A traditional Chinese medicine compound for treating Sjogren's syndrome, a preparation method and application thereof

Abstract

The application discloses a traditional Chinese medicine compound for treating Sjogren's syndrome and a preparation method and application thereof. The traditional Chinese medicine compound is prepared from Huangqi, Danshen, Maidong and Dendrobium officinale. The traditional Chinese medicine for treating Sjogren's syndrome has the functions of tonifying qi, generating fluid, nourishing yin, moistening dryness, promoting blood circulation and removing blood stasis. The traditional Chinese medicine for treating Sjogren's syndrome has unique advantages, can obviously improve the dry mouth, anti-fatigue, submandibular gland lymphocyte infiltration and T cell subpopulation proportion disorder of Sjogren's syndrome, and is an effective pure traditional Chinese medicine preparation.

Description

Technical Field

[0001] The present invention relates to a traditional Chinese medicine compound for treating autoimmune diseases, and in particular to a traditional Chinese medicine compound for treating Sjögren's syndrome, and a preparation method and application thereof. Background Art

[0002] Sjögren's syndrome ( Sjögren's syndrome (SS) is a chronic systemic autoimmune disease characterized by lymphocyte infiltration of exocrine glands, leading to secondary glandular dysfunction and resulting in dry mouth and eye symptoms. Globally, the incidence of primary Sjögren's syndrome reaches 0.5%, with a higher incidence in women, with a female-to-male ratio of 8:1. The pathogenesis of Sjögren's syndrome is unclear, but may involve epithelial cells, viruses, genetic susceptibility, and environmental triggers. These factors trigger an abnormal response to autoantigens, leading to massive lymphocyte infiltration of exocrine glands and ultimately causing chronic inflammation and secretory dysfunction.

[0003] Currently, there is no effective cure for Sjögren's syndrome in clinical practice. Treatments primarily focus on symptom relief to reduce glandular complications, primarily targeting ocular and oral symptoms, and immunosuppressive therapy for systemic symptoms. Artificial tears are used to alleviate dry eyes; oral dryness treatments typically include increasing daily water intake, chewing sugar-free gum, and rinsing with artificial saliva; and systemic symptoms are typically treated with immunosuppressants such as corticosteroids and hydroxychloroquine. However, long-term use of immunosuppressants can lead to various adverse reactions. Summary of the Invention

[0004] The present invention aims to address the deficiencies of the prior art and provide a traditional Chinese medicine compound for treating Sjögren's syndrome, as well as its preparation method and application. The present invention has conducted in-depth research on the pharmacological activity of the compound and its application in the treatment of Sjögren's syndrome, confirming that the compound preparation can significantly alleviate Sjögren's syndrome, increase salivary flow rate, relieve fatigue, regulate the Th17 / TREG cell ratio, and reduce inflammatory factor levels. Furthermore, in experiments on human submandibular gland cells treated with TNF-α and IFN-γ, it was found that the compound can increase cell viability, inhibit cell apoptosis, reduce intracellular ROS and related cytokine levels, and reduce the chemotaxis of T cells to the compound. The compound has achieved excellent therapeutic effects in both animals and cells.

[0005] To achieve the above object, the present invention provides a traditional Chinese medicine compound for treating Sjögren's syndrome, wherein the traditional Chinese medicine compound is mainly prepared by mixing and concentrating the extracts of multiple traditional Chinese medicines;

[0006] The aforementioned multi-flavor Chinese medicine comprises the following components in the following mass fraction ratios:

[0007] Astragalus 30%-40%,

[0008] Salvia miltiorrhiza 30%-40%,

[0009] Ophiopogon japonicus 20%-25%,

[0010] Dendrobium officinale 10%-15%.

[0011] To achieve the above object, the present invention also provides a method for preparing a traditional Chinese medicine compound for treating Sjögren's syndrome, comprising the following steps:

[0012] (1) Place Astragalus, Salvia miltiorrhiza, Ophiopogon japonicus, and Dendrobium officinale into an extraction tank, add water, and soak for half an hour;

[0013] (2) heating the water in the extraction tank to boiling, boiling for 1 hour, and filtering to obtain decoction A;

[0014] (3) Add water to the extraction tank, boil over low heat, repeat step (2), and filter to obtain decoction B;

[0015] (4) Combine the filtered decoction A and decoction B, centrifuge at 3000 rpm for 10 min, and collect the supernatant;

[0016] (5) Cooling, packaging, and sterilizing the supernatant obtained in step (4) for later use; and obtaining the Chinese herbal compound.

[0017] To achieve the above object, the present invention also provides a use of a traditional Chinese medicine compound for treating Sjögren's syndrome in preparing a medicine for treating Sjögren's syndrome.

[0018] Furthermore, the dosage form of the Chinese herbal compound is tablets, capsules, granules or sprays.

[0019] Furthermore, the Chinese herbal compound is added with a medically acceptable drug carrier to prepare a preparation.

[0020] The beneficial effects of the present invention are:

[0021] (1) The Chinese herbal compound of the present invention can improve the symptoms of Sjögren's syndrome;

[0022] (2) According to the theory of traditional Chinese medicine, the compound can firstly replenish Qi and raise the clearness, transform it into essence and fine particles, so that the blood and body fluids can be transported and distributed; secondly, it can generate body fluids to control dryness and fire, so that the essence, blood and body fluids are sufficient and the dryness symptoms can be moistened; thirdly, it can remove blood stasis and dredge the meridians, so that the Qi flow is unobstructed and the body fluids can be distributed; the four herbs are used together to achieve the effects of "replenishing Qi, generating body fluids and removing blood stasis", so that the dryness symptoms can be cured;

[0023] (3) Increase salivary flow rate, reduce submandibular gland lymphocyte infiltration, restore salivary gland function, and improve fatigue symptoms;

[0024] (4) Reduce related inflammatory factors in the submandibular gland and the ratio of Th17 / TREG cells, which has anti-inflammatory and improves immune imbalance in the body;

[0025] (5) reduce the levels of anti-SSA / RO and anti-SSB / LA autoantibodies in serum;

[0026] (6) Reduce ROS levels in the submandibular gland, increase submandibular gland cell activity, and inhibit submandibular gland cell apoptosis. BRIEF DESCRIPTION OF THE DRAWINGS

[0027] Figure 1 The figure is a schematic diagram showing the effect of the compound on the submandibular gland index, spleen index and salivary flow rate of mice; Figure 1 (A) is the submandibular gland index, Figure 1 (B) is the spleen index, Figure 1 (C) in the figure is the salivary flow rate;

[0028] Figure 2 Schematic diagram of the effect of the compound on the histomorphology and pathology of the submandibular gland in mice; Figure 2 (A) is HE staining of mouse submandibular gland tissue. Figure 2 (B) in the middle is the submandibular gland lesion score;

[0029] Figure 3 Schematic diagram of the effect of the compound on the levels of anti-SSA / RO and anti-SSB / LA in mouse serum; Figure 3 (A) is the level of anti-SSA / RO in mouse serum. Figure 3 (B) in the middle is the anti-SSB / LA level in mouse serum;

[0030] Figure 4 The figure is a schematic diagram showing the effect of the compound on the salivary flow rate and the levels of autoantibodies anti-SSA / RO and anti-SSB / LA in mice; Figure 4 (A) in the figure is the saliva flow rate of mice at 6-18 weeks. Figure 4 (B) is the level of anti-SSA / RO in mouse serum. Figure 4 (C) in the middle is the anti-SSB / LA level in mouse serum;

[0031] Figure 5 The figure is a schematic diagram showing the effect of the compound on the forced swimming time, serum lactate and urea nitrogen levels in mice; Figure 5 (A) is the forced swimming time of mice, Figure 5 (B) is the serum lactate level, Figure 5 (C) in the figure is the serum urea nitrogen level;

[0032] Figure 6 The figure shows the effect of the compound on the submandibular gland pathology, lesion score and AQP5 expression in mice; Figure 6 (A) is HE staining of mouse submandibular gland tissue. Figure 6 (B) in the figure is the submandibular gland lesion score. Figure 6 (C) is the score of AQP5 level in mouse submandibular gland. Figure 6 (D) in the middle is AQP5 IHC staining of mouse submandibular gland;

[0033] Figure 7 Schematic diagram of the effect of the compound on the ratio of Th17 / TREG cells in mouse spleen; Figure 7 (A) is the flow cytometry of Th17 cells in mouse spleen. Figure 7 (B) is the proportion of Th17 cells in mouse spleen. Figure 7 (C) is the flow cytometry of Treg cells in mouse spleen. Figure 7 (D) is the proportion of Treg cells in mouse spleen. Figure 7 (E) in the middle is the ratio of Th17 / Treg cells in mouse spleen;

[0034] Figure 8 The figure is a schematic diagram showing the effect of the compound on related cytokines in the submandibular gland of mice; Figure 8 (A) shows the levels of inflammatory factors TNF-α, IFN-γ, IL-1β, IL-6, and IL-18 in the submandibular gland of mice. Figure 8 (B) shows the levels of chemokines CXC9 and CXCL10 in the submandibular gland of mice;

[0035] Figure 9 Schematic diagram of the effect of the compound on cell viability and apoptosis of human submandibular gland cells; Figure 9 (A) in the middle is the cell viability of human submandibular gland cells, Figure 9 (B) is the apoptosis rate of human submandibular gland cells, Figure 9 (C) is a flow cytometric analysis of apoptosis in human submandibular gland cells;

[0036] Figure 10 Schematic diagram of the effect of the compound on ROS in human submandibular gland cells; Figure 10 (A) is the ROS level in human submandibular gland cells. Figure 10 (B) is a flow cytometric graph of ROS in human submandibular gland cells;

[0037] Figure 11 This is a schematic diagram of the effect of the compound on cytokines related to human submandibular gland cells;

[0038] Figure 12 Schematic diagram of the effect of the compound on T cell chemotaxis. DETAILED DESCRIPTION

[0039] The present invention provides a traditional Chinese medicine compound for treating Sjögren's syndrome, which is mainly prepared by mixing and concentrating the extracts of multiple traditional Chinese medicines;

[0040] The aforementioned multi-flavor Chinese medicine comprises the following components in the following mass fraction ratios:

[0041] Astragalus 30%-40%,

[0042] Salvia miltiorrhiza 30%-40%,

[0043] Ophiopogon japonicus 20%-25%,

[0044] Dendrobium officinale 10%-15%.

[0045] The preparation method of the above-mentioned traditional Chinese medicine compound for treating Sjögren's syndrome comprises the following steps:

[0046] (1) Place Astragalus, Salvia miltiorrhiza, Ophiopogon japonicus, and Dendrobium officinale into an extraction tank, add water, and soak for half an hour;

[0047] (2) heating the water in the extraction tank to boiling, boiling for 1 hour, and filtering to obtain decoction A;

[0048] (3) Add water to the extraction tank, boil over low heat, repeat step (2), and filter to obtain decoction B;

[0049] (4) Combine the filtered decoction A and decoction B, centrifuge at 3000 rpm for 10 min, and collect the supernatant;

[0050] (5) Cooling, packaging, and high-temperature sterilizing the supernatant obtained in step (4) for standby use, wherein the sterilization temperature is 121° C. and the sterilization time is 15 min; and obtaining the Chinese herbal compound.

[0051] The use of the above-mentioned traditional Chinese medicine compound for treating Sjögren's syndrome in the preparation of a drug for treating Sjögren's syndrome includes: the traditional Chinese medicine compound is in the form of tablets, capsules, granules or sprays, and the traditional Chinese medicine compound is added with a medically acceptable drug carrier to prepare a preparation.

[0052] Example 1

[0053] The present invention provides a traditional Chinese medicine compound for treating Sjögren's syndrome, which is prepared from the following medicinal materials in parts by weight: 30g of astragalus, 30g of salvia miltiorrhiza, 15g of ophiopogon, and 12g of dendrobium officinale.

[0054] 30 g of Astragalus, 30 g of Salvia miltiorrhiza, 15 g of Ophiopogon japonicus, and 12 g of Dendrobium officinale were taken by weight, added with pure water 10 times the total weight of the Chinese medicinal materials, soaked for half an hour, and decocted three times for 1 hour each time, and the decoctions were combined to prepare an extract (decoction liquid).

[0055] Furthermore, the extract prepared above is freeze-dried, added with powdered sugar and dextrin, mixed evenly, dried, sieved, and granulated to prepare granules.

[0056] Furthermore, the extract prepared above is added with corn starch and calcium stearate, mixed evenly, granulated and dried, and compressed into tablets.

[0057] Furthermore, the extract prepared above is added with an appropriate amount of pure water, stirred evenly, filtered, sterilized and filled to obtain a spray.

[0058] Example 2: Experiment on the function of treating Sjögren's syndrome using the present invention

[0059] 1. Take 5.46g of Astragalus, 2.73g of Ophiopogon, 5.46g of Salvia miltiorrhiza, and 2.184g of Dendrobium officinale respectively, put them into 200ml of pure water and boil them twice for 1h. Combine the two decoctions and concentrate them using a rotary evaporator to adjust the total crude drug concentration to 565.5mg / mL.

[0060] Experiments on the function of treating Sjögren's syndrome were carried out using the present invention:

[0061] 2.8-week animal experiment: The animals were divided into 4 groups, with 10 mice in each group. The blank group consisted of 10 ICR mice. The other 3 groups consisted of 30 NOD / LTJ mice, namely the model group, the hydroxychloroquine administration group, and the Chinese medicine compound administration group (226.2 mg / 20 g).

[0062] 3. The salivary flow rate of mice was measured, the submandibular gland and spleen were weighed and the organ index was calculated, the submandibular gland was pathologically evaluated, and the mouse serum was collected for the determination of relevant indicators.

[0063] 4. Experimental Results

[0064] (1) Test for determination of submandibular gland index, spleen index and salivary flow rate of mice: After anesthesia, mice were injected with pilocarpine, and their salivary flow rate was measured. The submandibular gland and spleen were weighed and the organ index was calculated.

[0065] like Figure 1 As shown in (A), compared with the blank group, the submandibular gland index of the model group mice showed a significant decrease; compared with the model group, the submandibular gland index of the compound group was significantly increased. Figure 1As shown in (B), compared with the blank group, the spleen index of the model group increased significantly; compared with the model group, the spleen index of the compound group decreased significantly. Figure 1 As shown in (C), compared with the blank group, the salivary flow rate of the mice in the model group was significantly decreased, and compared with the model group, the salivary flow rate of the mice in the compound group was significantly increased.

[0066] (2) Pathological investigation of the submandibular gland of mice: The submandibular gland of mice was stained with H&E, and the pathological changes were observed under a microscope and the lesion points were counted.

[0067] like Figure 2 As shown in (A) and (B) in the figure, the submandibular gland alveoli of mice in the blank group had normal structure, with no necrosis of the epithelium or ducts, no interstitial edema or congestion, and no lymphocytic infiltration or germinal centers. Compared with the blank group, the intercellular spaces in the submandibular gland sections of the model group were larger, the alveoli were heterogeneous in size, and the glands showed significant atrophy in some areas, accompanied by periluminal lymphocytic infiltration or the formation of germinal centers. Compared with the model group, the degree of ductal dilatation in the submandibular gland sections of the compound group was reduced, the homogeneity of alveolar size was improved, and the periluminal lymphocytic infiltration was reduced. Submandibular gland sections of mice in the blank group had virtually no lesions. Compared with the blank group, the lesion scores of mice in the model group were significantly increased, with more lesions. Compared with the model group, the pathological scores and lesion counts in the compound group were reduced.

[0068] (3) Determination of anti-SSA / RO and anti-SSB / LA antibody levels in mouse serum: Mouse serum samples were collected and incubated, washed, reacted with substrates, and terminated according to the kit instructions. After completion, the samples were read using a microplate reader, and a standard curve was prepared to calculate the concentration of each well.

[0069] like Figure 3 As shown in (A) and (B), compared with the blank group, the levels of anti-SSA / RO and anti-SSB / LA antibodies in the serum of mice in the model group were increased; compared with the model group, the levels of anti-SSA / RO and anti-SSB / LA in the serum of mice in the compound group were significantly decreased.

[0070] Example 2: Experiment on the function of treating Sjögren's syndrome using the present invention

[0071] 1. Take 5.46g of Astragalus, 2.73g of Ophiopogon, 5.46g of Salvia miltiorrhiza, and 2.184g of Dendrobium officinale respectively, put them into 200ml of pure water and boil them for 1h twice, combine the two decoctions, and concentrate them using a rotary evaporator. Adjust the total crude drug concentration to 282.75mg / ml in the low-dose group, 565.5mg / ml in the medium-dose group, and 847.5mg / ml in the high-dose group.

[0072] 2.10-week animal experiment: The animals were divided into 6 groups, with 10 mice in each group, including 10 ICR mice in the blank group; the other 5 groups, totaling 50 NOD / LTJ mice, were model group, tofacitinib tablet group, low-dose Chinese medicine compound group (113.1 mg / 20 g), medium-dose Chinese medicine compound group (226.2 mg / 20 g), and high-dose Chinese medicine compound group (339.3 mg / 20 g).

[0073] 3. The mice were subjected to salivary flow rate measurement, fatigue test, pathological evaluation of the submandibular gland, serum samples from the mice were collected for determination of relevant indicators, the Th17 / TREG cell ratio in the spleen of the mice was measured, and the submandibular gland of the mice was collected for detection of relevant proteins.

[0074] 4. Experimental Results

[0075] (1) Detection of saliva flow and autoantibodies in mice: After anesthesia, mice were injected with pilocarpine to measure saliva flow rate, and serum was collected from mice to detect autoantibodies using ELISA kits.

[0076] like Figure 4 As shown in (A), compared with the blank group, the saliva flow of the model group mice was lower at the sixth week, but not significantly. By the tenth week, the saliva flow of the model group and the compound group mice was significantly lower than that of the blank group, while the saliva flow of the tofacitinib tablet group was significantly increased compared with the model group. At the fourteenth week, the saliva flow of the mice in the medium-dose and high-dose compound groups increased to varying degrees, which was significantly different from the tenth week. At the eighteenth week, the saliva flow of the mice in the compound group was significantly increased compared with the model group. Figure 4 As shown in (B), compared with the blank group, the anti-SSA / RO level in the serum of the mice in the model group was significantly increased; compared with the model group, the anti-SSA / RO level in the serum of the mice in the compound group was significantly decreased to varying degrees; Figure 4 As shown in Figure (C), compared with the blank group, the model group showed a significant increase in serum anti-SSB / LA levels; compared with the model group, the compound group showed significant decreases to varying degrees. These results indicate that the compound can increase salivary flow in NOD / LTJ mice and reduce serum levels of related autoantibodies.

[0077] (2) Mouse fatigue evaluation: A lead weight of 10% of the mouse's body weight was tied to the mouse's tail. The mouse was forced to swim and its swimming time was recorded until the mouse could no longer float to the surface. Serum was collected for the determination of lactic acid and urea nitrogen.

[0078] like Figure 5 As shown in (A), compared with the blank group, the forced swimming time of the mice in the model group was significantly reduced; compared with the model group, the swimming time of the mice in the Buqi Shengjin Quyu Decoction group was significantly increased; Figure 5As shown in (B), there was no significant difference in serum lactic acid content between the blank group and the model group; compared with the model group, the lactic acid content in the compound group was significantly reduced; Figure 5 As shown in (C), compared with the blank group, the urea nitrogen level in the serum of the model group was significantly increased; compared with the model group, the urea nitrogen level in the serum of the compound group was significantly decreased; the results showed that the compound can alleviate the fatigue symptoms of NOD / LTJ mice.

[0079] (3) Pathological investigation of the submandibular gland of mice and detection of AQP5 protein: The submandibular gland of mice was taken for H&E staining. The pathological changes were observed under a microscope and the lesions were counted. Immunohistochemical staining of AQP5 was also performed.

[0080] like Figure 6 As shown in (A) and (B) in the figure, compared with the blank group, the number of lesions caused by lymphocyte infiltration in the submandibular gland in the model group was more, the lesion area was larger, and the lesion score in the model group was higher; compared with the model group, the number and size of lesions in the submandibular gland in the compound group were reduced, and the lesion score was significantly reduced; Figure 6 (C) and (D) show that compared with the blank group, the expression of AQP5 in the submandibular gland of the model group was significantly decreased; compared with the model group, the expression of AQP5 in the submandibular gland of the medium and high dose compound groups was significantly increased to varying degrees in a dose-dependent manner; the results showed that the compound could improve the pathological condition of the submandibular gland and increase the expression of AQP5.

[0081] (4) Determination of the ratio of Th17 / TREG cells in mouse spleen: The mouse spleen was ground and cells were extracted, and cell determination was performed using flow cytometry.

[0082] like Figure 7 As shown in (A)-(E) in the figure, compared with the blank group, the number of Th17 cells in the model group increased significantly, the number of Treg cells decreased significantly, and the Th17 / Treg cell ratio decreased significantly; compared with the model group, the number of Th17 cells in the spleen of mice in the low-dose compound group decreased significantly, the Treg cells decreased but not significantly, and the Th17 / Treg cell ratio decreased significantly; compared with the model group, the number of Th17 cells in the medium-dose compound group decreased significantly, the Treg cells decreased significantly, and the Th17 / Treg cell ratio decreased significantly; compared with the model group, the number of Th17 cells in the high-dose compound group decreased significantly, the Treg cells decreased significantly, and the Th17 / Treg cell ratio decreased significantly; the results showed that the compound can reduce the Th17 / Treg cell ratio in the spleen of mice.

[0083] (5) Determination of inflammatory factors such as TNF-α, IFN-γ, IL-1β, IL-6, IL-18, CXCL9 and CXCL10 in the submandibular gland of mice: The submandibular gland of mice was ground and the protein supernatant was extracted. The relevant indicators were measured using ELISA kits.

[0084] like Figure 8 As shown in (A) and (B) in the figure, compared with the blank group, the levels of TNF-α, IFN-γ, IL-1β, IL-6, IL-18, CXCL9 and CXCL10 in the submandibular gland of the mice in the model group were significantly increased; compared with the model group, the levels of IFN-γ, IL-1β, CXCL9 and CXCL10 in the submandibular gland of the mice in the low-dose compound group were significantly decreased, and the levels of TNF-α, IL-6 and IL-18 showed a downward trend but no significance; compared with the model group, the levels of TNF-α, IFN-γ, IL-1β, IL-6, CXCL9 and CXCL10 in the submandibular gland of the mice in the medium-dose compound group were significantly decreased, and the level of IL-18 was decreased but no significance; compared with the model group, the levels of TNF-α, IFN-γ, IL-1β, IL-6, IL-18, CXCL9 and CXCL10 in the submandibular gland of the mice in the high-dose compound group were significantly decreased; the results showed that the compound can reduce the levels of inflammatory factors in the submandibular gland.

[0085] Example 3: Experiment on the function of treating Sjögren's syndrome using the present invention

[0086] 1. Extraction of Drug-Containing Serum: Forty SD rats were randomly divided into a blank control group and low-, medium-, and high-dose compound dosage groups, with 10 rats in each group. The drug solution was decocted according to the previously described method. The low-, medium-, and high-dose Buqi Shengjin Quyu Decoction groups were administered 1 ml of the drug daily by oral gavage. The control group was given an equal volume of water by oral gavage for 7 consecutive days. Serum was collected after the last dose.

[0087] 2. Using human submandibular gland cells as a model, the cells were stimulated with TNF-α and IFN-γ, and then treated with compound drug-containing serum to measure cell viability and apoptosis rate, intracellular ROS levels, related cytokines, and T cell chemotaxis to the compound-treated cell supernatant.

[0088] 3. Experimental Results

[0089] (1) Determination of cell viability and apoptosis rate of human submandibular gland cells: 10% CCK-8 was added to the drug-treated human submandibular gland cells, and the results were measured using an enzyme-labeled instrument; the drug-treated human submandibular gland cells were incubated with flow cytometry reagents, and the apoptosis rate was determined using flow cytometry.

[0090] like Figure 9 As shown in (A), (B) and (C), compared with the blank group, TNF-α and IFN-γ can significantly reduce the viability of human submandibular gland cells in the model group and increase the cell apoptosis rate; compared with the model group, low, medium and high doses of the compound drug-containing serum can significantly alleviate the decline in cell viability and the increase in proliferation rate of human submandibular gland cells; and after the addition of PI3K agonist, the effect of high dose was reversed; the results showed that the compound group can increase the cell viability of human submandibular gland cells and inhibit cell apoptosis.

[0091] (2) Determination of ROS in human submandibular gland cells: Human submandibular gland cells treated with the drug were incubated with flow cytometry reagents, and the intracellular ROS level was determined by flow cytometry.

[0092] like Figure 10 As shown in (A) and (B), compared with the blank group, the ROS level of human submandibular gland cells in the model group was significantly increased; compared with the model group, the ROS level of human submandibular gland cells was significantly decreased after treatment with the compound drug-containing serum in a dose-dependent manner; the results showed that the compound can reduce the increase in ROS in A-253 cells caused by TNF-α and IFN-γ stimulation.

[0093] (3) Determination of related cytokine levels in human submandibular gland cells: The cell supernatant of the drug-treated human submandibular gland cells was taken and the levels of related cytokines were determined using ELISA.

[0094] like Figure 11 As shown in the results, compared with the blank group, the levels of IL-1β, IL-6, CXCL9, and CXCL10 in human submandibular gland cells in the model group were significantly increased; compared with the model group, the compound-containing serum could significantly inhibit the levels of IL-1β, IL-6, CXCL9, and CXCL10 in human submandibular gland cells; the results showed that the compound-containing serum could inhibit the decrease in IL-1β, IL-6, CXCL9, and CXCL10 levels. The results showed that the compound could inhibit the decrease in IL-6, CXCL9, and CXCL10 levels.

[0095] (4) Determination of T cell chemotaxis to human submandibular gland cell supernatant: The cell supernatant of human submandibular gland cells treated with drug was collected and the chemotaxis of T cells was evaluated using Transwell.

[0096] like Figure 12 As shown in the results, after stimulation with TNF-α and IFN-γ, the supernatant of human submandibular gland cells in the model group significantly increased the chemotaxis of Jurkat T cells; compared with the model group, the serum containing the compound drug significantly inhibited the chemotaxis of Jurkat T cells. The results showed that the compound can inhibit the chemotaxis of Jurkat T cells induced by the supernatant of human submandibular gland cells stimulated by TNF-α and IFN-γ.

[0097] The above embodiments are used to explain the present invention rather than to limit the present invention. Any modifications and changes made to the present invention within the spirit of the present invention and the protection scope of the claims shall fall within the protection scope of the present invention.

Claims

1. A Chinese medicine composition for treating Sjögren's syndrome, characterized in that: The Chinese medicine composition is prepared by mixing and concentrating the extracts of multiple Chinese medicines; The aforementioned multiple Chinese medicinal materials are each composed of the following components in the following mass fraction ratios: Astragalus 30%-40%, Salvia miltiorrhiza 30%-40%, Ophiopogon japonicus 20%-25%, Dendrobium officinale 10%-15%.

2. The method for preparing the Chinese medicine composition for treating Sjögren's syndrome according to claim 1, characterized in that: The following steps are involved: (1) Place Astragalus, Salvia miltiorrhiza, Ophiopogon japonicus, and Dendrobium officinale into an extraction tank, add water, and soak for half an hour; (2) heating the water in the extraction tank to boiling, boiling for 1 hour, and filtering to obtain decoction A; (3) Add water to the extraction tank, boil over low heat, repeat step (2), and filter to obtain decoction B; (4) Combine the filtered decoction A and decoction B, centrifuge at 3000 rpm for 10 min, and collect the supernatant; (5) Cooling, packaging, and sterilizing the supernatant obtained in step (4) for later use; and obtaining the Chinese medicine composition.

3. Use of the traditional Chinese medicine composition for treating Sjögren's syndrome according to claim 1 in preparing a medicine for treating Sjögren's syndrome.

4. The use according to claim 3, characterized in that The dosage form of the medicine is tablet, capsule, granule or spray.

5. The use according to claim 3, characterized in that The traditional Chinese medicine composition is added with a medically acceptable drug carrier to prepare a preparation.

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