Traditional Chinese medicine compound for treating sicca syndrome as well as preparation method and application of traditional Chinese medicine compound

Abstract

The invention provides a traditional Chinese medicine compound for treating sicca syndrome and a preparation method and application thereof, and belongs to the technical field of traditional Chinese medicine compounds. The traditional Chinese medicine compound provided by the invention has the effects of eliminating dampness and heat, removing blood stasis and promoting salivation, can safely and effectively treat sicca syndrome by improving functions of exocrine glands and regulating immunity, belongs to a pure traditional Chinese medicine preparation, and has no toxic or side effect. The traditional Chinese medicine compound is proved to be capable of remarkably relieving the sicca syndrome, relieving dry mouth and eye symptoms of a patient, relieving fatigue, regulating and controlling a plurality of signals taking RAGE / TLR4 as a core, inhibiting excessive activation of an HMGB1-RAGE / TLR4-NF-kappa B pro-inflammatory pathway from gene and protein levels, inhibiting generation of pro-inflammatory cytokines, reducing inflammatory response and improving the immunity of the patient. The effect of treating the sicca syndrome is achieved. By acting on STAT3 phosphorylation, Tfh proliferation is inhibited, IL-21 secretion is reduced, humoral immune disorder is relieved, and the sicca syndrome is treated, so that the condition of the sicca syndrome is relieved, and the prognosis of the sicca syndrome is improved.

Description

Technical Field

[0001] The present invention relates to the technical field of Chinese medicinal compound prescriptions, and in particular to a Chinese medicinal compound prescription for treating Sjögren's syndrome, and a preparation method and application thereof. Background Art

[0002] Sjögren's syndrome is an autoimmune disease that invades the exocrine glands. It is characterized by high lymphocytic infiltration of the lacrimal glands and salivary glands, and clinically manifested by dry mouth and eyes. It may also be accompanied by systemic involvement symptoms such as arthritis, rash, rampant dental caries, interstitial lung disease and other symptoms. The prevalence of this disease in the Chinese population is 0.33% to 0.77%, and its incidence rate has increased year by year, seriously affecting the quality of life of patients. However, most of the drugs on the market currently use symptomatic treatment or immunosuppressants, and there are no drugs specifically for this disease. However, due to uncertain efficacy or large toxic side effects, Sjögren's syndrome has become "incurable". Therefore, finding a safe and effective way to improve Sjögren's syndrome is what clinicians are looking forward to.

[0003] Sjögren's syndrome belongs to the category of "dryness and numbness" in traditional Chinese medicine. Most clinical doctors believe that the root cause of this disease is the loss of yin and fluid. However, with the deepening of modern doctors' understanding of this disease, the pathogenesis of "dryness" and "dampness" has received more and more attention. The "dryness" symptoms of Sjögren's syndrome are closely related to the metabolism of body fluids, and are closely related to the important organs involved in the movement and distribution of water, such as the spleen, lungs, kidneys, liver, triple energizer and bladder. Unfavorable metabolism of body fluids will produce "dampness". "Dampness" as a pathological product and pathogenic factor further hinders the movement of body fluids, resulting in the manifestation of "dryness". "Dampness" turns from cold to heat. From cold to cold and dampness, it is easy to hurt yang, and there is no warming to promote the distribution of body fluids, which will cause "dryness"; dampness stays and turns into heat, dampness and heat accumulate, block the qi mechanism, and consume body fluids, which will cause "dryness". Clinical diagnosis and treatment should carefully distinguish the severity of dampness and body fluid deficiency, distinguish the dryness and dampness in different organs and parts, and the emphasis of cold and heat, so as to accurately treat different pathogenesis. Understanding the pathogenesis of Sjögren's syndrome from the perspective of "dryness and dampness as co-morbidities" theory can guide the treatment of Sjögren's syndrome with traditional Chinese medicine, and help traditional Chinese medicine to exert greater advantages in the clinical diagnosis and treatment of Sjögren's syndrome. There are certain difficulties in the treatment of Sjögren's syndrome with Western medicine, and there is currently a lack of effective drugs proven by high-quality evidence-based medicine. Traditional Chinese medicine has unique advantages in the treatment of Sjögren's syndrome.

[0004] In the prior art, the Chinese patent database discloses several compound Chinese medicines for treating Sjögren's syndrome, such as CN101804174B, CN102813832B, CN102526596A, CN102772671A, CN103055034A, etc. The drugs disclosed in the above patents are all based on the principle of invigorating qi and nourishing yin. Even if they are also used to nourish dryness and heat, their effects are minimal. There is no Chinese patent medicine for intervening Sjögren's syndrome on the market. Therefore, it is necessary to provide a Chinese medicine compound for treating Sjögren's syndrome that can clear dampness and heat, remove blood stasis and promote the production of body fluid, and is safe and effective. Summary of the invention

[0005] The purpose of the present invention is to provide a Chinese medicine compound for treating Sjögren's syndrome and its preparation method and application. The Chinese medicine compound provided by the present invention has the functions of clearing away damp-heat, blood stasis and promoting body fluid, and can treat both the symptoms and the root cause of Sjögren's syndrome safely and effectively.

[0006] In order to achieve the above-mentioned object of the invention, the present invention provides the following technical solutions:

[0007] The invention provides 10 parts of Artemisia annua, 10 parts of white peony root, 10 parts of scutellaria baicalensis, 10 parts of dried tangerine peel, 10 parts of dendrobium and 10 parts of salvia miltiorrhiza.

[0008] The present invention also provides a method for preparing the Chinese medicinal compound for treating Sjögren's syndrome according to the above technical solution, comprising the following steps:

[0009] (1) In an extraction tank, Artemisia annua, white peony root, scutellaria baicalensis, dried orange peel, dendrobium officinale, salvia miltiorrhiza and pure water are mixed, and the drugs are immersed in the pure water for soaking pretreatment to obtain a pretreated mixture;

[0010] (2) heating the pretreated mixture to boiling, and sequentially performing boiling treatment and filtering to obtain a first decoction liquid, and retaining the solid mixture in an extraction tank;

[0011] (3) adding pure water to the extraction tank containing the solid mixture in step (2), slowly heating it to boiling, and repeating the operation of step (2) to obtain a second decoction liquid;

[0012] (4) combining the first decoction obtained in step (2) and the second decoction obtained in step (3), and sequentially performing centrifugation, cooling, subpackaging and sterilization to obtain a Chinese medicine compound for treating Sjögren's syndrome.

[0013] Preferably, the amount of pure water used in step (1) is sufficient to cover the raw materials including Artemisia annua, white peony root, scutellaria baicalensis, dried tangerine peel, dendrobium and salvia miltiorrhiza.

[0014] Preferably, the soaking pretreatment in step (1) is soaking at room temperature for 20 to 40 minutes.

[0015] Preferably, the boiling treatment time in step (2) is 0.5 to 3 hours.

[0016] Preferably, the centrifugal speed in step (4) is 2000-3500 r / min, and the centrifugal time is 8-14 min.

[0017] Preferably, the concentration of the total effective drug ingredients in the traditional Chinese medicine compound for treating Sjögren's syndrome in step (4) is 0.4-0.8 g / mL.

[0018] The present invention also provides the use of the traditional Chinese medicine compound for treating Sjögren's syndrome described in the above technical solution in preparing a medicine for treating Sjögren's syndrome.

[0019] Preferably, the dosage form of the Chinese herbal compound for treating Sjögren's syndrome is tablets, capsules, granules or sprays.

[0020] Preferably, the Chinese herbal compound for treating Sjögren's syndrome is prepared as a preparation using a drug carrier.

[0021] The invention provides a Chinese medicine compound for treating Sjögren's syndrome, which has the effects of clearing dampness and heat, removing blood stasis and promoting body fluid, improving exocrine gland function, regulating immunity, and safely and effectively treating Sjögren's syndrome, belonging to a pure Chinese medicine preparation with no toxic side effects. The Chinese medicine compound of the invention uses Artemisia annua and White Peony Root as the main medicine. Artemisia annua is cold but does not hurt the spleen and stomach, and is good at clearing dampness and heat in the liver and gallbladder; White Peony Root restrains yin and softens the liver to prevent yin injury, is slightly bitter and can replenish yin, is slightly sour and can astringe, treats blood deficiency and promotes nutrient qi, and is good at nourishing liver yin and replenishing liver blood; Artemisia annua and White Peony Root are compatible, clearing dampness and heat and astringing yin and nourishing blood are carried out in parallel, and both attack and supplement are used, which can dredge water channels and replenish the source of body fluid. Scutellaria baicalensis is bitter and cold, clears heat and dries dampness, is good at clearing dampness and heat in the upper and middle two jiaos, Artemisia annua and Scutellaria baicalensis are compatible, Artemisia annua clears the stagnant heat of Shaoyang, and Scutellaria baicalensis clears the evil heat of the gallbladder, and the two medicines are used together to strengthen the power of clearing heat and removing dampness. Tangerine peel is warm in nature, can invigorate the spleen and replenish qi, and can regulate qi and remove stagnation. Dendrobium is sweet, light and slightly cold, and is good at clearing the stomach and promoting fluid production. It is used with Salvia miltiorrhiza to promote blood circulation and remove blood stasis. The whole prescription clears heat without promoting dampness, removes dampness without hurting yin, nourishes yin without nourishing greasiness, and plays the role of clearing dampness and heat, removing blood stasis and promoting fluid production. The present invention conducts in-depth research on the pharmacological activity of the Chinese medicine compound and the treatment of Sjögren's syndrome, and confirms that the Chinese medicine compound can significantly relieve Sjögren's syndrome, relieve the symptoms of dry mouth and dry eyes of patients, relieve fatigue, regulate multiple signals with RAGE / TLR4 as the core, inhibit the overactivation of the HMGB1-RAGE / TLR4-NF-κB pro-inflammatory pathway from the gene and protein levels, inhibit the production of pro-inflammatory cytokines, reduce inflammatory response, and play a therapeutic effect on Sjögren's syndrome. By acting on STAT3 phosphorylation to inhibit Tfh proliferation, reduce IL-21 secretion, alleviate humoral immune disorders, treat Sjögren's syndrome, thereby alleviating the condition of Sjögren's syndrome and improving the prognosis of Sjögren's syndrome. BRIEF DESCRIPTION OF THE DRAWINGS

[0022] Figure 1 The results of the effects of different concentrations of Qingshao Yiquan Drink on the cell viability of THP-1 macrophages in the present invention are shown in FIG. Figure 1 As shown, Figure 1 From left to right, they are 12h, 24h and 48h.

[0023] Figure 2 It is a bar graph of the relative expression of RAGE, TLR4, NF-κB p65, IL-1β, TNF-α, and IL-6 mRNA in each group of THP-1 macrophages in the present invention, wherein Figure 2 A to F in the middle are RAGE, TLR4, NF-κB p65, IL-1β, TNF-α, and IL-6 mRNA in each group, respectively;

[0024] Figure 3 The electrophoresis diagram of each group of THP-1 macrophage proteins in the present invention, wherein: Figure 3 A in the middle is the electrophoresis diagram of RAGE and TLR4 proteins in each group. Figure 3 Middle B is the electrophoresis diagram of p-NF-κBp65 and NF-κB p65 proteins in each group;

[0025] Figure 4 This is a bar graph comparing the relative expression of THP-1 cell proteins in each group of the present invention, wherein: Figure 4 A to D are comparisons of the expression levels of RAGE, TLR4, p-NF-κB p65, and p-NF-κB p65 / NF-κB p65 proteins in each group of cells, respectively;

[0026] Figure 5 It is a bar graph of the concentration of pro-inflammatory cytokines in the cell culture supernatant of each group in the present invention, wherein, Figure 5 A to C in the middle are the levels of IL-1β, TNF-α, and IL-6 in the cell culture supernatant of each group;

[0027] Figure 6 This is a statistical bar chart showing the difference in Tfh expression after intervention with different doses of Qingshao Yiquan Decoction in the present invention.

[0028] Figure 7 The flow cytometric diagram of the intervention of Tfh by Qingshao Yiquan Decoction described in the present invention

[0029] Figure 8 The results of the effects of different doses of Qingshao Yiquan Decoction on IL-21 expression in the present invention are shown in FIG.

[0030] Fig. 9 This is a graph showing the effect of different doses of Qingshao Yiquan Decoction on the expression of STAT3 and pSTAT3 in the present invention. Fig. 9From left to right in the middle are STAT3 percentage, pSTAT3 percentage, and STAT3 phosphorylation rate;

[0031] Fig.10 This is a flow cytometric graph of the Qingshao Yiquan Decoction described in the present invention intervening in STAT3 and pSTAT3 expression. DETAILED DESCRIPTION

[0032] The present invention provides a Chinese medicinal compound for treating Sjögren's syndrome. The Chinese medicinal compound comprises the following raw materials, in parts by weight, which are prepared by extraction and decoction:

[0033] 10 parts of Artemisia annua, 10 parts of the white peony root, 10 parts of scutellaria baicalensis, 10 parts of dried tangerine peel, 10 parts of dendrobium, 10 parts of salvia miltiorrhiza.

[0034] In the present invention, unless otherwise specified, the raw materials used are conventional commercial products in the art.

[0035] The present invention also provides a method for preparing the Chinese medicinal compound for treating Sjögren's syndrome according to the above technical solution, comprising the following steps:

[0036] (1) In an extraction tank, Artemisia annua, white peony root, scutellaria baicalensis, dried orange peel, dendrobium officinale, salvia miltiorrhiza and pure water are mixed, and the drugs are immersed in the pure water for soaking pretreatment to obtain a pretreated mixture;

[0037] (2) heating the pretreated mixture to boiling, and sequentially performing boiling treatment and filtering to obtain a first decoction liquid, and retaining the solid mixture in an extraction tank;

[0038] (3) adding pure water to the extraction tank containing the solid mixture in step (2), slowly heating it to boiling, and repeating the operation of step (2) to obtain a second decoction liquid;

[0039] (4) combining the first decoction obtained in step (2) and the second decoction obtained in step (3), and sequentially performing centrifugation, cooling, subpackaging and sterilization to obtain a Chinese medicine compound for treating Sjögren's syndrome.

[0040] In the present invention, the amount of pure water used is sufficient to cover the raw materials including Artemisia annua, white peony root, scutellaria baicalensis, dried orange peel, dendrobium and salvia miltiorrhiza. In the present invention, the soaking pretreatment is preferably soaked at room temperature for 20 to 40 minutes. The present invention can soften the medicinal materials through soaking pretreatment, which is helpful to improve the release of effective ingredients during decoction.

[0041] In the present invention, the boiling treatment time is preferably 0.5 to 3 hours, more preferably 1 to 2 hours. The present invention uses boiling treatment to improve the absorption and utilization of active ingredients in the body.

[0042] The present invention has no particular limitation on the filtering method, and the solid-liquid separation can be achieved by adopting the technical solutions well known in the art.

[0043] In the present invention, the rotation speed of the centrifuge is preferably 2000-3500 r / min, and the time of the centrifuge is preferably 8-14 min. The present invention realizes solid-liquid separation by centrifugation, and removes solid particles and impurities in the combined decoction liquid.

[0044] The present invention has no particular limitation on the cooling, packaging and sterilization methods, and any technical solution known in the art may be used.

[0045] In the present invention, the concentration of the total effective drug components in the traditional Chinese medicine compound for treating Sjögren's syndrome is preferably 0.4-0.8 g / mL.

[0046] The present invention also provides the use of the traditional Chinese medicine compound for treating Sjögren's syndrome described in the above technical solution in preparing a medicine for treating Sjögren's syndrome.

[0047] In the present invention, the dosage form of the Chinese herbal compound for treating Sjögren's syndrome is preferably tablets, capsules, granules or sprays.

[0048] In the present invention, the Chinese medicinal compound for treating Sjögren's syndrome is preferably prepared by using a drug carrier.

[0049] The technical solutions in the present invention will be described clearly and completely below in conjunction with the embodiments of the present invention. Obviously, the described embodiments are only a part of the embodiments of the present invention, not all of them. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0050] Example 1

[0051] A Chinese medicinal compound for treating Sjögren's syndrome, wherein the Chinese medicinal compound comprises the following raw materials by weight, which are prepared by extraction and decoction:

[0052] Artemisia annua 10g, white peony root 10g, scutellaria baicalensis 10g, tangerine peel 10g, dendrobium 10g, salvia miltiorrhiza 10g.

[0053] The preparation method of the above-mentioned Chinese medicinal compound for treating Sjögren's syndrome comprises the following steps:

[0054] (1) In an extraction tank, mix Artemisia annua, white peony root, scutellaria baicalensis, dried orange peel, dendrobium officinale, salvia miltiorrhiza and 1000 mL of pure water, immerse the drugs in the pure water, and perform soaking pretreatment for 30 minutes to obtain a pretreated mixture;

[0055] (2) heating the pretreated mixture to boiling and concentrating it to 100 mL, boiling it for 1 h and filtering it in sequence to obtain a first decoction solution, and the solid mixture is retained in the extraction tank, and the final solid mixture concentration is 0.6 g / mL;

[0056] (3) adding pure water to the extraction tank containing the solid mixture in step (2), slowly heating it to boiling, and repeating the operation of step (2) to obtain a second decoction liquid;

[0057] (4) combining the first decoction obtained in step (2) and the second decoction obtained in step (3), and sequentially performing centrifugation, cooling, subpackaging and sterilization to obtain a Chinese herbal medicine for treating Sjögren's syndrome, wherein the concentration of total active drug ingredients in the Chinese herbal medicine for treating Sjögren's syndrome is 0.6 g / mL;

[0058] The centrifugal speed is 3000 r / min, and the centrifugal time is 10 min.

[0059] The Chinese medicine compound for treating Sjögren's syndrome prepared in Example 1 is prepared into a Chinese medicine compound freeze-dried powder by the following method, which is also named Qingshao Yiquanyin: The Chinese medicine compound for treating Sjögren's syndrome prepared in Example 1 is precooled at 4°C, cooled and filtered, and then divided into freeze-dried bottles, covered with plastic wrap, pierced with holes on the surface, and placed in a -80°C refrigerator for freezing. The vacuum freeze dryer is precooled to -50°C, and the condensed liquid medicine is placed in the freeze dryer. After vacuum low-temperature drying for 48 hours, Qingshao Yiquanyin freeze-dried powder is prepared (finally 7.14g of freeze-dried powder is obtained, and each gram of freeze-dried powder is equivalent to about 8.4g of the original medicinal material). The prepared freeze-dried powder is divided and stored, and when used, it is fully dissolved with complete culture medium and filtered with a 0.22μm filter.

[0060] In vitro experiment 1

[0061] The above-prepared Chinese medicine compound freeze-dried powder was used in an in vitro experiment for treating Sjögren's syndrome, and the specific method was as follows:

[0062] (1) PMA was used to induce THP-1 cells to differentiate into macrophages; recombinant human HMGB1 (rhHMGB1) was used to induce macrophages to establish an inflammatory model;

[0063] (2) The CCK-8 method was used to detect the effects of Qingshao Yiquanyin at concentrations of 0, 3.25 μg / mL, 62.5 μg / mL, 125 μg / mL, 250 μg / mL, 500 μg / mL, and 1000 μg / mL on the viability of THP-1 macrophages at 12 h, 24 h, and 48 h, respectively, to determine the optimal induction time and concentration range; the effects of different concentrations of Qingshao Yiquanyin on the viability of THP-1 macrophages were detected as shown in the figure. Figure 1 As shown, Figure 1From left to right, they are 12h, 24h and 48h. Figure 1 It can be seen that compared with the blank group, the cell survival rate of THP-1 macrophages stimulated by 1000μg / mL Qingshao Yiquanyin freeze-dried powder solution was significantly reduced after 24h and 48h (P<0.01, P<0.05); compared with the blank group, the cell survival rate of THP-1 macrophages stimulated by 500μg / mL Qingshao Yiquanyin freeze-dried powder solution had a downward trend after 24h and 48h (P>0.05). Compared with the blank group, the 125μg / mL Qingshao Yiquanyin freeze-dried powder solution stimulated THP-1 macrophages for 48h and showed a significant stimulating effect on cell proliferation (P<0.01), while the intervention for 24h did not show a stimulating effect on cell proliferation; the concentration and time range of Qingshao Yiquanyin freeze-dried powder solution stimulating THP-1 macrophages within 24h with a concentration lower than 125μg / mL is relatively safe.

[0064] (3) THP-1 macrophages were divided into a blank group, a model group, and low-, medium-, and high-dose Qingshao Yiquanyin groups; the blank group was not treated with any treatment, the model group was added with 500 ng / mL rhHMGB1 culture medium, and the dosages of the low-, medium-, and high-dose Qingshao Yiquanyin groups were 31.25 μg / mL, 62.5 μg / mL, and 125 μg / mL, respectively;

[0065] The PCR method was used to detect the expression of RAGE, TLR4, NF-κB p65, IL-1β, TNF-α, and IL-6 mRNA in each group. The WB method was used to detect the expression of RAGE, TLR4, NF-κB p65, and phosphorylated NF-κB p65 proteins in each group. The ELISA method was used to detect the levels of IL-1β, TNF-α, and IL-6 in the cell culture supernatant of each group.

[0066] The test results of in vitro experiment 1 are:

[0067] (1) Figure 2 The relative expression levels of RAGE, TLR4, NF-κB p65, IL-1β, TNF-α, and IL-6 mRNA in THP-1 macrophages of each group are shown in the bar graph. Figure 2 A to F are the mRNAs of RAGE, TLR4, NF-κB p65, IL-1β, TNF-α and IL-6 in each group, respectively. Figure 2It can be seen that the relative expression levels of RAGE, TLR4, NF-κB p65, IL-1β, TNF-α, and IL-6 mRNA in each group of cells were determined by PCR: the relative expression levels of RAGE, TLR4, NF-κB p65, IL-1β, TNF-α, and IL-6 mRNA in the model group were higher than those in the blank group (P<0.01, P<0.05); the relative expression levels of RAGE, TLR4, and TNF-α mRNA in the low, medium, and high dose groups of Qingshao Yiquan Drink were lower than those in the model group (P<0.01, P<0.05); the relative expression levels of NF-κB p65, IL-1β, and IL-6 mRNA in the medium and high dose groups of Qingshao Yiquan Drink were lower than those in the model group (P<0.05, P<0.01).

[0068] (2) Figure 3 The electrophoresis diagrams of THP-1 macrophage proteins in each group are shown in Figure 2. Figure 3 A in the middle is the electrophoresis diagram of RAGE and TLR4 proteins in each group. Figure 3 Middle B is the electrophoresis diagram of p-NF-κBp65 and NF-κB p65 proteins in each group. Figure 4 It is a bar chart comparing the relative expression of THP-1 cell proteins in each group, among which, Figure 4 A to D are the comparison of the expression levels of RAGE, TLR4, p-NF-κB p65, and p-NF-κB p65 / NF-κB p65 proteins in each group of cells. Figure 3 and Figure 4 It can be seen that the Western blot method was used to determine the expression of RAGE, TLR4, NF-κB p65 and phosphorylated NF-κB p65 proteins in THP-1 macrophages of each group: the relative expression levels of RAGE, TLR4 and phosphorylated NF-κB p65 proteins in the model group were higher than those in the blank group (P<0.01); the relative expression levels of RAGE, TLR4 and phosphorylated NF-κB p65 proteins in the medium and high dose groups of Qingshao Yiquan Drink were lower than those in the model group (P<0.01); the p-NF-κB p65 / NF-κB p65 protein ratios in the model group and the low and medium dose groups of Qingshao Yiquan Drink were higher than those in the blank group (P<0.01, P<0.05); the p-NF-κB p65 / NF-κB p65 protein ratio in the high dose group of Qingshao Yiquan Drink was lower than that in the model (P<0.01).

[0069] (3) Figure 5 It is a bar graph of the concentration of pro-inflammatory cytokines in the cell culture supernatant of each group, among which, Figure 5 A to C are the levels of IL-1β, TNF-α, and IL-6 in the cell culture supernatant of each group. Figure 5It can be seen that the concentrations of IL-1β, TNF-α and IL-6 in the cell culture supernatant were detected by ELISA: ① IL-1β: the IL-1β levels in the model group and the low-dose group of the compound were higher than those in the blank group (P<0.01, P<0.05); the IL-1β levels in the low, medium and high-dose groups of the Qingshao Yiquan Drink were lower than those in the model group (P<0.01); ② TNF-α: the TNF-α levels in the model group and the low and medium-dose groups of the compound were higher than those in the blank group (P<0.05); the TNF-α level in the high-dose group of the Qingshao Yiquan Drink was lower than that in the model group (P<0.05); ③ IL-6: the IL-6 levels in the model group and the low-dose group of the compound were higher than those in the blank group (P<0.05, P<0.01); the IL-6 levels in the medium and high-dose groups of the Qingshao Yiquan Drink were lower than those in the model group (P<0.01, P<0.05).

[0070] The test results of the above-mentioned in vitro experiment 1 show that: (1) The principle of medication for the syndrome of dryness and dampness in Sjögren's syndrome is to clear away dampness and heat, and to remove blood stasis and promote the production of body fluid. The Chinese medicine compound prepared in Example 1 of the present invention is an empirical basic prescription for treating the syndrome of dryness and dampness in Sjögren's syndrome; (2) The compound may exert a therapeutic effect on Sjögren's syndrome by regulating multiple signal pathways with RAGE / TLR4 as the core; (3) HMGB1-mediated inflammatory response is involved in the pathological process of Sjögren's syndrome, and inhibiting HMGB1-mediated inflammatory response is a new target for the treatment of Sjögren's syndrome; (4) The compound can inhibit the overactivation of the HMGB1-RAGE / TLR4-NF-κB pro-inflammatory pathway at the gene and protein levels, inhibit the production of pro-inflammatory cytokines, reduce the inflammatory response, and exert a therapeutic effect on Sjögren's syndrome.

[0071] In vitro experiment 2

[0072] The present invention explores the mechanism of action of the Chinese herbal compound prepared in Example 1 in intervening with Tfh in the treatment of Sjögren's syndrome, specifically: extracting peripheral blood PBMCs from the normal control group and patients with Sjögren's syndrome with dryness and dampness, isolating naive CD4+T cells, inducing differentiation of Tfh, observing the Tfh differentiation rate of the normal control group and Sjögren's syndrome with dryness and dampness, the control group is used as the naive CD4+T cell group as the undifferentiated Tfh cells and the cells differentiated into Tfh for comparison, the experimental group is used as the Tfh group, and the high, medium and low doses of the Qingshao Yiquan Drink are 125 μg / mL, 62.5 μg / mL and 31.25 μg / mL, respectively, to intervene in Tfh, as the high, medium and low dose groups of Qingshao Yiquan Drink, observing the effect of the Chinese herbal compound prepared in Example 1 on the proliferation of Tfh and exploring its mechanism.

[0073] The test results of the above in vitro experiment 2 are:

[0074] (1) Figure 6It is a statistical bar chart showing the difference in Tfh expression after intervention with different doses of Qingshao Yiquan Decoction. Figure 7 This is the flow cytometric diagram of the Qingshao Yiquan Decoction intervening in Tfh. Figure 6 and Figure 7 It can be seen that compared with the Tfh group, the high, medium and low doses of Qingshao Yiquan Drink can effectively inhibit Tfh proliferation (P < 0.05), and the higher the dose of the compound, the higher the Tfh inhibition rate;

[0075] (2) Figure 8 The figure is a result diagram showing the effect of different doses of Qingshao Yiquan Decoction on IL-21 expression. Figure 8 It can be seen that compared with the Tfh group, the expression of IL-21 in the high, medium and low dose groups of Qingshao Yiquan Drink was also significantly reduced (P < 0.05), and the difference in IL-21 expression between the high, medium and low dose groups of Qingshao Yiquan Drink was also statistically significant (P < 0.05), and the higher the dose of Qingshao Yiquan Drink, the lower the IL-21 expression;

[0076] (3) Fig. 9 The results of the effects of different doses of Qingshao Yiquan Decoction on the expression of STAT3 and pSTAT3 are shown in FIG. Fig. 9 From left to right in the figure are STAT3 percentage, pSTAT3 percentage and STAT3 phosphorylation rate. Fig.10 The figure is a flow cytometric graph of the Qingshao Yiquan Decoction intervening in STAT3 and pSTAT3 expression. Fig. 9 and Fig.10 It can be seen that the expression of STAT3 and pSTAT3 in the Tfh group was higher than that in the naive CD4+T cell group (P<0.05) and the different dose groups of the compound (P<0.05), and there was no significant difference in the expression of STAT3 and pSTAT3 in the high, medium and low dose groups of the Qingshao Yiquan Drink (P>0.05). The pSTAT3 / STAT3 phosphorylation rate of the medium and high dose groups of the Qingshao Yiquan Drink was inhibited compared with the Tfh group.

[0077] The test results of the above-mentioned in vitro experiment 2 show that: (1) the Qingshao Yiquan Decoction can treat Sjögren's syndrome by inhibiting the abnormal proliferation of Tfh and reducing the secretion of IL-21 to regulate humoral immune disorders; (2) the Qingshao Yiquan Decoction can inhibit the proliferation of Tfh by reducing STAT3 phosphorylation, alleviate humoral immune disorders, treat Sjögren's syndrome, thereby alleviating the symptoms of Sjögren's syndrome and improving the prognosis of Sjögren's syndrome.

[0078] In summary, the Chinese medicine compound provided by the present invention can improve the symptoms of Sjogren's syndrome; in the theory of traditional Chinese medicine, the compound can clear away dampness and heat and nourish yin and blood at the same time, attack and supplement at the same time, both dredge the water channel and replenish the source of body fluid, and secondly, tonify the spleen and replenish qi, regulate qi and remove stagnation, transform essence and micro, so that the blood circulation and body fluid can be moisturized; thirdly, it can remove blood stasis and dredge the meridians, and the qi mechanism is unobstructed, so that the body fluid can be applied; the whole prescription clears away heat without helping dampness, removes dampness without hurting yin, nourishes yin without nourishing greasiness, and plays the role of "clearing dampness and heat, removing blood stasis and promoting body fluid". The Chinese medicine compound of the present invention can achieve the relief of Sjogren's syndrome by inhibiting abnormal proliferation of Tfh, reducing IL-21 secretion, and regulating immune disorders. The Chinese medicine compound of the present invention can reduce STAT3 phosphorylation to inhibit Tfh proliferation, alleviate humoral immune disorders, and treat Sjogren's syndrome dryness and dampness syndrome, thereby relieving Sjogren's syndrome and improving the prognosis of Sjogren's syndrome. The Chinese medicinal compound of the present invention can inhibit the overactivation of the HMGB1-RAGE / TLR4-NF-κB pro-inflammatory pathway at the gene and protein levels, inhibit the production of pro-inflammatory cytokines, reduce the inflammatory response, and exert a therapeutic effect on Sjögren's syndrome.

[0079] The above is only a preferred embodiment of the present invention. It should be pointed out that for ordinary technicians in this technical field, several improvements and modifications can be made without departing from the principle of the present invention. These improvements and modifications should also be regarded as the scope of protection of the present invention.

Claims

1. A Chinese medicinal compound for treating Sjögren's syndrome, characterized in that: The Chinese medicine compound comprises the following raw materials by weight, which are prepared by extraction and decoction: 10 parts of Artemisia annua, 10 parts of the white peony root, 10 parts of scutellaria baicalensis, 10 parts of dried tangerine peel, 10 parts of dendrobium, 10 parts of salvia miltiorrhiza.

2. A method for preparing the Chinese medicinal compound for treating Sjögren's syndrome according to claim 1, characterized in that: The following steps are involved: (1) In an extraction tank, Artemisia annua, white peony root, scutellaria baicalensis, dried orange peel, dendrobium officinale, salvia miltiorrhiza and pure water are mixed, and the drugs are immersed in the pure water for soaking pretreatment to obtain a pretreated mixture; (2) heating the pretreated mixture to boiling, and sequentially performing boiling treatment and filtering to obtain a first decoction liquid, and retaining the solid mixture in an extraction tank; (3) adding pure water to the extraction tank containing the solid mixture in step (2), slowly heating it to boiling, and repeating the operation of step (2) to obtain a second decoction liquid; (4) combining the first decoction obtained in step (2) and the second decoction obtained in step (3), and sequentially performing centrifugation, cooling, subpackaging and sterilization to obtain a Chinese medicine compound for treating Sjögren's syndrome.

3. The preparation method according to claim 2, characterized in that: The amount of pure water used in step (1) is sufficient to cover the raw materials including Artemisia annua, white peony root, scutellaria baicalensis, dried tangerine peel, dendrobium and salvia miltiorrhiza.

4. The preparation method according to claim 2, characterized in that: The soaking pretreatment in step (1) is performed at room temperature for 20 to 40 minutes.

5. The preparation method according to claim 2, characterized in that: The boiling treatment time in step (2) is 0.5 to 3 hours.

6. The preparation method according to claim 2, characterized in that: The centrifugal speed in step (4) is 2000-3500 r / min, and the centrifugal time is 8-14 min.

7. The preparation method according to claim 2, characterized in that: The concentration of the total effective drug components in the traditional Chinese medicine compound for treating Sjögren's syndrome in step (4) is 0.4-0.8 g / mL.

8. Use of the Chinese medicinal compound for treating Sjögren's syndrome according to claim 1 in preparing a drug for treating Sjögren's syndrome.

9. The use according to claim 8, characterized in that: The dosage form of the traditional Chinese medicine compound for treating Sjögren's syndrome is tablets, capsules, granules or sprays.

10. The use according to claim 8, characterized in that: The traditional Chinese medicine compound for treating Sjögren's syndrome is prepared into a preparation by using a drug carrier.

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