CN111840358A - Application of jingulian capsule in preparing anti-breast cancer medicine - Google Patents
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Abstract
The invention provides application of a tropaeolum capsule in preparing an anti-breast cancer medicament, and relates to the technical field of biological pharmacy. The application of the invention particularly relates to the preparation of a pharmaceutical preparation for inhibiting the activity of breast cancer cells, inhibiting the proliferation of the breast cancer cells and promoting the apoptosis of the breast cancer cells. The invention provides a medicinal preparation for treating breast cancer. The invention proves that the Jingulian capsule can inhibit the activities of MDA-MB-231 cells and 4T1 cells and has concentration dependence; can inhibit MDA-MB-231 cell proliferation, and is concentration and time dependent; can alter MDA-MB-231 cell morphology; the human breast cancer cell MDA-MB-231 has obvious apoptosis influence; human breast cancer cells MDA-MB-23 arrest the cell cycle in S phase. And the jingu lotus capsule is a clinically used Miao medicine, can be directly applied to a human body without clinical safety evaluation, and has good application prospect.
Description
Technical Field
The invention belongs to the technical field of biological pharmacy, and particularly relates to an application of a tropaeolum capsule in preparation of an anti-breast cancer medicament.
Background
The Jingulian capsule is a kind of Miao medicine prepared with Yibai medicine from Guizhou province company, and consists of Tou Guxiang, Chinese peach leaf, sargentgloryvine stem, Chinese alangium and psammosilene tunicoides as main material and corn starch as supplementary material. The product is hard capsule, and the content is yellowish brown to reddish brown powder, with light smell and slightly bitter and astringent taste. The capsule has the effects of expelling wind, removing dampness, relieving swelling and alleviating pain. The medicine can be used for treating joint diseases clinically, particularly rheumatism, rheumatoid and other reasons, and joint swelling, pain and difficulty in flexion and extension caused by the medicine can be treated by the Jingulian capsule. The medicine has the effect of promoting the repair of cartilage in joints, can well improve circulation and promote blood circulation to remove blood stasis, has great benefit for improving the life quality of patients, but has no record of using the medicine for preventing and treating breast cancer.
Disclosure of Invention
In view of the above, the present invention aims to provide an application of a tropaeolum capsule in the preparation of an anti-breast cancer drug, which can significantly inhibit the activity and proliferation of breast cancer cells, promote cancer cell apoptosis, and have a potential value in the prevention and treatment of breast cancer or the inhibition of breast cancer cell proliferation.
In order to achieve the above object, the present invention provides the following technical solutions:
the invention provides application of a tropaeolum capsule in preparing a medicinal preparation for treating breast cancer.
The invention provides application of a tropaeolum capsule in preparing a medicinal preparation for inhibiting breast cancer cell activity.
Preferably, the breast cancer cells comprise human breast cancer cells MDA-MB-231 and mouse breast cancer cells 4T 1.
Preferably, the pharmaceutical preparation comprises injection, powder, tablet, granule, capsule, solution, emulsion, suspension, spray, aerosol, powder spray, ointment, plaster, patch, suppository, drop or dripping pill.
The invention also provides application of the jingu lotus capsule in preparing a medicinal preparation for inhibiting breast cancer cell proliferation.
Preferably, the breast cancer cells comprise human breast cancer cells MDA-MB-231 and mouse breast cancer cells 4T 1.
The invention also provides application of the jingu lian capsule in preparing a medicinal preparation for promoting breast cancer cell apoptosis.
Preferably, the breast cancer cells comprise human breast cancer cells MDA-MB-231 and mouse breast cancer cells 4T 1.
The invention also provides a medicinal preparation for treating breast cancer, which comprises the active ingredients of the tropaeolum capsule or the tropaeolum capsule.
The invention also provides a preparation method of the pharmaceutical preparation, which comprises the following steps: the Jingu lotus capsule is directly prepared into a medicinal preparation for treating breast cancer, or the Jingu lotus capsule and other medicines for resisting breast cancer are compounded to prepare a compound medicinal preparation, or the effective components of the Jingu lotus capsule and a pharmaceutically acceptable carrier are prepared into a medicinal preparation for treating breast cancer.
The invention provides application of a jingu lian capsule in preparing a medicinal preparation for treating breast cancer, and particularly relates to an application of a jingu lian capsule in inhibiting activity of breast cancer cells, inhibiting proliferation of the breast cancer cells, promoting apoptosis of the breast cancer cells and changing forms of the breast cancer cells, so that an effect of treating the breast cancer is achieved. In the embodiment of the invention, the human breast cancer cell MDA-MB-231 and the mouse breast cancer cell 4T1 are tested by using the physiological saline solution of the contents of the jingu lotus capsule, and samples with different concentrations are provedThe solution can inhibit the activity of MDA-MB-231 cells and 4T1 cells, and has concentration dependence and IC500.13 plus or minus 0.02mg/ml and 0.59 plus or minus 0.26mg/ml respectively; the sample solutions with different concentrations can inhibit the proliferation of the MDA-MB-231 cells, and the growth rate of the MDA-MB-231 cells is gradually reduced along with the increase of the concentration and is in a concentration and time dependence; the number of MDA-MB-231 cells is obviously reduced along with the increase of the concentration of the sample solution, and after the sample solution acts on the MDA-MB-231 cells, the cell morphology is changed, namely, the long fusiform is changed into the round shape; sample solutions with different concentrations have obvious influence on apoptosis of human breast cancer cells MDA-MB-231; the sample solutions with different concentrations have obvious influence on the MDA-MB-231 cycle of human breast cancer cells, block the cell cycle in the S phase, and the higher the drug concentration is, the more obvious the block is. Therefore, the jingu lian capsule has potential value in preparing medicine for preventing and treating breast cancer or breast cancer cell proliferation inhibiting medicine preparation. And the jingu lotus capsule is a clinically used Miao medicine, can be directly applied to a human body without clinical safety assessment, has a good application prospect, and can rapidly improve the market share of the product.
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FIG. 1 shows the inhibition rate of Jingulian capsule with different concentrations on human breast cancer cell MDA-MB-231;
FIG. 2 shows the inhibition detection rate of Jingulian capsule at different concentrations on mouse breast cancer cell 4T 1;
FIG. 3 is a graph showing the effect of different concentrations of the tropaeolum capsule on the growth of human breast cancer cell MDA-MB-231;
FIG. 4 is a graph showing the effect of different concentrations of the tropaeolum capsule on the morphology of human breast cancer cells MDA-MB-231;
FIG. 5 shows the result of apoptosis detection by flow cytometry after different concentrations of tropaeolum capsules act on human breast cancer MDA-231 cells for 24h and 48 h;
FIG. 6 shows the results of cycle detection by flow cytometry after different concentrations of the tropaeolum capsule act on human breast cancer cell MDA-231 cells for 24h and 48 h.
Detailed Description
The invention also provides application of the jingu lotus capsule in preparing a medicinal preparation for treating breast cancer.
The Jingulian capsule is manufactured by Guizhou Yibai pharmaceutical products GmbH. The experiment with human breast cancer cell MDA-MB-231 and mouse breast cancer cell 4T1 is preferred in the examples of the present invention, but it is not considered as the specific protection scope of the present invention. The sources of the human breast cancer cell MDA-MB-231 and the mouse breast cancer cell 4T1 are not particularly limited, and the human breast cancer cell MDA-MB-231 and the mouse breast cancer cell 4T1 can be purchased commercially or frozen in a laboratory, and in the embodiment of the invention, the human breast cancer cell MDA-MB-231 and the mouse breast cancer cell 4T1 are purchased from American ATCC. The embodiment of the invention proves that after the two breast cancer cells are treated by normal saline solution (0.125mg/ml, 0.25mg/ml, 0.5mg/ml, 1mg/ml and 2mg/ml) of the active ingredients of the tropaeolum capsule for 72 hours, the MTT method detects the inhibition rate of sample solutions with different concentrations on MDA-MB-231 cells and 4T1 cells, and the result shows that the sample solutions with different concentrations can inhibit the activities of the MDA-MB-231 cells and 4T1 cells, and have concentration dependence and IC500.13 plus or minus 0.02mg/ml and 0.59 plus or minus 0.26mg/ml respectively.
The dosage form of the pharmaceutical preparation is not particularly limited, and preferably comprises injection, powder, tablets, granules, capsules, solutions, emulsions, suspensions, sprays, aerosols, powders, ointments, plasters, patches, suppositories, drops or dripping pills; the tablet preferably comprises a sublingual tablet.
The invention provides application of a tropaeolum capsule in preparing a medicinal preparation for inhibiting breast cancer cell activity. In the application of the present invention, the sources of the jingu lian capsules, the sources of the breast cancer cells, the dosage forms of the pharmaceutical preparations, and the like are preferably the same as those described above, and are not described herein again.
The invention also provides application of the jingu lotus capsule in preparing a medicinal preparation for inhibiting breast cancer cell proliferation. In the application of the present invention, the sources of the jingu lian capsules, the sources of the breast cancer cells, the dosage forms of the pharmaceutical preparations, and the like are preferably the same as those described above, and are not described herein again.
The invention also provides application of the jingu lian capsule in preparing a medicinal preparation for promoting breast cancer cell apoptosis. In the application of the present invention, the sources of the jingu lian capsules, the sources of the breast cancer cells, the dosage forms of the pharmaceutical preparations, and the like are preferably the same as those described above, and are not described herein again.
The invention also provides a medicinal preparation for treating breast cancer, which comprises the active ingredients of the tropaeolum capsule or the tropaeolum capsule. In the application of the present invention, the sources of the jingu lian capsules, the sources of the breast cancer cells, the dosage forms of the pharmaceutical preparations, and the like are preferably the same as those described above, and are not described herein again.
The invention also provides a preparation method of the pharmaceutical preparation, which comprises the following steps: the Jingu lotus capsule is directly prepared into a medicinal preparation for treating breast cancer, or the Jingu lotus capsule and other medicines for resisting breast cancer are compounded to prepare a compound medicinal preparation, or the effective components of the Jingu lotus capsule and a pharmaceutically acceptable carrier are prepared into a medicinal preparation for treating breast cancer.
The application of the jingu lian capsule in preparing anti-breast cancer drugs provided by the present invention is described in detail with reference to the following examples, but they should not be construed as limiting the scope of the present invention.
Preparing a sample solution: the contents of the jingu lotus capsule are prepared into sample solutions with the concentrations of 0.125mg/ml, 0.25mg/ml, 0.5mg/ml, 1mg/ml and 2mg/ml by using physiological saline respectively.
Preparation of MTT solution: 0.2g of MTT powder is weighed, 40ml of sterilized double distilled water is added, the mixture is dissolved at 60 ℃, the concentration is 5mg/ml, the mixture is filtered and sterilized by a 0.2 mu m filter membrane, and the mixture is stored at 4 ℃.
Cell line: human breast cancer cells MDA-MB-231, mouse breast cancer cells 4T1, were purchased from ATCC in the United states. Cells at 37 ℃ and 5% CO2The culture was performed in a culture box using DMEM medium containing 5% FBS.
Example 1
Cell viability assay
Logarithmic phase cells were collected, MDA-MB-231 cells and 4T1 cells were packed at 6X 103Inoculating the cells in 96-well plate at a density per well, culturing overnight, and adding sample solutions with different concentrations for treatment after the cells adhere to the wallPhysiological saline was used as a control group. And after the drug acts for 72 hours, adding 10 mu l of MTT with the concentration of 5mg/ml into each hole, incubating for 4 hours at 37 ℃, discarding the supernatant, adding 160 mu l of DMSO into each hole for dissolving, and measuring the absorbance value OD (490nm) by using an enzyme-linked immunosorbent assay (ELISA) after the blue-purple crystal formazan is completely dissolved.
The calculation method of the inhibition rate comprises the following steps: inhibition rate (control OD value-treatment OD value)/control OD value. Median inhibitory rate (IC)50) The calculating method of (2): obtaining corresponding IC by using forecast function50。
As shown in FIGS. 1 and 2, after MDA-MB-231 cells and 4T1 cells were treated with sample solutions of 0.125mg/ml, 0.25mg/ml, 0.5mg/ml, 1mg/ml and 2mg/ml for 72 hours, the MTT method examined the inhibition rates of the sample solutions of different concentrations on MDA-MB-231 cells and 4T1 cells. The sample solutions with different concentrations can inhibit the activity of MDA-MB-231 cells and 4T1 cells, and have concentration dependence and IC500.13 plus or minus 0.02mg/ml and 0.59 plus or minus 0.26mg/ml respectively.
Example 2
Cell growth curve assay
Logarithmic phase MDA-MB-231 cells were collected according to 6X 103The density of each hole is inoculated in a 96-hole plate, the culture is carried out overnight, after the cells adhere to the wall, sample solutions with different concentrations (0.25mg/ml, 0.5mg/ml, 1mg/ml and 2mg/ml) are added for treatment, the OD 490nm is measured in 12h, 24h, 36h, 48h and 72h respectively, and a growth curve is drawn.
As shown in FIG. 3, the sample solutions with different concentrations all inhibited the proliferation of MDA-MB-231 cells, and the growth rate of MDA-MB-231 cells gradually decreased with increasing concentration, indicating that the sample solutions were able to inhibit the proliferation of MDA-MB-231 and were concentration-and time-dependent.
Example 3
Cell morphology observation
Logarithmic phase MDA-MB-231 cells were collected according to 6X 103The density of each well is inoculated in a 96-well plate, the culture is carried out overnight, after the cells are attached to the wall, sample solutions (0.25mg/ml, 0.5mg/ml and 1mg/ml) with different concentrations are added for treatment, and normal saline is used as a control group. After 24h, the cells were observed for morphological changes.
As shown in FIG. 4, the number of MDA-MB-231 cells decreased significantly as the concentration of the sample solution increased. 1mg/ml of the sample solution obviously inhibits the cell growth, and after the sample solution acts on MDA-MB-231 cells, the cell morphology is changed, and the cell morphology is changed from a long fusiform shape to a round shape.
Example 4
Apoptosis detection
MDA-MB-231 cells were plated at 2X 105The amount of each hole is inoculated in a circular culture dish of 60mm for culture, after the cells are attached to the wall, sample solutions (0.5mg/ml and 1mg/ml) with different concentrations are added for 24 hours or 48 hours, and then the cells are collected. Washing with PBS 3 times, adding 500 μ l 1 × Binding Buffer to resuspend cells, adding 5 μ l annexin V-FITC and 10 μ l propidium iodide, incubating at room temperature in dark for 15min, and detecting with flow cytometer to calculate the apoptosis rate.
The result is shown in fig. 5, the jingu lotus capsule has significant effect on human breast cancer cell MDA-MB-231 apoptosis.
Example 5
Cell cycle assay
MDA-MB-231 cells were plated at 2X 105The amount of each hole is inoculated in a circular culture dish of 60mm for culture, after the cells are attached to the wall, sample solutions (0.5mg/ml and 1mg/ml) with different concentrations are added for 24 hours or 48 hours, and then the cells are collected. Centrifuge at 1000rpm for 5min, wash once with pre-chilled PBS, resuspend cells with 70% pre-chilled ethanol, and fix overnight at-20 ℃. Centrifuging at 1000rpm for 5min, discarding the supernatant, washing with precooled PBS once, centrifuging at 1000rpm for 5min, and collecting the cells. The cells were resuspended in 500. mu.l PBS, 5. mu.l RNaseA, 25. mu.l PI and 1000.25. mu.l TritonX were added, mixed, centrifuged at 1000rpm for 5min, the dye was discarded, resuspended in 200. mu.l PBS and cell cycle analysis was performed using flow cytometry.
The result is shown in fig. 6, when the Jingu lotus capsule is added with medicine for 24h and 48h, the human breast cancer cell MDA-MB-231 cycle is obviously influenced, the cell cycle is blocked in the S phase, and the higher the medicine concentration is, the more obvious the blocking is.
In the above examples, each experiment was independently repeated three times, all data were expressed as mean ± SD, Student's t-test was performed with GraphPadPrism 7, P <0.05 was considered statistically significant, P <0.05, P < 0.01.
From the above results, the jingu lian capsule has potential value in preparing a medicine for preventing and treating breast cancer or a medicine preparation for inhibiting breast cancer cell proliferation.
The foregoing is only a preferred embodiment of the present invention, and it should be noted that, for those skilled in the art, various modifications and decorations can be made without departing from the principle of the present invention, and these modifications and decorations should also be regarded as the protection scope of the present invention.
Claims (10)
1. Application of JINGULIAN Capsule in preparing medicinal preparation for treating breast cancer is provided.
2. Application of JINGULIAN Capsule in preparing medicinal preparation for inhibiting breast cancer cell activity is provided.
3. The use of claim 2, wherein the breast cancer cells comprise human breast cancer cells MDA-MB-231 and mouse breast cancer cells 4T 1.
4. The use according to claim 2, wherein the pharmaceutical preparation comprises an injection, powder, tablet, granule, capsule, solution, emulsion, suspension, spray, aerosol, powder spray, ointment, plaster, patch, suppository, drop or dripping pill.
5. Application of JINGULIAN Capsule in preparing medicinal preparation for inhibiting breast cancer cell proliferation is provided.
6. The use of claim 5, wherein the breast cancer cells comprise human breast cancer cells MDA-MB-231 and mouse breast cancer cells 4T 1.
7. Application of JINGULIAN Capsule in preparing medicinal preparation for promoting breast cancer cell apoptosis is provided.
8. The use of claim 7, wherein the breast cancer cells comprise human breast cancer cells MDA-MB-231 and mouse breast cancer cells 4T 1.
9. The medicinal preparation for treating breast cancer is characterized by comprising a tropaeolum capsule or an effective component of the tropaeolum capsule.
10. A method of preparing a pharmaceutical formulation according to claim 9, comprising the steps of: the Jingu lotus capsule is directly prepared into a medicinal preparation for treating breast cancer, or the Jingu lotus capsule and other medicines for resisting breast cancer are compounded to prepare a compound medicinal preparation, or the effective components of the Jingu lotus capsule and a pharmaceutically acceptable carrier are prepared into a medicinal preparation for treating breast cancer.
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Cited By (2)
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| CN114869977A (en) * | 2022-03-28 | 2022-08-09 | 贵州省中国科学院天然产物化学重点实验室(贵州医科大学天然产物化学重点实验室) | Application of lung cough in preparation of product for preventing and treating breast cancer |
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| CN113230299A (en) * | 2021-04-28 | 2021-08-10 | 贵州省中国科学院天然产物化学重点实验室(贵州医科大学天然产物化学重点实验室) | Application of Jingan capsule in preparing medicine for treating breast cancer |
| CN114869977A (en) * | 2022-03-28 | 2022-08-09 | 贵州省中国科学院天然产物化学重点实验室(贵州医科大学天然产物化学重点实验室) | Application of lung cough in preparation of product for preventing and treating breast cancer |
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