CN117045628A - Research and application of alkaloid compound in prevention and treatment of abdominal aortic aneurysm - Google Patents
Research and application of alkaloid compound in prevention and treatment of abdominal aortic aneurysm Download PDFInfo
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Classifications
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/16—Amides, e.g. hydroxamic acids
- A61K31/165—Amides, e.g. hydroxamic acids having aromatic rings, e.g. colchicine, atenolol, progabide
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- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/18—Magnoliophyta (angiosperms)
- A61K36/88—Liliopsida (monocotyledons)
- A61K36/896—Liliaceae (Lily family), e.g. daylily, plantain lily, Hyacinth or narcissus
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- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K49/00—Preparations for testing in vivo
- A61K49/0004—Screening or testing of compounds for diagnosis of disorders, assessment of conditions, e.g. renal clearance, gastric emptying, testing for diabetes, allergy, rheuma, pancreas functions
- A61K49/0008—Screening agents using (non-human) animal models or transgenic animal models or chimeric hosts, e.g. Alzheimer disease animal model, transgenic model for heart failure
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P35/00—Antineoplastic agents
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Abstract
The invention discloses an application of alkaloid compounds in preventing and treating abdominal aortic aneurysm, and a research method thereof comprises the following steps: step 1: firstly, performing mouse AAA molding; step 2: injecting colchicine 0.1mg/kg into the abdominal cavity of the mouse while molding the mouse AAA; step 3: after 2 weeks, the abdominal aorta of the mice was isolated, and the extent of aortic expansion and injury were assessed by staining the blood vessels HE/EVG/Masson; step 4: finally, the abdominal aortic tissue of the mice is detected, and the possible molecular mechanism of the colchicine for improving the AAA injury of the mice is explored. The colchicine with definite anti-inflammatory action is provided for the first time, the mouse AAA injury can be improved by inhibiting the infiltration of the adventitia inflammatory cells of the vascular wall, inhibiting the conversion of vascular smooth muscle cells to synthesis, the expression of matrix metalloproteinase and the like, the novel application of the colchicine which is a common clinical medicine is disclosed, and a novel intervention strategy and a research and development idea are provided for the prevention and treatment of abdominal aortic aneurysm.
Description
Technical Field
The invention relates to the technical field of medicines, in particular to a study and application of an alkaloid compound in preventing and treating abdominal aortic aneurysm.
Background
Abdominal aortic aneurysms (abdominal aortic aneurysm, AAA) are an aortic dilation disorder in which the aortic diameter irreversibly expands, exceeding 50% of normal diameter, for a number of reasons. In humans, AAA is considered to occur when the aortic diameter (especially the diameter of the aortic segment below the renal artery bifurcation) >3cm, AAA is one of the most risky vascular degenerative diseases in vascular surgery. The disease of AAA is hidden, along with the continuous expansion of the aorta, the tumor body is continuously enlarged, and the risk of rupture is gradually increased; once broken, its mortality rate is as high as 80%. Statins, angiotensin converting enzyme inhibitors, angiotensin II receptor antagonists and the like can delay AAA progression to some extent, but their exact efficacy is to be clarified. No clear indication is shown for AAA, and searching for new targets and new therapeutic drugs for AAA is needed.
Colchicine is a neutral lipophilic tricyclic alkaloid extracted from seeds and bulbs of colchicine, a plant of the family liliaceae, and was first obtained by crude extraction in the first year of 1820 by Pelletier and Caventou. Corrodi and Hardegger in 1955 revealed the chemical structure of colchicine, widening its clinical applications. Colchicine is approved by the united states food and drug administration (american's Food and Drug Administration, FDA) for the treatment of familial mediterranean fever, gout, arthritis, biliary cirrhosis, and the like. Colchicine plays an anti-inflammatory and immune-suppressing role mainly by suppressing neutrophil and macrophage functions; and simultaneously, by blocking microtubule formation, inhibiting cell mitosis and playing an anti-fibrosis role. The FDA published an announcement in 2023: a tablet of colchicine 0.5mg can be used to treat patients with existing atherosclerotic disease, or adults with multiple cardiovascular risk factors. Also, a study in the European journal of prevention cardiology has shown that the use of colchicine at low doses can benefit patients who have been treated with antihypertensive and lipid-lowering drugs.
The research finds that: hypertension and hyperlipidemia are important risk factors for inducing AAA to cause tumor rupture. However, there is currently no report on colchicine for the treatment of AAA.
Disclosure of Invention
1. Technical problem to be solved
The invention aims to solve the problem that colchicine is not used for treating AAA in the prior art, and provides a research and application of an alkaloid compound in preventing and treating abdominal aortic aneurysm.
2. Technical proposal
In order to achieve the above purpose, the present invention adopts the following technical scheme:
an alkaloid compound, wherein the alkaloid compound is colchicine.
The invention also provides an application of the alkaloid compound in improving pathological injury of abdominal aortic aneurysm.
The invention also provides an application of the alkaloid compound in preventing and treating abdominal aortic aneurysm.
The invention also provides an application of the alkaloid compound in preparing the medicine for preventing and treating the abdominal aortic aneurysm.
The invention also provides a research method of alkaloid compounds in preventing and treating abdominal aortic aneurysm, comprising the following steps:
step 1: firstly, performing mouse AAA molding;
step 2: injecting colchicine 0.1mg/kg into the abdominal cavity of the mouse while molding the mouse AAA;
step 3: after 2 weeks, the abdominal aorta of the mice was isolated, and the extent of aortic expansion and injury were assessed by staining the blood vessels HE/EVG/Masson;
step 4: finally, the abdominal aortic tissue of the mice is detected, and the possible molecular mechanism of the colchicine for improving the AAA injury of the mice is explored.
Preferably, in the step 1, the mouse AAA molding process is to incubate with porcine trypsin PPE and combine drinking water to administer aminopropionitrile fumaric acid BAPN to induce the mouse to form abdominal aortic aneurysm.
Preferably, in the step 4, the abdominal aortic tissue of the mouse is detected by Western blot, immunofluorescent staining and multicolor flow cytometry sorting.
3. Advantageous effects
Compared with the prior art, the invention has the advantages that:
in the invention, colchicine with definite anti-inflammatory action is provided for the first time, mouse AAA injury can be improved by inhibiting vascular wall adventitia inflammatory cell infiltration, inhibiting vascular smooth muscle cell to synthetic transformation, matrix metalloproteinase expression and other aspects, the novel application of the colchicine which is a common clinical medicine is disclosed, and a novel intervention strategy and development thought are provided for preventing and treating abdominal aortic aneurysm.
Drawings
FIG. 1 is a diagram showing the successful construction of a mouse AAA model using PPE+BAPN as proposed in the present invention;
FIG. 2 is a graph showing experimental results of the colchicine proposed in the present invention to slow aortic dilation in a mouse AAA model;
FIG. 3 is a graph showing experimental results of the inhibition of VSMCs to synthetic transformation and the expression of MMPs by colchicine as proposed in the present invention;
FIG. 4 is a graph showing experimental results of colchicine as proposed in the present invention for reducing the level of AAA vascular wall inflammation and oxidative stress;
FIG. 5 is a graph showing the experimental results of colchicine proposed in the present invention for reducing the infiltration of inflammatory cells in the adventitia of the vascular wall.
Detailed Description
The following description of the embodiments of the present invention will be made clearly and completely with reference to the accompanying drawings, in which it is apparent that the embodiments described are only some embodiments of the present invention, but not all embodiments.
Example 1:
an alkaloid compound, wherein the alkaloid compound is colchicine.
In this embodiment, a research method of alkaloid compounds in preventing and treating abdominal aortic aneurysm includes the following steps:
step 1: firstly, performing mouse AAA molding, wherein the mouse AAA molding comprises the following steps of incubating by using porcine trypsin PPE and administering aminopropionitrile fumarate BAPN to induce the mouse to form abdominal aortic aneurysm;
step 2: injecting colchicine 0.1mg/kg into the abdominal cavity of the mouse while molding the mouse AAA;
step 3: after 2 weeks, the abdominal aorta of the mice was isolated, and the extent of aortic expansion and injury were assessed by staining the blood vessels HE/EVG/Masson;
step 4: finally, detecting the abdominal aortic tissue of the mice by Western blot, immunofluorescence staining and multicolor flow cell sorting, and exploring the possible molecular mechanism of colchicine for improving the AAA injury of the mice.
In the embodiment, colchicine with definite anti-inflammatory action is provided for the first time, mouse AAA injury can be improved by inhibiting vascular wall adventitia inflammatory cell infiltration, inhibiting vascular smooth muscle cell to synthetic transformation, matrix metalloproteinase expression and other aspects, the novel application of the colchicine which is a common clinical medicine is disclosed, and a novel intervention strategy and development thought are provided for preventing and treating abdominal aortic aneurysm.
Example 2:
the mice used in this example were C57BL/6J mice (hereinafter abbreviated as C57 mice), purchased from Shandong Marek laboratory animals Inc., and all animal manipulations involved in the laboratory were humanized.
In this example, construction of mouse AAA model:
(1) After C57 mice were anesthetized with 1% sodium pentobarbital (40 mg/kg), the abdomen was facing upward and fixed on a foam plate;
(2) Sterilizing the abdomen and surrounding area of the mouse with 75% alcohol, gently shearing the skin of the abdomen of the mouse with sterilized scissors and forceps, and opening the abdomen layer by layer;
(3) Slightly pulling out small intestine, large intestine, etc. by using cotton swab, and placing on gauze strip soaked in physiological saline;
(4) Gently freeing the mouse abdominal aorta, especially the aortic segment above the iliac artery and below the renal artery branch, using a cotton swab and forceps;
(5) Soaking the cosmetic cotton sliver cut in advance in PPE solution, and then wrapping the cosmetic cotton sliver on free aortic tissue;
(6) Incubation time was 40min, wherein about 5 μl of PPE solution was added once for 10 min;
(7) After the incubation time is over, taking out the cosmetic cotton sliver, and lightly wiping the cosmetic cotton sliver for 3 times after the cotton swab is used for dipping physiological saline;
(8) Closing abdomen layer by layer, suturing the mice, and placing the mice back into the animal room after waking up;
(9) Mice were given 0.4% BAPN drinking water, normal state control group was the abdominal aortic group of mice incubated with normal saline, and drinking water without BAPN was given. All mice drink water and eat freely, and can freely move or rest in the rearing cage. After 2 weeks, the mouse AAA formation was judged comprehensively by taking the materials and combining HE staining (using Image software) with an electronic vernier caliper.
In this embodiment, as shown in fig. 1: the PPE incubates the abdominal aorta, and the abdominal aorta is obviously distended and raised in the mice after drinking 2 weeks in combination with the BAPN, and the abdominal aorta is obviously distended. The combination of HE staining (Image software measurement) with electronic vernier calipers is as follows: the mean value of the abdominal aorta diameter of the model group is about 1.0mm, and the mean value of the abdominal aorta of the normal control group is about 0.4mm. The conversion shows that the abdominal aorta diameter of the model group exceeds the average value of the diameter of the normal control group by more than 1.5 times, which proves that the PPE+BAPN is utilized to successfully construct the mouse AAA model.
Example 3:
the mouse AAA model used in this example was obtained using PPE+BAPN induction, and the procedure was the same as in example 2.
In this example, colchicine (0.1 mg/kg) was administered simultaneously with AAA molding to mice, i.e. once a day, for 2 weeks. After the end of the dosing, after anesthetizing the mice with pentobarbital, the aortic tissue of the mice was obtained for subsequent experiments.
In this embodiment, after the molding is completed, the mouse is perfused with physiological saline under an anesthetic state to obtain an aorta; removing the peripheral adhesion tissues of the aorta by using a biological microscope, measuring the maximum aortic diameter by combining an electronic vernier caliper with HE staining (Image software measurement), detecting the expression of VSMCs phenotype-related proteins and MMPs by using Western blot, analyzing the change of inflammatory factors and oxidative stress level in the vascular wall by using immunofluorescence staining, and detecting the infiltration condition of inflammatory cells in the adventitia of the vascular wall by using flow cytometry.
(1) Antibodies to VSMCs phenotype associated proteins and MMPs were purchased from CST corporation, and operated according to the specification, α -SMA was a contractile VSMCs marker, OPN was a synthetic VSMCs marker, MMP2 and MMP9 were two MMPs with significantly elevated expression in AAA disease.
(2) The antibody against the inflammatory factor-related protein TNF-alpha/IL-1 beta/IL-8 was purchased from abcam, and the antibody against the oxidative stress-related protein NOX2/NOX4 was purchased from santa cruz. The stronger the immunofluorescence, the higher the expression level of the protein.
(3) Antibody for multicolor flow cell sorting: CD45 antibodies, CD11b antibodies, F4/80 antibodies, CD86 antibodies, and CD163 antibodies were purchased from affnity corporation.
In this embodiment, as shown in fig. 2: colchicine can obviously slow down the expansion of the abdominal aorta in AAA; fig. 3 shows: colchicine can inhibit the conversion of VSMCs from contractile to synthetic forms in aortic tissue diseased from AAA mice, inhibiting the expression of MMP2 and MMP 9;
in this embodiment, fig. 4 shows: colchicine can reduce the production of inflammatory factors and oxidative stress in aortic tissue of AAA mice lesions. Fig. 5 shows: colchicine can reduce infiltration of adventitia inflammatory cells around aortic tissue diseased in AAA mice. These results demonstrate that colchicine can ameliorate AAA damage and exert vascular protection.
The foregoing is only a preferred embodiment of the present invention, but the scope of the present invention is not limited thereto, and any person skilled in the art, who is within the scope of the present invention, should make equivalent substitutions or modifications according to the technical scheme of the present invention and the inventive concept thereof, and should be covered by the scope of the present invention.
Claims (7)
1. An alkaloid compound, characterized in that the alkaloid compound is colchicine.
2. An alkaloid compound according to claim 1, characterized in that it is used for improving pathological lesions of abdominal aortic aneurysm.
3. An alkaloid compound according to claim 1, characterized by its use for the prevention and treatment of abdominal aortic aneurysms.
4. The use of an alkaloid compound according to claim 1 for the preparation of a medicament for the prevention and treatment of abdominal aortic aneurysm.
5. The method for studying alkaloid compounds for preventing and treating abdominal aortic aneurysm according to claim 1, comprising the following steps:
step 1: firstly, performing mouse AAA molding;
step 2: injecting colchicine 0.1mg/kg into the abdominal cavity of the mouse while molding the mouse AAA;
step 3: after 2 weeks, the abdominal aorta of the mice was isolated, and the extent of aortic expansion and injury were assessed by staining the blood vessels HE/EVG/Masson;
step 4: finally, the abdominal aortic tissue of the mice is detected, and the possible molecular mechanism of the colchicine for improving the AAA injury of the mice is explored.
6. The method for studying alkaloid compounds for preventing and treating abdominal aortic aneurysm according to claim 5, wherein the mouse AAA molding process in step 1 is to use porcine trypsin PPE to incubate combined drinking water to administer aminopropionitrile fumarate BAPN to induce mice to form abdominal aortic aneurysm.
7. The method for studying alkaloid compounds for preventing and treating abdominal aortic aneurysm according to claim 5, wherein in the step 4, the mouse abdominal aortic tissue is detected by Western blot, immunofluorescence staining and multicolor flow cytometry sorting.
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| CN202311126327.9A CN117045628A (en) | 2023-09-04 | 2023-09-04 | Research and application of alkaloid compound in prevention and treatment of abdominal aortic aneurysm |
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| CN202311126327.9A CN117045628A (en) | 2023-09-04 | 2023-09-04 | Research and application of alkaloid compound in prevention and treatment of abdominal aortic aneurysm |
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