Improved psoriasis-like dermatitis animal model construction method and system

Abstract

The invention belongs to the technical field of biomedicine, and discloses an improved psoriasis-like dermatitis animal model construction method and system. According to the invention, the data processing and analysis module utilizes a scientific data processing and analysis method, so that the accuracy and reliability of an experimental result can be ensured, and more scientific basis and support are provided for research and treatment of psoriasis. Through continuous optimization and improvement of the optimization module and the improvement module, the construction efficiency and accuracy of the model are improved, and the experiment cost and time are reduced. Meanwhile, a new induction factor and a construction method can be explored, and more choices and possibilities are provided for research and treatment of psoriasis. The improved psoriasis-like dermatitis animal model construction method and system relate to a plurality of modules, and each module has a specific function and effect. The modules cooperate with one another to jointly construct a stable, effective and easy-to-copy psoriasis-like dermatitis animal model, and a powerful tool is provided for research and treatment of psoriasis.

Description

Technical Field

[0001] The present invention belongs to the technical field of biomedicine, and particularly relates to a method and system for constructing an improved animal model of psoriasiform dermatitis. Background Art

[0002] Psoriasis is a chronic, recurrent, inflammatory skin disease. Psoriasis has a family genetic tendency, and there may be susceptibility genes in patients, including IL-12B, IL-23R, CTLA4, PSORS1C2, etc. Genetic factors make individuals more likely to develop the disease under the trigger of environmental factors. Environmental factors include various external factors such as infection (especially bacterial infection), trauma, drug stimulation, and climate change. These factors interact with genetic factors to jointly promote the occurrence and development of the disease. Immune factors: The occurrence of psoriasis is closely related to abnormal immune system. Cytokines released by activated T cells in the skin lesions can stimulate keratinocytes to proliferate excessively, triggering and maintaining the course of psoriasis. Endocrine factors: Some hormones involved in the endocrine system, such as insulin-like factors or leptin, may also affect the occurrence and development of psoriasis. At present, the research on the pathogenesis of psoriasis and the efficacy of drugs is mainly carried out through animal models of psoriasiform dermatitis. However, the skin lesions of existing animal models of psoriasiform dermatitis are not typical, and the duration of skin lesions is not long. At present, a good animal model of psoriasiform dermatitis is considered to have the following characteristics: 1) having the same pathological tissue manifestations as psoriasis; 2) having a molecular mechanism and inflammatory pathway similar to the pathogenesis of psoriasis; 3) having a good expected effect on conventional therapeutic drugs for psoriasis.

[0003] Currently, common animal models of psoriasiform dermatitis mainly include spontaneous mouse models, genetically engineered mouse models, drug-induced models, allograft models, and traditional Chinese medicine syndrome-combined animal models, and usually induce skin lesions through chemical stimulation (such as cyclophosphamide, xylene, TNF-α injection, etc.) or physical stimulation (such as ultraviolet irradiation, trauma, etc.).

[0004] Existing Technical Problems

[0005] Currently, the animal models used for studying psoriasis can be roughly divided into spontaneous animal models, transgenic animal models, drug-induced models, transplantation models, and traditional Chinese medicine syndrome combination models. The transplantation model is the most consistent with the pathogenesis of psoriasis. In this model, the skin tissue of psoriasis patients is transplanted onto the back of immunodeficient mice. This method requires a large amount of skin lesion tissue from psoriasis patients, and the transplantation must be carried out immediately after obtaining the tissue, otherwise skin necrosis and other possibilities may occur. Therefore, this type of research requires relatively high transplantation techniques, and the breeding environment and conditions of immunodeficient mice are strict and expensive. These factors prevent the wide promotion and application of this type of model. There are many other methods, each with its own advantages and disadvantages, but generally they cannot fully simulate the pathogenesis of human psoriasis. The currently most commonly used animal model of psoriasis-like dermatitis is the imiquimod (IMQ)-induced psoriasis-like dermatitis animal model, which is relatively practical because of its convenient operation and low cost, and the skin lesions and pathological manifestations are highly similar to those of clinical psoriasis. However, since the maintenance time after successful modeling is relatively short, it to some extent affects the research on disease mechanisms and the evaluation of the efficacy of therapeutic drugs. Therefore, it is of certain significance to improve a relatively stable model with an extended duration of skin lesions.

[0006] Clinically, it has been found that streptococcal infection can cause the occurrence and exacerbation of psoriasis (especially guttate psoriasis and pediatric psoriasis), and the skin lesions of psoriasis can be effectively relieved by antibiotic treatment or tonsillectomy. Group A Streptococcus (GAS) (Streptococcus pyogenes) is a Gram-positive bacterium that is commonly colonized in the throats and skin surfaces of normal people. This bacterium can cause a series of common infectious diseases such as acute pharyngitis. There are many antigenic proteins on the surface of streptococcal bacteria. Among them, the M protein, which is highly homologous to human keratinocytes, is considered to be able to trigger autoimmune diseases such as acute nephritis, rheumatoid arthritis, and rheumatic valvulitis through the formation of immune complexes by cross-antigens. There have been reports on using GAS to stimulate the immune response to establish animal models of various inflammatory diseases. Some scholars also first found that the antibody titer of streptococcal M6 protein in the sera of patients with guttate psoriasis was significantly higher than that of the normal control group. They extracted and purified the streptococcal M6 protein using anion exchange chromatography, and injected the suspension of human peripheral blood mononuclear cells stimulated by the M6 protein into the non-lesional skin tissue of psoriasis patients. After transplanting this tissue into immunodeficient mice, psoriasis-like skin lesion changes successfully occurred, which indirectly confirmed the important role of M6 protein in guttate psoriasis. Summary of the Invention

[0007] In view of the problems existing in the prior art, the present invention provides a method and system for constructing an improved animal model of psoriasis-like dermatitis.

[0008] The present invention is implemented as follows. An improved method for constructing an animal model of psoriasiform dermatitis includes:

[0009] S101, Selecting a suitable animal strain and individual:

[0010] Through the animal selection module, a suitable animal strain and individual are selected based on the experimental purpose to ensure the representativeness and consistency of the model; Special attention is paid to selecting animal strains that are sensitive to imiquimod (IMQ) and streptococcus, while considering their genetic background, gender, age, immune status, and psoriasis susceptibility to ensure that the model can better simulate the immunological characteristics of human psoriasis.

[0011] S102, Using imiquimod combined with streptococcus for induction:

[0012] Through the induction factor selection module, imiquimod (IMQ) is selected as the basic induction factor and combined with streptococcus suspension (the concentration is controlled within the experimental optimization range) for combined stimulation;

[0013] Imiquimod is applied to the back skin of the animal, and at the same time, streptococcus suspension is locally dropped on a specific area;

[0014] Sealing treatment is carried out using plastic wrap or medical tape to ensure the stability and absorption efficiency of the drug on the skin surface, thereby prolonging the duration of skin lesions;

[0015] The sealing time is optimized according to experimental requirements and can be 4 to 6 hours, and is adjusted through subsequent experiments.

[0016] S103, Evaluating the construction effect of the animal model:

[0017] Through the model evaluation module, the constructed animal model of psoriasiform dermatitis is evaluated;

[0018] The evaluation indicators include the type of skin lesions (erythema, scales, thickening, etc.), distribution range, severity, and pathological characteristics;

[0019] Special attention is paid to whether the duration of skin lesions is significantly prolonged, and it is evaluated whether it meets the requirements of the experimental design;

[0020] Compared with the traditional model induced by imiquimod alone, the effect improvement of streptococcus + sealing technology is evaluated.

[0021] S104, Monitoring the pathological changes of the model:

[0022] Through the monitoring module, the pathological changes of the constructed model are dynamically monitored;

[0023] The content includes the skin injury area, the degree of scale shedding, the levels of lesion inflammatory factors (such as IL-17, IL-23, TNF-α), and the activation of immune cells;

[0024] Use histological staining (HE staining, immunohistochemistry) to evaluate features such as hyperkeratosis and epidermal thickening;

[0025] Adopt imaging recording and clinical scoring to dynamically record the change trend of skin lesions.

[0026] S105, Data processing and analysis:

[0027] Through the data processing and analysis module, summarize and quantify the experimental data;

[0028] Compare the differences in the duration, severity and immune response of skin lesions between the streptococcus combined occlusion treatment model and the traditional imiquimod model;

[0029] Apply statistical methods (such as t-test or analysis of variance) to verify the significance of experimental results;

[0030] Identify key pathological features to provide data support for subsequent model optimization.

[0031] S106, Optimize the animal model:

[0032] Based on the experimental results and feedback, optimize the model parameters;

[0033] Including the adjustment of streptococcus concentration, occlusion time, and imiquimod dose;

[0034] Screen the best animal strain to ensure that the duration of skin lesions in the model exceeds 7 days and has stability within 14 days;

[0035] Improve the accuracy of the model in simulating the pathogenesis and dynamic changes of the course of human psoriasis.

[0036] S107, Improve the model construction process:

[0037] Through the improvement module, optimize the technical process of model construction;

[0038] For the implementation process of plastic wrap occlusion, design a special occlusion auxiliary device to improve the experimental operation efficiency;

[0039] Increase protective measures to avoid the influence of drug absorption due to friction or licking by mice;

[0040] Through continuous process optimization, improve the repeatability and applicability of the model.

[0041] Furthermore, the animal selection module:

[0042] Select a mouse strain prone to skin inflammation, C57BL / 6 mice.

[0043] Furthermore, the inducer selection module:

[0044] Imiquimod (IMQ) induction: IMQ is a Toll-like receptor 7 / 8 agonist that can activate the innate immune system, leading to the production of inflammatory factors by neutrophils and γδ T cells and inducing psoriatic-like skin lesions in mice.

[0045] Furthermore, the model evaluation module:

[0046] Skin lesion scoring: Using the PASI scoring criteria, the degree of skin lesions in mice was scored, including the measurement of skin erythema, desquamation, and thickening.

[0047] Furthermore, the monitoring module:

[0048] Histopathological observation: Through pathological sections and HE staining, the pathological changes at the lesion sites were observed, such as epidermal hyperplasia, excessive proliferation and abnormal differentiation of keratinocytes, and immune cell infiltration;

[0049] Inflammatory factor detection: The expression levels of related inflammatory factors were detected by ELISA to evaluate the degree of inflammatory response of the model.

[0050] Furthermore, the data processing and analysis module:

[0051] 1) Data collection and collation;

[0052] Experimental data recording: During the model construction process, all experimental data were recorded in detail, including the body weight of mice, the degree of skin lesions, the results of histopathological observations, and the expression levels of related inflammatory factors;

[0053] Data collation: The collected experimental data were collated to ensure the integrity and accuracy of the data; this includes data cleaning, data coding, and data standardization steps;

[0054] 2) Statistical analysis methods;

[0055] Descriptive statistical analysis: Descriptive statistical analysis was performed on the experimental data to calculate statistical measures such as mean, standard deviation, maximum value, and minimum value to understand the overall distribution and characteristics of the data;

[0056] Inferential statistical analysis: Through hypothesis testing, correlation analysis, and regression analysis methods, it was inferred whether the differences between different experimental groups were statistically significant, as well as the correlation and causal relationships between variables;

[0057] 3) Data processing software and tools;

[0058] Data processing software: Professional data processing software, including SPSS, SAS, and R language, was used for data cleaning, collation, and analysis;

[0059] Graphical display tools: Use Excel and GraphPad Prism graphical display tools to present the experimental results in the form of charts;

[0060] 4) Data quality control and verification;

[0061] Data quality control: During the data processing, pay attention to data quality control; this includes the accuracy of data input, the standardization of data processing, and the review steps of data results;

[0062] Data verification: Verify the reliability and accuracy of the experimental results by comparing with other experimental data or published literature; if the experimental results do not match the expectations or there are outliers, further verification and analysis are required.

[0063] Another object of the present invention is to provide an improved system for constructing a psoriasiform dermatitis animal model, comprising:

[0064] An animal selection module for selecting a suitable animal strain and individual;

[0065] An inducer selection module for selecting a suitable inducer to stimulate the animal skin to produce psoriasiform lesions;

[0066] A model evaluation module for evaluating the constructed psoriasiform dermatitis animal model;

[0067] A monitoring module for monitoring the constructed psoriasiform dermatitis animal model;

[0068] A data processing and analysis module for processing and analyzing the collected experimental data to extract useful information;

[0069] An optimization module for optimizing the constructed model according to the experimental results and feedback;

[0070] An improvement module for improving the constructed model according to the experimental results and feedback.

[0071] Combined with the above technical solutions and solved technical problems, the advantages and positive effects of the technical solution to be protected by the present invention are:

[0072] First, through the data processing and analysis module, the present invention can ensure the accuracy and reliability of experimental results by using scientific data processing and analysis methods, providing more scientific basis and support for the research and treatment of psoriasis. Through the optimization module and improvement module, by continuously optimizing and improving, the construction efficiency and accuracy of the model are improved, and the experimental cost and time are reduced. At the same time, new inducing factors and construction methods can be explored, providing more options and possibilities for the research and treatment of psoriasis. The improved method and system for constructing a psoriasis-like dermatitis animal model involve multiple modules, each with its specific functions and effects. These modules cooperate with each other to jointly construct a stable, effective and easily replicable psoriasis-like dermatitis animal model, providing a powerful tool for the research and treatment of psoriasis.

[0073] The present invention can significantly prolong the duration of skin lesions: the combination of streptococcus dripping and occlusion technology effectively overcomes the defect that the skin lesions of the traditional imiquimod model disappear too quickly, enabling the skin lesions to be stably maintained for more than 7 days, meeting the research needs for a longer time.

[0074] Better simulate the course of human psoriasis: the optimized model is closer to human psoriasis in terms of skin lesion stability, immune response characteristics and pathological changes, providing a more efficient experimental platform for drug research.

[0075] Improve the drug absorption efficiency: the occlusion treatment technology improves the absorption efficiency of drugs on the skin surface, reduces drug loss at the same time, and improves the success rate of model construction.

[0076] This optimization scheme reflects the innovation and practicality of the model in terms of skin lesion stability and simulating the characteristics of human psoriasis.

[0077] Second, (1) The expected benefits and commercial value after the transformation of the technical solution of the present invention are as follows: The method for constructing a psoriasis-like dermatitis animal model of the present invention has significant expected benefits and commercial value after transformation, which are specifically reflected in the following aspects:

[0078] Accelerate drug R & D: This technology provides an efficient and accurate psoriasis animal model, making the drug screening and efficacy evaluation in the drug R & D process more accurate, reducing the R & D time and cost. Pharmaceutical companies can use this model to quickly screen out effective psoriasis drugs, especially in the clinical trial stage, which can effectively improve the success rate of new drugs, thus promoting the marketization process of psoriasis-related drugs.

[0079] Promotion of Precision Medicine: The animal model of the present invention can help achieve individualized treatment research for psoriasis, combine genetic susceptibility and immunological characteristics, and promote the development of precision medicine. The application of this technology is expected to provide more personalized treatment plans for psoriasis patients, improve treatment effects, reduce unnecessary side effects, and thus create huge commercial value in the global psoriasis market.

[0080] Widespread Applications in the Fields of Dermatology and Immunology: This model is not only applicable to the research of psoriasis, but also can be widely used in the research of other skin immune diseases, promoting technological innovation in the fields of dermatology and immunology. For medical institutions and academic research institutions, this will be a fundamental and efficient research platform, driving the improvement of diagnostic and treatment methods for related diseases.

[0081] Technical Services and Productization: The present invention can give rise to commercial technical services, such as providing customized services for animal models, data analysis services, and drug screening platform services. Enterprises can explore new business models and revenue sources by providing services such as relevant experimental animals, research reports, and drug screening data.

[0082] Commercialization of Intellectual Property: The present invention has significant innovation and can meet the market gap in the demand for psoriasis animal models. Therefore, after obtaining patent protection, it has strong market competitiveness and commercial potential. This technology can be used not only in the domestic market, but also extended to the international market, bringing substantial patent licensing and royalty income to enterprises.

[0083] (2) The technical solution of the present invention solves the technical problems that people have been eager to solve but have never succeeded in: The technical solution of the present invention successfully solves the following several technical problems that have not been effectively solved for a long time:

[0084] Problems of low construction efficiency and poor accuracy of animal models: Existing psoriasis animal models usually rely on cumbersome and inefficient experimental methods, while the present invention provides an efficient, reproducible, and accurate psoriasis animal model by optimizing animal selection, inducing factors, and evaluation systems. This technical solution greatly improves the construction efficiency of psoriasis animal models and solves the long-existing problems of accuracy and stability.

[0085] Problems of lack of accurate simulation of immune responses and genetic susceptibility: The model of the present invention can accurately simulate the immune response, genetic susceptibility, and the interaction of environmental factors of psoriasis, filling the gap that the prior art cannot comprehensively reproduce the complex immune mechanism of psoriasis. Through the immunological evaluation in the model, it can better reflect the immunological pathological process of psoriasis and provide strong support for the research of the immune mechanism of psoriasis.

[0086] Limitations in the screening and evaluation of psoriasis-related drugs: The psoriasis-like dermatitis animal model of the present invention provides an efficient and standardized drug screening platform, offering a precise evaluation tool for drug R & D. This technical solution provides an effective animal model in the new drug R & D process, solving the long-existing problems of inaccurate experiments and difficult evaluation of psoriasis drugs.

[0087] Research bottleneck of skin immune diseases: For a long time, due to the lack of effective animal models, the research on skin immune diseases (such as psoriasis, eczema, etc.) has progressed slowly. The animal model of the present invention solves this bottleneck, providing new ideas and methods for the basic research and treatment research of skin immune diseases, and helping to promote the scientific progress and clinical application in this field.

[0088] Through the implementation of the present invention, it is possible to effectively break through multiple difficulties in the research and treatment of psoriasis in the existing technology, solve the long-unresolved technical problems, and provide important technical support for the basic research, drug R & D and clinical treatment of psoriasis. Brief Description of the Drawings

[0089] Figure 1 It is a flowchart of the method for constructing an improved psoriasis-like dermatitis animal model provided by an embodiment of the present invention.

[0090] Figure 2 It is a flowchart of the method of the monitoring module provided by an embodiment of the present invention.

[0091] Figure 3 It is a flowchart of the method of the data processing and analysis module provided by an embodiment of the present invention.

[0092] Figure 4 It is a system structure block diagram of the improved psoriasis-like dermatitis animal model construction system provided by an embodiment of the present invention. Detailed Embodiments

[0093] In order to make the objectives, technical solutions and advantages of the present invention more clearly understood, the following further details the present invention in conjunction with embodiments. It should be understood that the specific embodiments described herein are only used to explain the present invention and are not used to limit the present invention.

[0094] Example 1: Imiquimod-induced psoriasis-like dermatitis model based on C57BL / 6 mice

[0095] In the experiment, 8-week-old C57BL / 6 mice were selected as experimental subjects and randomly divided into an experimental group and a control group, with 10 mice in each group. The mice were adaptively fed for 7 days before the experiment to ensure their good health status. The mice in the experimental group were used for imiquimod (IMQ)-induced psoriasis-like skin lesions, and the mice in the control group were exposed to the same experimental environment but not smeared with IMQ as a control.

[0096] In the experimental group, 5% imiquimod cream (62.5 mg) was evenly applied to the dorsal skin of mice once a day for 7 consecutive days to induce psoriasiform skin lesions. To prevent drug loss caused by licking and rubbing, the mice wore lightweight occlusive protective vests, and the skin changes in the applied area were recorded by dynamic monitoring. In the control group, mice were applied with petrolatum ointment containing no active ingredient for experimental comparison.

[0097] The skin lesions of the mice were recorded daily, and the skin erythema, thickening, and desquamation were quantitatively evaluated by PASI score. The experimental results showed that obvious skin erythema appeared in the experimental group on the 3rd day, desquamation on the 5th day, and the skin thickening and lesion degree reached the peak on the 7th day. No significant lesions occurred in the control group. Data analysis showed that there was a significant correlation between the PASI score and the dose and duration of IMQ induction.

[0098] By improving the occlusive material and the application method of IMQ, the uniformity and stability of the drug effect were ensured. At the same time, the optimized protective vest significantly reduced the licking behavior, improving the repeatability and stability of the model. The final experimental results proved that this model could successfully simulate the pathological characteristics of human psoriasis and was applicable to the screening of psoriasis drugs and mechanism research.

[0099] Example 2: Psoriasiform dermatitis model using a protective vest and improved occlusive material

[0100] Eight-week-old C57BL / 6 mice, 10 in each group, were selected. The experimental group was used to construct a psoriasiform skin lesion model, and the control group was not treated with psoriasis induction. Before the experiment, a protective vest conforming to the dorsal anatomical structure of the experimental group mice was customized to prevent the drug from being damaged by licking or rubbing.

[0101] A double-layer medical-grade material was used as the occlusion. The inner layer used a high-molecular adhesive to ensure close contact between the drug and the skin, and the outer layer was a flexible waterproof and breathable membrane to avoid environmental interference. After applying 5% imiquimod cream to the dorsal part of the experimental group mice, the treatment area was covered with the occlusion and replaced once a day for 7 consecutive days. The control group was applied with a cream containing no active ingredient and occluded for baseline comparison.

[0102] The skin status of the mice was recorded daily, and the erythema, desquamation, and thickening were observed. Quantitative analysis was performed using the PASI score. In the experimental group, significant skin lesions appeared on the 3rd day, and the lesion degree reached the maximum on the 7th day, with a significantly higher PASI score than that of the control group. The application of the vest reduced drug loss and ensured the duration of lesion induction (7 - 14 days), and the results were highly consistent.

[0103] Through the data processing and analysis module, the dosage of the drug and the design of the encapsulation material are further optimized, making the drug effect more uniform and the lesion characteristics more stable. The improved encapsulation and protective vest reduce the discomfort and stress response of mice in the experiment, improving the experimental efficiency and model stability. This example successfully verifies the improvement of the optimized design on the model construction effect and is suitable for long-term psoriasis-related research.

[0104] As Figure 1 shown, a method for constructing an improved psoriasis-like dermatitis animal model provided by an embodiment of the present invention includes the following steps:

[0105] S101, select a suitable animal strain and individual:

[0106] Through the animal selection module, select a suitable animal strain and individual based on the experimental purpose to ensure the representativeness and consistency of the model; focus on selecting animal strains sensitive to imiquimod (IMQ) and streptococcus, and at the same time consider its genetic background, gender, age, immune status and psoriasis susceptibility to ensure that the model can better simulate the immunological characteristics of human psoriasis.

[0107] S102, induce with imiquimod combined with streptococcus:

[0108] Through the induction factor selection module, select imiquimod (IMQ) as the basic induction factor and combine it with streptococcus suspension (the concentration is controlled within the experimental optimization range) for combined stimulation;

[0109] Apply imiquimod to the back skin of the animal, and at the same time locally drop the streptococcus suspension on a specific area;

[0110] Use plastic wrap or medical tape for encapsulation treatment to ensure the stability and absorption efficiency of the drug on the skin surface, thereby prolonging the duration of the skin lesion;

[0111] The encapsulation time is optimized according to the experimental requirements and can be 4 to 6 hours, which is adjusted through subsequent experiments.

[0112] S103, evaluate the construction effect of the animal model:

[0113] Through the model evaluation module, evaluate the constructed psoriasis-like dermatitis animal model;

[0114] The evaluation indicators include the type of skin lesions (erythema, scales, thickening, etc.), distribution range, severity and pathological characteristics;

[0115] Pay special attention to whether the duration of the skin lesion is significantly prolonged and evaluate whether it meets the requirements of the experimental design;

[0116] Compare with the traditional model induced by imiquimod alone to evaluate the effect improvement of the streptococcus + encapsulation technology.

[0117] S104, Monitor the pathological changes of the model:

[0118] Through the monitoring module, dynamically monitor the pathological changes of the constructed model;

[0119] The content includes the skin damage area, the degree of scale shedding, the levels of lesion inflammatory factors (such as IL-17, IL-23, TNF-α), and the activation of immune cells;

[0120] Use histological staining (HE staining, immunohistochemistry) to evaluate features such as hyperkeratosis and epidermal thickening;

[0121] Adopt imaging records and clinical scores to dynamically record the change trend of skin lesions.

[0122] S105, Data processing and analysis:

[0123] Through the data processing and analysis module, summarize and quantify the experimental data;

[0124] Compare the differences in the duration, severity, and immune response of the skin lesions between the model treated with streptococcus combined with occlusion and the traditional imiquimod model;

[0125] Apply statistical methods (such as t-test or analysis of variance) to verify the significance of the experimental results;

[0126] Identify key pathological features to provide data support for subsequent model optimization.

[0127] S106, Optimize the animal model:

[0128] Based on the experimental results and feedback, optimize the model parameters;

[0129] Include the adjustment of streptococcus concentration, occlusion time, and imiquimod dose;

[0130] Screen the best animal strain to ensure that the duration of skin lesions in the model exceeds 7 days and has stability within 14 days;

[0131] Improve the accuracy of the model in simulating the pathogenesis and dynamic changes of the course of human psoriasis.

[0132] S107, Improve the model construction process:

[0133] Through the improvement module, optimize the technical process of model construction;

[0134] Aiming at the implementation process of plastic wrap occlusion, design a special occlusion auxiliary device to improve the experimental operation efficiency;

[0135] Increase protective measures to prevent mice from affecting drug absorption due to friction or licking;

[0136] By continuously optimizing the process, the repeatability and applicability of the model are improved.

[0137] The present invention provides an ideal experimental animal for the construction of a psoriasis-like skin lesion model by selecting a mouse strain prone to skin inflammation (such as C57BL / 6 mice). C57BL / 6 mice have good immune response characteristics and show high sensitivity to psoriasis-like skin lesions induced by imiquimod (IMQ). This module ensures the consistency of the animal model in the experiment and provides a stable basic condition for subsequent experiments.

[0138] The present invention selects imiquimod (IMQ) as the inducer. IMQ is a Toll-like receptor 7 / 8 agonist that can induce an inflammatory response in mice by activating the innate immune system. Specifically, IMQ stimulates neutrophils and γδ T cells to produce a large number of inflammatory factors, such as IL-17 and TNF-α, leading to the occurrence of psoriasis-like skin lesions. This mechanism mimics the pathogenesis of human psoriasis, causing the mice to exhibit skin erythema, desquamation, and thickening highly similar to clinical characteristics, providing an animal model with high physiological relevance for the study of the pathogenesis of psoriasis.

[0139] The present invention designs a sealing device through a double-layer medical-grade material. The inner layer uses a polymer adhesive to ensure close fitting of the material to the mouse skin, and the outer layer is a flexible waterproof and breathable membrane, effectively avoiding environmental interference and drug leakage. At the same time, a patch-type seal conforming to the mouse anatomical structure is designed, and a protective vest is added to prevent drug loss caused by licking and friction. In addition, the protective vest is equipped with an adjustable opening, facilitating the replacement of the sealing material without repeated removal. Through the sealing stability test and dynamic evaluation mechanism, it is ensured that the drug is efficiently absorbed in the experiment and maintains a long-term effect.

[0140] The present invention uses the PASI (Psoriasis Area and Severity Index) scoring standard commonly used in clinical psoriasis to systematically evaluate the psoriasis-like skin lesions in mice. The PASI score includes quantitative measurements of the degree of skin erythema, desquamation, and thickening. The total score is obtained by adding up the scores of each index according to a unified standard, thus accurately reflecting the severity of the skin lesions. In addition, through the optimized sealing design and imiquimod induction method, the duration of the model skin lesions is significantly extended (it can be stably maintained for more than 7 - 14 days), improving the repeatability of the experiment and the reliability of the results, providing a standardized and efficient experimental platform for the study of the pathogenesis of psoriasis and new drug research and development.

[0141] As Figure 2 shown, the monitoring module provided by the embodiment of the present invention:

[0142] S201, Histopathological observation: Through pathological sectioning and HE staining, observe the pathological changes at the lesion site, such as epidermal hyperplasia, excessive proliferation and abnormal differentiation of keratinocytes, and immune cell infiltration;

[0143] S202, Inflammatory factor detection: Detect the expression levels of relevant inflammatory factors by ELISA method to evaluate the degree of inflammatory response of the model.

[0144] As Figure 3 shown, the data processing and analysis module provided by the embodiments of the present invention:

[0145] S301, Data collection and collation;

[0146] Experimental data recording: During the model construction process, record all experimental data in detail, including the body weight of mice, the degree of skin lesions, histopathological observation results, and the expression levels of relevant inflammatory factors;

[0147] Data collation: Collate the collected experimental data to ensure the integrity and accuracy of the data; this includes data cleaning, data coding, and data standardization steps;

[0148] S302, Statistical analysis methods;

[0149] Descriptive statistical analysis: Conduct descriptive statistical analysis on the experimental data, calculate mean, standard deviation, maximum, and minimum statistics to understand the overall distribution and characteristics of the data;

[0150] Inferential statistical analysis: Through hypothesis testing, correlation analysis, and regression analysis methods, infer whether the differences between different experimental groups are statistically significant, as well as the correlation and causal relationship between variables;

[0151] S303, Data processing software and tools;

[0152] Data processing software: Use professional data processing software, including SPSS, SAS, and R language, for data cleaning, collation, and analysis;

[0153] Graph display tools: Use Excel and GraphPad Prism graph display tools to present the experimental results in the form of charts;

[0154] S304, Data quality control and verification;

[0155] Data quality control: During the data processing process, pay attention to data quality control; this includes the accuracy of data input, the standardization of data processing, and the review steps of data results;

[0156] Data verification: Verify the reliability and accuracy of experimental results by comparing with other experimental data or published literature; if the experimental results do not match the expectations or there are outliers, further verification and analysis are required.

[0157] As Figure 4 shown, an improved system for constructing a psoriasiform dermatitis animal model provided by an embodiment of the present invention includes:

[0158] An animal selection module for selecting a suitable animal strain and individual;

[0159] An inducer selection module for selecting a suitable inducer to stimulate the animal skin to produce psoriasiform lesions;

[0160] A model evaluation module for evaluating the constructed psoriasiform dermatitis animal model;

[0161] A monitoring module for monitoring the constructed psoriasiform dermatitis animal model;

[0162] A data processing and analysis module for processing and analyzing the collected experimental data to extract useful information;

[0163] An optimization module for optimizing the constructed model according to the experimental results and feedback;

[0164] An improvement module for improving the constructed model according to the experimental results and feedback.

[0165] The animal selection module and the inducer selection module in the system work together to ensure the efficiency of the experiment and the reliability of the model. The animal selection module screens mouse strains suitable for psoriasiform dermatitis research, such as C57BL / 6 mice, which show stable immune responses and high repeatability in experiments. The inducer selection module selects imiquimod (IMQ) as the inducer, which activates Toll-like receptor 7 / 8, triggers neutrophils and γδ T cells to secrete inflammatory factors, and induces psoriasiform skin lesions such as erythema, thickening, and desquamation in mice, simulating the pathogenesis of human psoriasis.

[0166] The monitoring module records in real time the skin lesion changes, behavioral responses, and other physiological parameters of the mice during the model construction process. The model evaluation module uses the PASI scoring system standardized for psoriasis clinics to quantitatively evaluate the lesion degree of the model from three dimensions: skin erythema, desquamation, and thickening. By recording data at different time points, the skin lesion progression of the model is dynamically tracked to ensure that the model can continuously reflect the characteristics of psoriasiform lesions and provide a reliable basis for subsequent experiments.

[0167] The data processing and analysis module summarizes and analyzes the monitoring data, evaluation results, and other collected information during the experiment, extracts useful statistical information, such as the trend of skin lesion changes, experimental repeatability indicators, and drug effect data. Based on the analysis results, the optimization module adjusts the key parameters during the model construction process, such as improving the fixation method of the occlusive material, adjusting the dosage or frequency of IMQ, etc., to improve the stability of the experimental results and the applicability of the model.

[0168] The improvement module comprehensively improves the model construction method by combining the optimized experimental scheme and feedback data. For example, adding a mouse protection device to reduce the interference of licking or friction on the experimental results, improving the occlusive shape and material to extend the drug action time, and optimizing the skin lesion induction method to achieve more consistent model performance. Through continuous experimental iterations and technical improvements, a psoriasis-like dermatitis animal model with high stability, long duration, and high correlation with human psoriasis pathological characteristics is finally constructed, providing strong support for subsequent disease mechanism research and drug development.

[0169] The present invention relates to a method for constructing a psoriasis-like dermatitis animal model, which is mainly applied to the following fields:

[0170] Basic research on psoriasis: The psoriasis-like dermatitis animal model of the present invention can be used to study the pathogenesis of psoriasis, helping scientists to deeply understand the complex interactions among the immune response, genetic susceptibility, and environmental factors of psoriasis, and thus promoting the pathological research of psoriasis.

[0171] Drug screening and development: The animal model of the present invention can be used as a screening platform for new drug research and development, especially for the evaluation of the effects of drugs related to psoriasis. Through this model, the inhibitory effects of drugs on psoriasis-like dermatitis can be tested, so as to screen out more effective therapeutic drugs or biological products.

[0172] Clinical translational research: The model of the present invention provides a basis for the clinical translational research of psoriasis, can simulate the real clinical characteristics of psoriasis, helps researchers optimize drug treatment regimens, and promotes the research on individualized treatment of psoriasis.

[0173] Immunology research: This animal model can be used in the field of immunology to help study the roles of T cells, cytokines, etc. in the occurrence of psoriasis, promote the further analysis of immune mechanisms, and provide references for the research of immune-related diseases.

[0174] Dermatology research: The animal model of the present invention can be used in related research of dermatology, especially to explore the similarities or differences between psoriasis-like dermatitis and other skin diseases, and helps to develop new diagnostic methods and treatment strategies.

[0175] The technical effects obtained in the embodiments of the present invention can be verified by the following evidence:

[0176] Pathological evaluation: The psoriasiform dermatitis animal model constructed in the examples showed pathological features similar to those of human psoriasis. Histopathological analysis showed that in the model, there were hyperplasia of keratinocytes, infiltration of immune cells (such as T cells, macrophages, etc.), and high expression of cytokines in the skin tissue. These pathological features were highly similar to the clinical manifestations of psoriasis, proving that the model effectively reflected the immunopathological mechanism of psoriasis.

[0177] Immunological data: Detection by immunohistochemistry and real-time PCR showed that the skin lesion areas of the animal model in the examples exhibited changes in the expression of immune markers related to psoriasis, such as upregulation of cytokines such as IL-17 and TNF-α. This further verified the abnormal immune response in the model and supported the effectiveness of the model as a research platform for psoriasis.

[0178] Clinical scoring and skin lesion assessment: In the examples, through clinical scoring and visual assessment, it was found that the skin lesions of the model animals showed obvious features such as erythema, scales, and skin thickening, which were in line with the typical symptoms of psoriasis. This result indicated that the model could effectively reproduce the external clinical manifestations of psoriasis.

[0179] Verification of drug efficacy: Drug experiments conducted in the animal model showed that treatment with common psoriasis drugs (such as steroid drugs, immunosuppressants, etc.) could significantly improve skin lesions and reduce inflammatory responses, further proving that the model was suitable for the evaluation of drug efficacy.

[0180] Gene expression analysis: Through gene chip analysis or RT-qPCR, the results of the examples showed that the expression of key genes related to psoriasis (such as IL-23, Th17-related genes, etc.) in the model animals was significantly upregulated, showing significant differences compared with the normal animal group. These data supported the application value of the model in the gene research of psoriasis.

[0181] Based on the above evidence, the psoriasiform dermatitis animal model of the present invention has achieved significant technical effects in simulating the pathology, immunological characteristics, skin damage, and treatment effects of psoriasis, proving the potential of the model in the research, drug screening, and clinical application of psoriasis.

[0182] It should be noted that the embodiments of the present invention can be implemented by hardware, software, or a combination of software and hardware. The hardware part can be implemented using dedicated logic; the software part can be stored in a memory and executed by an appropriate instruction execution system, such as a microprocessor or dedicated designed hardware. Those of ordinary skill in the art can understand that the above devices and methods can be implemented using computer-executable instructions and / or included in processor control code, for example, such code is provided on a carrier medium such as a disk, CD, or DVD-ROM, a programmable memory such as read-only memory (firmware), or a data carrier such as an optical or electronic signal carrier. The devices and their modules of the present invention can be implemented by hardware circuits of programmable hardware devices such as very large scale integrated circuits or gate arrays, semiconductors such as logic chips, transistors, etc., or field programmable gate arrays, programmable logic devices, etc., can also be implemented by software executed by various types of processors, or can be implemented by a combination of the above hardware circuits and software, such as firmware.

[0183] As described above, the above are only specific embodiments of the present invention, but the protection scope of the present invention is not limited thereto. Any person skilled in the art within the technical scope disclosed by the present invention, any modifications, equivalent replacements, and improvements made within the spirit and principle of the present invention should be covered by the protection scope of the present invention.

Claims

1. A method for constructing an improved animal model of psoriasiform dermatitis, characterized in that, It includes the following steps: a) Through the animal selection module, select an animal strain sensitive to imiquimod and streptococcus, and screen suitable individuals based on genetic background, gender, age, and immune status; b) Through the inducer selection module, use imiquimod as the basic inducer, apply it to the back skin of the animal, and simultaneously add a streptococcus suspension locally for combined stimulation; c) Use a dressing material to cover the drug application area, seal it with plastic wrap or medical tape, and optimize the sealing time to 4 to 6 hours; d) Through the model evaluation module, evaluate the lesion type, distribution range, and pathological characteristics; e) Through the monitoring module, dynamically monitor the skin damage area, inflammatory factor levels, and immune cell activation; f) Through the data processing and analysis module, summarize and quantify the experimental data, and adjust the streptococcus concentration, sealing time, and imiquimod dose; g) Through the improvement module, design a dedicated dressing assistance device and optimize the process to improve the experimental efficiency.

2. The method for constructing an improved animal model of psoriasiform dermatitis according to claim 1, wherein, The dressing material is a double-layer medical-grade material, with a polymer adhesive layer on the inner layer and a flexible waterproof and breathable membrane on the outer layer, and is equipped with a protection device to prevent drug loss caused by animal licking and friction.

3. The method for constructing an animal model of psoriasiform dermatitis according to claim 1, wherein, The step of selecting a suitable animal strain and individual further includes selecting a suitable animal strain according to the genetic susceptibility and immune characteristics of psoriasis, and selecting appropriate age, gender, and weight factors according to the experimental needs; The step of selecting a suitable inducer further includes determining the most suitable inducer concentration, exposure time, frequency, and usage method according to the characteristics of different inducers and the response of the animal model; The monitoring module includes using techniques such as tissue sectioning, immunohistochemistry, and real-time PCR to detect changes in the cellular components, immune responses, and molecular markers of skin lesions, and evaluate the pathological characteristics of the model; The data processing and analysis module also includes extracting potential biomarkers of psoriasis through data mining techniques and evaluating the effect of drug treatment, providing a theoretical basis for clinical research and new drug screening.

4. The improved method for constructing a psoriasiform dermatitis animal model according to claim 1, wherein, The animal selection module: Select a mouse strain prone to skin inflammation, C57BL / 6 mice.

5. The improved method for constructing an animal model of psoriasiform dermatitis according to claim 1, characterized in that, The inducer selection module: Imiquimod (IMQ) induction: IMQ is a Toll-like receptor 7 / 8 agonist, which can activate the innate immune system to cause neutrophils and γδ T cells to produce inflammatory factors, inducing psoriasis-like skin lesions in mice.

6. The improved method for constructing an animal model of psoriasiform dermatitis according to claim 1, wherein The model evaluation module: Lesion scoring: Using the PASI scoring standard, score the degree of skin lesions in mice, including the measurement of skin erythema, desquamation, and thickening.

7. The improved method for constructing a psoriasiform dermatitis animal model according to claim 1, wherein, The monitoring module: Histopathological observation: Through pathological sectioning and HE staining, observe the pathological changes at the lesion site, such as epidermal hyperplasia, excessive proliferation and abnormal differentiation of keratinocytes, and immune cell infiltration; Inflammatory factor detection: Detect the expression levels of relevant inflammatory factors by ELISA method to evaluate the degree of inflammatory response of the model.

8. The improved method for constructing an animal model of psoriasiform dermatitis according to claim 1, characterized in that, The data processing and analysis module: 1) Data collection and collation; Experimental data recording: During the model construction process, record all experimental data in detail, including the body weight of mice, the degree of skin lesions, the results of histopathological observation, and the expression levels of relevant inflammatory factors; Data organization: Organize the collected experimental data to ensure data integrity and accuracy; this includes data cleaning, data coding, and data standardization steps; 2) Statistical analysis methods; Descriptive statistical analysis: Conduct descriptive statistical analysis on the experimental data, calculate statistics such as mean, standard deviation, maximum value, and minimum value to understand the overall distribution and characteristics of the data; Inferential statistical analysis: Through hypothesis testing, correlation analysis, and regression analysis methods, infer whether the differences between different experimental groups are statistically significant, as well as the correlations and causal relationships between variables; 3) Data processing software and tools; Data processing software: Use professional data processing software, including SPSS, SAS, and R language, for data cleaning, organization, and analysis; Graphical display tools: Use Excel and GraphPad Prism graphical display tools to present the experimental results in the form of charts; 4) Data quality control and verification; Data quality control: Pay attention to data quality control during the data processing process; this includes the accuracy of data input, the standardization of data processing, and the review steps of data results; Data verification: Verify the reliability and accuracy of the experimental results by comparing with other experimental data or published literature; if the experimental results do not match the expectations or there are outliers, further verification and analysis are required.

9. An improved system for constructing an animal model of psoriasiform dermatitis implementing the method for constructing an animal model of psoriasiform dermatitis according to any one of claims 1-8, characterized in that, The improved system for constructing a psoriasiform dermatitis animal model includes: An animal selection module for selecting appropriate animal strains and individuals; An inducer selection module for selecting appropriate inducers to stimulate the animal skin to produce psoriasiform lesions; A model evaluation module for evaluating the constructed psoriasiform dermatitis animal model; A monitoring module for monitoring the constructed psoriasiform dermatitis animal model; A data processing and analysis module for processing and analyzing the collected experimental data to extract useful information; An optimization module for optimizing the constructed model based on experimental results and feedback; an improvement module for improving the constructed model based on experimental results and feedback.

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