Evaluation method for the establishment of a non-diagnostic mouse back psoriasis model

Abstract

The present invention provides an evaluation method after the construction of a non-diagnostic mouse back psoriasis model, comprising the following steps: S1, determining the percentage of the mouse skin lesion area to the skin lesion area of ​​the coating area, and scoring according to the skin lesion area ratio B with 0-2 points, wherein 0%=0 point, 1%~30%=1 point, and 31%~100%=2 points; S2, determining the erythema score E, E is divided into 0-2 points according to the depth of the erythema color; S3, determining the scale score D, scoring according to the skin lesion area covered by scales, wherein no scale coverage=0 point, 30%~100%=2 points. Lesions covered with scales = 1 point, 60% lesions covered with scales = 2 points, 90% and above lesions covered with scales = 3 points; S4. Determine the plaque score I, which is scored from 0 to 3 points according to the size of the plaque, with none = 0 points, average plaque diameter <2 mm = 1 point, average plaque diameter 2-4 mm = 2 points, and average plaque diameter >4 mm = 3 points; S5. Determine the severity of the lesions S, S = B+E+D+I. The S score can more accurately describe and quantitatively compare the severity of psoriasis inflammation on the back skin of mice, which is convenient for intuitive and accurate statistical analysis.

Description

Technical Field

[0001] The present invention relates to the research field of psoriasis, and in particular to an evaluation method after a non-diagnostic purpose mouse back psoriasis model is constructed. Background Art

[0002] Psoriasis is an immune-mediated chronic inflammatory skin disease. In the study of psoriasis, the construction of animal models of psoriasis mainly relies on local drug induction and is applied to research at the genetic level. We chose imiquimod for local application to the back skin of mice. After induction, erythema and scaling were produced phenotypically; histologically, it led to excessive proliferation of keratinocytes, new blood vessel formation, and immune cell infiltration, with phenotypes and pathological characteristics similar to human psoriasis. Clinically, the patient's current condition is evaluated by scoring the area and severity of the skin lesions, namely the PASI score. The method is to divide the human body into four parts: head and neck, trunk, upper limbs and lower limbs. First, the scores of the skin lesions in each part are calculated separately, and then the scores of the type and severity of the skin lesions are added together to finally obtain the total PASI score. However, due to actual needs and conditions during the research process, we only chose to score the phenotypic changes in the mouse back skin. In addition, considering that the induced mouse pathogenicity cycle is short, usually reaching the phenotypic peak in about five days, it is quite different from the PASI score used by clinical patients. Therefore, in order to make our research data more effective and more accurately reflect the changes in the pathological phenotype of mice, we invented a new scoring method for the experimental mouse back psoriasis model. Summary of the invention

[0003] The present invention provides an evaluation method for constructing a non-diagnostic mouse back psoriasis model. The evaluation method can be used to more efficiently and intuitively evaluate the quality of the animal model construction and determine the severity of skin lesions, which is conducive to in-depth and systematic research on psoriasis.

[0004] The technical solution of the present invention is a method for evaluating a non-diagnostic mouse back psoriasis model after it is constructed, which is characterized by comprising the following steps:

[0005] S1. Determine the percentage of the mouse skin lesion area to the skin lesion area of ​​the drug application area, and score it with 0-2 points according to the skin lesion area ratio B, where 0% = 0 points, 1%-30% = 1 point, and 31%-100% = 2 points;

[0006] S2. Determine the erythema score E, which is divided into 0-2 points according to the color depth of the erythema, where no erythema is 0 points, light red is 1 point, and red and darker is 2 points;

[0007] S3. Determine the scaling score D, which is divided into 0-3 points based on the percentage of the lesion surface covered with scaling, where no scaling = 0 point, 30% scaling = 1 point, 60% scaling = 2 points, and 90% or more scaling = 3 points;

[0008] S4. Determine the plaque score I, which is scored from 0 to 3 points according to the size of the plaque, with no plaque = 0 points, average plaque diameter < 2 mm = 1 point, average plaque diameter 2 to 4 mm = 2 points, and average plaque diameter > 4 mm = 3 points;

[0009] S5. Determine the severity of skin lesions S, S = B + E + D + I.

[0010] Furthermore, the skin lesion area ratio B=skin lesion area / total experimental skin area.

[0011] Furthermore, the proportion of the surface area of ​​the skin lesions covered with scales is the surface area of ​​the skin lesions covered with scales / total surface area of ​​the skin lesions.

[0012] Furthermore, the plaque size is the maximum diameter of all plaques / the number of plaques.

[0013] The present invention also relates to the application of the evaluation method in describing and quantifying the severity of skin inflammation in a mouse back psoriasis model.

[0014] Furthermore, the S score segmentation standard is: S<4 is mild inflammation, S between 4 and 7 is moderate inflammation, and S>7 is severe inflammation.

[0015] The present invention has the following beneficial effects:

[0016] 1. The existing PASI score for psoriasis is mainly calculated based on the skin lesion area of ​​the limbs, trunk and head and neck, which involves a wide range of skin and has a long disease cycle, and is not suitable for in vivo research in mice. The present invention preferably constructs a mouse back psoriasis model, which has a larger and more intact skin area than other parts, is easy to apply and evenly, and is easy to obtain; the phenotype appears quickly and is typical and intuitive.

[0017] 2. The mouse model is used in the pathological basis research of psoriasis in the present invention. The method for evaluating the severity of skin inflammation in the mouse back psoriasis model provided by the present invention is more in line with the skin phenotype and disease development cycle after psoriasis is induced in mice, and is targeted and practical, and can accurately describe the severity of skin inflammation on the back of mice in a short period of time and accurately compare the subsequent severity.

[0018] 3. This evaluation method can observe the entire course of the psoriasis mouse model and provide dynamic monitoring data for subsequent research, which will help to conduct in-depth research on the pathogenic mechanism and clinical treatment of psoriasis and provide real and effective theoretical data.

[0019] 4. Compared with the multiplication mode of skin lesion area and index in PASI, the addition evaluation method is adopted in this application, the purpose of which is to quantify the skin inflammation in the mouse back psoriasis model, so as to more accurately and intuitively evaluate the degree of skin inflammation. In the PASI scoring, the key indicators are skin lesions, erythema, infiltration and desquamation, while the main indicators of the present invention, skin lesions, erythema, scales, and plaques, are more typical in the mouse back psoriasis model. Since the onset cycle of the mouse back psoriasis model is short, the disease develops and disappears quickly, the quantification of skin inflammation using the evaluation method in this application can better reflect the differences in the disease at different stages. BRIEF DESCRIPTION OF THE DRAWINGS

[0020] Figure 1 Graphs showing phenotypic changes in the back psoriasis model of mice in the example, wherein A is a graph showing the back after mild hair removal; B, C, D, E, and F are graphs showing phenotypic changes after five consecutive days of imiquimod induction.

[0021] Figure 2 This is a quantitative score diagram of the mouse back psoriasis model in the example. DETAILED DESCRIPTION

[0022] The embodiments of the present invention will be described in detail below with reference to examples, but those skilled in the art will appreciate that the following examples are only used to illustrate the present invention and should not be construed as limiting the scope of the present invention.

[0023] The evaluation method of the non-diagnostic mouse back psoriasis model after construction includes the following steps:

[0024] S1. Determine the percentage of the mouse skin lesion area to the skin lesion area of ​​the drug application area, and score it with 0-2 points according to the skin lesion area ratio B, where 0% = 0 points, 1%-30% = 1 point, and 31%-100% = 2 points;

[0025] S2. Determine the erythema score E, which is divided into 0-2 points according to the color depth of the erythema, where no erythema is 0 points, light red is 1 point, and red and darker is 2 points;

[0026] S3. Determine the scaling score D, which is divided into 0-3 points according to the percentage of the surface area of ​​the lesion covered with scales, where no scaling = 0 point, 30% lesions covered with scales = 1 point, 60% lesions covered with scales = 2 points, and 90% or more lesions covered with scales = 3 points; the scaling coverage of lesions is the area of ​​lesions covered with scales / total area of ​​lesions.

[0027] S4. Determine the plaque score I, and score it on a scale of 0-3 points according to the plaque size, with none = 0 points, average plaque diameter <2 mm = 1 point, average plaque diameter 2-4 mm = 2 points, and average plaque diameter >4 mm = 3 points; the plaque size is the maximum diameter of all plaques / number of plaques.

[0028] S5. Determine the severity of skin lesions S, S = B + E + D + I, and quantify the severity of inflammation in the back skin of the psoriasis mouse model according to the S score.

[0029] By scoring the skin lesion area on the back of mice and the typical clinical symptoms of psoriasis, such as erythema, scaling, and plaques, the severity of psoriasis inflammation on the back of mice can be comprehensively evaluated, which is conducive to more accurate and in-depth research on the psoriasis model on the back of mice. At the same time, it can also be conducive to the basic research related to psoriasis.

[0030] The method for constructing a mouse back psoriasis model is to first perform mild hair removal on the experimental skin of the mouse back, and then evenly apply 62.5 mg of imiquimod on the experimental skin of the mouse back for five consecutive days. Figure 1 As shown, A is a picture of the back of mice after mild hair removal, and B, C, D, E, and F are pictures of the phenotypic changes of mice after imiquimod induction for five consecutive days. Figure 1 The composition of the S score for each figure is as follows:

[0031] Figure A is a picture of the back of a mouse after gentle hair removal;

[0032] Figure B is the phenotype of mice one day after imiquimod was applied to the back, where the B value is 0, the E value is 1, the D value is 0, the I value is 0, and the S value is 1;

[0033] Figure C is the phenotype of mice two days after imiquimod was applied to the back, where the B value is 1, the E value is 1, the D value is 1, the I value is 0, and the S value is 3;

[0034] Figure D is the phenotype of mice three days after imiquimod was applied to the back, where the B value is 1, the E value is 1, the D value is 2, the I value is 1, and the S value is 5;

[0035] Figure E is the phenotype of mice two days after imiquimod was applied to the back, where the B value is 2, the E value is 2, the D value is 2, the I value is 2, and the S value is 8;

[0036] Figure F is the phenotype of mice two days after imiquimod was applied to the back, where the B value is 2, the E value is 2, the D value is 3, the I value is 3, and the S value is 10.

[0037] The above are only preferred embodiments of the present invention, and do not limit the patent scope of the present invention. For those skilled in the art, the present invention may have various changes and modifications. Any modification, equivalent replacement, improvement, etc. made within the spirit and principle of the present invention shall be included in the patent protection scope of the present invention.

Claims

1. A method for evaluating a non-diagnostic mouse back psoriasis model after construction, characterized in that: It includes the following steps: S1. Determine the percentage of mouse skin lesion area to the applied area, and score the lesion area B on a scale of 0-2, where 0% = 0 points, 1%-30% = 1 point, and 31%-100% = 2 points. S2. Determine the erythema score E, which is 0-2 points based on the color depth of the erythema, where no erythema is 0 points, light red is 1 point, and red or darker is 2 points; S3. Determine the scaling score D, which is 0-3 points based on the percentage of the lesion surface covered with scales, where no scale coverage = 0 point, 30% scale coverage = 1 point, 60% scale coverage = 2 points, and 90% or more scale coverage = 3 points. S4. Determine the plaque score I, which is scored on a scale of 0-3 based on plaque size: no plaque = 0, average plaque diameter <2 mm = 1, average plaque diameter 2-4 mm = 2, and average plaque diameter >4 mm = 3. S5. Determine the severity of the skin lesions (S = B + E + D + I).

2. The evaluation method according to claim 1, wherein: The skin lesion area ratio B=skin lesion area / total experimental skin area.

3. The evaluation method according to claim 1, wherein: The ratio of the surface area of ​​the skin lesions covered by scales is the surface area of ​​the skin lesions covered by scales / the total surface area of ​​the skin lesions.

4. The evaluation method according to claim 1, wherein: The plaque size is the maximum diameter of all plaques divided by the number of plaques.

5. Use of the evaluation method according to any one of claims 1 to 4 in describing and quantitatively comparing the severity of skin inflammation in a mouse back psoriasis model.

6. The use according to claim 5, characterized in that: The S score segmentation standard is: S<4 is mild inflammation, S between 4 and 7 is moderate inflammation, and S>7 is severe inflammation.

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