Plant extract for improving immune function of senescent dendritic cells and application thereof
By using apigenin from artemisia annua extract, the problem of decreased immune function in senescent dendritic cells was addressed, cell viability and CD86 gene expression were improved, and the immune activation capacity of senescent dendritic cells was enhanced.
Patent Information
- Application Number
- CN202610371267.4
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2026-03-25
- Publication Date
- 2026-06-16
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Abstract
Description
Technical Field
[0001] This invention belongs to the field of biotechnology, specifically relating to a plant extract that improves the immune function of senescent dendritic cells and its applications. Background Technology
[0002] With the aging of the global population, age-related health issues have become a major public health challenge. Immunosenogenesis is an important physiological function decline characteristic of the aging process, manifested as a decreased ability of the immune system to respond to antigens and increased susceptibility to disease infection and tumors. Dendritic cells, as the most potent professional antigen-presenting cells in the body, are a key bridge connecting innate and adaptive immunity. Dendritic cells initiate specific immune responses by taking up and processing antigens and expressing MHC molecules and co-stimulatory molecules CD40 and CD86, transmitting activation signals to T lymphocytes. However, aging leads to a decline in the immune function of dendritic cells, particularly a significant decrease in the expression of co-stimulatory molecules CD40 and CD86, resulting in an inability to effectively activate naive T cells and thus a decline in immune response. In addition, senescent dendritic cells secrete high levels of inflammatory factors (such as IL-12), further disrupting the homeostasis of the immune microenvironment.
[0003] Currently, there are extremely limited clinical interventions for immune aging, and there is an urgent need to develop drugs that can improve the functional decline of aging immune cells. Summary of the Invention
[0004] The main problem this invention aims to solve is how to improve the decreased immune activation capacity of senescent dendritic cells.
[0005] To address the above problems, this invention provides the application of artemisia annua extract in the following:
[0006] 1. Application of Artemisia annua extract in improving the immune function of senescent dendritic cells.
[0007] 2. Application of Artemisia annua extract in improving immune aging.
[0008] 3. Application of Artemisia annua extract in the preparation of products that improve the immune function of senescent dendritic cells.
[0009] 4. Application of Artemisia annua extract in the preparation of products that improve the body's immune aging.
[0010] Further, the preparation method of the artemisia annua extract can be as follows: weigh 50g of artemisia annua, mix it with 75% ethanol at a ratio of 1:15 (g / mL), extract it by ultrasonication at 50℃ for 0.5 h, filter and collect the supernatant, and extract the remaining residue again at a material-to-liquid ratio of 1:10 (g / mL). Combine the two supernatants, and obtain the artemisia annua extract by rotary evaporation and freeze drying.
[0011] The senescent dendritic cells described are a model of senescent dendritic cells successfully induced by the addition of 75 mM D-galactose. The morphology of these senescent dendritic cells undergoes significant changes, including cell shrinkage, increased nuclear volume, and decreased adhesion.
[0012] In this article, the active ingredient of the artemisia annua extract is apigenin.
[0013] In this article, the improvement of immune function of senescent dendritic cells is manifested as follows: 1) Increase the vitality of the senescent dendritic cells and improve cell survival rate; 2) Reduce or inhibit the secretion of pro-inflammatory cytokines by the senescent dendritic cells; 3) Increase the expression of the STAT3 gene in the senescent dendritic cells and decrease the expression of the IL1B gene in the senescent dendritic cells.
[0014] 4) Enhance the expression of CD86 gene, a co-stimulatory molecule related to the immune activation function of senescent dendritic cells.
[0015] In one specific embodiment, the concentration of the artemisia extract used is 10 μg / mL.
[0016] This invention also provides the application of the aforementioned apigenin in the preparation of products that improve the immune function of aging dendritic cells or in the preparation of products that improve the body's immune aging.
[0017] This invention also provides the application of apigenin, as described above, in enhancing the immune activation capacity of senescent dendritic cells.
[0018] Furthermore, the concentration of apigenin used is 10 μM.
[0019] This invention, combining network pharmacology, molecular docking technology, and cell molecular biology methods, systematically elucidates that Artemisia annua can be used to improve the immune function of senescent dendritic cells, providing a scientific basis for improving the body's immune aging. This invention has the following beneficial effects: 1) Key active ingredients for function are clearly identified: This invention identified 70 chemical components in Artemisia annua extract and determined that apigenin is the key active ingredient in Artemisia annua's senescent immune regulation; 2) Targeting is clearly defined: This invention discovered that the active ingredient apigenin in Artemisia annua extract has the characteristic of "bidirectional regulation" of the function of senescent dendritic cells, both increasing the expression of the co-stimulatory molecule CD86 gene downregulated due to aging and reducing the level of the inflammatory factor IL-12 increased due to cellular aging; 3) Low safety risk: As a plant with both medicinal and edible properties, Artemisia annua has good long-term safety. Attached Figure Description
[0020] Figure 1This is the total ion chromatogram of Artemisia annua extract.
[0021] Figure 2 Venn diagram of active ingredients of Artemisia annua and targets for regulating immune aging.
[0022] Figure 3 This is a network diagram illustrating the interaction between artemisinin active ingredients and potential target genes regulating immune aging.
[0023] Figure 4 The effect of different concentrations of D-galactose on the viability of senescent dendritic cells.
[0024] Figure 5 This is a staining image of β-galactosidase (left is the control group, right is the D-galactose group).
[0025] Figure 6 The effect of Artemisia annua extract on the vitality of senescent dendritic cells.
[0026] Figure 7 The effect of Artemisia annua extract on the secretion of pro-inflammatory cytokines in senescent dendritic cells.
[0027] Figure 8 The effect of Artemisia annua extract on the expression of target genes regulated by senescent dendritic cells.
[0028] Figure 9 This is a network diagram of artemisia annua, active ingredients, and overlapping targets.
[0029] Figure 10 This is a docking diagram of apigenin with its core target molecules. A) Docking diagram of apigenin with TNF; B) Docking diagram of apigenin with AKT1; C) Docking diagram of apigenin with STAT3; D) Docking diagram of apigenin with IL1B.
[0030] Figure 11 The effect of apigenin on the vitality of senescent dendritic cells.
[0031] Figure 12 The effect of apigenin on the secretion of pro-inflammatory cytokines in senescent dendritic cells.
[0032] Figure 13 The effect of apigenin on the expression of genes related to immune activation in senescent dendritic cells. Detailed Implementation
[0033] The present invention will now be described in further detail with reference to specific embodiments. The given embodiments are merely illustrative of the invention and not intended to limit its scope. The embodiments provided below can serve as a guide for further improvements by those skilled in the art and do not constitute a limitation on the invention in any way.
[0034] Unless otherwise specified, the experimental methods used in the following examples are conventional methods, performed according to the techniques or conditions described in the literature in this field or according to the product instructions. Unless otherwise specified, the materials and reagents used in the following examples are commercially available.
[0035] Unless otherwise specified, the quantitative experiments in the following examples are all repeated three times, and the results are averaged.
[0036] The following examples use statistical software to process the data. The experimental results are expressed as mean ± standard deviation. One-way ANOVA was used, and different letters between groups indicate significant differences.
[0037] Example 1: Preparation and component identification of artemisia annua extract Accurately weigh 50g of Artemisia annua (Beijing Tongrentang), mix it with 75% ethanol at a ratio of 1:15 (g / mL), and extract it by ultrasonication at 50℃ for 0.5 h. Filter and collect the supernatant. Extract the remaining residue again at a material-to-liquid ratio of 1:10 (g / mL). Combine the two supernatants and obtain Artemisia annua extract by rotary evaporation and freeze-drying.
[0038] Sample preparation: Accurately weigh 5 mg of Artemisia annua extract and add it to a 1.5 mL centrifuge tube, then prepare a 10 mg / mL solution with 50% methanol. Vortex to mix and then sonicate for 30 min. Centrifuge at 14000 rpm (4 ℃) for 10 min, and transfer 200 μL of the supernatant to a liquid chromatography vial for LC-MS detection.
[0039] Liquid chromatography conditions: Chromatographic separation was performed on a Vanquish UHPLC System. Column: ACQUITY UPLC HSS T3 column (2.1 × 100 mm, 1.8 µm); column temperature: 35 ℃; mobile phase elution conditions are shown in Table 1; sample chamber temperature: 4 ℃; flow rate: 0.3 mL / min; injection volume: 2 μL.
[0040] Table 1. Gradient elution conditions of Artemisia annua extract using mobile phase
[0041] Mass spectrometry conditions: High-energy electrospray ionization (HESI) source was used; scanning mode was Full-ms / dd-ms2, with simultaneous acquisition of positive and negative ions; scan range was 100-1500 Da, resolution was 60000; spray voltage (SV): 3kV; capillary temperature (CT): 320 ℃; auxiliary device temperature (AT): 350 ℃; fragmentation energy (NCE): 10, 30, 50 V; sheath gas: 35; aux gas: 10.
[0042] The total ion chromatogram of the Artemisia annua extract sample solution is shown below. Figure 1 As shown, the annotations on the right indicate Positive (+) / Negative (-) patterns. Through comparison with reference standards, precise molecular weight matching, and deduction of secondary fragment ions, a total of 70 compounds were identified in the Artemisia annua extract. Flavonoids (such as apigenin and vitexin) and coumarins were abundant among these compounds, as detailed in Table 2.
[0043] Table 2. Identification of components in Artemisia annua extract
[0044] Example 2: Predicting the regulatory effects and targets of artemisinin extract on immune aging based on network pharmacology. 1. Screening of active components and their regulatory targets in Artemisia annua extract Based on the component identification results, the chemical components of Artemisia annua extract were imported into the Swiss AMDE database, and effective active ingredients were screened based on gastrointestinal absorption efficiency and drug-likeness. The chemical structure and SMILES number of Artemisia annua alcohol extract were obtained from the PubChem database and imported into the SwissTargetPrediction platform to predict its potential targets. Finally, all predicted results with a probability value greater than 0 were selected and retained.
[0045] Through screening, a total of 36 active ingredients derived from Artemisia annua extract were obtained. These were imported into the SwissTarget Prediction database, yielding 36 potential active ingredients with corresponding targets (see Table 3). After screening and deduplication of all targets, 564 regulatory targets of the active ingredients were identified.
[0046] Table 3. Active ingredients in Artemisia annua extract
[0047] 2. Screening of regulatory targets related to immune aging Using "Immunity" and "Immunosenescence" as keywords, immune-related targets were collected from the OMIM and GeneCards databases. In GeneCards, targets with a relevance score ≥5.0 were retained. All retrieved targets were summarized and sorted, and duplicate targets were removed.
[0048] Immune-related targets were obtained from the GeneCards and OMIM databases, with 191 targets collected from the OMIM database and 1827 targets collected from the GeneCards database. After removing non-standard and duplicate targets, a total of 1870 targets regulating immune aging were collected.
[0049] 3. Intersection of target genes regulating "artemisinin active ingredients - immune aging" By intersecting the 564 regulatory targets of artemisinin's active components with the 1870 regulatory targets of immunosenescence, 190 commonly regulated targets were obtained. Figure 2 These targets are considered potential targets for artemisia annua extract to regulate immune aging.
[0050] 4. Construct a protein-protein interaction network diagram. The intersection target gene set obtained in step 3 was imported into the STRING database for analysis. The minimum confidence level for protein-protein interactions in the database was set to 0.4, resulting in a protein-protein interaction network diagram. In Cytoscape, the degree values of the protein nodes were sorted from largest to smallest. Combined with network analysis, the core targets of cellular immune regulation were identified. The darker the color of a target node, the more proteins that can interact with it, and the more important the protein at that node.
[0051] Depend on Figure 3 It is known that tumor necrosis factor (TNF) TNF ), AKT serine / threonine kinase 1 ( AKT1 ), signal transduction and transcription activator 3 ( STAT3 ), interleukin-1β ( IL1B ) is the core hub gene that regulates the network.
[0052] Example 4: The regulatory effect of artemisia annua extract on the immune function of senescent dendritic cells 1. Construction of a dendritic cell senescence model Using the dendritic cell line (DC2.4, Xiamen Yimo Biotechnology Co., Ltd.) as the research object, we established a dendritic cell senescence model to verify the regulatory effect of artemisia annua extract on the physiological function of senescent immune cells.
[0053] Dendritic cells in logarithmic growth phase were seeded into 96-well plates at 100 μL per well. Cells were cultured at 37°C and 5% CO2 for 12 h until cell adhesion. After discarding the supernatant, the cells were washed twice with PBS, and five concentration gradients of D-galactose (IG0540, Solarbio) were added to the culture medium: 0, 25, 50, 75, and 100 mM. Cell viability was assessed using the CCK-8 assay after 48 h of culture.
[0054] The results are as follows Figure 4 As shown, cell viability exhibited a significant negative correlation with D-galactose concentration. Compared to the blank control group, cell viability decreased in a concentration-dependent manner as the D-galactose concentration gradually increased from 25 mM to 100 mM. When the D-galactose concentration reached 75 mM, cell viability significantly decreased to 58.33%, approaching the half-maximal inhibitory concentration (IC50).
[0055] Cells from each group were fixed in fixative for 15 min, washed with PBS, and then 1 mL of cell senescence β-galactosidase staining reagent (C0602, Beyotime, containing 10 μL of β-galactosidase staining solutions A and B, 930 μL of β-galactosidase staining solution C, and 50 μL of X-galactose) was added. After incubating the cells overnight at 37°C in the dark, they were observed under an inverted optical microscope to be stained blue-green, and then photographed. The specific experimental procedures were strictly performed according to the kit instructions.
[0056] The senescence status of dendritic cells in the blank control group and the 75 mM D-galactose treatment group was assessed using β-galactosidase staining. The results are as follows: Figure 5 As shown, compared with the blank control group, the number of β-galactosidase-positive (blue-green) cells in the model group was significantly increased, indicating an increased proportion of senescent cells. Simultaneously, the cell morphology in this group showed significant changes, including cell shrinkage, increased nuclear volume, and decreased adhesion. These typical senescence-related changes confirm that the addition of 75 mM D-galactose successfully induced dendritic cell senescence, and the dendritic cell senescence model was successfully established.
[0057] 2. Effects of Artemisia annua extract on the viability of senescent dendritic cells Control group: Only DC2.4 dendritic cell culture medium (IM-M035, Xiamen Yimo Biotechnology Co., Ltd.) was used, without any added treatment factors.
[0058] Model group: 75 mM D-galactose was added to DC2.4 dendritic cell-specific culture medium to induce cell senescence.
[0059] Artemisia annua extract group: 75 mM D-galactose and 10 μg / mL Artemisia annua extract were added to DC2.4 dendritic cell-specific culture medium to observe the effect of Artemisia annua on senescent dendritic cells.
[0060] Dendritic cells in logarithmic growth phase were seeded into 96-well plates at 100 μL per well and cultured for 12 h until cell adhesion. After discarding the supernatant, the cells were washed twice with PBS. Then, 75 mM D-galactose (IG0540, Solarbio) solution prepared in DC2.4 dendritic cell culture medium was added, and the cells were cultured for 48 hours. Finally, 10 μg / mL of Artemisia annua extract was added, and cell viability was assessed using the CCK-8 assay.
[0061] Depend on Figure 6 It can be seen that there are significant differences in cell viability among the groups. P <0.05). Compared with the control group, the cell viability of the model group decreased by 46.87%, and the addition of Artemisia annua extract increased the viability of senescent dendritic cells by 16.16%, indicating that Artemisia annua extract can increase the activity of senescent dendritic cells and maintain the normal growth state of cells.
[0062] 3. Effects of Artemisia annua extract on the secretion of pro-inflammatory cytokines by senescent dendritic cells Supernatants from dendritic cells treated with various drug concentrations were collected and centrifuged at 1500 r / min for 20 min to remove impurities. Subsequently, the ELISA kit (ml037868, Shanghai Enzyme-Link Biotechnology Co., Ltd.) was strictly followed according to its instructions. The simplified steps are as follows: Prepare standards by adding the test supernatant and standards, and incubate at 37℃ for 1.5 h; after washing, add biotinylated antibody and enzyme conjugate sequentially, washing thoroughly after each incubation; add chromogenic buffer for 15 min to terminate the reaction, and immediately measure the absorbance of each well at 450 nm using a microplate reader. Then, calculate the IL-12 level based on the standard curve.
[0063] Depend on Figure 7 The significant increase in IL-12 levels in the model group compared to the control group (P < 0.05) indicates that senescence induces dendritic cells into a dysfunctional pro-inflammatory state, leading to disordered immune activation. Conversely, the IL-12 levels in the artemisia annua extract group were significantly lower than those in the model group (P < 0.05), suggesting that artemisia annua extract can inhibit the secretion of inflammatory factors from senescent dendritic cells and improve immune homeostasis. Further research is needed to investigate its specific targets and molecular mechanisms.
[0064] 4. Effects of Artemisia annua extract on gene expression in senescent dendritic cells To verify the regulatory effect of artemisia annua extract on senescent dendritic cells, the expression of STAT3 and IL1B genes was detected. RNA was extracted from senescent dendritic cells using a rapid RNA extraction kit (RE241-01, Beijing Aipuruisheng Technology Co., Ltd.), and the extracted RNA was reverse transcribed into cDNA using an UltraScript First-Strand cDNA Synthesis RT-PCR Kit (RT317-02, Beijing Aipuruisheng Technology Co., Ltd.).
[0065] Next, using cDNA as a template, real-time quantitative PCR (qPCR) was performed according to the AipMix 2× SYBR Green qPCR SuperMix kit (FP301-02, Beijing Aipruisheng Technology Co., Ltd.) instructions. The regulatory target genes predicted by network pharmacology were used... STAT3 , IL1B ( ) is the target gene, and the relative expression level of the target gene is calculated. The primer sequence is shown in Table 4.
[0066] Table 4. Primer sequence information
[0067] The results are as follows Figure 8 As shown, D-galactose-induced dendritic cell senescence leads to STAT3 Gene expression was significantly reduced, while IL1B Gene expression was significantly increased ( P <0.05); while artemisia annua extract improves senescent dendritic cells. STAT3 Gene expression was reduced, and senescent dendritic cells were significantly decreased. IL1B Gene expression ( P <0.05), maintaining the physiological homeostasis of senescent dendritic cells.
[0068] Example 5: Screening of key active ingredients in artemisia annua extract for regulating the immune function of senescent dendritic cells 1. Construction of the "Artemisia annua-active ingredient-intersecting target" network diagram Using Cytoscape 3.9.1 software, the names, active ingredients, and targets of Artemisia annua were imported to construct a network diagram of "Artemisia annua-active ingredients-intersecting targets". Network Analyzer results showed that the network had 227 nodes, 701 edges, a network density of 0.027, a network diameter of 4, a network radius of 2, a network centrality of 0.187, a network heterogeneity of 1.396, and a characteristic path length of 3.231. (See [link to relevant documentation]). Figure 9Based on the Degree values sorted from highest to lowest, the top five active ingredients identified were apigenin, vitexin, citric acid, artemisinin lactone, and osthol. Therefore, these compounds may be important active components in artemisia annua that regulate the immune function of senescent dendritic cells.
[0069] 2. Molecular docking analysis of active ingredients and target genes First, the structure of the active ingredient was downloaded from the PubChem database. Then, the protein structures of the key targets were downloaded from the Uniprot protein structure database and saved in PDB format. Pymol software was used to remove water molecules and small ligands from the core targets, and AutoDock Tools was used for hydrogenation and charge processing before saving the results as pdbqt format. TNF , AKT1 , STAT3 , IL1B Using apigenin, vitexin, citric acid, artemisinin, and osthol as ligands, molecular docking was performed using pyrx software.
[0070] The results are shown in Table 5. The molecular docking binding energy of all combinations is less than -5 kcal / mol, indicating that the active ingredients of Artemisia annua can bind stably to the core target site.
[0071] The docking results were visualized using PyMOL, as shown below. Figure 10 As shown, apigenin has a low binding energy to the core target, indicating that apigenin is a key active ingredient in artemisia annua extract that regulates the function of senescent dendritic cells.
[0072] Table 5. Molecular docking analysis of active components of Artemisia annua with target genes
[0073] 3. The regulatory effect of apigenin on the immune function of senescent dendritic cells 1) Effects of apigenin on the viability of senescent dendritic cells Dendritic cells in the logarithmic growth phase were seeded at 100 μL / well in sterile 96-well plates. After 12 h, the culture medium was removed from the wells, and 75 mM D-galactose was added. The cells were cultured for 48 h, the culture medium was discarded, and then different concentrations of complete culture medium containing apigenin (IA0400, Solarbio) were added. The relative cell viability was calculated according to the above method. The experiment was divided into 6 groups, and the specific procedures for each group are as follows: Control group: Only culture medium was used, without any treatment factors added.
[0074] Model group: 75 mM D-galactose was added to the culture medium to induce cell senescence.
[0075] Positive control group: 75 mM D-galactose and 0.1 mM metformin hydrochloride (D9351, Solarbio) were added to the culture medium.
[0076] Low concentration of apigenin: 75 mM D-galactose and 5 μM apigenin were added to the culture medium.
[0077] Medium concentration group of apigenin: 75 mM D-galactose and 10 μM apigenin were added to the culture medium.
[0078] High concentration of apigenin: 75 mM D-galactose and 20 μM apigenin were added to the culture medium.
[0079] Depend on Figure 11 It can be seen that, compared with the control group, the cell survival rate in the model group was significantly reduced, indicating that senescence modeling leads to a decrease in cell viability. After the addition of apigenin, the cell survival rate in the apigenin group (5 μM, 10 μM) was higher than that in the model group ( P The concentration of apigenin (<0.05) indicates that apigenin can improve the viability of senescent dendritic cells. Specifically, cell viability initially increased and then decreased with increasing apigenin concentration. Therefore, 10 μM was selected as the appropriate dosage for subsequent experiments.
[0080] 2) Effects of apigenin on the secretion of pro-inflammatory cytokines by senescent dendritic cells Supernatants from dendritic cells treated with various drug concentrations were collected and centrifuged at 1500 r / min for 20 min to remove impurities. Subsequently, the ELISA kit (ml037868, Shanghai Enzyme-Link Biotechnology Co., Ltd.) was strictly followed according to its instructions. The simplified steps are as follows: Prepare standards by adding the test supernatant and standards, and incubate at 37℃ for 1.5 h; after washing, add biotinylated antibody and enzyme conjugate sequentially, washing thoroughly after each incubation; add chromogenic buffer for 15 min to terminate the reaction, and immediately measure the absorbance of each well at 450 nm using a microplate reader. Then, calculate the IL-12 level based on the standard curve.
[0081] Depend on Figure 12 It can be seen that, compared with the control group, the IL-12 content of senescent dendritic cells in the model group was significantly increased ( P <0.05 indicates that dendritic cells have entered a dysfunctional pro-inflammatory state, leading to disordered immune activation. Compared with the model group, the levels of IL-12 secreted by senescent dendritic cells in the positive control group and the apigenin group were significantly decreased ( P The result was <0.05, indicating that apigenin plays a role in inhibiting the secretion of pro-inflammatory cytokines from senescent dendritic cells and improving immune activation.
[0082] 3) Effects of apigenin on the expression of genes related to immune activation in senescent dendritic cells RNA was extracted from samples using a rapid RNA extraction kit (RE241-01, Beijing Aipuruisheng Technology Co., Ltd.). Then, following the instructions of the UltraScript First-Strand cDNA Synthesis RT-PCR Kit (RT317-02, Beijing Aipuruisheng Technology Co., Ltd.), the extracted RNA was reverse transcribed to synthesize first-strand cDNA. After cDNA preparation, real-time quantitative PCR was performed using the cDNA as a template, following the instructions of the AipMix 2× SYBR Green qPCR SuperMix Kit (FP301-02, Beijing Aipuruisheng Technology Co., Ltd.). The experiment selected co-stimulatory molecules for dendritic cell activation of T cells. CD40 and CD86 As the target gene, the relative expression level of the target gene was calculated using internal reference genes. Primer sequence information is detailed in Table 4.
[0083] Depend on Figure 13 It was found that, compared with the control group, the expression levels of CD40 and CD86, co-stimulatory genes of dendritic cells, were significantly decreased in the model group (P < 0.05), indicating that the function of senescent dendritic cells in activating T cells was reduced, thereby affecting the cellular immune response. Apigenin significantly upregulated the expression level of CD86 gene in senescent dendritic cells (P < 0.05), improving the immune activation function of senescent dendritic cells, demonstrating that apigenin is a key active ingredient in Artemisia annua that improves the immune function of senescent dendritic cells.
[0084] The above results indicate that Artemisia annua extract can improve the immune function of senescent dendritic cells and enhance their ability to activate immune responses. Combining network pharmacology, molecular docking techniques, and in vitro cell experiments, apigenin was identified as the key active substance in Artemisia annua extract that regulates the immune function of senescent dendritic cells.
[0085] The present invention has been described in detail above. Those skilled in the art will recognize that the invention can be practiced in a wide range of ways with equivalent parameters, concentrations, and conditions without departing from its spirit and scope, and without requiring unnecessary experiments. While specific embodiments have been provided, it should be understood that further modifications can be made to the invention. In summary, according to the principles of the invention, this application is intended to include any changes, uses, or improvements to the invention, including changes made using conventional techniques known in the art that depart from the scope disclosed herein.
Claims
1. Application of Artemisia annua extract in improving the immune function of senescent dendritic cells.
2. Application of Artemisia annua extract in improving immune aging.
3. Application of Artemisia annua extract in the preparation of products that improve the immune function of senescent dendritic cells.
4. Application of Artemisia annua extract in the preparation of products that improve the body's immune aging.
5. The application according to any one of claims 1-4, characterized in that, The preparation method of the Artemisia annua extract is as follows: Weigh 50g of Artemisia annua, mix it with 75% ethanol at a ratio of 1:15 (g / mL), and extract it by ultrasonic extraction at 50℃ for 0.5 h. Filter and collect the supernatant. Extract the remaining residue again at a material-to-liquid ratio of 1:10 (g / mL). Combine the two supernatants and obtain the Artemisia annua extract by rotary evaporation and freeze drying.
6. The application according to any one of claims 1-4, characterized in that, The active ingredient in the artemisia annua extract is apigenin.
7. The application according to claim 1 or 3, characterized in that, The improvement in the immune function of senescent dendritic cells is manifested in the following ways: 1) Increase the vitality of the senescent dendritic cells and improve cell survival rate; 2) Reduce or inhibit the secretion of pro-inflammatory cytokines by the senescent dendritic cells; 3) Increase the expression of the STAT3 gene in the senescent dendritic cells and decrease the expression of the IL1B gene in the senescent dendritic cells; 4) Enhance the expression of CD86 gene, a co-stimulatory molecule related to the immune activation function of senescent dendritic cells.
8. The application according to any one of claims 1-4, characterized in that, The concentration of the artemisia extract used is 10 μg / mL.
9. The use of apigenin as described in claim 4 in the preparation of products that improve the immune function of aging dendritic cells or in the preparation of products that improve the body's immune aging.
10. The application of apigenin according to claim 4 in enhancing the immune activation ability of senescent dendritic cells.