A method for extracting medicinal components from tung oil and use thereof in treating psoriasis

Abstract

A method for extracting tung oil as a medicinal substance comprises the following steps: Step 1: Selecting mature tung seeds, removing the outer shell, leaving the oil-containing embryo, drying and pulverizing it into tung seed kernel powder; Step 2: Mixing and stirring the tung seed kernel powder with ethanol to obtain crude tung oil; Step 3: Allowing the crude tung oil to stand at low temperature to remove the aqueous phase, gum, and pigments, obtaining refined tung oil; Step 4: Detoxifying the refined tung oil using a mild saponification process; Step 5: Re-extracting the detoxified tung oil with ethyl acetate to obtain a tung oil extract enriched with active ingredients. This invention also proposes a psoriasis treatment drug based on natural tung oil extract, aiming to overcome the limitations of existing therapies in terms of cost, administration method, side effects, and insufficient coverage of complex pathological mechanisms. It achieves comprehensive treatment of psoriasis through multi-dimensional regulation of inflammation, cell proliferation, and metabolism using a tung oil extraction method and its application in treating psoriasis.

Description

Technical Field

[0001] This invention relates to the field of drug preparation and application, specifically to a method for extracting tung oil and its application in treating psoriasis. Background Technology

[0002] Psoriasis is a common, chronic, relapsing, inflammatory skin disease with a complex pathological mechanism involving multiple levels, including abnormal activation of the immune system, excessive proliferation and disordered differentiation of keratinocytes, and metabolic imbalances. Mild to moderate plaque psoriasis is the most common clinical subtype of psoriasis, belonging to chronic immune-mediated inflammatory skin diseases. It typically presents as well-defined, red, infiltrated plaques covered with multiple layers of silvery-white scales, commonly occurring on the scalp, elbows, knees, and other extensor surfaces of the limbs. Its core pathological features include excessive epidermal proliferation, parakeratosis, and infiltration of immune cells in the dermis; patients with this type of psoriasis have lesions covering <10% of the body surface area, and a Psoriasis Area and Severity Index (PASI) <10. Various treatments exist for psoriasis, but many methods have limited efficacy, significant side effects, or a high relapse rate. Therefore, developing novel, highly effective, and safe therapeutic drugs remains a research hotspot and challenge in this field.

[0003] In existing technologies, the core pathological mechanisms of psoriasis have been extensively studied. Among these, the overactivation of the IL-17 / IL-23 inflammatory axis is considered a key driver of the inflammatory response in psoriasis. This axis involves various cytokines, such as IL-17 and IL-23, which can activate downstream signaling pathways, leading to the release of large amounts of pro-inflammatory factors (such as IL-6, IL-1β, CXCL1, and CCL20), thereby recruiting and activating immune cells, forming persistent inflammatory infiltration, and stimulating abnormal activation of keratinocytes. Clinically, monoclonal antibodies targeting IL-17 or IL-23 (such as secukinumab and gusejinumab) have become important biologic therapies, showing good efficacy. However, these biologics are expensive, require injection, and have potential side effects such as increased risk of infection, limiting their widespread application. Furthermore, not all patients respond to existing biologics; some patients experience secondary failure. Therefore, finding natural products or small molecule drugs that can effectively target and inhibit the IL-17 / IL-23 inflammatory axis, but are less expensive, easier to administer, and safer, has important clinical significance and application value.

[0004] Besides the inflammatory response, abnormal proliferation and disordered differentiation of keratinocytes are another core pathological feature of psoriasis. In psoriatic lesions, the cell cycle of keratinocytes is significantly shortened, their proliferation rate is abnormally accelerated, and their differentiation process is inhibited, leading to typical pathological changes in the epidermis such as hyperkeratosis, acanthosis, and thinning or disappearance of the granular layer. This process is precisely regulated by multiple intracellular signaling pathways. Existing research shows that the Wnt signaling pathway, Notch signaling pathway, PI3K-Akt signaling pathway, and p53 signaling pathway all play key roles in the proliferation, differentiation, and apoptosis of keratinocytes. In psoriasis, these pathways often become dysfunctional; for example, overactivation of the Wnt pathway may promote cell proliferation, while inactivation of the p53 pathway may weaken the ability to clear abnormally proliferating cells. Currently, some topical drugs (such as vitamin D3 derivatives and retinoids) and systemic drugs (such as methotrexate) partially inhibit keratinocyte proliferation by affecting these pathways, but they often have side effects such as skin irritation, hepatotoxicity, or teratogenicity, and their long-term efficacy and safety still need further evaluation. Therefore, developing drugs that can synergistically regulate keratinocyte homeostasis through multiple pathways and targets, thereby more safely and effectively improving the pathological characteristics of psoriasis skin, is an important direction in current drug development.

[0005] In recent years, increasing research has focused on the role of metabolic disorders in the development and progression of psoriasis. Psoriasis patients often exhibit lipid metabolism abnormalities, including fatty acid metabolism disorders and increased cholesterol synthesis. These metabolic disorders are not merely accompaniments to the disease but can actively exacerbate inflammatory responses. For example, abnormal lipid metabolism can produce pro-inflammatory lipid mediators such as prostaglandins and leukotrienes, which can further activate immune cells, creating a vicious cycle of metabolism and inflammation. Furthermore, metabolic reprogramming may also affect the energy supply and biosynthesis of keratinocytes, thereby promoting their excessive proliferation. Currently, there are relatively few interventions specifically targeting metabolic disorders in existing psoriasis treatments. While lifestyle modifications (such as dietary control) may benefit some patients, there is a lack of specific drug targets. Therefore, addressing the metabolic level by identifying natural products that can correct psoriasis-related metabolic disorders, thereby indirectly or directly alleviating inflammation and cell proliferation, provides new insights and potential breakthroughs for psoriasis treatment.

[0006] The shortcomings of existing technologies can be summarized as follows: 1. Existing psoriasis treatment drugs (mild to moderate plaque psoriasis), especially biologics targeting the IL-17 / IL-23 inflammatory axis, are expensive, require injection, have a high potential risk of infection, and have poor response or secondary failure in some patients. 2. Existing drugs that treat mild to moderate plaque psoriasis by regulating the proliferation and differentiation of keratinocytes (such as vitamin D3 derivatives, retinoids, methotrexate, etc.) generally have side effects such as skin irritation, hepatotoxicity or teratogenicity, and their long-term efficacy and safety need to be improved. 3. Current treatment options for mild to moderate plaque psoriasis lack interventions specifically targeting metabolic disorders such as lipid metabolism that accompany the disease, and there is a lack of effective drug targets to break the vicious cycle of "metabolism-inflammation". 4. Existing natural products used in the treatment of psoriasis suffer from unclear mechanisms of action, unknown active ingredients, and insufficient clinical evidence. Many studies on natural extracts remain at the level of phenomenological observation, lacking a precise elucidation of their molecular mechanisms regulating the core inflammatory pathways of psoriasis, making it difficult to develop them into standardized and highly effective drugs. The seeds of the tung tree (Vernicia fordii), also known as tung seed or oil tung seed, are the core utilized part of plants in the genus Vernicia of the family Euphorbiaceae. They are highly toxic and have an extremely high oil content, making them an important raw material for industrial and traditional medicinal use. Summary of the Invention

[0007] I. Technical problems to be solved This invention addresses the shortcomings of existing technologies by proposing a novel, highly effective, and safe treatment for mild to moderate plaque psoriasis. Based on natural tung oil extract, this drug aims to overcome the limitations of existing therapies in terms of cost, administration method, side effects, and insufficient coverage of complex pathological mechanisms. It achieves comprehensive treatment of psoriasis through multi-dimensional regulation of inflammation, cell proliferation, and metabolism via a tung oil extraction method and its application in treating psoriasis.

[0008] II. Specific Technical Solutions A method for extracting tung oil, comprising the following steps: Step 1: Select mature tung seeds, remove the outer shell, leaving the oil-containing embryo, dry and then grind them into tung seed powder; Step 2: Mix and stir the tung seed kernel powder with ethanol to obtain crude tung seed oil; Step 3: Let the crude tung oil stand at a low temperature to remove the aqueous phase, gums and pigments, and obtain refined tung oil; Step 4: Use a mild saponification process to detoxify the refined tung oil; Step 5: The attenuated tung oil is re-extracted using ethyl acetate to obtain a tung oil extract enriched with active ingredients.

[0009] As a preferred option, a sixth step is also included, in which a water bath depressurization solvent and α-tocopherol are added to protect the active ingredients and prevent oxidative degradation.

[0010] As a preferred option, step one specifically involves drying the oil-containing embryo at 35-45℃ for 4–6 hours until the water content is <8% to reduce rancidity, and then pulverizing it into a 40–60 mesh powder to improve extraction efficiency.

[0011] As a preferred embodiment, step two specifically involves mixing tung seed powder and ethanol at a material-to-liquid ratio of 1:6 (g / mL), stirring and extracting in a water bath at 40-60℃ for 2-4 hours, repeating the extraction 2-3 times, combining the extracts, filtering to remove residues, and obtaining crude tung seed oil containing fatty acids, active ingredients, and impurities.

[0012] As a preferred option: Step 3 specifically involves letting the crude tung oil stand for 24 hours to allow suspended impurities to settle naturally, taking the supernatant, then adding 2%-3% hot water of the oil volume, stirring, letting it stand to separate into layers, discarding the aqueous phase, removing the gum, and finally adding 1%-2% activated carbon of the oil volume, stirring at 45-55℃, filtering to remove pigments and residual impurities, and obtaining a clarified refined oil.

[0013] As a preferred embodiment, step four specifically involves contacting the oil phase with NaOH at 40-60℃ to convert the irritating components into soaps. After saponification, the soaps are washed with an equal volume of warm water to allow them to enter the aqueous phase. The washing process is repeated until the aqueous phase is clear to ensure that the irritating components are completely removed. Subsequently, HPLC fingerprinting is used to monitor the process and ensure that the active ingredients are retained during the detoxification process.

[0014] As a preferred option, the obtained tung oil is used as a topical skin medication.

[0015] As a preferred option, the obtained tung oil is used to treat psoriasis.

[0016] The beneficial effects of this invention are as follows: The drug based on tung oil extraction provided by this invention exhibits significant therapeutic effects in animal models of psoriasis, effectively reducing erythema, scaling, and thickening of skin lesions. Histological examination also confirms that it can reduce stratum corneum thickening and inflammatory cell infiltration. Its beneficial effects stem from a clear multi-target mechanism of action. RNA-seq transcriptome analysis confirms that the extract can significantly downregulate the gene expression of pro-inflammatory cytokines (such as IL-17A) in key inflammatory pathways of psoriasis (such as the IL-17 signaling pathway), thereby inhibiting excessive inflammatory responses. Simultaneously, its action involves intervention in the regulation of cell signaling complexes and the expression of inflammation-related genes. Compared with existing biological agents, this invention is derived from natural plants, is expected to have fewer side effects, and is a topical preparation, making it convenient to use. Compared with existing topical chemical drugs, it exerts its effects by regulating the core pathological pathways of psoriasis, with a more clearly defined and cutting-edge mechanism. Therefore, this invention provides a potential drug option for the treatment of mild to moderate plaque psoriasis that combines effectiveness, safety, and innovation.

[0017] In the extraction method, a mixture of tung seed powder and ethanol is used to improve extraction efficiency and maximize the dissolution of fat-soluble active substances. A mild saponification process is used for detoxification, which promotes ester bond hydrolysis and avoids the degradation of heat-sensitive active ingredients (such as polyphenols). A trace amount of alkali concentration ensures that the reaction is limited to surface esters, without destroying the main structure of triglycerides and preserving the oil phase skeleton. After saponification, the mixture is immediately washed with an equal volume of warm water to allow the soap to enter the aqueous phase. The washing is repeated until the aqueous phase is clear to ensure thorough removal of irritating components. Attached Figure Description

[0018] Figure 1 Images of a mouse psoriasis model group are shown in this embodiment of the invention.

[0019] Figure 2 The image shown is from the mouse psoriasis treatment group in an embodiment of the present invention.

[0020] Figure 3 This is a schematic diagram of erythema scores for the model group and the treatment group in an embodiment of the present invention.

[0021] Figure 4 This is a schematic diagram of the scaling scores for the model group and the treatment group in an embodiment of the present invention.

[0022] Figure 5 This is a schematic diagram of the infiltration thickness scoring for the model group and the treatment group in an embodiment of the present invention.

[0023] Figure 6 This is a schematic diagram of the PASI scores for the model group and the treatment group in an embodiment of the present invention.

[0024] Figure 7 This is a pathological image of HE staining of the back skin tissue of mice in the control group and the model group in an embodiment of the present invention.

[0025] Figure 8 This is a pathological image of HE staining of the back skin tissue of mice in the model group and the treatment group, as shown in this embodiment of the invention.

[0026] Figure 9 This is a schematic diagram of the functional enrichment analysis of the differentially expressed gene KEGG in an embodiment of the present invention.

[0027] Figure 10 This is a GSEA enrichment analysis diagram of the IL-17 signaling pathway in an embodiment of the present invention.

[0028] Figure 11 This is a heatmap of differentially expressed genes in the IL-17 signaling pathway between the treatment group and the model group in an embodiment of the present invention. Detailed Implementation

[0029] The preferred embodiments of the present invention will now be described in detail with reference to the accompanying drawings, so that the advantages and features of the present invention can be more easily understood by those skilled in the art, thereby providing a clearer and more explicit definition of the scope of protection of the present invention.

[0030] The specific steps for extracting tung oil are as follows: S1. Select mature tung oil tree (Vernicia fordii) seeds, ensuring they are of pure variety, with plump kernels, a glossy appearance, free from mold and insect infestation, and meet the standards for active ingredient content, to facilitate better preservation of the active ingredients in subsequent extraction processes. Remove the hard shell from the selected tung oil tree seeds, leaving the oil-containing embryo. Dry at 40℃ for 4–6 hours until the moisture content is <8% to reduce rancidity. After drying, pulverize to an appropriate particle size (40–60 mesh) to improve extraction efficiency.

[0031] S2. Using edible ethanol as the extraction solvent, mix tung seed powder with ethanol at a material-to-liquid ratio of 1:6 (g / mL), and extract by stirring in a water bath at 40-60℃ for 2-4 hours. Repeat the extraction 2-3 times, and combine the extracts. Filter to remove residues to obtain crude tung seed oil, containing fatty acids, active ingredients, and impurities. This step aims to maximize the dissolution of fat-soluble active substances.

[0032] S3. Crude oil often contains suspended solids, plant pigments, waxes, and primary free fatty acids, which can cause irritation and oxidation. Further extraction is required. Let the crude oil stand for 24 hours to allow suspended impurities to settle naturally. Take the supernatant, then add a small amount of 50°C hot water (2%-3% of the oil volume), stir, and let it stand to separate into layers. Discard the aqueous phase and remove the gum. Finally, add activated carbon (1%-2% of the oil volume), stir at 50°C for 30 minutes, and filter to remove pigments and residual impurities to obtain clarified refined oil.

[0033] S4. The main irritant components in tung oil include phorbol esters (containing ester bonds) and free fatty acids. Phraphorbol esters can undergo ester bond hydrolysis under alkaline conditions, generating low-toxicity phorbol and fatty acids; free fatty acids react with alkali to form soaps, which can be removed by washing with water. Based on this, this study employed a mild saponification process for detoxification. At 40-60℃, a trace amount of NaOH (0.8%, w / v) was introduced into the oil phase to convert the irritant components into soaps. This temperature selection promotes ester bond hydrolysis while avoiding the degradation of heat-sensitive active ingredients (such as polyphenols); the trace alkali concentration ensures the reaction is limited to surface esters, without damaging the main triglyceride structure, thus preserving the oil phase skeleton. Immediately after saponification, the oil was washed with an equal volume of warm water to allow the soaps to enter the aqueous phase. Washing was repeated until the aqueous phase was clear, ensuring thorough removal of the irritant components. Subsequent HPLC fingerprint monitoring ensured the retention of active ingredients during the detoxification process.

[0034] S5. The attenuated oil still contains a large number of components. In order to increase the proportion of anti-inflammatory and fat-soluble active substances, the attenuated oil (oil: ethyl acetate = 1:2) was first extracted with a small amount of ethyl acetate. The solvent layer was enriched by using a separatory funnel and the solvent was recovered to obtain the "enriched active ingredient extract".

[0035] S6. Place in a 40–45℃ water bath under reduced pressure to evaporate the solvent, while adding α-tocopherol (vitamin E) to protect the active ingredients and prevent oxidative degradation.

[0036] Specific extraction method of tung oil extract 1. Select mature tung oil tree (Vernicia fordii) seeds, ensuring they are of pure variety, with plump kernels, a glossy appearance, free from mold and insect infestation, and meet the standards for active ingredient content, to facilitate better preservation of the active ingredients in subsequent extraction processes. Remove the hard shell from the selected tung oil tree seeds, leaving the oil-containing embryo. Dry at 40℃ for 4–6 hours until the moisture content is <8% to reduce rancidity. After drying, pulverize to an appropriate particle size (40–60 mesh) to improve extraction efficiency.

[0037] 2. Using edible ethanol as the extraction solvent, mix the tung seed powder with ethanol at a material-to-liquid ratio of 1:6 (g / mL), and extract by stirring in a water bath at 40-60℃ for 2-4 hours. Repeat the extraction 2-3 times, and combine the extracts. Filter to remove residues to obtain crude tung seed oil (containing fatty acids, active ingredients, and impurities). This step aims to maximize the dissolution of fat-soluble active substances.

[0038] 3. Crude oil often contains suspended solids, plant pigments, waxes, and primary free fatty acids (which can cause irritation and oxidation), requiring further extraction. Let the crude oil stand at a low temperature for 24 hours to allow suspended impurities to settle naturally. Collect the supernatant, then add a small amount of hot water (2%-3% of the oil volume), stir, and let it stand to separate into layers. Discard the aqueous phase and remove the gums. Finally, add activated carbon (1%-2% of the oil volume), stir at 50°C for 30 minutes, and filter to remove pigments and residual impurities, obtaining a clarified refined oil.

[0039] 4. The main irritant components in tung oil include phorbol esters (containing ester bonds) and free fatty acids. Phraphorbol esters can undergo ester bond hydrolysis under alkaline conditions, generating low-toxicity phorbol and fatty acids; free fatty acids react with alkali to form soaps, which can be removed by washing with water. Based on this, this study employed a mild saponification process for detoxification. At 40-60℃, a trace amount of NaOH (0.8%, w / v) was introduced into the oil phase to convert the irritant components into soaps. This temperature selection promotes ester bond hydrolysis while avoiding the degradation of heat-sensitive active ingredients (such as polyphenols); the trace alkali concentration ensures the reaction is limited to surface esters, without damaging the main triglyceride structure, thus preserving the oil phase skeleton. Immediately after saponification, the oil was washed with an equal volume of warm water to allow the soaps to enter the aqueous phase. Washing was repeated until the aqueous phase was clear, ensuring thorough removal of the irritant components. Subsequent HPLC fingerprint monitoring ensured the retention of active ingredients during the detoxification process.

[0040] 5. The attenuated oil still contains a large number of components. In order to increase the proportion of anti-inflammatory and fat-soluble active substances, the attenuated oil (oil: ethyl acetate = 1:2) was first extracted with a small amount of ethyl acetate. The solvent layer was enriched by using a separatory funnel and the solvent was recovered to obtain the "enriched active ingredient extract".

[0041] 6. Place in a 40–45 ℃ water bath under reduced pressure to evaporate the solvent, while adding α-tocopherol (vitamin E) to protect the active ingredients and prevent oxidative degradation.

[0042] The efficacy and mechanism of tung seed oil extracted using the above method in a mouse model of psoriasis were verified as follows: 1. Preparation and Grouping of Experimental Animals: Thirty healthy, 8-week-old male BALB / c mice of similar weight were selected. All mice were acclimatized for one week in a standard experimental animal room (temperature 22±2℃, humidity 50±10%, 12-hour light-dark cycle). Subsequently, the 30 mice were randomly divided into three groups of 10 mice each: a normal control group, a psoriasis model group, and a tung oil extract treatment group. After grouping, the mice in each group were numbered, and their initial weight and dorsal skin condition were recorded.

[0043] 2. Induction of Psoriasis-like Lesions: In addition to the normal control group, psoriasis-like lesions were induced in mice in the model group and treatment group. Specifically, 62.5 mg of 5% imiquimod (IMQ) cream was weighed daily using a precision electronic balance. The operator, wearing sterile gloves, used a sterile cotton swab to evenly apply the weighed IMQ cream to a carefully shaved area (approximately 2 cm × 3 cm) on the back of the mice. This induction process was repeated daily for 7 consecutive days to stably establish an animal model mimicking key pathological features of human psoriasis (including epidermal hyperplasia, parakeratosis, and dermal inflammatory infiltration). An equal amount of petroleum jelly was applied daily to the same area on the back of the normal control mice as a control. Figure 1 and Figure 2 As shown.

[0044] 3. Drug Intervention Treatment: On day 3 of the induction period, when the model group mice began to show visible mild erythema and rough skin, the treatment intervention was initiated. Approximately 30 minutes after applying IMQ daily, the tung oil extract described in this invention was applied to the same lesion area. Application was once daily, with each application being approximately 20 mg, ensuring complete coverage of the lesion area. The model group and normal control group mice received only an equal amount of placebo at the corresponding time points. The entire experimental period (induction and treatment were synchronized) lasted 7 days.

[0045] 4. Efficacy Assessment and Sample Collection: On day 7 of the experiment, after a final status assessment of all mice, they were humanely euthanized and back skin tissue samples were immediately collected. Efficacy assessment included: (a) Macroscopic scoring: Two researchers, unaware of the group assignments, independently scored the psoriasis area and severity index (PASI) on the back skin lesions of the mice. Assessment indicators included the severity of erythema, infiltration, and scaling (0-4 points), and the total score was calculated. (b) Histopathological analysis: Partial back skin tissue was taken, fixed in 4% paraformaldehyde for 24 hours, then embedded in paraffin, sectioned (5 μm thick), and stained with hematoxylin and eosin (HE). Observation and photography were performed under an optical microscope. Epidermal thickness (vertical distance from the bottom of the granular layer to the top of the stratum corneum, with at least 10 randomly selected fields of view averaged) was quantitatively measured using image analysis software. The degree of infiltration of inflammatory cells (mainly lymphocytes and neutrophils) in the dermis was assessed using a semi-quantitative scoring method. Figures 3-6 As shown, in addition, to further visualize the histological pathological changes, Figure 7 The images show HE section pathological images of the back skin of mice in the control group and the model group. The model group showed significant epidermal thickening and increased inflammatory cell infiltration. Figure 8The images show HE sections of the skin on the backs of mice in the model group and the treatment group. The treatment group showed reduced epidermal thickness and improved inflammatory infiltration.

[0046] 5. Transcriptomics (RNA-seq) Analysis: Another portion of the collected skin tissue was immediately flash-frozen in liquid nitrogen and then transferred to a -80°C cryogenic freezer for RNA extraction. Total RNA was extracted from each sample using the TRIzol reagent method, and the concentration, purity, and integrity of the RNA were detected using Nanodrop and Agilent Bioanalyzer (RIN value > 8.0 required). For RNA samples meeting the requirements, high-throughput sequencing was performed using the Illumina platform to construct cDNA libraries and perform paired-end sequencing. After quality control and alignment to a reference genome, the extracted data were used for quantitative gene expression analysis.

[0047] 6. Bioinformatics Analysis and Mechanism Elucidation: First, using the model group as a control, differential expression analysis was performed on the gene expression data of the treatment group (e.g., using DESeq2 software). A significance threshold was set as an adjusted p-value < 0.05 and an absolute fold change > 1.5, screening for differentially expressed genes (DEGs) regulated by tung oil extract. Subsequently, a systematic functional analysis was conducted on these DEGs: (i) KEGG pathway enrichment analysis: used to reveal signaling pathways with significantly enriched differentially expressed genes. Results are presented in bubble chart or bar chart format, as shown in the appendix. Figure 9 As shown, pathways closely related to the core inflammatory mechanism of psoriasis, such as the "IL-17 signaling pathway," "TNF signaling pathway," and "MAPK signaling pathway," are significantly enriched and their gene expression shows an overall downregulation trend. This directly proves that the composition of the present invention acts on the key pathological links of the disease.

[0048] (ii) GO functional enrichment analysis: Annotation was performed at three levels: cellular component, biological process, and molecular function. The analysis results were expected to show that downregulated genes were significantly enriched in cellular components such as the membrane raft (GO:0045121) and the nuclear transcription regulator complex (GO:0005667), as well as in biological processes such as the positive regulation of inflammatory response (GO:0050729) and cytokine production (GO:0001816). This reveals, from the perspectives of subcellular structure and biological function, that tung oil extract may regulate the expression network of downstream inflammation-related genes by affecting signal transduction platforms on the cell membrane and gene transcription complexes in the cell nucleus.

[0049] (iii) GSEA analysis and IL-17 signaling pathway inhibition: as attached Figure 10-11 As shown, GSEA analysis revealed a significant downregulation of the IL-17 signaling pathway in the treatment group, indicating that tung oil extract can inhibit the activation of this core inflammatory pathway. Furthermore, the heatmap showed differences in the expression of key genes in the IL-17 signaling pathway (such as IL-17A, IL-6, and CXCL1) between the treatment and model groups, further validating the anti-inflammatory effect of this drug at the molecular level.

[0050] 7. Key Molecular Validation: To verify the reliability of the RNA-seq results, several key pro-inflammatory cytokine genes located in the core enrichment pathway were selected from the differentially expressed gene list, such as IL-17A, TNF-α, and IL-23p19. Real-time quantitative reverse transcription polymerase chain reaction (RT-qPCR) was used, with GAPDH or β-actin as internal reference genes, to perform absolute or relative quantification of the mRNA expression levels of these genes in the skin tissues of three groups of mice. The experiment required three technical replicates. Expected results showed that, compared with the normal control group, the mRNA expression levels of these cytokines were significantly increased in the model group; while compared with the model group, their expression levels were significantly reduced by tung oil extract in the treatment group. This trend is highly consistent with the RNA-seq data, thus confirming the anti-inflammatory effect of the drug of this invention at the molecular level.

[0051] Through the above implementation scheme, this invention specifically and comprehensively demonstrates how to construct an animal model of psoriasis, apply the described tung oil extract composition for intervention, and verify its efficacy and mechanism of action at multiple levels from macroscopic pathological improvement and microscopic tissue repair to genomics and molecular biology. This embodiment confirms that the pharmaceutical composition of this invention can effectively alleviate mild to moderate plaque psoriasis-like symptoms. Its mechanism lies in multi-target regulation of key inflammatory signaling pathways such as IL-17 and TNF, affecting related cellular components and biological processes, thereby inhibiting excessive inflammatory responses and abnormal epidermal proliferation. This provides a solid and complete chain of experimental evidence for the effectiveness and beneficial effects of the described technical solution. Although embodiments of the invention have been shown and described, it will be understood by those skilled in the art that various changes, modifications, substitutions and alterations can be made to these embodiments without departing from the principles and spirit of the invention, the scope of which is defined by the appended claims.

Claims

1. A method for extracting tung seed oil as a medicinal substance, characterized in that, The specific steps include: Step 1: Select mature tung seeds, remove the outer shell, leaving the oil-containing embryo, dry and then grind them into tung seed powder; Step 2: Mix and stir the tung seed kernel powder with ethanol to obtain crude tung seed oil; Step 3: Let the crude tung oil stand to remove the aqueous phase, gum, and pigments, thus obtaining refined tung oil. Oil; Step 4: Use a mild saponification process to detoxify the refined tung oil; Step 5: The attenuated tung oil is re-extracted using ethyl acetate to obtain enriched active ingredients. Tung oil extract.

2. The method for extracting tung oil according to claim 1, characterized in that: A sixth step is also included, in which a water bath is used to evaporate the solvent and α-tocopherol, in order to protect the active ingredients and prevent oxidative degradation.

3. The method for extracting tung oil according to claim 1, characterized in that: Step one specifically involves drying the oil-containing embryo at 35-45℃ for 4-6 hours until the water content is <8% to reduce rancidity, and then pulverizing it into 40-60 mesh powder to improve extraction efficiency.

4. The method for extracting tung oil according to claim 1, characterized in that: Step two specifically involves mixing tung seed powder and ethanol at a material-to-liquid ratio of 1:6 (g / mL), stirring and extracting in a water bath at 40-60℃ for 2-4 hours, repeating the extraction 2-3 times, combining the extracts, filtering to remove residues, and obtaining crude tung seed oil containing fatty acids, active ingredients, and impurities.

5. The method for extracting tung oil according to claim 1, characterized in that: Step three specifically involves letting the crude tung oil stand for 24 hours to allow suspended impurities to settle naturally, taking the supernatant, then adding 2%-3% water based on the oil volume, stirring, letting it stand to separate into layers, discarding the aqueous phase, removing the gum, and finally adding 1%-2% activated carbon based on the oil volume, stirring at 45-55℃, filtering to remove pigments and residual impurities, and obtaining a clarified refined oil.

6. The method for extracting tung oil according to claim 1, characterized in that: Step four specifically involves contacting the oil phase with NaOH at 40-60℃ to convert the irritating components into soaps. After saponification, the soaps are washed with an equal volume of warm water to allow them to enter the aqueous phase. The washing process is repeated until the aqueous phase is clear to ensure that the irritating components are completely removed. Subsequently, HPLC fingerprinting is used to monitor the process and ensure that the active ingredients are retained during the detoxification process.

7. The method for extracting tung oil according to any one of claims 1-6, characterized in that: The obtained tung oil was used as a topical skin medication.

8. The topical skin medication according to claim 7, characterized in that: Used to treat psoriasis.

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