 9, Caspase-9, Bax, MMP9, Bcl-2, and β -actin) (from Biyun Biotech, Inc.) were used to incubate for 2h at room temperature. PBS-T washing the membrane 3 times, 10 min/time. Secondary antibodies (Caspase-3, Caspase-8, cleared-Caspase-3, cleared-Caspase 8, cleared-Caspase 9, Bax, MMP9, Bcl-2 and beta-actin) (from Biyun Biotech, Inc.) labeled with horseradish peroxidase were then incubated for 1h at room temperature. The membrane was washed 3 times with phosphate buffer solution for 10 min/time. Absorbing phosphate buffer solution completely, adding ECL developer (purchased from Biyuntian biotechnology limited) in a volume ratio of 1:1, exposing, and marking as a Control group, wherein the Control group is represented in the figure;

second group: western Blot detection was performed in a similar manner to the first panel, except that: after the growth of PANC-1 cells was treated with 5. mu.g/mL of the Tartary Buckwheat Flavone (TBF) prepared in example 1, total protein was extracted with RIPA cell protein lysate (from Biyutian Biotechnology Co., Ltd.) and subjected to subsequent treatment, which is shown as 5. mu.g/mL of TBF;

third group: western Blot detection was performed in a similar manner to the second panel, except that: the PANC-1 cells after growth were treated with 10. mu.g/mL of the Tartary Buckwheat Flavone (TBF) prepared in example 1, which is shown as 10. mu.g/mL of TBF.

The results of the expression of Caspase-3, Caspase-8, cleared-Caspase-3, cleared-Caspase 8, cleared-Caspase 9, Bax, MMP-9, Bcl-2 and beta-actin by the first to third groups of PANC-1 cells are shown in FIG. 1, and the results show that 5. mu.g/mL and 10. mu.g/mL of buckwheat flavonoids up-regulate the expression of Caspase-3, Caspase-8, cleared-Caspase-3, cleared-Caspase 8, cleared-Caspase 9 and Bax apoptotic proteins and down-regulate the expression of Bcl-2, compared with the control group. Therefore, the tartary buckwheat flavone has the activity function of promoting the apoptosis of PANC-1 cells. The data result also indicates that the tartary buckwheat flavone can play an anti-pancreatic cancer activity role by promoting the apoptosis of PANC-1 cells.

Example 4

TUNEL apoptosis assay

To explore the effect of buckwheat flavonoids on the apoptosis ability of PANC-1 cells, the PANC-1 cells cultured in example 2 were incubated as follows:

dividing the PANC-1 cells obtained by culturing in the example 2 into 3 groups on average, digesting each group by 0.25% pancreatin, inoculating the cells into a 96-well plate, and inoculating 6 multiple wells in each group, and performing TUNEL apoptosis detection experiments respectively according to the following steps when the PANC-1 cells grow to the density of 50-60%:

the first group: the PANC-1 cells after growth were not treated with any drugs, and 500. mu.L of 4% paraformaldehyde (purchased from Biyuntian Biotech Co., Ltd., the same shall apply hereinafter) was added to fix the PANC-1 cells for 30 min. After that, the cells were washed once with a sterilized phosphate buffer solution (1 XPBS), 500. mu.L of a 0.3% TritonX-100 penetrant solution (purchased from Biyuntian Biotech Co., Ltd., the same applies hereinafter) was added, and then incubated at room temperature for 5 min. Washing with 1 XPBS twice, adding 50 μ L of TUNEL assay (from Biyuntian Biotechnology Co., Ltd., the same below) to each well, and keeping out light. Then incubated at 37 ℃ in the dark for 1 h. Photographs were taken after three washes with 1 × PBS. This group was designated as a Control group and indicated by Control in the figure;

the second group: the PANC-1 cells after growth were treated with 5. mu.g/mL of the Tartary Buckwheat Flavone (TBF) prepared in example 1, and then fixed with 500. mu.L of 4% paraformaldehyde for 30 min. After which it was washed once with sterile phosphate buffered saline (1 XPBS), 500. mu.L of 0.3% Triton X-100 penetrant solution was added, followed by incubation at room temperature for 5 min. Wash twice with 1 XPBS, add 50. mu.L of TUNEL assay to each well, and care away from light. Then incubated at 37 ℃ in the dark for 1 h. Photographs were taken after three washes with 1 × PBS. Expressed as 5. mu.g/mL TBF;

group III: TUNEL apoptosis assay was performed in a similar manner to group ii except that: the PANC-1 cells after growth were treated with 10. mu.g/mL of the Tartary Buckwheat Flavone (TBF) prepared in example 1, which is shown as 10. mu.g/mL of TBF.

The experimental photographs of TUNEL apoptosis detection of the first to third groups are shown in FIG. 2, and the results show that, compared with the control group, 5. mu.g/mL and 10. mu.g/mL of buckwheat flavonoids promote apoptosis of PANC-1 cells, so that DNA fragmentation is more obvious. Wherein, when the weight concentration of the tartary buckwheat flavone is 10 mug/mL, the apoptosis promotion effect of the PANC-1 cell is more obvious.

Example 5

CCK-8(Cell Counting Kit-8) Cell proliferation assay

To explore the effect of buckwheat flavonoids on the proliferation activity of PANC-1 cells, the PANC-1 cells cultured in example 2 were incubated as follows:

dividing the PANC-1 cells obtained by culturing in example 2 into 7 groups on average, digesting each group with 0.25% pancreatin, inoculating the digested cells into a 96-well plate, and inoculating 6 multiple wells in each group, and detecting the proliferation activity of the PANC-1 cells when the PANC-1 cells grow to 50-60% of density according to the following steps:

group I: the PANC-1 cells after growth are not treated by any drug, 10 mu L of CCK-8 reagent (purchased from Biyunstian biotechnology, Inc., the same below) is respectively added into each well, after incubation for 2h, the absorbance value of A450 is measured by an enzyme-labeling instrument, A650 is used as a reference wavelength for dual-wavelength measurement, then the data is subjected to statistical analysis and is marked as a Control group, and the Control is represented in the figure;

group II: using dimethyl sulfoxide (DMSO) culture solution as negative control, specifically, treating PANC-1 cells which have finished growth by using DMSO culture solution with the concentration of 100%, continuing to culture for 24h, then respectively adding 10 mu L of CCK-8 reagent into each hole, incubating for 2h, then measuring the absorbance value of A450 by using an enzyme labeling instrument, taking A650 as reference wavelength to carry out dual-wavelength measurement, and then carrying out statistical analysis on the data, wherein DMSO is used as the reference wavelength in the figure;

group III: proliferation activity of PANC-1 cells was examined in a similar manner to group ii, except that: the PANC-1 cells after growth are treated with 1. mu.g/mL of the Tartary Buckwheat Flavone (TBF) prepared in example 1, which is expressed as 1. mu.g/mL of TBF;

group IV: proliferation activity of PANC-1 cells was examined in a similar manner to group ii, except that: the PANC-1 cells after growth were treated with 5. mu.g/mL of the Tartary Buckwheat Flavone (TBF) prepared in example 1, which is shown as 5. mu.g/mL of TBF;

group V: proliferation activity of PANC-1 cells was examined in a similar manner to group ii, except that: the PANC-1 cells after growth were treated with 10. mu.g/mL of the Tartary Buckwheat Flavone (TBF) prepared in example 1, which is 10. mu.g/mL of TBF;

group VI: proliferation activity of PANC-1 cells was examined in a similar manner to group ii, except that: the PANC-1 cells after growth are treated with 20 μ g/mL of the Tartary Buckwheat Flavone (TBF) prepared in example 1, which is expressed as 20 μ g/mL of TBF;

group VII: proliferation activity of PANC-1 cells was examined in a similar manner to group ii, except that: treating PANC-1 cells with 40 μ g/mL of Tartary Buckwheat Flavone (TBF) prepared in example 1, wherein the TBF is 40 μ g/mL;

the results of the statistical analysis of the 7 experiments are shown in FIG. 3 (wherein the mean. + -. standard error of the mean:. about. p <0.001), and show that the proliferation activity of PANC-1 cells is decreased as the weight concentration of the flavonoids is increased. When the weight concentration of the tartary buckwheat flavone is 5 mug/mL, the tartary buckwheat flavone has obvious effect of inhibiting the cell proliferation activity of PANC-1, and statistical analysis shows that the tartary buckwheat flavone has significant difference. The effect of inhibiting the proliferation activity of PANC-1 cells is most obvious when the weight concentration of the tartary buckwheat flavone is 40 mug/mL.

From the above results, it was found that tartary buckwheat flavonoids have the function of inhibiting the proliferation activity of PANC-1 cells. Analysis shows that the inhibition effect is concentration-dependent, namely the larger the weight concentration of the tartary buckwheat flavone is, the more remarkable the effect of inhibiting proliferation is. In addition, the tartary buckwheat flavone is also suggested to play an anti-tumor activity role by inhibiting the proliferation of PANC-1 cells.

The invention has been described in detail with reference to the preferred embodiments and illustrative examples. It should be noted, however, that these specific embodiments are only illustrative of the present invention and do not limit the scope of the present invention in any way. Various modifications, equivalent substitutions and alterations can be made to the technical content and embodiments of the present invention without departing from the spirit and scope of the present invention, and these are within the scope of the present invention. The scope of the invention is defined by the appended claims.

Claims

1. The pharmaceutical composition for preventing, treating or improving pancreatic tumors is characterized by containing the tartary buckwheat flavone, wherein the content of total flavone in the tartary buckwheat flavone is more than 80 wt%.

2. The pharmaceutical composition of claim 1, wherein said total flavonoids comprise rutin and quercetin, wherein rutin comprises greater than 75 wt% of the total flavonoids and quercetin comprises greater than 1 wt% of the total flavonoids.

3. The pharmaceutical composition according to claim 1 or 2, wherein the effective dose of the tartary buckwheat flavone is 1-100 μ g/mL.

4. The pharmaceutical composition of claim 3, wherein the formulation of the pharmaceutical composition comprises tablets, capsules, pills, granules, oral liquid, and injections.

5. Application of Tartary buckwheat flavone in preparing medicine for preventing, treating or improving pancreatic tumor is provided.

6. The use according to claim 5, wherein the tartary buckwheat flavone is used for preparing a medicament for preventing, treating or improving pancreatic cancer.

7. The use of claim 6, wherein the tartary buckwheat flavone can regulate the expression of pancreatic cancer-related proteins, up-regulate the expression of apoptosis proteins Caspase-3, Caspase-8, cleared-Caspase-3, cleared-Caspase 8, cleared-Caspase 9 and Bax, and down-regulate the expression of Bcl-2.

8. The use according to one of claims 5 to 7, wherein the pancreatic cancer comprises ductal cell carcinoma, acinar cell carcinoma, islet cell carcinoma and the rare types of pancreatic cancer comprising undifferentiated carcinoma, carcinosarcoma, pancreatic blast.

9. The use of claim 8, wherein the content of total flavonoids in the tartary buckwheat flavonoids is up to 80 wt%.

10. The use according to one of claims 5 to 9, wherein the buckwheat flavonoids is prepared from buckwheat powder by alcohol extraction and resin purification.