Application of apigenin in preparation of product for treating sjogren syndrome

Abstract

The invention discloses application of apigenin in preparation of a product for treating sjogren syndrome, and belongs to the technical field of medicines. The apigenin is used for remarkably regulating and controlling the immune system of the body, namely enhancing the immunosuppression function of MDSCs of the body, further inhibiting activation of dendritic cells DCs, resulting in reduction of the expression level of B cell activating factors BAFF, finally achieving the purposes of inhibiting B cell differentiation and reducing immunocompetence, and relieving the symptoms of SS.

Description

Technical Field

[0001] The invention belongs to the field of medical technology, and in particular relates to application of apigenin in preparing a product for treating Sjögren's syndrome. Background Art

[0002] Sjögren's syndrome Spleen and stomach syndrome (SS) is a systemic autoimmune disease with a specific tendency to cause inflammation of the exocrine glands, mainly the salivary glands and lacrimal glands, but also the nose, upper respiratory tract, oropharynx, etc. Studies have shown that abnormal activation of B lymphocytes plays an important role in SS disease. In-depth research on the function of B cells provides a new strategy for the effective treatment of SS by regulating the abnormal function of B cells in SS disease. At present, immunosuppressive drugs are often used in the clinical treatment of SS disease. However, this treatment is prone to side effects and relapse. In recent years, with the continuous deepening of research, it has been shown that traditional Chinese medicine has good immunosuppressive properties and is often used to treat a variety of immune diseases.

[0003] Apigenin is a flavonoid compound with unique biological activity and widely distributed in nature. It is mainly found in plants of the Thymelaeaceae, Verbenaceae, and Selaginellaceae. At present, many studies at home and abroad have shown that apigenin has anti-inflammatory, antioxidant and anti-tumor activities, and plays an important role in lowering blood pressure, preventing atherosclerosis and inhibiting tumor growth. Summary of the invention

[0004] One of the purposes of the present invention is to provide an application of apigenin in preparing a product for treating Sjögren's syndrome, wherein the CAS number of the apigenin is 520-36-5.

[0005] Preferably, the product is a medicine.

[0006] More preferably, the dosage form of the drug is any one of powder, tablet, granule, capsule, solution, emulsion and suspension.

[0007] More preferably, apigenin is the only active ingredient in the product.

[0008] More preferably, the medicine further contains medically acceptable excipients.

[0009] The second object of the present invention is to provide a traditional Chinese medicine composition for treating Sjögren's syndrome, which is composed of the following components in parts by weight: 15-25 parts of apigenin, 1-3 parts of nobiletin, 1-2 parts of rutin, 1-2 parts of gallic acid, 1-2 parts of neohesperidin, 1-2 parts of curcumin, 1-2 parts of paeoniflorin, 1-2 parts of phloretin, 1-2 parts of pterostilbene, and 1-2 parts of curcumin.

[0010] Preferably, the traditional Chinese medicine composition is composed of the following components in parts by weight: 25 parts of apigenin, 1 part of nobiletin, 1 part of rutin, 1 part of gallic acid, 1 part of neohesperidin, 1 part of curcumene, 1 part of paeoniflorin, 1 part of phloretin, 1 part of pterostilbene, and 1 part of curcumin.

[0011] The third object of the present invention is to provide a method for preparing a Chinese medicinal compound for treating Sjögren's syndrome, the method comprising the following steps:

[0012] (1) crushing the above-mentioned Chinese medicine composition;

[0013] (2) After adding water, ultrasonic treatment was performed at 45-60°C for 30-60 min;

[0014] (3) Filter and collect the filtrate, and spray-dry the filtrate to obtain a solid powder or vacuum concentrate the filtrate to obtain a spray preparation.

[0015] Compared with the prior art, the present invention has the following beneficial effects:

[0016] (1) Apigenin has many advantages, including its widespread distribution in nature, wide range of sources, and easy collection.

[0017] (2) The present invention utilizes apigenin to significantly regulate the body's immune system, that is, to enhance the body's MDSCs cell immunosuppressive function, thereby inhibiting the activation of dendritic cells DCs, leading to a decrease in the expression level of B cell activating factor BAFF, and ultimately achieving the purpose of inhibiting B cell differentiation and reducing immune activity, thereby alleviating the symptoms of SS. BRIEF DESCRIPTION OF THE DRAWINGS

[0018] Figure 1 This is the detection of clinical SS patient-related indicators in Example 1.

[0019] Figure 2 This is the correlation between SS disease score and BAFF level in Example 1.

[0020] Figure 3 This is the identification of human peripheral blood B cells isolated in Example 1.

[0021] Figure 4 This is the determination of the content of antibodies secreted by B cells in Example 1.

[0022] Figure 5 This is the saliva flow rate test of ESS mice in Example 2 (to verify whether the ESS modeling is successful).

[0023] Figure 6 The expression level of autoantibodies in the serum of ESS mice in Example 2 (to verify whether the ESS modeling is successful).

[0024] Figure 7The normal mouse MDSC (CD11b + Gr1 + ) Separation flow cytometry diagram.

[0025] Figure 8 The mRNA levels of iNOS and Arg-1 in MDSC cells were detected by RTPCR in Example 3, wherein the ESS group refers to model mice.

[0026] Fig. 9 The ELISA method in Example 3 was used to detect the levels of cytokines iNOS and Arg-1 secreted by MDSC cells, wherein the ESS group refers to model mice.

[0027] Fig.10 This is a flow cytometry diagram of the separation of mouse DC cells in Example 4.

[0028] Fig.11 This is the determination of the in vitro proliferation activity of mouse DC cells in Example 4.

[0029] Fig.12 This is the determination of the content of cytokine BAFF released by DC cells in Example 4.

[0030] Fig.13 This is a flow cytometry diagram of the separation of mouse B cells in Example 5.

[0031] Fig.14 This is the determination of the in vitro proliferation activity of mouse B cells in Example 5.

[0032] Fig.15 This is the determination of the content of antibodies (IgG, IgA and IgM) released by B cells in Example 5.

[0033] Fig.16 This is the determination of the proliferation activity of apigenin on mouse MDSC cells in Example 6.

[0034] Fig.17 This is the effect of apigenin on the apoptosis of MDSC in Example 6.

[0035] Fig.18 The mRNA levels of iNOS and Arg-1 were measured before and after apigenin treatment of MDSC cells in Example 6.

[0036] Fig.19 The protein levels of iNOS and Arg-1 were measured before and after apigenin treatment of MDSC cells in Example 6.

[0037] Fig. 20 This is the effect of MDSCs pretreated with apigenin in Example 7 on the mRNA expression level of BAFF in DC cells.

[0038] Fig.21 This is the in vitro detection of the effect of DC cells pre-treated with apigenin and MDSC on B cell activity in Example 8.

[0039] Fig. 22 This is the HE staining detection of submandibular gland tissue before and after apigenin treatment in the ESS model in Example 2.

[0040] Fig.23 This is the saliva flow rate detection of ESS mice after apigenin treatment in Example 2.

[0041] Fig.24 This is the expression level of autoantibodies in the serum of ESS mice after apigenin treatment in Example 2.

[0042] Fig.25 The mRNA levels of iNOS and Arg-1 in MDSC cells were measured after ESS was treated with apigenin for 7 days in Example 2.

[0043] Fig.26 This is the determination of BAFF levels in DC cells after 7 days of apigenin treatment in Example 2.

[0044] Fig. 27 The proliferation activity of B cells extracted after 7 days of apigenin treatment in Example 2 was detected. DETAILED DESCRIPTION

[0045] Example 1 Detection of clinical sample related indicators

[0046] 1.1 Detection of relevant cytokines. After whole blood was collected from patients with Sjögren's syndrome (SS) and healthy controls (HC), the serum was immediately separated by centrifugation at 1400×g, and then the serum was aliquoted and stored at -80°C to avoid repeated freezing and thawing. All samples were placed at room temperature before cytokine detection, and the expression level of BAFF in the serum of the SS group and the HC group was detected by ELISA. The experimental method was referred to the instructions provided by the manufacturer. At the same time, this method was also used to measure the expression levels of the above cytokines in subsequent animal models.

[0047] The inclusion criteria for patients with Sjögren's syndrome were as follows:

[0048] Oral symptoms (at least one of the following is true): ① Daily dry mouth for more than 3 months ② Recurrent or persistent swelling of the parotid gland after adulthood ③ The need to use water or liquids to help swallow dry food.

[0049] Eye symptoms (at least one of the following must be met): ① Unbearable dry eyes for more than 3 months ② Recurrent gritty sensation (foreign body sensation) in the eyes ③ The need to use artificial tears 3 times or more per day.

[0050] Ocular signs (at least one of the following is met): ① Schirmer test ≤ 5 mm / 5 min ② Van Bijsterveld corneal fluorescein staining score ≥ 4.

[0051] Salivary gland damage (at least one of the following conditions is met): ① unstimulated salivary flow rate ≤ 1.5 ml / 15 min ② parotid gland radiography is positive ③ salivary gland isotope examination is positive

[0052] Histological examination: Labial gland pathology showed ≥1 lymphocyte focus (4 mm 2 A lymphocyte focus is defined as a collection of at least 50 lymphocytes in the labial gland stroma.

[0053] Autoantibody test: anti-SSA antibody or anti-SSB antibody is positive.

[0054] No other diseases.

[0055] 1.2 Correlation between SS score and BAFF level. The disease activity index of each patient was scored according to the EULAR 2010 Sjögren's syndrome disease activity index (ESSDAI) standard, and the correlation between BAFF content and ESSDAI score was statistically analyzed.

[0056] 1.3 Human B cell isolation and functional detection:

[0057] Isolation of B cells. PBMCs were isolated using Ficoll reagent and analyzed using EasySep TM Human B CellEnrichment Kit / EasySep TM Human B cell enrichment kit was used for magnetic bead sorting, and finally flow cytometry was used to detect the purity of sorted B cells.

[0058] Detection of antibodies secreted by B cells: The activity of the B cells sorted above was detected by ELISA, that is, the level of antibodies produced by the B cells was detected.

[0059] Implementation 2 Construction and functional verification of ESS animal model

[0060] 2.1 Model construction. 8-week-old female C57BL / 6 mice and 6-week-old male C57BL / 6 mice were purchased from the Experimental Animal Center of Nantong University. The mice were housed in a specific pathogen-free animal barrier, and all experiments were approved by the Animal Ethics Committee of Nantong University.

[0061] Bilateral salivary glands were isolated from female C57BL / 6 mice (8 weeks old) and homogenized in PBS to prepare submandibular gland (SG) protein. SG protein was emulsified to 2 mg / mL in an equal volume of Freund's complete adjuvant (CFA) and injected subcutaneously into naturally immunized mice (6-week-old male mice) at a dose of 100 μL / mouse, with injections in the neck on days 0 and 7, and booster injections on day 14 with 1 mg / mL SG protein emulsified in Freund's incomplete adjuvant (IFA).

[0062] 2.2 Detection of related diseases. Detection of mouse saliva flow rate (35 days after the above-mentioned booster injection of SG protein emulsified with 1 mg / mL Freund's incomplete adjuvant IFA): The above-mentioned modeled mice and an equal number of normal mice were anesthetized and injected with pilocarpine at a dose of 5 mg / kg body weight intraperitoneally. Saliva was collected from the mouse mouth using a 20 mL pipette tip for 15 minutes, and all mice were measured every 7 days. Detection of autoantibodies: ELISA kits were used to detect autoanti-M3R antibodies and antinuclear antibodies in serum.

[0063] 2.3 HE detection of ESS model mice and their submandibular gland tissue treated with apigenin in vivo. The ESS model constructed above was randomly divided into 2 groups, with 3 mice in each group. One group was gavaged at a dose of 100 mg / kg, and a normal saline treatment group of the same volume was set up at the same time. The drug was administered once every two days for a total of 5 weeks. After the experiment, the submandibular gland tissue of the mice was removed and soaked in 4% paraformaldehyde overnight, gradient dehydrated according to standard procedures, and the tissue was paraffin-embedded, sliced, and stained with hematoxylin and eosin according to the instructions, and HE staining images were collected for analysis. In addition, relevant cell function experiments were carried out, that is, MDSC, DC and B cells were isolated and extracted after 7 days of apigenin treatment of ESS mice, and relevant cell activity was detected.

[0064] Example 3 Isolation and extraction of mouse MDSC cells and functional verification

[0065] 3.1 Isolation, extraction and identification of MDSCs. First, EasySep TM Mouse MDSC (CD11b + Gr1 + )Isolation Kit / EasySep TM Mouse Myeloid-Derived Suppressor Cell (MDSC) Isolation Kit, which can quickly enrich CD11b from mouse spleen + Gr1 + Then, we used flow cytometry to detect the purity of the sorted MDSC cells.

[0066] 3.2 Verification of MDSC function. Detection of Arg-1 and iNOS mRNA levels: RTPCR was used to detect the gene transcription of Arg-1 and iNOS in the sorted MDSC cells. Detection of Arg-1 and iNOS protein levels: ELISA was used to detect the protein levels of Arg-1 and iNOS released by MDSC.

[0067] Example 4 Isolation and extraction of mouse DC cells and functional verification

[0068] 4.1 Isolation, extraction and identification of DC cells. We used EasySep TM Mouse Plasmacytoid DCIsolation Kit / EasySep TM Mouse pDC Dendritic Cell Isolation Kit was used to isolate pDCs from mouse spleen single cell suspension. Then, we used flow cytometry to detect the purity of the isolated DCs.

[0069] 4.2 DC function verification DC cell proliferation activity: CCK8 assay was used to further analyze the proliferation activity of DC cells. Detection of BAFF protein level released by DC cells: ELISA was used to detect the protein level of BAFF released by DC cells.

[0070] Example 5 Isolation and extraction of mouse B cells and functional verification

[0071] 5.1 Isolation, extraction and identification of B cells. Mouse B lymphocytes were sorted using the EasySep Mouse B Cell Isolation Kit. Specifically, mouse spleens were collected and prepared into single-cell suspensions, and B lymphocytes were sorted using a magnetic bead sorting kit. Subsequently, the purity of the extracted B cells was detected using a flow cytometer.

[0072] 5.2 Verification of B cell function. Cell proliferation activity: The proliferation activity of B cells was detected by CCK8 detection method. Determination of antibody content released by B cells: IgG, IgA and IgM in the supernatant were quantitatively analyzed by ELISA.

[0073] Example 6 Detection of cell activity of MDSCs treated with apigenin

[0074] 6.1 The MDSCs isolated above were cultured, plated in six-well plates, and treated with 0, 10 ng / mL, and 30 ng / mL apigenin for 12 h. The cells were collected, RNA was extracted, and the gene transcription levels of iNOS and Arg-1 before and after apigenin treatment of MDSCs were detected by RTPCR. The supernatant was collected, and the protein levels of iNOS and Arg-1 released before and after MDSC treatment were detected by ELISA.

[0075] 6.2 Detection of cell proliferation activity. Cells were treated with 0, 10 ng / mL, and 30 ng / mL apigenin, respectively, and plated in 96-well plates. CCK8 was detected at 0, 12 h, 24 h, 36 h, 48 h, and 60 h, respectively.

[0076] 6.3 Apoptosis detection: Flow cytometry was used to detect the apoptosis of MDSC cells treated with apigenin.

[0077] Example 7 Detection of the effect of pre-treated MDSC on DC cell function

[0078] 7.1 Treatment process: The MDSCs isolated above were cultured, treated with 30 ng / mL apigenin for 12 h, and the cells were collected.

[0079] 7.2 Co-culture process. MDSC and DC cells were co-cultured using the transwell experiment. Specific operation: First, the pre-treated MDSC cells were inoculated into a chamber with a pore size of 0.4 μm and cultured overnight. Then the DC cells were cultured in the lower chamber. The cells were cultured in RPMI 1640 complete medium. The co-culture ratio of MDSC and DC cells was 1:5. After 48 hours of co-culture, the DC cells and culture supernatant were harvested for functional verification.

[0080] 7.3 Functional detection The protein expression level of BAFF in DC cells by MDSCs pretreated with apigenin was detected by ELISA.

[0081] Example 8 Detection of the effect of pretreated DC cells on B cell activity

[0082] 8.1 Co-culture process. The transwell experiment was also used to co-culture the DCs and B cells treated with the above MDSC cells. Specific operation: First, the pre-treated DC cells were inoculated into a chamber with a pore size of 0.4 μm and cultured overnight. Then the B cells were cultured in the lower chamber. The cells were cultured in RPMI 1640 complete medium. The co-culture ratio of DC and B cells was 1:5. After 48 hours of co-culture, the B cells and culture supernatant were harvested for functional verification.

[0083] 8.2 Activity detection The cell activity of the collected B cells was detected in vitro using CCK8 assay.

[0084] In the early stage of this study, a large number of clinical SS patient samples were collected, and the protein expression level of BAFF in the serum of SS patients and healthy volunteers was measured by ELISA. In SS patients, the BAFF content was significantly higher than that in healthy subjects (HC) ( Figure 1 ). According to the EULAR 2010 Sjögren's Syndrome Disease Activity Index (ESSDAI) standard, it was found that the BAFF content level in SS patients was significantly positively correlated with the disease score ( Figure 2 ). In order to further analyze the pathogenesis of SS, we analyzed the proportion and subpopulation distribution of B cells in clinical samples. We first isolated B cells from peripheral blood. After magnetic bead sorting, the purity of B cells increased to more than 86% ( Figure 3 ), which can lay the foundation for subsequent B cell activity experiments. The antibody levels produced by B cells were then tested, and normal human B cells were set as a control group. The B cells sorted in SS patients can produce higher levels of antibodies, including IgG, IgA and IgM ( Figure 4 In this experiment, the mouse model used was experimental Sjögren's syndrome induced by immunization with salivary gland proteins. After modeling, the salivary flow rate of ESS mice was significantly reduced compared with the control group ( Figure 5 ), serum anti-muscarinic acetylcholine 3 receptor antibody (anti-M3R antibody) and antinuclear antibody levels were significantly increased ( Figure 6 ). Through the detection of the above indicators, it was finally confirmed that the ESS model was successfully established by immunizing C57BL / 6 mice with mouse salivary gland proteins. Subsequently, we selected the spleen of the mice, prepared a single cell suspension, and used the above magnetic bead sorting kit to isolate CD11b + Gr1 + Cells, the purity of MDSC cells obtained by flow cytometry sorting is about 90%. At the same time, without magnetic bead sorting, the positive rate of MDSC in flow cytometry is only about 18% ( Figure 7 ). We found through RT-PCR that the mRNA levels of Arg-1 and iNOS in the sorted MDSC cells were significantly lower than those in normal mice ( Figure 8 ). Subsequently, we detected by ELISA that the protein levels of Arg-1 and iNOS released by MDSCs in the ESS group were much lower than those in the normal group ( Fig. 9). It was further confirmed that in SS disease, the function of MDSC cells is greatly inhibited, thereby promoting the progression of SS symptoms. Subsequently, pDC cells were sorted from mouse spleen single cell suspension using flow cytometry. The flow cytometry results showed that the DC cells obtained by magnetic bead sorting had a high purity and could be used for subsequent cell function research ( Fig.10 We further analyzed the proliferation activity of DC cells through CCK8 experiments. The proliferation activity of DC cells isolated from ESS model mice was significantly higher than that of normal mice ( Fig.11 ). At the same time, the level of BAFF cytokine released by DC cells in ESS mice was significantly higher than that in the normal group ( Fig.12 ), these experiments showed that in ESS mice, DC cells were more active, especially releasing higher levels of BAFF, which further promoted the activity of B cells, thereby promoting the progression of SS disease. Subsequently, we extracted high-purity B cells after magnetic bead sorting, laying the foundation for subsequent mechanism research ( Fig.13 ). The CCK-8 assay was used to detect the proliferation activity of B cells after magnetic bead sorting in healthy mice and ESS model mice. Compared with the control group, the activity of B cells in the ESS model group was significantly higher ( Fig.14 ). We then tested the antibodies (IgG, IgA and IgM) secreted by B cells. The results showed that in ESS model mice, B cells were able to produce higher concentrations of antibodies ( Fig.15 ). Through the above experiments, it was further confirmed that in SS disease, MDSC, DC and B cells interact with each other, and their abnormal functions aggravate the disease state. Then we explored the mechanism of apigenin in treating SS. First, we used apigenin to treat the MDSC isolated from the above ESS model mice, and detected a significant increase in the proliferation ability of MDSC cells through CCK8 experiment ( Fig.16 ), flow cytometry detected that the apoptosis level gradually decreased with the increase of apigenin concentration ( Fig.17 ). RT-PCR assay detected that the mRNA levels of iNOS and Arg-1 in MDSC cells were significantly increased after apigenin treatment ( Fig.18 ), and the protein levels of iNOS and Arg-1 released by MDSCs after apigenin treatment were significantly increased by ELISA ( Fig.19 ), which indicates that apigenin can enhance the cell activity of MDSC and exert the function of immunosuppression. In the same way, DCs were co-cultured with MDSCs treated with apigenin. RT-PCR showed that MDSCs pretreated with apigenin could inhibit the transcription of BAFF gene in DCs ( Fig. 20). Next, a transwell experiment was used to co-culture DCs and B cells. The ELISA results showed that DCs treated with apigenin and MDSCs could significantly reduce the level of antibodies released by B cells ( Fig.21 ). Subsequently, we further verified the therapeutic effect of apigenin on SS disease in vivo in ESS model mice. First, HE staining showed that the submandibular gland tissue of ESS model mice was improved after apigenin treatment ( Fig. 22 ). After treating ESS mice with apigenin for a period of time, the salivary flow rate of the mice was increased ( Fig.23 ) and decreased secretion of autoantibodies ( Fig.24 ). MDSC cells, DC cells and B cells were sorted out by the above method. First, RT-PCR was used to detect that the mRNA levels of Arg-1 and iNOS in the MDSC cells of the apigenin treatment group were increased ( Fig.25 After 7 days of apigenin treatment, the BAFF level of DC cells decreased significantly ( Fig.26 ) and B cell activity was also inhibited ( Fig. 27 ), which reflects that apigenin has significant therapeutic effect on ESS mice.

[0085] In summary, apigenin can effectively inhibit the expression level of BAFF in the glandular area, effectively inhibit B cell activation, maturation, differentiation and proliferation, and ultimately the symptoms of Sjögren's syndrome are effectively and continuously relieved, laying a theoretical foundation for the further use of apigenin to treat Sjögren's syndrome.

[0086] Example 9 Preparation of Traditional Chinese Medicine Compound for Treating Sjögren's Syndrome

[0087] 1. Take the following Chinese medicinal components by weight: 25 parts of apigenin; 1 part of nobiletin; 1 part of rutin; 1 part of gallic acid; 1 part of neohesperidin; 1 part of curcumene; 1 part of paeoniflorin; 1 part of phloretin; 1 part of pterostilbene; 1 part of curcumin. Grind the above Chinese medicinal components in a mortar and mix them well, filter them through a 100-mesh sieve, and collect the mixed powder.

[0088] 2. Add 2 times the weight of water to the above powder, dissolve it at 60°C for 30 minutes under ultrasound, collect the filtrate after filtering, spray dry to obtain solid powder, make a granular preparation, and finally prepare a Chinese medicine combination for treating Sjögren's syndrome.

[0089] Example 10 Preparation of a Chinese medicinal compound for treating Sjögren's syndrome

[0090] 1. Take the following Chinese medicinal components by weight: 25 parts of apigenin; 1 part of nobiletin; 1 part of rutin; 1 part of gallic acid; 1 part of neohesperidin; 1 part of curcumene; 1 part of paeoniflorin; 1 part of phloretin; 1 part of pterostilbene; 1 part of curcumin. Grind the above Chinese medicinal components in a mortar and mix them well, filter them through a 100-mesh sieve, and collect the mixed powder.

[0091] 2. Add 2 times the weight of water to the above powder, dissolve it at 45°C under ultrasound for 60 minutes, collect the filtrate after filtering, spray dry to obtain solid powder, make a granular preparation, and finally prepare a Chinese medicine combination for treating Sjögren's syndrome.

[0092] Example 11 Preparation of a Chinese medicinal compound for treating Sjögren's syndrome

[0093] 1. Take the following Chinese medicinal components by weight: 25 parts of apigenin; 1 part of nobiletin; 1 part of rutin; 1 part of gallic acid; 1 part of neohesperidin; 1 part of curcumene; 1 part of paeoniflorin; 1 part of phloretin; 1 part of pterostilbene; 1 part of curcumin. Grind the above Chinese medicinal components in a mortar and mix them well, filter them through a 100-mesh sieve, and collect the mixed powder.

[0094] 2. Add 2 times the weight of water of the mixed raw materials to the above powder, dissolve it at 60°C under ultrasound for 30 minutes, collect the filtrate after filtration, and vacuum concentrate it to the same weight volume of the mixed raw materials to prepare a spray preparation, and finally prepare a Chinese medicine combination for treating Sjögren's syndrome.

[0095] Example 12 Preparation of Traditional Chinese Medicine Compound for Treating Sjögren's Syndrome

[0096] 1. Take the following Chinese medicinal components by weight: 25 parts of apigenin; 1 part of nobiletin; 1 part of rutin; 1 part of gallic acid; 1 part of neohesperidin; 1 part of curcumene; 1 part of paeoniflorin; 1 part of phloretin; 1 part of pterostilbene; 1 part of curcumin. Grind the above Chinese medicinal components in a mortar and mix them well, filter them through a 100-mesh sieve, and collect the mixed powder.

[0097] 2. Add 2 times the weight of water of the mixed raw materials to the above powder, dissolve it at 45°C under ultrasound for 60 minutes, collect the filtrate after filtration, and vacuum concentrate it to the same weight volume of the mixed raw materials to prepare a spray preparation, and finally prepare a Chinese medicine combination for treating Sjögren's syndrome.

[0098] The embodiments described above are only descriptions of the preferred modes of the present invention, and are not intended to limit the scope of the present invention. Without departing from the design spirit of the present invention, various modifications and improvements made to the technical solutions of the present invention by ordinary technicians in this field should all fall within the protection scope determined by the claims of the present invention.

Claims

1. Application of apigenin in the preparation of products for treating Sjögren's syndrome.

2. The use according to claim 1, characterized in that: The product described is a drug.

3. The use according to claim 2, characterized in that: The dosage form of the drug is any one of powder, tablet, granule, capsule, solution, emulsion and suspension.

4. The use according to any one of claims 1 to 3, characterized in that: Apigenin is the only active ingredient in the product.

5. The use according to claim 2 or 3, characterized in that: The medicine also contains medically acceptable auxiliary materials.

6. The use according to claim 4, characterized in that: The medicine also contains medically acceptable auxiliary materials.

7. A Chinese medicine composition for treating Sjögren's syndrome, characterized in that: The traditional Chinese medicine composition is composed of the following components in parts by weight: 15-25 parts of apigenin, 1-3 parts of nobiletin, 1-2 parts of rutin, 1-2 parts of gallic acid, 1-2 parts of neohesperidin, 1-2 parts of curcumin, 1-2 parts of paeoniflorin, 1-2 parts of phloretin, 1-2 parts of pterostilbene, and 1-2 parts of curcumin.

8. The Chinese medicine composition according to claim 7, characterized in that: The traditional Chinese medicine composition consists of the following components in parts by weight: 25 parts of apigenin, 1 part of nobiletin, 1 part of rutin, 1 part of gallic acid, 1 part of neohesperidin, 1 part of curcumin, 1 part of paeoniflorin, 1 part of phloretin, 1 part of pterostilbene, and 1 part of curcumin.

9. A method for preparing a Chinese medicinal compound for treating Sjögren's syndrome, characterized in that: The preparation method comprises the following steps: (1) crushing the Chinese medicine composition according to claim 7 or 8; (2) After adding water, ultrasonic treatment was performed at 45-60°C for 30-60 min; (3) Filter and collect the filtrate, and spray-dry the filtrate to obtain a solid powder or vacuum concentrate the filtrate to obtain a spray preparation.

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