CN117257808A - Application of rutaecarpine in preparation of product for promoting wound healing - Google Patents
Application of rutaecarpine in preparation of product for promoting wound healing Download PDFInfo
- Publication number
- CN117257808A CN117257808A CN202311566333.6A CN202311566333A CN117257808A CN 117257808 A CN117257808 A CN 117257808A CN 202311566333 A CN202311566333 A CN 202311566333A CN 117257808 A CN117257808 A CN 117257808A
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- China
- Prior art keywords
- rutaecarpine
- wound healing
- mice
- cells
- product
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- ACVGWSKVRYFWRP-UHFFFAOYSA-N Rutecarpine Chemical compound C1=CC=C2C(=O)N(CCC=3C4=CC=CC=C4NC=33)C3=NC2=C1 ACVGWSKVRYFWRP-UHFFFAOYSA-N 0.000 title claims abstract description 113
- BCPAKGGXGLGKIO-UHFFFAOYSA-N Pseudorutaecarpin Natural products C1=CC=C2C(=O)N(CCC3=C4C5=CC=CC=C5N3)C4=NC2=C1 BCPAKGGXGLGKIO-UHFFFAOYSA-N 0.000 title claims abstract description 53
- 230000029663 wound healing Effects 0.000 title claims abstract description 46
- 230000001737 promoting effect Effects 0.000 title claims abstract description 28
- 238000002360 preparation method Methods 0.000 title abstract description 6
- 206010012601 diabetes mellitus Diseases 0.000 claims abstract description 28
- 206010052428 Wound Diseases 0.000 claims description 15
- 208000027418 Wounds and injury Diseases 0.000 claims description 15
- 238000001727 in vivo Methods 0.000 claims description 5
- 210000001339 epidermal cell Anatomy 0.000 claims description 3
- 210000000130 stem cell Anatomy 0.000 claims description 3
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- 241000699670 Mus sp. Species 0.000 abstract description 46
- 230000000694 effects Effects 0.000 abstract description 28
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- 239000003814 drug Substances 0.000 abstract description 17
- 238000002474 experimental method Methods 0.000 abstract description 17
- 241000699666 Mus <mouse, genus> Species 0.000 abstract description 16
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- 238000013508 migration Methods 0.000 abstract description 9
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- 210000001161 mammalian embryo Anatomy 0.000 abstract description 6
- 239000002547 new drug Substances 0.000 abstract description 3
- 210000004027 cell Anatomy 0.000 description 35
- 150000001875 compounds Chemical class 0.000 description 21
- IAZDPXIOMUYVGZ-UHFFFAOYSA-N Dimethylsulphoxide Chemical compound CS(C)=O IAZDPXIOMUYVGZ-UHFFFAOYSA-N 0.000 description 18
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- 206010072170 Skin wound Diseases 0.000 description 9
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- ZSJLQEPLLKMAKR-UHFFFAOYSA-N Streptozotocin Natural products O=NN(C)C(=O)NC1C(O)OC(CO)C(O)C1O ZSJLQEPLLKMAKR-UHFFFAOYSA-N 0.000 description 5
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- KRKNYBCHXYNGOX-UHFFFAOYSA-N citric acid Chemical compound OC(=O)CC(O)(C(O)=O)CC(O)=O KRKNYBCHXYNGOX-UHFFFAOYSA-N 0.000 description 3
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- 229920001285 xanthan gum Polymers 0.000 description 3
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- NCTDBYQRELKCRJ-UHFFFAOYSA-N 1h-indole;quinazoline Chemical class C1=CC=C2NC=CC2=C1.N1=CN=CC2=CC=CC=C21 NCTDBYQRELKCRJ-UHFFFAOYSA-N 0.000 description 1
- NHBKXEKEPDILRR-UHFFFAOYSA-N 2,3-bis(butanoylsulfanyl)propyl butanoate Chemical compound CCCC(=O)OCC(SC(=O)CCC)CSC(=O)CCC NHBKXEKEPDILRR-UHFFFAOYSA-N 0.000 description 1
- 108010005094 Advanced Glycation End Products Proteins 0.000 description 1
- 241000894006 Bacteria Species 0.000 description 1
- 206010006895 Cachexia Diseases 0.000 description 1
- 241000195649 Chlorella <Chlorellales> Species 0.000 description 1
- 206010009900 Colitis ulcerative Diseases 0.000 description 1
- 229920000742 Cotton Polymers 0.000 description 1
- 239000006144 Dulbecco’s modified Eagle's medium Substances 0.000 description 1
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 1
- 102000018233 Fibroblast Growth Factor Human genes 0.000 description 1
- 108050007372 Fibroblast Growth Factor Proteins 0.000 description 1
- 206010020710 Hyperphagia Diseases 0.000 description 1
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- 102100037486 Reverse transcriptase/ribonuclease H Human genes 0.000 description 1
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 1
- 206010042220 Stress ulcer Diseases 0.000 description 1
- FZWLAAWBMGSTSO-UHFFFAOYSA-N Thiazole Chemical compound C1=CSC=N1 FZWLAAWBMGSTSO-UHFFFAOYSA-N 0.000 description 1
- 201000006704 Ulcerative Colitis Diseases 0.000 description 1
- 238000002835 absorbance Methods 0.000 description 1
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- RNFNDJAIBTYOQL-UHFFFAOYSA-N chloral hydrate Chemical compound OC(O)C(Cl)(Cl)Cl RNFNDJAIBTYOQL-UHFFFAOYSA-N 0.000 description 1
- 229960002327 chloral hydrate Drugs 0.000 description 1
- 238000010276 construction Methods 0.000 description 1
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- 238000012258 culturing Methods 0.000 description 1
- 231100000135 cytotoxicity Toxicity 0.000 description 1
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- 238000013118 diabetic mouse model Methods 0.000 description 1
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- 230000010339 dilation Effects 0.000 description 1
- LOKCTEFSRHRXRJ-UHFFFAOYSA-I dipotassium trisodium dihydrogen phosphate hydrogen phosphate dichloride Chemical compound P(=O)(O)(O)[O-].[K+].P(=O)(O)([O-])[O-].[Na+].[Na+].[Cl-].[K+].[Cl-].[Na+] LOKCTEFSRHRXRJ-UHFFFAOYSA-I 0.000 description 1
- 230000035619 diuresis Effects 0.000 description 1
- 231100000673 dose–response relationship Toxicity 0.000 description 1
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- 210000003195 fascia Anatomy 0.000 description 1
- 229940126864 fibroblast growth factor Drugs 0.000 description 1
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- 230000002439 hemostatic effect Effects 0.000 description 1
- 238000011534 incubation Methods 0.000 description 1
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- 206010033675 panniculitis Diseases 0.000 description 1
- 244000052769 pathogen Species 0.000 description 1
- 239000000546 pharmaceutical excipient Substances 0.000 description 1
- 229940124531 pharmaceutical excipient Drugs 0.000 description 1
- 230000000144 pharmacologic effect Effects 0.000 description 1
- 239000002953 phosphate buffered saline Substances 0.000 description 1
- 239000000902 placebo Substances 0.000 description 1
- 229940068196 placebo Drugs 0.000 description 1
- 238000007747 plating Methods 0.000 description 1
- 208000022530 polyphagia Diseases 0.000 description 1
- 229920000136 polysorbate Polymers 0.000 description 1
- 230000009090 positive inotropic effect Effects 0.000 description 1
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- 230000001105 regulatory effect Effects 0.000 description 1
- 230000008439 repair process Effects 0.000 description 1
- 229940109850 royal jelly Drugs 0.000 description 1
- -1 rutaecarpine compound Chemical class 0.000 description 1
- 239000012679 serum free medium Substances 0.000 description 1
- 239000011780 sodium chloride Substances 0.000 description 1
- 210000005070 sphincter Anatomy 0.000 description 1
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- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 1
- 230000003442 weekly effect Effects 0.000 description 1
- 230000037314 wound repair Effects 0.000 description 1
Classifications
-
- 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
- A61K31/33—Heterocyclic compounds
- A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
- A61K31/495—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with two or more nitrogen atoms as the only ring heteroatoms, e.g. piperazine or tetrazines
- A61K31/505—Pyrimidines; Hydrogenated pyrimidines, e.g. trimethoprim
- A61K31/519—Pyrimidines; Hydrogenated pyrimidines, e.g. trimethoprim ortho- or peri-condensed with heterocyclic rings
-
- A—HUMAN NECESSITIES
- 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)
- A61K36/185—Magnoliopsida (dicotyledons)
- A61K36/75—Rutaceae (Rue family)
- A61K36/754—Evodia
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P17/00—Drugs for dermatological disorders
- A61P17/02—Drugs for dermatological disorders for treating wounds, ulcers, burns, scars, keloids, or the like
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P3/00—Drugs for disorders of the metabolism
- A61P3/08—Drugs for disorders of the metabolism for glucose homeostasis
- A61P3/10—Drugs for disorders of the metabolism for glucose homeostasis for hyperglycaemia, e.g. antidiabetics
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- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Medicinal Chemistry (AREA)
- Pharmacology & Pharmacy (AREA)
- Animal Behavior & Ethology (AREA)
- General Health & Medical Sciences (AREA)
- Public Health (AREA)
- Veterinary Medicine (AREA)
- Natural Medicines & Medicinal Plants (AREA)
- Diabetes (AREA)
- Engineering & Computer Science (AREA)
- General Chemical & Material Sciences (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Epidemiology (AREA)
- Organic Chemistry (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Alternative & Traditional Medicine (AREA)
- Dermatology (AREA)
- Mycology (AREA)
- Biotechnology (AREA)
- Microbiology (AREA)
- Medical Informatics (AREA)
- Botany (AREA)
- Emergency Medicine (AREA)
- Endocrinology (AREA)
- Hematology (AREA)
- Obesity (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
- Acyclic And Carbocyclic Compounds In Medicinal Compositions (AREA)
Abstract
The invention belongs to the technical field of biological medicines, and particularly relates to application of rutaecarpine in preparation of a product for promoting wound healing. Experiments prove that the rutaecarpine can remarkably promote migration of human normal skin immortalized keratinocytes, mouse embryo fibroblasts and human foreskin fibroblasts, has a certain promotion effect on wound healing of diabetic mice and non-diabetic mice, and provides a new drug choice for wound healing.
Description
Technical Field
The invention belongs to the technical field of biological medicines, and particularly relates to application of rutaecarpine in preparation of a product for promoting wound healing.
Background
Skin is the largest organ in the human body and is also the first line of defense of the human body against external pathogens, and once the skin is subjected to a large area or severe wound, the skin can lose the ability to repair itself, so that the wound is difficult to recover. The accumulation of advanced glycation end products in skin tissues of diabetics can cause oxidative damage to fibroblasts, thereby thinning the dermis, leading to vulnerable skin and less healing of the wound. Furthermore, in the early stages of diabetic wound healing, the expression and content of fibroblast growth factor is reduced, thus affecting wound healing.
At present, various medicaments for promoting wound healing exist, but the application range, the medicament effect, the cost and the toxic effects are different, common medicaments include antibiotics, wound repair gel or gel, lotion, wound dressing and the like, wherein the wound dressing can simulate better environment and also can prevent external bacterial invasion, has better effects of stopping bleeding, resisting bacteria, repairing skin, promoting wound healing and the like, and is the most popular medicament for promoting wound healing in the technical field of biological medicines.
The rutaecarpine is an indole quinazoline alkaloid extracted from the rutaecarpa, has positive inotropic effect, positive time-varying effect and heart protecting effect on heart, has vascular dilation and blood pressure reduction effects, can relax internal sphincter, inhibit gastrointestinal movement, and has therapeutic effect on stress ulcer and ulcerative colitis. In addition, rutaecarpine also has pharmacological effects such as anti-tumor effect, however, the prior art does not have research on the promotion of wound healing by rutaecarpine.
The patent publication No. CN112426562A discloses a purely natural medical dressing for promoting wound healing, which consists of chlorella powder, royal jelly and reed flowers, does not contain any biochemical products such as antibiotics, protease, metal ions and the like, does not generate obvious stimulation effect on wounds or secondary injury on human bodies in the using process of the dressing, and has simple processing technology and low cost.
Patent publication CN110090224a discloses a colloidal dressing for promoting wound healing, the composition comprising beta-glucan and xanthan gum for preparing a medicament for promoting wound healing. The xanthan gum and the beta-glucan can synergistically promote wound healing, and meanwhile, the xanthan gum and the beta-glucan have a synergistic thickening effect after being combined, so that the use amount of a thickening agent in a preparation or auxiliary materials can be reduced, and the cost is reduced.
In view of the above, there is no description in the prior art about rutaecarpine in promoting wound healing.
Disclosure of Invention
The invention aims to provide an application of rutaecarpine in preparing a product for promoting wound healing, and provides a new drug choice for promoting wound healing. Experiments prove that the rutaecarpine can remarkably promote migration of human normal skin immortalized keratinocytes, mouse embryo fibroblasts and human foreskin fibroblasts, and has a certain promotion effect on wound healing of diabetic mice and non-diabetic mice.
In order to achieve the above object, the technical scheme of the present invention is as follows:
the invention provides a new application of rutaecarpine in preparing a product for promoting wound healing.
Further, the wound healing promoting product is used for treating diabetic wounds.
Further, the wound healing promoting product is used for treating non-diabetic wounds.
Further, the concentration range of the rutaecarpine on the stem cells of the epidermal cells is 1-5 mu mol/L.
Further, the concentration range of the rutaecarpine on the stem cells of the epidermal cells is 2-4 mu mol/L.
Further, the in-vivo application concentration of the rutaecarpine is 0.5-1 mg/kg.
Further, the in-vivo application concentration of the rutaecarpine is 0.6-1 mg/kg.
The invention also provides a pharmaceutical composition for promoting wound healing, which comprises rutaecarpine.
Further, the pharmaceutical composition for promoting wound healing also comprises one or more pharmaceutically acceptable carriers or pharmaceutical excipients.
Furthermore, the pharmaceutical composition for promoting wound healing is one of an application gel, an oral preparation and an injection.
The applicant of the invention discovers through in vitro experimental research that rutaecarpine can obviously promote migration of human normal skin immortalized keratinocytes, and lower concentration can obviously promote migration of mouse embryo fibroblasts and human foreskin fibroblasts; the in vivo experimental study shows that the rutaecarpine has certain promotion effect on wound healing of diabetic mice and non-diabetic mice.
According to the invention, the influence of the medicine on the cell growth is measured through MTT, the half-inhibition rate concentration of the rutaecarpine on the mouse embryo fibroblasts, the human normal skin immortalized keratinocytes, the human foreskin fibroblasts and the mouse embryo fibroblasts is obtained, the action concentration of the rutaecarpine in an in-vitro test is determined, then the influence of the rutaecarpine on the cell migration capacity is measured through a cell scratch experiment, and the influence of the rutaecarpine on the cell migration capacity is measured through a Transwell experiment. In addition, the invention also constructs a BALB/c type diabetes mouse model, and applies rutaecarpine to diabetic mice and non-diabetic mice, and observes the influence of the rutaecarpine on the wound healing degree of the mice.
Compared with the prior art, the new application of rutaecarpine in preparing the product for promoting wound healing has the following technical advantages:
(1) The first experiment proves that the rutaecarpine can effectively promote cell movement, has the effect of accelerating wound healing, and provides a new drug choice for promoting wound healing;
(2) Experiments prove that the rutaecarpine compound provided by the invention can promote migration of human normal skin immortalized keratinocytes, mouse embryo fibroblasts and human foreskin fibroblasts, and has good safety;
(3) The compound rutaecarpine provided by the invention has remarkable effect on healing skin wounds of diabetic mice and non-diabetic mice, can be used for preparing medicaments for promoting wound healing, and has extremely high medicinal value.
Drawings
FIG. 1 is a graph showing the effect of rutaecarpine on 3T3 cell activity;
FIG. 2 is a graph showing the effect of rutaecarpine on HACAT cell activity;
FIG. 3 is the effect of rutaecarpine on HFF-1 cell activity;
FIG. 4 is a graph showing the effect of rutaecarpine on L-929 cell activity;
FIG. 5 is a graph of rutaecarpine 0h, 24h cell confluence;
FIG. 6 is a graph showing the effect of rutaecarpine on promoting 3T3 cell migration;
FIG. 7 is a graph showing the experimental effect of rutaecarpine on promoting HFF-1 cell migration;
FIG. 8 is an effect of rutaecarpine on skin wound healing in diabetic mice;
FIG. 9 is a graph showing the comparison of rutaecarpine to diabetic mice skin wound healing rate;
FIG. 10 is an effect of skin wound healing in a normoglycemic mice placebo (solvent) group;
FIG. 11 is an effect of rutaecarpine on skin wound healing in normoglycemic mice;
FIG. 12 shows the effect of rutaecarpine on skin wound healing rate in normoglycemic mice.
Detailed Description
The present invention is further described below by way of specific embodiments, but the present invention is not limited to the following examples. Various modifications may be made by those skilled in the art in light of the basic idea of the invention, but are within the scope of the invention without departing from the basic idea of the invention.
The reagents used in this embodiment are all commonly used reagents and can be purchased from conventional reagent manufacturing and selling companies.
Test example one, compound cytotoxicity assay
The experimental object: mouse embryonic fibroblasts (3T 3); human normal skin immortalized keratinocytes (HACAT); human foreskin fibroblasts (HFF-1); mouse embryonic fibroblasts (L-929).
Experimental drugs: a compound: rutaecarpine (TS-0296); compound solvent: dimethyl sulfoxide (DMSO).
The experimental method comprises the following steps: the first pm plating: the log phase cells were collected, the cell suspension concentration was adjusted, and 90uL, 10000 cells were added per well. Dosing the next morning: adding 10 μl of the drug with concentration gradient into each well, arranging 3 compound wells for each drug concentration, and placing in 5% CO 2 Incubate at 37 ℃. Plate collection after 48h of dosing: the reaction was stopped after incubation for 4h by first visual inspection under an inverted microscope, then adding 20uL of thiazole blue solution (MTT, 5mg/mL, i.e., 0.5% MTT) per well, carefully sucking off the culture medium in the well, adding 100 uL of DMSO per well, and shaking at low speed on a shaker for 10min to allow the crystals to dissolve well. OD value detection: the absorbance of each well was measured at 490nm wavelength of the microplate reader, the relative cell viability or drug inhibition was calculated, and the drug concentration-response curve was plotted using graphpadprism 7.0. In the experiment, a culture medium without cells is used as a blank control group, a DMSO solution with the same dilution ratio as the compound is added into each hole to be used as a negative control group, and an experiment group containing the compound TS-0296 is provided. Relative survival = (compound experimental group OD value-blank group OD value)/(negative control group OD value-blank group OD value) ×100%; drug inhibition = 1-relative survival.
Experimental results: IC50 is the half-inhibitory concentration, i.e. the concentration of drug that results in half of the cells dying after a certain period of drug treatment. As shown in FIGS. 1-4, the IC50 of the compound rutaecarpine of the invention was 28.98. Mu. Mol/L in 3T3 cells, 132.8. Mu. Mol/L in HACAT cells, 35.67. Mu. Mol/L in HFF-1 cells, and 34.07. Mu. Mol/L in L-929 cells for 48 h. Based on the IC50 results of each cell strain, 10 mu mol/L is taken as the highest concentration in the subsequent functional experiments, so that the rutaecarpine is ensured not to generate obvious cytotoxicity.
Test example two, cell scratch test
The experimental object: human normal skin immortalizes keratinocytes (HACAT).
Experimental drugs: a compound: rutaecarpine (TS-0296); compound solvent: dimethyl sulfoxide (DMSO).
The experimental method comprises the following steps: the thin head of the Mark pen is used to draw horizontal lines on the back of the 6-hole plate before the plate is paved, and the horizontal lines are evenly drawn by a ruler, approximately every 0.5-1 cm, three lines are drawn in sequence after the horizontal lines cross the through holes, and the lines are named as a line, b line and c line. The line b crosses the midpoint, and the other two lines are equally spaced on both sides of the line b. Six-well plates were plated, 2mL of complete medium containing 10% FBS (fetal bovine serum) was added to each well, 2X 10 6 The cells were each plated with 2 duplicate wells and incubated for about 24 hours. The cell number is preferably regulated so that the cells can be covered with the wall over night by more than 90%. The lid of the well plate is opened the next day, the old culture medium is sucked off, the ruler is perpendicular to the b-line frame on the well plate, the 200 uL gun head is tightly attached to the ruler to uniformly move up and down to manufacture cell scratch lines, and two parallel lines are drawn on two sides of the line at equal intervals, and the parallel lines are named as line 1, line 2 and line 3 from left to right respectively. Washing cells with sterile 1 XPBS (phosphate buffer solution) for 3 times, removing the scraped cells, adding complete medium containing 20% FBS, adding compound TS-0296 (working concentration of 5 μmol/L), and adding 37deg.C and 5% CO 2 Culturing in an incubator. Taking samples according to the dosing time of 0 and 24 hours, and taking photos.
Experimental results: as can be seen from FIG. 5, compound TS-0296 at a concentration of 5. Mu. Mol/L significantly promoted migration of cellular HACAT after 24 hours of action.
Test example III, migration test
The experimental object: mouse embryonic fibroblasts (3T 3), human foreskin fibroblasts (HFF-1).
Experimental drugs: a compound: rutaecarpine (TS-0296); compound solvent: dimethyl sulfoxide (DMSO).
The experimental method comprises the following steps: preparing a cell suspension: centrifuging after digestion of digested cells, removing culture solution, washing with PBS (phosphate buffer solution) for 1 time, re-suspending in serum-free medium, and adjusting cell density to appropriate concentration (plate densities of 3T3 cells and HFF-1 cells are 4×10) 5 and/mL). Splicing jointA cell: each cell was set with a negative control group (DMSO was added at the same dilution ratio as compound TS-0296), the dosing group (i.e., working concentrations of compound TS-0296,3T3 and HFF-1 were each set at 1, 5, 10. Mu. Mol/L), each group was 2 multiplex wells, cell suspensions were taken according to cell density (3T 3 cells plated at 80000/200. Mu.L/well, HFF-1 cells plated at 80000/200. Mu.L/well), 10% BSA (bovine serum albumin) was added to give a final percentage of 0.1%, compound TS-0296 was added, and finally DMEM (high glucose) was used to make the total volume of each well 200. Mu.L, and after mixing, the cells were gently and evenly added to the Transwell upper chamber. Immediately, 800. Mu.L of medium containing 20% FBS (fetal bovine serum) was typically added to the 24-well plate lower chamber, and plates were harvested about 24 hours after administration. Cell staining: the Transwell chamber was removed, the culture medium in the well was discarded, and the cells not migrated in the upper chamber were gently scraped off with a cotton swab and placed in a clean 24-well plate. Washing with 1 XPBS (phosphate buffered saline) for 1 time, fixing with methanol for 30 minutes, blotting methanol, air drying the chamber in a fume hood, staining with 0.1% crystal violet for 20 minutes, blotting off recovered crystal violet, washing with PBS for 1 time. The PBS was blotted and the fume hood dried. And (3) result statistics: cells were observed under a 5X microscope using an advanced research microscope, randomly five fields under a 10X microscope, photographed, counted and statistically plotted.
Experimental results: from FIGS. 6 and 7, it is clear that the compound TS-0296 can significantly promote the migration of fibroblasts and human foreskin fibroblasts at low concentrations (1 and 5. Mu. Mol/L) and can only significantly promote the migration of human foreskin fibroblasts at higher concentrations (10. Mu. Mol/L).
Test example four influence of Compounds on wound healing in mice skin
The experimental method comprises the following steps:
1. high fat diet in combination with STZ established a diabetic mouse model:
(1) Mouse ordering: animals were ordered 3 weeks old, male, BALB/c mice and kept in SPF rearing room for 5 days while isolation observations were made. Randomly extracting 5 mice to enter a modeling module, extracting 10 mice as non-modeling modules, feeding high-fat feed to the modeling module, and feeding common feed to the non-modeling module;
(2) The first day of the experiment was the first day when the model mice began to eat high fat diet. Body weight was recorded weekly, hair, feeding and urination were observed, feed was removed at night on day 28, and no water was allowed to pass overnight;
(3) Preparing Streptozotocin (STZ) injection: 20 mg of STZ powder was dissolved in 4 mL citric acid buffer to prepare 5mg/mL of an injection. All operations were performed on ice, protected from light. The STZ solution is ready for use.
(4) Injection of STZ: the following day mice were measured for body weight and fasting blood glucose: the mice with the model group are injected with STZ solution (40 mg/kg/d) intraperitoneally, once a day for 5 consecutive days, the first day of injection is continuous with fasting, and the second day of injection is started to recover high-fat feeding; continuously observing the feeding, drinking and urination conditions of the mice;
(5) Two groups of mice were tested for fasting blood glucose 2 weeks after dosing:
in the model building mice, a diabetic mouse which is stable at more than 11.1 mmoL/L and has polydipsia, diuresis, polyphagia and emaciation is defined as a model building successful mouse, a subsequent experiment is included, a mouse which does not accord with a model standard exits the experiment, and the model building success rate is counted;
(6) During the experiment, the sanitary nursing of the model-making mice is paid attention to, and if the model-making mice show polydipsia and urination, the drinking water needs to be supplemented in time, so that the interval for replacing padding is shortened.
2. Construction of full-thickness wound on back skin of mouse and observation of influence of compound on skin wound healing
(1) Mouse ordering: animals were ordered from the laboratory animal center for 13 week old, male, BALB/c mice and kept in the SPF rearing room for 5 days while isolation observations were made. The mice are randomly divided into a negative control group, a diabetes building module and a normal blood sugar group, wherein 5 mice are in each group; the diabetic modeling group mice are modeled according to the modeling method, and the normal blood sugar group mice and the negative control group mice are fed normally;
(2) After 2 weeks of successful modeling, diabetic mice, normal glycemic mice and negative control mice were moved to the operating room and anesthetized by intraperitoneal injection of 5% chloral hydrate (100 ul/20 g);
(3) Placing the fully anesthetized mice on a pad in prone position, and removing back hair with depilatory cream;
(4) Sterilizing back skin with 75% alcohol, drilling 2 circular skin full-layer wounds with diameters of about 15 mm on two sides of the highest position of the central line of the back by using a trephine with diameters of 15 mm, shaving subcutaneous tissues by using surgical scissors and forceps, and exposing fascia on the surface of muscle;
(5) Hemostatic and disinfectant wound, and open wound; taking a ruler as a control, and photographing to record the shape and the size of the wound;
(6) Administration: each mouse of the negative control group was intraperitoneally injected with 0.1mL containing 10% DMSO-40% PEG400-5% tween 80-45% saline; diabetes mice and non-diabetes mice were intraperitoneally injected with 0.1mL of TS-0296, with an amount of TS-0296 of 1 mg/kg/d; the day of mice wounding was the first day (d 1), and each group of mice was injected at d1, 3, 5, 7, 9, 11, respectively, and the wound healing rate of diabetic mice d3, 5, 7, 9, 11, 13, 15 and the wound healing rate of non-diabetic mice d5, 7 were recorded.
(7) The anesthetized mice are resuscitated under the temperature control plate and transported back to the EVC cage of the clean rearing room.
Experimental results: the effect of rutaecarpine on skin wound of mice with diabetes is shown in fig. 8 and 9, and the effect of rutaecarpine on skin wound healing of mice with normal blood sugar is shown in fig. 10, 11 and 12. As can be seen from fig. 8, 9, 10, 11 and 12, rutaecarpine has a good effect of promoting wound healing in both diabetic mice and normal mice.
The above embodiments are merely illustrative of the principles of the present invention and its effectiveness, and are not intended to limit the invention. Modifications and variations may be made to the above-described embodiments by those skilled in the art without departing from the spirit and scope of the invention. Accordingly, it is intended that all equivalent modifications and variations of the invention be covered by the claims, which are within the ordinary skill of the art, be within the spirit and scope of the present disclosure.
Claims (7)
1. Application of rutaecarpine in preparing product for promoting wound healing is provided.
2. The use according to claim 1, wherein the wound healing promoting product is for the treatment of diabetic wounds.
3. The use according to claim 1, wherein the wound healing promoting product is for the treatment of non-diabetic wounds.
4. The use according to claim 1, wherein the rutaecarpine has a concentration in the range of 1-5 μmol/L on epidermal stem cells.
5. The use according to claim 4, wherein the rutaecarpine has a concentration of 2-4 μmol/L on the stem cells of the epidermal cells.
6. The use according to claim 1, wherein the in vivo application concentration of rutaecarpine is 0.5-1 mg/kg.
7. The use according to claim 6, wherein the in vivo application concentration of rutaecarpine is 0.6-1 mg/kg.
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Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20090247635A1 (en) * | 2008-03-31 | 2009-10-01 | Eli Ehrenpreis | Method for treating anal pruritis and other perianal disorders |
| CN101953838A (en) * | 2010-08-13 | 2011-01-26 | 李岱 | Application of rutaecarpine in treating psoriasis |
| CN102342926A (en) * | 2011-07-27 | 2012-02-08 | 华东师范大学 | Application of ferulic acid in preparation of medicine used for promoting skin wound healing |
| US20160081959A1 (en) * | 2014-09-24 | 2016-03-24 | Glanbia Plc | Method of using arginine silicate inositol complex for wound healing or repair |
| CN108904504A (en) * | 2018-06-20 | 2018-11-30 | 暨南大学 | Application of the rutaecarpin in the drug of preparation enhancing NLRP3 inflammation corpusculum activation |
-
2023
- 2023-11-23 CN CN202311566333.6A patent/CN117257808B/en active Active
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20090247635A1 (en) * | 2008-03-31 | 2009-10-01 | Eli Ehrenpreis | Method for treating anal pruritis and other perianal disorders |
| CN101953838A (en) * | 2010-08-13 | 2011-01-26 | 李岱 | Application of rutaecarpine in treating psoriasis |
| CN102342926A (en) * | 2011-07-27 | 2012-02-08 | 华东师范大学 | Application of ferulic acid in preparation of medicine used for promoting skin wound healing |
| US20160081959A1 (en) * | 2014-09-24 | 2016-03-24 | Glanbia Plc | Method of using arginine silicate inositol complex for wound healing or repair |
| CN108904504A (en) * | 2018-06-20 | 2018-11-30 | 暨南大学 | Application of the rutaecarpin in the drug of preparation enhancing NLRP3 inflammation corpusculum activation |
Non-Patent Citations (3)
| Title |
|---|
| MEILING WANG等: "Rutaecarpine prevented ox-LDL-induced VSMCs dysfunction through inhibiting overexpression of connexin 43", EJP, vol. 853, pages 84 - 92, XP085673011, DOI: 10.1016/j.ejphar.2019.03.028 * |
| 学堂在线健康网: "吴茱萸外敷的功效", pages 1, Retrieved from the Internet <URL:http://mip.xuetangzaixian.com/zhongyao/118085.html> * |
| 肖培根等: "中华医学百科全书", 中国协和医科大学出版社, pages: 197 * |
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