Use of alpha-linolenic acid in treatment of psoriasis by modulation of gamma delta T cells

Abstract

The invention provides application of alpha-linolenic acid to treatment of psoriasis by adjusting gamma delta T cells, and belongs to the technical field of biological medicine. The invention provides an application of alpha-linolenic acid (ALA) in preparation of a medicine for treating psoriasis, and after ALA intervention, the skin lesion severity of a psoriasis animal model is obviously reduced, the spleen index is obviously reduced, proinflammatory factors are down-regulated, and immunoregulatory factors are up-regulated. The ALA has a direct treatment effect on local skin lesions, is highly in line with the core requirements of'improving skin lesion appearance and relieving epidermal hyperplasia 'of psoriasis patients in clinic, improves the overall treatment effect through'local and whole body' dual anti-inflammatory effects, and is expected to be developed into an external or systemic medicine in the future to be used for treating mild to moderate psoriasis.

Description

Technical Field

[0001] This invention belongs to the field of biomedical technology, specifically relating to the use of α-linolenic acid in treating psoriasis by regulating γδT cells. Background Technology

[0002] Psoriasis is a persistent and recurring inflammatory skin disease affecting nearly 3% of the US population. Although patients' health and lifespan are impacted by psoriasis, there is currently no cure. Several treatment options exist to alleviate skin symptoms, with topical therapies, phototherapy, oral systemic therapies, and biological therapies being standard treatments for psoriasis. However, these treatments can cause various side effects, impacting patient satisfaction. Therefore, there is a growing need to understand the pathogenesis of psoriasis in order to identify novel molecular targets and develop new therapies to reduce side effects.

[0003] γδT cells are a non-traditional population of T lymphocytes that play an indispensable role in host defense, immune surveillance, and immune system homeostasis. They exhibit unique developmental, distribution, and functional patterns, rapidly responding to various attacks and contributing to a variety of diseases. Although γδT cells constitute only a small fraction of the total T cell population, emerging evidence suggests that aberrantly activated γδT cells may play a role in the pathogenesis of psoriasis. Dermal γδT cells are the main IL-17-producing cells in the skin and respond to IL-23 stimulation. Furthermore, γδT cells exhibit memory cell-like characteristics and mediate recurrent flare-ups of psoriatic inflammation, but their exact mechanisms of action in psoriasis remain unclear.

[0004] Alpha-linolenic acid (ALA) is an essential fatty acid that the human body cannot synthesize and must be obtained through dietary intake. As a precursor to two important long-chain omega-3 fatty acids (EPA, 20:3n-5 and DHA, 22:3n-6), ALA has been reported to have various nutritional, health-promoting, and pharmacological effects, primarily including cardiovascular protection, neuroprotection, anti-cancer, anti-osteoporosis, anti-inflammatory, and antioxidant effects. Currently, there are no studies investigating the association between ALA, γδT cells, and psoriasis. Summary of the Invention

[0005] This invention provides the use of α-linolenic acid (ALA) in treating psoriasis by regulating γδT cells. ALA provides a new target for the treatment of psoriasis, and is safe, effective, and inexpensive, with very broad application prospects.

[0006] This invention provides the application of α-linolenic acid in the preparation of medicaments for treating psoriasis.

[0007] In a preferred embodiment of the present invention, the therapeutic effect of the drug includes at least one of the following: (1) alleviating skin phenotype;

[0008] (2) Reduce systemic inflammatory response;

[0009] (3) Regulate the expression of serum inflammatory factors.

[0010] In a preferred embodiment of the present invention, the relief of skin phenotypes includes relieving erythema, scaling, and significant skin thickening caused by psoriasis.

[0011] In a preferred embodiment of the present invention, the regulation of serum inflammatory factor expression includes inhibiting TNFα expression in serum and increasing IFNγ content.

[0012] The present invention also provides a medicament for treating psoriasis, the active ingredient of which includes alpha-linolenic acid, and also includes pharmaceutically acceptable excipients.

[0013] In a preferred embodiment of the present invention, the working concentration of α-linolenic acid in the drug, calculated in mice, is 2 mL / kg / d.

[0014] The present invention also provides the use of α-linolenic acid in the preparation of drugs that regulate γδT cells and improve the pathological manifestations of psoriasis.

[0015] In a preferred embodiment of the present invention, the regulation of γδT cells includes significantly reducing the proportion of γδT cells in skin tissue and increasing the proportion of Treg cells in plasma.

[0016] This invention also provides the application of α-linolenic acid in the preparation of products that improve skin condition.

[0017] In a preferred embodiment of the present invention, the type of product includes pharmaceuticals, food, or cosmetics.

[0018] Beneficial Effects: This invention utilizes an IMQ-induced mouse model of psoriasis, in which typical psoriasis symptoms, including erythema, scaling, and significant skin thickening, are observed on the back. After ALA intervention, the severity of skin lesions is significantly reduced, suggesting that ALA can improve the external clinical manifestations of psoriasis. Furthermore, ALA treatment significantly reduces the spleen index in the model mice, indicating that ALA can alleviate systemic inflammatory responses. Compared to the model mice, ALA intervention significantly reduces TNF-α expression (downregulation of pro-inflammatory factors) and significantly increases IFN-γ content (upregulation of immunomodulatory factors). These embodiments of the invention demonstrate that ALA has a direct therapeutic effect on local skin lesions, highly aligning with the core clinical needs of psoriasis patients to "improve the appearance of skin lesions and reduce epidermal hyperplasia." Through its dual anti-inflammatory effects of "local + systemic," it enhances the overall therapeutic effect and holds promise for future development as a topical or systemic drug for the treatment of mild to moderate psoriasis. Attached Figure Description

[0019] Figure 1The figures show the results of targeted fatty acid metabolomics analysis in clinical psoriasis lesions. In the figures, A: total ion current chromatogram of mixed standard; B: QC overlapping chromatogram; C: orthogonal-partial least squares discriminant analysis plot (OPLS-DA); D: volcano plot; E: ROC curve; F: box plot; BC in each figure represents the healthy control group, and MOD represents the psoriasis patient group.

[0020] Figure 2 The figure shows the results of targeted fatty acid metabolomics analysis in psoriatic lesions induced by IMQ in mice. In the figure, A: total ion current chromatogram of mixed standard; B: QC overlapping chromatogram; C: orthogonal-partial least squares discriminant analysis plot (OPLS-DA); D: volcano plot; E: ROC curve; F: box plot; BC in each figure represents the healthy control group, and MOD represents the psoriasis patient group.

[0021] Figure 3 Figure 1 shows the results of ALA alleviating IMQ-induced psoriatic dermatitis. A: Representative images of skin lesions in each group of mice on day 46 of modeling (n=8); B: H&E staining of skin tissue from each group of mice; magnification, 200×; C: Expression levels of TNF-α, IFN-γ, IL-1β, IL-6, IL-10, and IL-17A in the serum of mice in different treatment groups, detected using the BDCBA mouse inflammation kit. Data are expressed as mean ± standard deviation; *P<0.05, **P<0.01.

[0022] Figure 4 The figures show the results of GO and KEGG enrichment analysis of DEGs among the groups. In the figures, A: PCA analysis between samples; B: statistical graph of differentially expressed genes among the groups; C: differentially expressed gene cluster analysis; D: statistical graph of the number of DEGs enriched in KEGG metabolites; E: the top 20 KEGG enriched pathways; F: relative mRNA expression levels.

[0023] Figure 5 The figure shows the effects of ALA on Treg cells, γδT cells, and related inflammatory factors in psoriatic mice. Figure A: Flow cytometry staining and statistical graph of Treg cells in whole blood PBMCs; B: Flow cytometry staining and statistical graph of T cells in skin tissue; C: Flow cytometry staining and statistical graph of γδT cells in skin tissue; D: ELISA detection of IFN-γ and IL-17A in plasma; *P<0.05, **P<0.01, ns: no statistical difference; CON / NS represents the blank control group; CON / ALA represents the ALA control group; IMQ / NS represents the model group; IMQ / ALA represents the ALA intervention group.

[0024] Figure 6This is a schematic diagram of the modeling process. In the diagram, CON / NS refers to the normal control group; CON / ALA refers to the normal control group treated with ALA intervention; IMQ / NS refers to the model group; and IMQ / ALA refers to the model group treated with ALA intervention. Detailed Implementation

[0025] This invention provides the application of α-linolenic acid in the preparation of medicaments for treating psoriasis.

[0026] In this invention, a psoriasis mouse model was induced by IMQ. Mice in the IMQ group exhibited typical erythema, scaling, and thickening of the skin on their backs. The application of ALA significantly alleviated the severity of psoriatic lesions. Subsequently, the spleen index, reflecting the severity of inflammation, was statistically analyzed, showing a decrease in the spleen index in ALA-treated mice (Figure 1). This preliminarily demonstrates that ALA has an inhibitory effect on IMQ-induced psoriatic inflammation, and that ALA can reduce the clinical manifestations and spleen index in psoriatic mice. Histopathological results showed that ALA treatment improved epidermal thickening in the skin tissue of the IMQ mouse model, thus ALA can improve the pathological manifestations of psoriatic mice. This invention also detected the expression levels of psoriasis-related inflammatory molecules in the serum of mice in different treatment groups. It was found that the serum TNFα content was significantly reduced and the IFNγ content was significantly increased in the ALA group, while the expression levels of IL-1β, IL-6, IL-10, and IL-17A showed no significant difference, indicating that ALA can reduce the levels of psoriasis-related inflammatory factors in the serum of psoriatic mice.

[0027] The present invention also provides a medicament for treating psoriasis, the active ingredient of which includes alpha-linolenic acid, and also includes pharmaceutically acceptable excipients.

[0028] The drug described in this invention can be an oral medication or a topical medication.

[0029] In a preferred embodiment of the present invention, the working concentration of α-linolenic acid in the drug, calculated in mice, is 2 mL / kg / d.

[0030] The present invention also provides the use of α-linolenic acid in the preparation of drugs that regulate γδT cells and improve the pathological manifestations of psoriasis.

[0031] The regulation of γδT cells described in this invention includes significantly reducing the proportion of γδT cells in skin tissue and increasing the proportion of Treg cells in plasma.

[0032] This invention also provides the application of α-linolenic acid in the preparation of products that improve skin condition.

[0033] The types of products described in this invention include pharmaceuticals, food, or cosmetics.

[0034] To further illustrate the present invention, the following detailed description, in conjunction with embodiments, illustrates the use of α-linolenic acid provided by the present invention in treating psoriasis by regulating γδT cells, but these descriptions should not be construed as limiting the scope of protection of the present invention.

[0035] Example 1

[0036] 1. Experimental Design

[0037] First, we verified that ALA was deficient in clinical psoriasis patients and IMQ-induced psoriasis mice, clarifying its association with the disease. Second, through animal experiments, we determined that ALA can increase the production of anti-inflammatory cytokines and reduce the expression of pro-inflammatory cytokines during the process of improving psoriasis. By detecting relevant inflammatory factors in blood and skin tissues, such as TNF-α, IFN-γ, IL-17A, IL-1β, IL-6, and IL-10, and transcription factors such as Foxp3, RORγt, Klrg1, and ST2, we preliminarily clarified the mechanism by which ALA improves psoriasis by regulating the Treg / γδT cell axis.

[0038] (1) Animal experiment preparation:

[0039] 1) Six- to eight-week-old C57BL / 6 mice were housed in a constant temperature and humidity environment at the Experimental Animal Center of Ningxia Medical University. A 12h:12h light / dark cycle was maintained daily, with routine water and feeding. Bedding was changed every three days to maintain cleanliness and dryness. Mice were randomly divided into four groups of five mice each. After two days of environmental acclimatization, subsequent experiments were conducted. The skin on the backs of the mice was shaved using a shaver, with a shaved area of ​​approximately 2×3 cm. 2 .

[0040] 2) Preparation of the medications and gavage solution:

[0041] ALA gavage solution: ALA solution was prepared at a rate of 2 mL / kg / d based on the mouse's body weight; the control group was gavaged with an equal volume of DMSO solution; gavage was continued for 46 days.

[0042] (2) Group processing (e.g.) Figure 6 ):

[0043] 1) CON / NS group: Adaptive feeding for one week, hair removal cream was applied to the back of the mice 3 days in advance, and petroleum jelly cream was applied from day 0 for 8 consecutive days, stopped for 30 days, applied for another 8 days, and placebo was administered by gavage for 46 consecutive days.

[0044] 2) IMQ / NS group: After one week of acclimatization, the mice were locally dehaired on their backs with hair removal cream 3 days in advance. Starting from day 0, approximately 62.5 mg of imiquimod cream was applied to establish a psoriasis model for 8 consecutive days, followed by a 30-day break, and then another 8 days of application. A placebo was administered by gavage for 46 consecutive days.

[0045] 3) CON / ALA group: After one week of acclimatization, the mice were locally dehaired on their backs with hair removal cream 3 days in advance. Vaseline cream was applied from day 0 for 8 consecutive days, followed by another 8 days of application. ALA solution was then administered by gavage for 46 consecutive days.

[0046] 4) IMQ / ALA group: After one week of acclimatization, the mice were locally dehaired on their backs with hair removal cream 3 days in advance. Starting from day 0, approximately 62.5 mg of imiquimod cream was applied to establish a psoriasis model for 8 consecutive days, followed by a 30-day break, and then another 8 days of application. ALA solution was administered by gavage for 46 consecutive days.

[0047] Note: Mice weight and PASI scores were recorded daily during the experiment. On day 46, mice were sacrificed and their blood, skin, and spleen were collected for further analysis.

[0048] (3) PASI scoring criteria

[0049] The PASI (Psoriasis Area and Severity Index) score evaluates the clinical symptoms of psoriasis based on the degree of erythema, scaling, and thickening at the lesion site, and is used to determine the severity of the IMQ-induced psoriasis mouse model. Starting from the day of model establishment, two dermatology students familiar with the PASI evaluation criteria scored the mice daily before each operation, without knowing the treatments for each group. Each PASI feature was scored out of 5 points (0-4 points). The scores from the two students were recorded according to the standards in Table 1-3, and the scores of the three features were summed to obtain the total PASI score. The average score from the two students was taken as the final score.

[0050] (4) HE staining and immunohistochemical staining of skin tissue:

[0051] 1) HE staining:

[0052] Mouse skin tissue from the modeling area was fixed in 4% paraformaldehyde solution, dehydrated by low to high concentrations of alcohol, embedded in paraffin, and cut into 5-8 μm thin sections using a microtome. After drying, sections were prepared. Before staining, dewaxing was performed. Cell nuclei and ribosomes were stained blue-purple with the basic dye hematoxylin (H), and cytoplasmic components were stained red with the acidic dye eosin (E). After mounting, histopathological changes were observed under a light microscope. Magnification was adjusted to 4x, 10x, and 20x to observe and record the degree of epidermal keratinization and hyperplasia, dermal capillary dilation, and the number of leukocytes infiltrating the skin tissue.

[0053] 2) Immunohistochemical staining:

[0054] ① Dewaxing and hydration: The skin tissue sections were baked in a 55℃ oven for 15 min; then soaked in xylene for 10 min, and repeated twice; then soaked in anhydrous ethanol, 95% ethanol and 70% ethanol for 5 min respectively; finally washed with PBS for 5 min, and repeated twice.

[0055] ② Antigen retrieval: Place the slide in sodium citrate / EDTA antigen retrieval solution and microwave boil for 10 minutes; the purpose of this step is to fully expose the antigen epitopes.

[0056] ③ Washing: Place the slides in PBS and wash for 5 minutes, repeat 3 times;

[0057] ④ Blocking: Use 5% goat serum for blocking at room temperature for 1 hour to reduce nonspecific background;

[0058] ⑤ Primary antibody incubation: After removing excess blocking solution, add 50 μl of primary antibody solution and incubate at room temperature for 1 h or at 4°C overnight;

[0059] ⑥ Secondary antibody incubation: After repeating the washing step in ③, add 50 μl of secondary antibody solution and incubate at room temperature for 1 h;

[0060] ⑦ DAB staining: Incubate with DAB staining solution for 5-10 minutes and observe the degree of staining under a microscope;

[0061] ⑧ Hematoxylin counterstaining: Repeat the washing step in ③ and counterstain with hematoxylin for 2 minutes; after dehydration and clearing, mount and examine under a microscope.

[0062] (5) Real-time quantitative PCR reaction: qRT-PCR: Primers were designed and synthesized using Prime5. After total RNA was extracted from the sample using Trizol, cDNA was synthesized by reverse transcription. Using cDNA as a template, SYBR Green PCR was used to detect the transcriptional expression level of the above pathway molecules. The operation steps were followed according to the kit instructions.

[0063] (6) Western blot: Detection of GPR120 and transcription factors: Foxp3, RORγt, Klrg1, ST2 protein expression. Procedure: Extract total protein strictly according to the whole protein extraction kit, and measure protein concentration using the BCA method. Next, load an equal volume of the extracted protein into a 0.5 ml EP tube, add 10 μl of 5×SDS loading buffer, and mix well. Incubate the mixture at 95℃ for 5 min. Aliquot the sample in 20 μl units and store at -80℃. 8% SDS-PAGE gel electrophoresis: After loading, perform electrophoresis at 80V until the sample exceeds the stacking gel, then switch to 120V and continue electrophoresis until the sample reaches the bottom of the separating gel, then terminate the electrophoresis. One gel is used for Coomassie brilliant blue staining, and the other gel is used for membrane transfer. Semi-dry transfer: Immerse the PVDF membrane completely in methanol for 1 min; then transfer from bottom to top using thick filter paper-gel-membrane-thick filter paper. Transfer conditions: 230 mA, transfer for 40 min. Blocking: Prepare TBST blocking buffer with 5% skim milk powder and block at room temperature for 1 h. Primary antibody incubation: Add 1:500 diluted primary antibody and internal control Rabbit anti-mouse Actin (Beijing Zhongshan Jinqiao Biotechnology Co., Ltd.), and incubate overnight at 4℃. Wash 5 times with TBST buffer. Secondary antibody incubation: Add 1:2000 diluted secondary antibody (Beijing Zhongshan Jinqiao Biotechnology Co., Ltd.), and incubate at room temperature for 1 h. Wash 10 times with TBST buffer. Development: Add a mixture of enhanced chemiluminescence (ECL) solution A and solution B, and immediately acquire images using a ChemiDoc XRS+ chemiluminescence imaging system (Bio-Rad Laboratories, USA), and perform quantitative analysis using ImageJ.

[0064] (7) Flow cytometry multiplex protein quantification (CBA) for detecting inflammatory factor levels in mouse skin and serum: Detection steps: Preparation of various cytokine standards in the kit: Open a tube of lyophilized standard, transfer the microspheres to a 15ml centrifuge tube, and label this tube as the highest concentration standard. Gently resuspend the standard with 2ml of experimental diluent (do not shake violently), and equilibrate at room temperature for at least 15min. Subsequently, take out 9 12×75mm flow cytometry sample loading tubes and label them with serial dilution factors of 1:2, 1:4, 1:8, 1:16, 1:32, 1:64, 1:128, and 1:256, respectively. Add 300μl of experimental diluent to each tube. Note: Mix gently during dilution; do not shake violently. Mixing the appropriate cytokine-capturing microspheres: Determine the number of experimental tubes (including standard and control tubes); before mixing, prevent microsphere deposition by vortexing thoroughly for 3-5 seconds; to ensure each experimental tube contains 7 types of microspheres, use 10 μl of each type of capture microsphere; then vortex thoroughly to mix. Centrifuge the mixed capture microspheres at 200×g for 5 min; carefully aspirate the supernatant, add an equal volume of serum enhancement buffer, vortex thoroughly to mix, and incubate at room temperature in the dark for 30 min. Note: The amount of capture microspheres must be accurately calculated, as they cannot be reused once taken out. Dilution of the test samples: The detection range of CBA is 20-5000 pg / ml. To ensure reliable experimental results, serum samples in this project are diluted 1:4, and liver samples need to be diluted 1:100. Setting up the analytical BDFACSCelesta flow cytometer using the prepared microspheres: This was completed by a designated person from the applicant's institution with the assistance of the BD engineer. Flow Cytometry Testing: Vortex the pre-mixed capture microspheres, adding 50 μl to each experimental tube; add 50 μl of the serially diluted standards to each standard tube. Then, add 50 μl of the sample to each sample tube. Add 50 μl of mouse cytokine PE assay reagent to all experimental tubes, incubate at room temperature for 3 hours, add 1 ml of wash buffer to each tube, centrifuge at 200×g for 5 minutes, carefully aspirate the supernatant, add 300 μl of wash buffer to each tube, and resuspend the microspheres. Note: Samples must be tested on the same day. First, test the standards, in ascending order of concentration; then, test the diluted samples in descending order of concentration. Data Analysis: Data analysis was performed using the licensed FCAPArray software from the flow cytometry laboratory of the applicant's institution.

[0065] (8) Flow cytometry: The proportions of CD3, γδT cells and Treg cells in the skin tissue of IMQ-induced psoriasis mice were analyzed by flow cytometry. First, 2 × 10⁶ cells were collected in 1.5 ml EP tubes, resulting in three types of tubes: isotype control tubes, monostained positive control tubes, and experimental tubes. Cells were centrifuged at 400 × g for 5 min at 4°C, and the supernatant was discarded. Cells were resuspended in 1 ml PBS wash buffer and centrifuged at 400 × g for 5 min at 4°C, and the supernatant was discarded. This washing was repeated once. Cells were resuspended in 50 μl PBS wash buffer. 2 μl of the corresponding antibody was added to each of the isotype control tubes, monostained positive control tubes, and experimental tubes, and the cells were incubated at 4°C for 20 min in the dark. Cells were washed twice with PBS. Cells were then resuspended in 500 μl PBS fixative containing 1% paraformaldehyde for flow cytometry analysis (BDFACSCelesta flow cytometer). Cell populations were selected using FSC and SSC. The isotype control tubes and monostained positive control tubes were used to divide the cells, and the compensation values ​​between different channels were adjusted to analyze the proportion of monocytes in the experimental tubes.

[0066] (9) Immunoprecipitation (IP): Immunoprecipitation was used to analyze downstream pathway molecules of GPR120 and identify interacting proteins. Experimental Procedure: a. After completing the cell experiments, harvest cells (6-well plates), add 200 μl of IP lysis buffer (containing protease inhibitor) to each well, lyse on ice or at 4°C for 30 min, centrifuge at 12,000 g for 30 min, and collect the supernatant; b. Take a small amount of lysis buffer for Western blot analysis (as input), add 1 μl of cAMP pathway target molecule antibody and 10-50 μl of protein A / G-beads to the remaining lysis buffer, and incubate overnight at 4°C with gentle shaking; c. After immunoprecipitation, centrifuge at 3,000 g for 5 min at 4°C, centrifuge the protein-protein interaction complex bound to protein A / G-beads to the bottom of the tube, carefully aspirate the supernatant, and wash 3-4 times with 1 ml of lysis buffer; d. Finally, add 15 μl of 2×SDS loading buffer and boil for 10 min; e. Perform SDS-PAGE and Western blot analysis. The Western blot procedure is as described above.

[0067] (10) Immunohistochemistry and immunofluorescence in situ: Analysis of changes in the Treg / γδT cell axis

[0068] Immunohistochemical methods: Granulocytes were labeled with FSHR as the primary antibody and incubated with HRP-labeled secondary antibody (Beijing Zhongshan Jinqiao Company) for 30 min, followed by DAB substrate colorimetric reaction. Immunofluorescence methods: Nuclei were stained with DAPI using fluorescently labeled FSHR and detected by fluorescence microscopy.

[0069] 2. Experimental Results

[0070] (1) It is clear that psoriasis is caused by ALA deficiency.

[0071] Targeted fatty acid metabolomics analysis was performed on skin lesions from clinical psoriasis and mouse psoriasis models. The stability of the method was evaluated by calculating the relative standard deviation (RSD) of all quality control (QC) parameters using the peak area of ​​the standards. The total ions chromatogram (TIC) and QC overlap chromatograms showed that the stability of each substance was less than 15%, indicating that the method is stable and reliable and suitable for sample detection. Figure 1 AB in the middle Figure 2 (AB).

[0072] Orthogonal Projections to Latent Structures Discriminant Analysis (OPLS-DA) can effectively reduce model complexity and enhance its interpretability without compromising predictive power, thus maximizing the observation of inter-group differences. The results show that the samples from different groups are far apart, suggesting significant differences in metabolites among them. Figure 1 C in the middle Figure 2 Specifically, compared with the healthy control group, the skin lesions of clinical patients showed a significant reduction in 18 fatty acids, including C16:0, C118:0, C18:1N12, C18:1N9C, C18:2N6, C22:6N3, C18:3N6, C20:1, C18:3N3, C22:5N3, C18:1N7, C17:0, C15:0, C18:1N9T, C19:1N9T, C20:0, C20:2, and C22:4 (P<0.05). Figure 1 (D). The levels of C16:1T, C17:1T, C18:3N3, and C19:1N12T in the psoriasis mouse model were significantly lower than those in the model group (P < 0.05). Figure 2 (D). The area under the ROC curve (AUC) was further used to evaluate the sensitivity and specificity of the biomarker in predicting the event; the closer the AUC value is to 1, the higher the predictive accuracy. The results suggest that ALA may serve as a potential biomarker for psoriasis. Figure 1 China E and Figure 2 (E). Compared with the control group, C18:3N3 (ALA) was significantly lower in the skin lesions of psoriasis patients (P < 0.05). Figure 1 The levels were significantly reduced in both the IF and mouse models (P < 0.05). Figure 2 (F).

[0073] (2) ALA reduced the severity of IMQ-induced chronic relapsing psoriasis in a mouse model.

[0074] 1) ALA reduced the clinical manifestations and spleen index in psoriatic mice.

[0075] like Figure 3 As shown in Figure A: Mice in the IMQ group exhibited typical erythema, scaling, and thickening of the skin on their backs, while the application of ALA significantly alleviated the severity of psoriatic lesions. Next, the spleen index (spleen weight to body weight ratio) of mice in different treatment groups was calculated. The results showed that the spleen index was reduced in mice treated with ALA (Figure A). Figure 3 (A). This initially reflects the inhibitory effect of ALA on IMQ-induced psoriatic inflammation.

[0076] 2): ALA improves the pathological manifestations of psoriatic mice.

[0077] Histopathological results such as Figure 3 As shown in Figure B, ALA treatment improved epidermal thickening in the skin tissue of the IMQ mouse model.

[0078] 3): ALA reduces the level of psoriasis-related inflammatory factors in the serum of psoriasis-affected mice.

[0079] The expression levels of psoriasis-related inflammatory molecules in the serum of mice in different treatment groups were detected. Serum samples from each group were collected and analyzed using the CBA inflammatory factor kit. The results showed that the ALA group had significantly decreased TNFα levels and significantly increased IFNγ levels, while the expression levels of IL-1β, IL-6, IL-10, and IL-17A showed no significant difference. Figure 3 (C)

[0080] (3) The role of ALA in psoriasis: targeting energy metabolism

[0081] To determine the primary target of ALA in alleviating IMQ-induced psoriatic dermatitis, we performed RNA-seq transcriptomic analysis on skin tissues from CON / NS, CON / ALA, IMQ / NS, and IMQ / ALA mice. Figure 4 Compared with the CON / NS control group, the IMQ / NS group had 2026 significantly upregulated genes and 1020 significantly downregulated genes; compared with the IMQ / ALA group, the IMQ / ALA group had 935 significantly upregulated genes and 1529 significantly downregulated genes. Figure 4 (C). Functional annotation analysis of pathways according to the Kyoto Encyclopedia of Genetics and Genomes (KEGG) indicates that a large number of DEGs are clustered in metabolic categories (C). Figure 4 (Middle D). Bubble chart based on the top 20 enriched pathways ( Figure 4The study further emphasized the role of ALA treatment in energy metabolism processes such as ATP biosynthesis, aerobic respiration, electron transport chain, and nucleotide triphosphate biosynthesis. Simultaneously, it downregulated the expression level of Trdc transcripts, further validating that ALA may act on energy metabolism-related pathways to regulate the frequency and number of γδT cells and improve the inflammatory response. Flow cytometry results further confirmed that Treg cells significantly increased after ALA intervention (…). Figure 5 In the model mouse skin, γδT cells were significantly downregulated (A), while γδT cells were significantly downregulated (A). Figure 5 (Brain BC), and may also affect the secretion of IL-17A and IFN-γ. Figure 5 (D).

[0082] Although the above embodiments have provided a detailed description of the present invention, they are only some embodiments of the present invention, and not all embodiments. People can obtain other embodiments based on these embodiments without creative effort, and these embodiments all fall within the protection scope of the present invention.

Claims

1. Application of α-linolenic acid in the preparation of drugs for treating psoriasis.

2. The application according to claim 1, characterized in that, The therapeutic effects of the drug include at least one of the following: (1) alleviating skin phenotype; (2) Reduce systemic inflammatory response; (3) Regulate the expression of serum inflammatory factors.

3. The application according to claim 2, characterized in that, The relief of skin phenotypes includes relief of erythema, scaling, and significant skin thickening caused by psoriasis.

4. The application according to claim 2, characterized in that, The regulation of serum inflammatory factor expression includes inhibiting TNFα expression in serum and increasing IFNγ levels.

5. A drug for treating psoriasis, characterized in that, The active ingredients include alpha-linolenic acid, as well as pharmaceutically acceptable excipients.

6. The drug according to claim 5, characterized in that, The working concentration of α-linolenic acid in the drug, calculated for mice, is 2 mL / kg / d.

7. Application of α-linolenic acid in the preparation of drugs that regulate γδT cells and improve the pathological manifestations of psoriasis.

8. The application according to claim 7, characterized in that, The regulation of γδT cells includes significantly reducing the proportion of γδT cells in skin tissue and increasing the proportion of Treg cells in plasma.

9. Application of α-linolenic acid in the preparation of products that improve skin condition.

10. The application according to claim 9, characterized in that, The types of products mentioned include pharmaceuticals, food, or cosmetics.

Images