CN114467858A - Method for inducing white hair model - Google Patents
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- CN114467858A CN114467858A CN202210161963.4A CN202210161963A CN114467858A CN 114467858 A CN114467858 A CN 114467858A CN 202210161963 A CN202210161963 A CN 202210161963A CN 114467858 A CN114467858 A CN 114467858A
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01K—ANIMAL HUSBANDRY; AVICULTURE; APICULTURE; PISCICULTURE; FISHING; REARING OR BREEDING ANIMALS, NOT OTHERWISE PROVIDED FOR; NEW BREEDS OF ANIMALS
- A01K67/00—Rearing or breeding animals, not otherwise provided for; New or modified breeds of animals
- A01K67/02—Breeding vertebrates
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01K—ANIMAL HUSBANDRY; AVICULTURE; APICULTURE; PISCICULTURE; FISHING; REARING OR BREEDING ANIMALS, NOT OTHERWISE PROVIDED FOR; NEW BREEDS OF ANIMALS
- A01K2227/00—Animals characterised by species
- A01K2227/10—Mammal
- A01K2227/105—Murine
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- Life Sciences & Earth Sciences (AREA)
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Abstract
The invention discloses a method for inducing a white hair model, which comprises the following steps: step 1, dissolving a stock solution; step 2, subpackaging the solution; step 3, injection dosage: injecting the liquid substance into the body of the experimental mouse according to the weight of the white mouse and the dosage of 60 mg/kg; step 4, preparing suspension working solution: taking 1 tube of 100uL stock solution, and adding 10-15 times of normal saline to prepare suspension working solution; and 5, paraffin unhairing: paraffin depilation is carried out on 8-9 weeks old mice on day 0; step 6, in vivo study: continuously injecting 200uL of working solution into abdominal cavity of a mouse of 25g and 8-9 weeks old for 2-3 weeks every day according to the injection dosage of 60 mg/kg; step 7, mouse hair growth recording. The Cabozantinib can be used as a VEGFR2 inhibitor to induce leukotrichia, and when a mouse is injected for twenty-four days, the leukotrichia grows more densely.
Description
Technical Field
The invention relates to the technical field of inhibitors, in particular to a method for inducing a white hair model.
Background
While white hair is a clear sign of tissue deterioration during aging, age-related hair graying due to melanocyte stem cell (McSCs) dysfunction, and physiologically, McSCs and their progeny can constitute the hair follicle pigment system, and the activation of McSCs coincides with the hair cycle and proliferates to replenish the McSC population, and then produces melanocytes, depositing pigments into the growing hair shaft, studies have shown that complete loss of McSCs or reduction of their function results in the production of pigment-deficient hair shafts, known as gray hair, and as our black hair, mice have black and bright hair, and depend on melanin synthesis by melanocytes (melanocyte) at the bottom of the hair follicle, and if melanin is absent, or not normally synthesized, the newly grown hair becomes white.
Cabozantinib is mainly used as a targeted inhibitor in known applications and is used as an anti-tumor drug. IN the IN vivo study, Cabozantinib treats RIP-Tag2 mice carrying spontaneous islet cell tumor at a dose of 30mg/kg, disturbs 83% tumor blood vessels, reduces pericytes and empty basal membrane sleeves, causes extensive intratumoral hypoxia and tumor cell apoptosis, delays tumor blood vessel regrowth after drug withdrawal, more remarkably inhibits VEGFR rather than c-Met compared with Tyrosinekinase-IN-1, causes blood vessel reduction by 43%, indicates that VEGFR inhibition and other function-related Receptor Tyrosine Kinase (RTK) inhibiting angiogenesis IN an amplification manner, Cabozantinib also reduces invasion and metastasis of primary tumor, Cabozantinib treats SCID mice at a dose of 30mg/kg every day, remarkably eliminates growth and metastasis of MPNST transplantation tumor of human body, Cabozantinib treats breast cancer, a lung cancer glioma model, inhibits tumor growth, and has dose dependence, reduces tumor and endothelial cell proliferation apoptosis, promotes tumor growth, cabozantinib treated mice bearing MDA-MB-231 tumor and rats bearing C6 tumor alone at 100mg/kg and 10mg/kg, respectively, continuously inhibited tumor growth.
We found that Cabozantinib can effectively induce the growth of white hair in mice, but the specific action mechanism is not clear.
The existing method for inducing white hair has the defects that:
1. patent document CN109758569A discloses a composition for correcting white hair by targeted regulation of genes and its application, "comprising the following components in parts by weight: 0.5-6 parts of betaine extract, 0.2-4 parts of flower bean extract, 1-8 parts of carrot extract, 0.5-5 parts of oyster peptide, 0.1-2 parts of alpha-linolenic acid and 0.1-0.8 part of mineral element supplement; the mineral element supplements include Cu2+, Fe2+, Zn2+, Mg2+, and Se2 +. The application promotes melanin deposition by relieving the activity inhibition of toxic molecules on key genes CAT, KROX20, MITF, IRF4, CYP450 and hair follicle stem cells of premature gray hair or senile white hair generation, thereby blackening the white hair. As shown in the examples, white hair is improved to different degrees in white hair volunteers of different ages after taking the composition. However, the method cannot improve the hair volume, the operation is complicated, and the period of effectiveness of the method for changing from white to black is variable.
Disclosure of Invention
The present invention is directed to a method for inducing a white hair model to solve the above-mentioned problems of the background art.
In order to achieve the purpose, the invention provides the following technical scheme: a method of inducing a white hair model comprising the steps of:
a method of inducing a white hair model comprising the steps of:
step 1, stock solution dissolution;
step 2, subpackaging the solution;
step 3, injection dosage: injecting the liquid substance into the body of the experimental mouse according to the weight of the white mouse and the dosage of 60 mg/kg;
step 4, preparing suspension working solution: taking 1 tube of 100uL stock solution, and adding 10-15 times of normal saline to prepare suspension working solution;
and 5, paraffin unhairing: paraffin depilation is carried out on 8-9 weeks old mice on day 0;
step 6, in vivo study: continuously injecting 200uL of working solution into the abdominal cavity of a mouse of 25g and 8-9 weeks old every day for 2-3 weeks according to the injection dose of 60 mg/kg;
step 7, mouse hair growth recording.
Preferably, in the step S1, when the stock solution is dissolved and dispensed, 1mL of ldmso is added to 100mg of cabozantinib, and the mixed solution is stirred at 40-60rpm to dissolve the two solutions sufficiently to make the concentration of 100 mg/mL.
Preferably, when the solution in the step 2 is dispensed, the mixed solution obtained in the step 1 is dispensed into 100uL/EP tubes, the specification of the EP tubes is 2ml, and then the dispensed liquid substance is stored in an environment at-10 to-30 ℃.
Preferably, in the step 4, a standard test tube is prepared, 1 tube of 100uL stock solution is placed in the standard test tube, then sterile normal saline with 10-15 times volume is added into the standard test tube in a sterile operation, then the standard test tube is slightly shaken, and then the standard test tube is kept still for 10-20min, so as to obtain the suspension working solution.
Preferably, in step 6, the two groups of mice obtained in step 5 are injected with 200uL of working solution per day for 2-3 weeks at an injection dose of 60mg/kg into the abdominal cavity of 25g mice aged 8-9 weeks.
Preferably, in the step 7, the worker selects the mice to perform hair recordings on the 10 th, 17 th and 24 th days after the injection of the working solution, and compares the hair recordings with three groups of mice in the control group injected with the physiological saline.
Compared with the prior art, the invention has the beneficial effects that:
1. according to the invention, 200uL of working solution is continuously injected into the abdominal cavity of a mouse 8-9 weeks old and 25g every day according to the injection dose of 60mg/kg, and the mouse can grow white hair after twenty-four days, and the hair of the mouse is changed from initial sparse to bright and dense.
Drawings
FIG. 1 is a schematic overall flow diagram of the present invention;
FIG. 2 is a graph of day 10 effects recorded for hair growth in mice of the present invention;
FIG. 3 is a graph of day 17 efficacy of hair growth recordings in mice of the present invention;
FIG. 4 is a graph of day 24 efficacy of hair growth recordings in mice of the present invention.
Detailed Description
The technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the drawings in the embodiments of the present invention, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all of the embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.
In the description of the present invention, it should be noted that the terms "upper", "lower", "inner", "outer", "front", "rear", "both ends", "one end", "the other end", and the like indicate orientations or positional relationships based on those shown in the drawings, and are only for convenience of description and simplicity of description, but do not indicate or imply that the referred device or element must have a specific orientation, be constructed in a specific orientation, and be operated, and thus, should not be construed as limiting the present invention. Furthermore, the terms "first" and "second" are used for descriptive purposes only and are not to be construed as indicating or implying relative importance.
In the description of the present invention, it is to be noted that, unless otherwise explicitly specified or limited, the terms "mounted," "disposed," "connected," and the like are to be construed broadly, such as "connected," which may be fixedly connected, detachably connected, or integrally connected; can be mechanically or electrically connected; they may be connected directly or indirectly through intervening media, or they may be interconnected between two elements. The specific meanings of the above terms in the present invention can be understood in specific cases to those skilled in the art.
Referring to fig. 1-4, an embodiment of the present invention is shown: a method of inducing a white hair model comprising the steps of:
a method of inducing a white hair model comprising the steps of:
step 1, dissolving a stock solution;
step 2, subpackaging the solution;
step 3, injection dosage: injecting the liquid substance into the body of the experimental mouse according to the weight of the white mouse and the dosage of 60 mg/kg;
step 4, preparing suspension working solution: taking 1 tube of 100uL stock solution, and adding 10-15 times of normal saline to prepare suspension working solution;
and 5, paraffin unhairing: paraffin depilation is carried out on 8-9 weeks old mice on day 0;
step 6, in vivo study: continuously injecting 200uL of working solution into abdominal cavity of a mouse of 25g and 8-9 weeks old for 2-3 weeks every day according to the injection dosage of 60 mg/kg;
step 7, mouse hair growth recording.
When the stock solution is dissolved and split charged in the step S1, 1mLDMSO is added into 100mgCabozantinib, and the mixed solution is stirred at the rotating speed of 40-60rpm, so that the two solutions are fully dissolved and the concentration of the two solutions is 100 mg/mL.
Further, when a worker stirs the mixed solution of 100mg Cabozantinib and 1mLDMSO at a rotating speed of 40rpm, the time for completely mixing the two is 8 min; when a worker stirs the mixed solution of 100mgCabozantinib and 1mLDMSO at the rotating speed of 50rpm, the time for completely mixing the two is 5min, and the mixed solution is not spilled; when a worker stirred the mixed solution of 100mg Cabozantinib and 1ml DMSO at 60rpm, the time for the two solutions to be completely mixed was 4min, and the mixed solution was slightly spilled.
And (3) when the solution in the step 2 is subpackaged, subpackaging the mixed solution obtained in the step 1 into 100uL/EP tubes with the specification of 2ml, and then storing the liquid substances after subpackaging in an environment with the temperature of-10-30 ℃.
Further, when the mixed solution is stored in an environment with the temperature of-10 ℃, the storage time is two weeks, and the phenomenon of freezing does not occur; when the mixed solution is stored in an environment at the temperature of minus 20 ℃, the storage time is four weeks, the phenomenon of freezing does not occur, and when the mixed solution is stored in an environment at the temperature of minus 30 ℃, a little ice crystal particles appear in the solution.
And 4, preparing a standard test tube, placing 1 tube of 100uL stock solution into the standard test tube, adding 10-15 times of sterile normal saline into the standard test tube in an aseptic operation, slightly shaking the standard test tube, and standing for 10-20min to obtain the suspension working solution.
In step 6, the two groups of mice obtained in step 5 are subjected to intraperitoneal injection of 200uL of working solution every day for 2-3 weeks for 8-9 weeks old and 25g of mice according to the injection dose of 60 mg/kg.
In step 7, the worker selected mice to be subjected to hair recording on the 10 th, 17 th and 24 th days after the injection of the working solution, and compared the mice in the three control groups injected with physiological saline.
Further, injecting 200uL of working solution into the abdominal cavity of a mouse of 8-9 weeks old and 25g per day for 2-3 weeks according to the injection dose of 60mg/kg, wherein the hair of the experimental mouse is sparse when the mouse is injected on the tenth day; when the mice are injected on the seventeenth day, the black hairs of the mice in the control group injected with the physiological saline begin to grow out, and the white hairs of the mice in the drug group injected with Cabozantinib grow out; when the mice are injected for twenty-four days, the black hair of the control group mice is bright, the white hair of the drug group mice is thick, and the hair length is the same as that of the control group mice.
It will be evident to those skilled in the art that the invention is not limited to the details of the foregoing illustrative embodiments, and that the present invention may be embodied in other specific forms without departing from the spirit or essential attributes thereof. The present embodiments are therefore to be considered in all respects as illustrative and not restrictive, the scope of the invention being indicated by the appended claims rather than by the foregoing description, and all changes which come within the meaning and range of equivalency of the claims are therefore intended to be embraced therein. Any reference sign in a claim should not be construed as limiting the claim concerned.
Claims (6)
1. A method of inducing a white hair model, comprising: the method comprises the following steps:
step 1, dissolving a stock solution;
step 2, subpackaging the solution;
step 3, injection dosage: injecting the liquid substance into the body of the experimental mouse according to the weight of the white mouse and the dosage of 60 mg/kg;
step 4, preparing suspension working solution: taking 1 tube of 100uL stock solution, and adding 10-15 times of normal saline to prepare suspension working solution;
and 5, paraffin unhairing: paraffin depilation is carried out on 8-9 weeks old mice on day 0;
step 6, in vivo study: continuously injecting 200uL of working solution into the abdominal cavity of a mouse of 25g and 8-9 weeks old every day for 2-3 weeks according to the injection dose of 60 mg/kg;
step 7, mouse hair growth recording.
2. A method of inducing a white hair model according to claim 1, wherein: when the stock solution is dissolved and split-charged in the step S1, 1mLDMSO is added into 100mgCabozantinib, and the mixed solution is stirred at the rotating speed of 40-60rpm, so that the two solutions are fully dissolved and the concentration of the two solutions is 100 mg/mL.
3. A method of inducing a white hair model according to claim 1, wherein: and (3) when the solution in the step (2) is subpackaged, subpackaging the mixed solution obtained in the step (1) into 100uL/EP tubes with the specification of 2ml, and then storing the subpackaged liquid substance in an environment at the temperature of-10-30 ℃.
4. A method of inducing a white hair model according to claim 1, wherein: in the step 4, a standard test tube is prepared, 1 tube of 100uL stock solution is placed in the standard test tube, then sterile normal saline with the volume 10-15 times that of the standard test tube is added into the standard test tube in a sterile operation, then the standard test tube is slightly shaken, and then the standard test tube is kept stand for 10-20min, so that suspended working solution is obtained.
5. A method of inducing a white hair model according to claim 1, wherein: in the step 6, the two groups of mice obtained in the step 5 are subjected to intraperitoneal injection of 200uL of working solution every day for 2-3 weeks for 8-9 weeks old and 25g of mice according to the injection dose of 60 mg/kg.
6. A method of inducing a white hair model according to claim 1, wherein: in step 7, the worker selected mice to be subjected to hair recording on the 10 th, 17 th and 24 th days after the injection of the working solution, and compared the mice in the three control groups injected with physiological saline.
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| CN202210161963.4A CN114467858A (en) | 2022-02-22 | 2022-02-22 | Method for inducing white hair model |
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Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1225575A (en) * | 1996-07-12 | 1999-08-11 | 庄臣消费者有限公司 | Method for althering hair growth and hair pigmentation and compositions therefor |
| JP2013146243A (en) * | 2012-01-23 | 2013-08-01 | Chube Univ | Model animal for age-associated graying |
| CN111838074A (en) * | 2020-07-15 | 2020-10-30 | 浙江大学 | Method for constructing iron death mouse model by using iron death inducer Erastin and application of method |
| CN112806322A (en) * | 2021-03-22 | 2021-05-18 | 石河子大学 | Method for constructing pigment dropout model |
-
2022
- 2022-02-22 CN CN202210161963.4A patent/CN114467858A/en active Pending
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1225575A (en) * | 1996-07-12 | 1999-08-11 | 庄臣消费者有限公司 | Method for althering hair growth and hair pigmentation and compositions therefor |
| JP2013146243A (en) * | 2012-01-23 | 2013-08-01 | Chube Univ | Model animal for age-associated graying |
| CN111838074A (en) * | 2020-07-15 | 2020-10-30 | 浙江大学 | Method for constructing iron death mouse model by using iron death inducer Erastin and application of method |
| CN112806322A (en) * | 2021-03-22 | 2021-05-18 | 石河子大学 | Method for constructing pigment dropout model |
Non-Patent Citations (2)
| Title |
|---|
| 丁增峰等: ""FLT4调控羊驼黑色素细胞中黑色素生成"", 《中国生物化学与分子生物学报》 * |
| 祝保艳等: ""卡博替尼联合PD-L1单抗对黑色素瘤小鼠移植瘤的抑制作用机制"", 《中山大学学报（医学版）》 * |
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