CN113116973B

CN113116973B - Novel traditional Chinese medicine compound for preventing and treating sicca syndrome as well as preparation method and application thereof

Info

Publication number: CN113116973B
Application number: CN202110491885.XA
Authority: CN
Inventors: 郝静超; 赵荣惠; 文凤
Original assignee: Kunming Medical University
Current assignee: Kunming Medical University
Priority date: 2021-05-06
Filing date: 2021-05-06
Publication date: 2023-01-24
Anticipated expiration: 2041-05-06
Also published as: CN113116973A

Abstract

The invention discloses a novel traditional Chinese medicine compound for preventing and treating sicca syndrome and a preparation method thereof, which are characterized in that Chinese herbal medicine components are reasonably matched, and the effective components of the formula comprise common selfheal fruit-spike, desmodium, spreading hedyotis herb, chrysanthemum and honeysuckle which are all Chinese herbal medicine components. The method comprises the following processes and steps: decocting Prunellae Spica, herba Lysimachiae Christinae, herba Hedyotidis Diffusae, flos Chrysanthemi, and flos Lonicerae with 5 times and 3 times of distilled water for 1 hr each time, mixing decoctions, and concentrating under reduced pressure. Animal experiment screening proves that the obtained concentrated solution has good effects of improving xerophthalmia, improving gland functions, reducing autoantibody level and resisting inflammation. The maximum dosage experiment proves that the concentrated solution has higher clinical safety. Can be used for preventing and treating xerosis syndrome mainly caused by xerophthalmia, lymphocyte invasion of secretory gland, and inflammation, and has no recurrence.

Description

Novel traditional Chinese medicine compound for preventing and treating sicca syndrome as well as preparation method and application thereof

Technical Field

The invention relates to a medical application of a traditional Chinese medicine compound for preventing and treating sicca syndrome, in particular to an application of the compound in treating the sicca syndrome.

Background

Sicca syndrome (A)

SS) is also known as sjogren's syndrome and is a chronic inflammatory autoimmune disease that involves primarily the exocrine glands. It is divided into primary and secondary categories. The typical clinical manifestations are that salivary gland and lacrimal gland are damaged, the function is reduced, and the symptoms of dry mouth and eyes are caused, and other exocrine glands and other organs outside gland body are involved to cause multi-system damage. There are a number of autoantibodies and hyperimmune globulin in the patient's serum. Sjogren's syndrome occurs well after menopause or in women between 30 and 40 years of age with an incidence of about 0.3 to 0.4% and up to 0.77% in the elderly population.

No radical cure method is available for the disease at present. In clinic, measures are mainly taken for improving symptoms, controlling and delaying the progress of tissue and organ damage caused by immune response and secondary infection. The treatment aims to relieve dry mouth and eyes. The western medicine clinical treatment mainly comprises substitution therapy and symptomatic treatment, wherein the substitution therapy adopts 'artificial tears', 'saliva substitutes' and the like as local treatment medicines for improving dry eyes and dry mouth. And symptomatic treatment such as systemic treatment with hydroxychloroquine sulfate, glucocorticoid, and immunosuppressant (such as cyclophosphamide). The use of biologies such as B lymphocyte monoclonal antibodies (e.g., CD20 and CD22 monoclonal antibodies) to increase residual salivary gland function stabilizes Ig G levels. Other approaches such as muscarinic agonists, stem cell therapies, etc. have also been investigated. But all are difficult to cure radically and easy to repeat.

Under the guidance of the holistic view and syndrome differentiation theory of traditional Chinese medicine, the traditional Chinese medicine has certain effect on preventing and treating sicca syndrome. Total glucosides of paeony is a classic and effective Chinese patent medicine for treating sicca syndrome at present, is an immunomodulator, and can adjust over-high or over-low immunity of a patient to a normal level. But when used alone, the traditional Chinese medicine composition has slow effect, can not quickly improve the dry mouth and eyes symptom, and can not quickly reduce the infiltration degree of gland lymphocytes and reduce the expression quantity of autoantibodies. In patent 201210296796.0, a traditional Chinese medicine compound taking radix paeoniae alba as one of main traditional Chinese medicines shows that the traditional Chinese medicine compound can effectively inhibit rheumatoid factors and antibody indexes, but has poor intervention effects on lymphocyte infiltration, autoantibody SSA, SSB, ANA, inflammation TNF-alpha, BAFFR factors and the like in gland with specificity to xerosis. In addition, some traditional Chinese medicine compound medicines are applied to the treatment of the sjogren syndrome, such as patents 201210345391.1 and 201610918771.8, but the traditional Chinese medicine compound medicines have the defects of more traditional Chinese medicine taste, more complex formula and preparation process, more dosage of single-day medicine and the like.

Disclosure of Invention

In view of the above situation, the present invention aims to provide an application of a traditional Chinese medicine compound for preventing and treating xerophthalmia, xerostomia, secretory gland function improvement and autoantibody level reduction and inflammation.

The invention relates to a traditional Chinese medicine compound, which is characterized in that the main active ingredients of the traditional Chinese medicine compound comprise common selfheal fruit-spike, desmodium, oldenlandia diffusa, chrysanthemum and honeysuckle.

The traditional Chinese medicine compound is characterized in that the main active ingredients comprise 2g-30g of selfheal, 2g-30g of desmodium, 2g-30g of spreading hedyotis herb, 2g-30g of chrysanthemum and 2g-30g of honeysuckle.

The preparation method of the traditional Chinese medicine compound is characterized by comprising the following steps of decocting 2g-30g of selfheal, 2g-30g of desmodium, 2g-30g of spreading hedyotis herb, 2g-30g of chrysanthemum and 2g-30g of honeysuckle in water, concentrating and mixing to obtain a finished product.

The Chinese herbal compound is a medicine for preventing or treating sjogren syndrome and a preparation method and application thereof.

The specific description is as follows:

the technical scheme for solving the problem is that the traditional Chinese medicine compound and the preparation method thereof are characterized by comprising the following steps:

the traditional Chinese medicine compound composition for treating sicca syndrome comprises the following raw medicines: prunellae Spica, herba Lysimachiae Christinae, herba Hedyotidis Diffusae, flos Chrysanthemi and flos Lonicerae.

The traditional Chinese medicine compound composition for treating sicca syndrome comprises the following raw material medicines in parts by weight: 2g to 30g of common selfheal fruit-spike, 2g to 30g of longhairy antenoron herb, 2g to 30g of spreading hedyotis herb, 2g to 30g of chrysanthemum and 2g to 30g of honeysuckle.

The traditional Chinese medicine compound composition for treating sicca syndrome is any one of dosage forms in pharmaceutics. The Chinese medicinal composition can be directly or indirectly added with pharmaceutically acceptable excipient to make into any dosage form in pharmacy.

Preferably, the dosage form is decoction, powder, pill, paste, tablet or effervescent tablet.

The preparation is prepared by the conventional process, such as:

the decoction is as follows: mixing the Chinese herbal pieces, soaking in 500mL water, decocting, filtering, mixing filtrates, concentrating to 100mL, and making into liquid. The raw materials are soaked for 20-30 minutes before being decocted generally, the medicines are decocted with normal water quality, each dose of the medicines is decocted for 2 times, 500mL is added for each time, and after the medicines are decocted, the filtrates are combined and concentrated.

The powder is as follows: pulverizing the raw materials, mixing, and making into powder.

The pill preparation comprises the following steps: the compound preparation is a spherical or sphere-like solid preparation prepared by adding any available adhesive or other auxiliary materials into the raw material powder or the raw material extract.

The paste is prepared by the following steps: decocting the raw materials in the compound with water or vegetable oil, filtering, and making into dosage form.

The preparation method comprises the following steps: pulverizing the raw materials, mixing, adding any binder or other adjuvants, and making into desired dosage form.

The effervescent tablet comprises: pulverizing the raw materials, mixing, adding binder or other adjuvants, effervescent disintegrant, and correctant, and making into desired dosage form.

The Chinese medicinal compound composition for treating sicca syndrome can also be prepared into teabags. And (3) subpackaging the coarse powder of the traditional Chinese medicine decoction pieces or part of the small pieces of the traditional Chinese medicine decoction pieces in the compound into cotton gauze tea bags or diamond tea bags according to the formula compatibility weight ratio. The tea bag is put into 100mL of boiled water with the temperature of 60-100 ℃ and soaked for 10-15 minutes, and then the tea bag can be drunk.

The pharmacological effects of the invention are as follows:

the formula of the invention selects the selfheal, the desmodium, the oldenlandia diffusa, the chrysanthemum and the honeysuckle, and is obtained by combining the traditional Chinese medicine theory and the modern pharmacology and repeatedly carrying out clinical verification. The effects of the traditional Chinese medicines are as follows:

spica Prunellae, pungent, bitter and cold in nature, can clear heat and purge fire, improve vision, dissipate nodulation and relieve swelling. Is used for treating goiter and lymphadenectasis, and also has antioxidant, anti-tumor, antiviral and immunity regulating effects;

oldenlandia diffusa, bitter, sweet, cold in flavor and non-toxic. Clear heat and remove toxicity, cure abscess, and remove dampness. Has antitumor, antibacterial, antiinflammatory, fibroblast proliferation inhibiting, immunity regulating, and nervous system protecting effects. For cough and asthma due to lung heat, tonsillitis, pharyngolaryngitis, venomous snake bite, etc., it is combined with jin Yin Hua to treat sore, furuncle, pyogenic infections, etc.

Honeysuckle flower, flos Lonicerae, with warm nature, bitter and sweet taste, is nontoxic. It enters lung and stomach meridians. Clearing away heat and toxic material, diminishing inflammation and relieving swelling. Can be used for treating wind-heat common cold or fever due to epidemic febrile disease, sunstroke, toxic heat, dysentery, carbuncle, furuncle, pharyngitis, and various infectious diseases. The extract has strong antiinflammatory, antibacterial and antipyretic effects.

Jin Qian Cao is slightly cold in nature, sweet and salty in flavor. It enters liver, gallbladder, kidney and bladder meridians. Has effects in clearing away heat and dampness, treating stranguria, relieving swelling, promoting diuresis, and eliminating jaundice. Has anti-inflammatory effect; has inhibitory effect on cellular immunity and humoral immunity; relaxing smooth muscle of blood vessel, and inhibiting platelet aggregation. The decoction and granule have antibacterial and antiinflammatory effects and are effective in inhibiting Staphylococcus aureus.

Chrysanthemum, slightly cold in nature, bitter and sweet in taste. It enters lung and liver meridians. Dispel wind and clear heat, pacify liver and improve vision, clear heat and remove toxicity. Can be used for treating wind-heat type common cold, headache, giddiness, conjunctival congestion, swelling and pain, dim eyesight, sore, carbuncle, and toxic swelling.

The compatibility of the selfheal, the oldenlandia diffusa and the honeysuckle in the composition enhances the effects of the prescription on improving xerophthalmia, improving gland functions, reducing autoantibody level and resisting inflammation.

Has the advantages that:

1. the main active ingredients in the traditional Chinese medicine compound of the invention are composed of selfheal, desmodium, oldenlandia diffusa, chrysanthemum and honeysuckle. In the formula, the selfheal is taken as a monarch drug and plays a role in treating main diseases; the oldenlandia diffusa is used as a ministerial drug to assist the main drug to play a role; the honeysuckle, the desmodium and the chrysanthemum are all adjuvant drugs, treat accompanying diseases and simultaneously eliminate the possible toxicity of monarch drugs and ministerial drugs; the herba Lysimachiae Christinae also serves as guiding drug, and has effects of guiding meridian downward and harmonizing Chinese medicinal compound properties.

The selfheal and the oldenlandia diffusa in the traditional Chinese medicine compound formula have the functions of obviously enhancing the treatment, eliminating stagnation, reducing swelling, resisting inflammation, detoxifying and regulating immunity. The 2 medicines have common property in performance and efficacy, mainly take the selfheal as the main component, take the oldenlandia diffusa as the auxiliary component, and can improve the effect and the curative effect of the selfheal.

The inventor finds that the Chinese herbal compound has better effect under the condition that the dosage of the selfheal and the oldenlandia is about 1, the proportion of other medicines can be changed, the influence on the drug effect is small, and the compound has the synergistic effect.

In conclusion, the invention conforms to the compatibility principle of the traditional Chinese medicines and has no incompatibility. No diet contraindication.

2. The traditional Chinese medicine used in the traditional Chinese medicine compound is common traditional Chinese medicine and has simple formula. It has small dosage and definite effect on preventing and treating sjogren's syndrome. Animal experiment screening proves that the obtained concentrated solution has good effects of improving xerophthalmia, improving gland functions, reducing autoantibody level and resisting inflammation. The maximum dosage experiment proves that the concentrated solution has the characteristics of higher clinical safety, long-term use and the like. It can be used for preventing and treating xerosis syndrome with clinical symptoms of xerophthalmia, lymphocyte invasion of secretory gland, and inflammation, and recurrence is not likely to occur. The formula of the invention has convenient and various taking methods, can be decocted into decoction for taking, and can also be prepared into tablets, pills and powder, thus being convenient for storage and carrying. Can be prepared into tea bags, buccal tablets, effervescent tablets and other dosage forms, and has obvious advantages in the aspect of preventing and treating the sicca syndrome.

3. The prescription of the invention has small dosage, safety and innocuity, is suitable for long-term use of patients with sicca syndrome, and has 15 days of treatment cycle. The invention has better anti-inflammatory effect, can maintain the structural integrity of gland tissues, reduce the invasion focus of lymphocytes in glands, reduce the expression level of autoantibodies in vivo and simultaneously can play a better anti-inflammatory role. Compared with the patent 201210296796.0, the formula disclosed by the invention can be used for remarkably reducing the lymphocyte infiltration degree in gland with xerosis specificity and remarkably reducing the expression quantity of autoantibody SSA, SSB, ANA and inflammation TNF-alpha and BAFFR.

Drawings

FIG. 1 is a graph showing the effect of the present invention on promoting the crystallization of mouse tear fluid during administration (200X);

FIG. 2 mouse spleen HE staining;

FIG. 3 standard curve for TNF- α in ELISA;

FIG. 4ELISA method SSA protein standard curve;

FIG. 5ELISA method SSB protein standard curve;

FIG. 6ELISA method ANA protein Standard Curve;

FIG. 7ELISA method BAFFR protein standard curve.

Detailed Description

The invention is further described in the following examples in connection with specific embodiments thereof, it is to be understood that these examples are included solely for the purpose of illustration and are not intended as limitations on the scope of the invention, as modifications of various equivalent forms of the invention which would occur to those skilled in the art upon reading the present disclosure are intended to be within the scope of the invention as defined in the appended claims.

2g-30g of selfheal, 2g-30g of desmodium, 2g-30g of spreading hedyotis herb, 2g-30g of chrysanthemum and 2g-30g of honeysuckle, and the five traditional Chinese medicinal materials are taken, extracted by three times of distilled water, filtered, combined with filtrate, and concentrated to 100mL.

Example 1

2g of selfheal, 2g of desmodium, 2g of spreading hedyotis herb, 2g of chrysanthemum and 2g of honeysuckle, the five traditional Chinese medicinal materials are taken, extracted by distilled water for three times, filtered, the filtrate is combined, and the filtrate is concentrated to 100mL.

Example 2

7.5g of selfheal, 4g of desmodium, 7.5g of spreading hedyotis herb, 12g of chrysanthemum and 18g of honeysuckle, the five traditional Chinese medicinal materials are taken, extracted by distilled water for three times, filtered, the filtrates are combined, and the filtrate is concentrated to 100mL.

Example 3

15g of selfheal, 12g of desmodium, 15g of spreading hedyotis herb, 10g of chrysanthemum and 15g of honeysuckle flower, the five traditional Chinese medicinal materials are taken, extracted by distilled water for three times, filtered, the filtrate is merged, and the filtrate is concentrated to 100ml.

Example 4

30g of selfheal, 30g of desmodium, 30g of spreading hedyotis herb, 30g of chrysanthemum and 30g of honeysuckle, the five traditional Chinese medicinal materials are taken, extracted by distilled water for three times, filtered, the filtrate is combined, and the filtrate is concentrated to 100mL.

Example 5

1. Experimental research on effect of traditional Chinese medicine compound extract on relieving dry eye

The specific implementation scheme is as follows: SPF grade Balb/C mice, 48, female, weighing 18-22g. The mice were randomly divided into 5 groups (n = 8) except the normal group, and the sicca syndrome mouse model was replicated using the scopolamine hydrochloride method with a model replication cycle of 10 days. The mice were divided into a normal control group, a negative control group, a positive drug group (total glucosides of paeony tablets, 0.182g/kg calculated by clinical dose), and groups of examples 1-3 in sequence. Treatment is given from d11, wherein the normal group and the negative control group are perfused with gastric saline, the positive drug group is perfused with gastric white peony root total glycoside liquid medicine, and the groups of examples 1-3 are perfused with test liquid with corresponding concentration. The gavage volume is 1mL/20g body weight, 1 time per day, and the gavage is continuously carried out for 15 days. During this period, tears were removed from the slide glass every 5 days and examined microscopically (200X) for tears crystallization. The weight change and water intake of the mice were measured every 2 days. The test results are shown in table 1, table 2 and fig. 1.

TABLE 1 weight gain in mice during dosing (g, mean + -SD)

Note: (vi) a set of negative models, ^* P<0.05or ^** P<0.01; vs. total glucosides of paeony group, ^# P<0.05or ^## P<0.01。

the results show (Table 1) that the body weight of mice in the negative model group was significantly reduced during the administration period compared with that in the normal group ( ^** P<0.05 ); mice in the total glucosides of paeony group and in the groups of examples 1-3 showed a significant weight-up trend from the 3 rd day (d 13) of administration, compared with the negative model group ( ^* P<0.05or ^** P<0.01 ); compared with the positive control group of total glucosides of paeony, the mice in the examples 1-2 have significantly lower body weight regain than the total glucosides of paeony group ( ^# P<0.05or ^## P<0.01 ); however, in example 3, there was no significant difference in the mouse body weight regain (P)>0.05 It shows that the effect of example 3 on the improvement of the body weight of the mice is equivalent to that of the positive control group.

TABLE 2 change in water intake of mice during dosing (mL, mean + -SD)

Note: (vi) a set of negative models, ^* P<0.05or ^** P<0.01；vs. total glucosides of paeony group, ^# P<0.05or ^## P<0.01。

the results show (Table 2) that the consumption of water by the mice in the negative model group was significantly increased during the administration period compared with the normal group ( ^* P<0.05or ^** P<0.01 ); the water intake of mice in the total glucosides of paeony group and the groups of examples 1-3 showed a significantly decreased trend after d13, compared to the negative model group; compared with the total glucosides of paeony positive group, the water intake of example 1 at d13 and d15 is significantly lower than that of the total glucosides of paeony group (45.0 + -4.3 mL and 35.0 + -3.9 mL), 32.4 + -3.8 mL and 29.0 + -3.4 mL, respectively: ( ^# P<0.05or ^## P<0.01). The water intake of the mice in the examples 2 and 3 is not obviously different from that of the white paeony root total glycoside group. It is demonstrated that examples 1-3 had an effect on reducing the water intake of mice comparable to that of the total glucosides of paeony group.

The results show that fern crystal form of model mouse tear crystals is lost (form IV) in irregular punctate form due to the lack of mucin. After 15 days of administration (d 25), the tear crystal forms were improved (both form II) in both the total glucosides of paeony group and the test group, except for the model group. Compared with the total glucosides of paeony positive group, the tear crystal forms of the test groups, the examples 2 and the examples 3 have no significant difference with the positive control group.

The test result shows that: the traditional Chinese medicine compound extract has the comprehensive effects of maintaining the weight of a mouse, reducing the water intake of the mouse, repairing the crystalline form of tears and relieving the dry eye disease or is superior to the total glucosides of paeony. The research result can reflect that the traditional Chinese medicine compound extract can prevent and treat dry eye diseases to a certain extent.

2. Experimental study on improving gland function of traditional Chinese medicine compound extract

The specific implementation scheme is as follows: SPF grade BALB/c mice 48, female, weighing 18-22g. Except for the normal group, the other mice were randomly divided into 5 groups (n = 8), and the sicca syndrome mouse model was replicated by a scopolamine hydrochloride method, with a model replication cycle of 10 days. The mice were divided into a normal control group, a negative control group, a positive drug group (total glucosides of paeony tablets, 0.182g/kg in terms of clinical dose) and groups of examples 1-3 in this order. The treatment is given from d11, wherein the normal group and the negative control group are filled with gastric lavage physiological saline, the positive control group is filled with gastric lavage total glucosides of paeony liquor, the test liquor with the corresponding concentration for the groups of the examples 1-3 is filled with gastric lavage, the gastric lavage volume is 1mL/20g of body weight, 1 time/day, and the gastric lavage is continuously carried out for 15 days. After the dosing period was over, the orbit was bled and the mice were sacrificed. Dissecting, respectively taking submaxillary gland tissue and salivary gland tissue, and placing in 4% neutral formaldehyde solution for 48h. Then, the slices are sliced after paraffin embedding treatment, and the thickness is 5 mu m; placing the slices in an oven for dry baking until paraffin is melted, immediately soaking in xylene, and repeating for 2 times; sequentially soaking in gradient ethanol (anhydrous, 95%, 90%, 85%, 80%, 75%, 70%) for 10min, and placing in double distilled water; boiling in citrate buffer solution with pH of 6.0 and concentration of 0.01mol/L for 10min for antigen retrieval; washing with flowing water, washing with distilled water for 3min, and soaking in hematoxylin dye solution for 5min; washing with flowing water, differentiating with 1% hydrochloric acid ethanol differentiation solution for 5s, and blue-staining in flowing water for 30min; soaking in 70% ethanol and 80% ethanol for 3min; staining with hematoxylin-eosin (HE), soaking in ethanol, and dehydrating for 3min; and sealing the piece by using xylene transparent neutral resin. Pathological changes such as spleen and the like are observed under a microscope, 5-8 pathological sections are randomly photographed under 100-fold and 200-fold visual fields respectively, and the lymphocyte infiltration degree is evaluated by adopting a Chisholm-Mason histological scoring standard. Grading the grading standard: 0 point, occasionally lymphocyte infiltration; 1 point = a small amount scattered in lymphocyte infiltrates; 2 points = moderate infiltration of lymphocytes (not foci) with mild parenchymal lesions; 3 points = 0 to 1 sporadic lymphocyte infiltration foci, 5 low power fields. With moderate parenchymal lesions; 4 points = 2-3 easily seen lymphocyte infiltrates, 5 low power fields with severe parenchymal lesions. Onset of disease is classified into more than 2 points. The results are shown in tables 3 and 4.

TABLE 3 mouse glandular tissue lymphocyte infiltration score (Mean + -SD)

Note: v. a set of negative models, ^* P<0.05or ^** P<0.01; vs. total glucosides of paeony group, ^# P<0.05or ^## P<0.01。

results show (Table 3)The infiltration degree of lymphocytes in submaxillary gland tissues of mice in the negative model group was 3.40. + -. 0.55 (compared with that in the normal group) ^** P<0.01 ); the infiltration degree of lymphocytes in the submaxillary gland tissues of the total glucosides of paeony group and the test group examples 1-3 was significantly reduced compared with that of the negative model group: ( ^* P<0.05or ^** P<0.01 2.40 +/-0.55, 2.60 +/-0.55, 1.80 +/-0.45 and 1.60 +/-0.55; the infiltration scores of the group of example 3 were significantly reduced compared to the total glucosides of paeony group ( ^# P<0.05). It is demonstrated that example 3 inhibits lymphocyte infiltration in submaxillary gland tissue better than the positive control total glucosides of paeony group.

The results show (Table 3) that the degree of infiltration of lymphocytes in salivary gland tissues of mice in the negative model group is 4.10. + -. 0.55 higher than that in the normal group ( ^** P<0.01 ); the degree of lymphocyte infiltration in salivary gland tissues of the total glucosides of paeony and the test groups of examples 1 to 3 was significantly reduced compared with that of the negative model group ( ^** P<0.01 2.80 + -0.57, 2.50 + -0.50, 1.80 + -0.57 and 1.40 + -0.42, respectively; the infiltration scores of the groups of examples 2-3 were all significantly reduced compared to the total glucosides of paeony group: ( ^# P<0.05or ^## P<0.01). The inhibition effect of the examples 2 and 3 on the lymphocyte infiltration of the salivary gland tissue is better than that of the positive control white paeony root total glycoside group.

The test results show that: the traditional Chinese medicine compound extract has better inhibiting effect on the infiltration degree of the lymph cells of submaxillary gland tissues and salivary gland tissues than positive total glucosides of paeony. The research result reflects that the traditional Chinese medicine compound extract has the functions of protecting the integrity of glandular tissues of secretory glands and maintaining the basic functions to a certain extent.

3. Test for reducing level of autoantibody by using compound traditional Chinese medicine extract

The specific implementation scheme is as follows: SPF grade BALB/c mice 48, female, weighing 18-22g. The mice were randomly divided into 5 groups (n = 8) except the normal group, and the sicca syndrome mouse model was replicated using the scopolamine hydrochloride method with a model replication cycle of 10 days. The mice were divided into a normal control group, a negative control group, a positive drug group (total glucosides of paeony tablets, 0.182g/kg calculated by clinical dose), and groups of examples 1-3 in sequence. The treatment is given from d11, wherein the normal group and the negative control group are perfused with gastric saline, the positive control group is perfused with gastric white paeony root total glycoside liquid medicine, the test liquid medicine with corresponding concentration is perfused in the groups of the examples 1-3, the perfused volume is 1mL/20g of body weight, the times are 1 time per day, and the continuous perfusion is carried out for 15 days. After the dosing cycle was completed, orbital blood was collected, peripheral blood was collected and upper serum was separated. And detecting the OD value of the light absorption value under the wavelength of 450nm by using an ELISA method. And calculating the expression quantities of the SSA, SSB and ANA proteins in the serum according to the standard curves of the SSA, SSB and ANA proteins in the ELISA kit. The results are shown in Table 5.

TABLE 5 SSA, SSB, ANA protein expression levels (ng/mL, mean. + -. SD) in peripheral serum of mice

the results showed that the peripheral serum of mice in the negative model group had significantly increased expression levels of SSA, SSB and ANA proteins, as compared with the normal group ( ^* P<0.05or ^** P<0.01 ); compared with the negative model group, the total glucosides of paeony group can obviously inhibit the expression quantity of the SSB protein and the ANA protein in serum ( ^* P<0.05)( ^** P<0.01 5.1245 +/-0.2769 ng/mL and 39.4812 +/-6.3827 pg/mL respectively, but the inhibition effect on the SSA protein is not good, and is 83.5872 +/-6.1035 ng/mL (P)>0.05). Compared with the negative model group, examples 2 to 3 all significantly inhibited the expression levels of SSA, SSB and ANA proteins in peripheral serum of mice (S) ((S)) ^* P<0.05)( ^** P<0.01 In example 2, the expression amounts of SSA, SSB and ANA proteins are 76.2292 + -7.0896 ng/mL,4.9715 + -0.1247 ng/mL and 34.4028 + -4.9162 pg/mL, respectively. In example 3, the expression levels of SSA, SSB and ANA proteins were 75.9714. + -. 6.0628ng/mL, 4.4812. + -. 0.1933ng/mL and 33.1823. + -. 7.2492pg/mL, respectively. Compared with the positive control group, the inhibition effect of the example 2 on the expression level of the SSA protein is better than that of the total glucosides of paeony (total glucosides of paeony), (A), (B) ^# P<0.05 But has no significant difference in the inhibitory effect on the expression level of SSB and ANA proteins; however, example 3 inhibition of the expression level of SSA, SSB and ANA proteinsThe effect is obviously better than that of the total glucosides of paeony ( ^# P<0.05). Demonstrating that example 3 inhibits autoantibody effects better than the positive control.

And (3) test results: the Chinese medicinal compound extract has better inhibiting effect on the expression quantity of SSA, SSB and ANA proteins in peripheral serum than total glucosides of paeony. The research result reflects that the traditional Chinese medicine compound extract can effectively reduce the expression quantity of the autoantibody to a certain extent.

1. Experimental research on anti-inflammatory effect of traditional Chinese medicine compound extract

The specific implementation scheme is as follows: SPF-grade BALB/c mice 48, female, weighing 18-22g. The mice were randomly divided into 5 groups (n = 8) except the normal group, and the sicca syndrome mouse model was replicated using the scopolamine hydrochloride method with a model replication cycle of 10 days. The mice were divided into a normal control group, a negative control group, a positive drug group (total glucosides of paeony tablets, 0.182g/kg in terms of clinical dose) and groups of examples 1-3 in this order. Treatment is given from d11, wherein the normal group and the negative model control group are subjected to intragastric administration of normal saline, the radix paeoniae alba total glycoside group is subjected to intragastric administration of total paeony glycoside liquid medicine, the test liquid medicine with corresponding concentration in the groups of examples 1-3 is subjected to intragastric administration, the intragastric administration volume is 1mL/20g of body weight, the time is 1 time per day, and the intragastric administration is continuously carried out for 15 days. After the dosing cycle was completed, orbital blood was collected, peripheral blood was collected and upper serum was separated. And detecting the OD value of the light absorption value under the wavelength of 450nm by using an ELISA method. And calculating the expression levels of the TNF-alpha and BAFFR proteins in the serum according to the standard curves of the TNF-alpha and BAFFR in the ELISA kit. The spleen was removed from the dissected mice, and the spleen coefficients (spleen weight (mg)/body weight (g)) were compared. The results are shown in Table 3. And (3) soaking the spleen in a 4% neutral formaldehyde solution for 48 hours to prepare pathological sections, and finishing HE staining. The HE stained pathological section was placed under an inverted microscope and spleen tissue was observed under 100 x and 200 x objective lens in 5 fields at random. The results are shown in table 6, table 7, fig. 2.

TABLE 6 mouse peripheral serum TNF- α, BAFFR protein expression level (Mean + -SD)

Note: v. a set of negative models, ^* P<0.05or ^** P<0.01。

the results show (Table 6) that the peripheral serum TNF-alpha expression level of mice in the negative model group is significantly increased compared with that in the normal group ( ^** P<0.01 ); compared with a negative model group, the total glucosides of paeony group and the tested group of examples 1-3 can obviously reduce the expression quantity of TNF-alpha in the peripheral serum of mice, the expression quantity of the TNF-alpha in the total glucosides of paeony group is 48.6327 +/-4.6231 pg/mL, and the expression quantities of the TNF-alpha in the examples 1-3 are 49.3328 +/-3.2545 pg/mL,48.5520 +/-4.3418 pg/mL and 44.5130 +/-1.2311 pg/mL respectively. Compared with the positive control group of total glucosides of paeony, the TNF-alpha expression level in the example 3 is obviously reduced, and the statistical difference is shown in ( ^# P<0.05). It is shown that example 3 has an improved effect on the inhibition of TNF-alpha expression over total glucosides of paeony.

The expression level of BAFFR in peripheral serum of mice in a negative model group is obviously increased compared with that in a normal group ( ^** P<0.01 ); compared with the negative model group, the test group, the example 1 and the example 3 can obviously reduce the BAFFR expression level in the peripheral serum of the mice, and the BAFFR expression level is respectively 9.3760 +/-0.4305 ng/mL and 9.3783 +/-0.3173 ng/mL. However, the total glucosides of paeony positive control group and the control group of the white paeony have no obvious inhibiting effect and are respectively 9.9608 +/-1.1633 ng/mL and 9.3918 +/-0.4612 ng/mL. The expression level of BAFFR in example 3 is significantly reduced compared with the positive control group of total glucosides of paeony (statistical difference) ((R)) ^# P<0.05). It is demonstrated that example 3 has an inhibitory effect on BAFFR expression superior to total glucosides of paeony.

TABLE 7 mouse spleen coefficient variation (Mean + -SD)

the results show (Table 7), in comparison with the normal groupIn comparison, spleen coefficient of mice in the negative model group is significantly reduced ( ^* P<0.05 1.3. + -. 0.4). Compared with the negative model group, the coefficients of the total glucosides of paeony and the spleen of the examples 1 to 3 are significantly higher than those of the model group, namely 2.2 +/-0.8, 2.1 +/-0.6, 2.5 +/-0.9 and 2.8 +/-0.8 respectively. The spleen coefficients of examples 1-3 were not statistically different compared to the total glucosides of paeony positive group. It is demonstrated that examples 1-3 have comparable maintenance of spleen coefficients in mice to total glucosides of paeony.

HE staining results showed (fig. 2) that spleen white marrow of mice in the negative model group was heavily proliferated and germinal center was protruded (grade 3) compared with the normal group (grade 0); compared with the negative model group, the white marrow hyperplasia degree of the white peony root total glycosides group is relieved (grade 2), and the spleen white marrow hyperplasia can be obviously reduced in the tested groups of examples 1-3 (grade 1). Compared with the total glucosides of paeony positive group, the tested groups of examples 1 to 3 have better effect of reducing the splenic leukoplakia hyperplasia than the total glucosides of paeony group.

And (3) test results: the traditional Chinese medicine compound extract has the effects of reducing the expression quantity of TNF-alpha and BAFFR in peripheral serum, reducing spleen myelogenous hyperplasia and improving the spleen coefficient of mice, and is obviously superior to the white paeony root total glycoside positive group. The research result reflects that the traditional Chinese medicine compound extract has good anti-inflammatory effect to a certain extent.

Example 6 examination and implementation-determination of maximum dose of Compound extract of Chinese medicinal herbs

The specific implementation scheme is as follows: SPF-grade BALB/c mice 20, female, weighing 18-22g. The groups were randomly divided into 2 groups of 10 individuals, each group was a normal control group (normal saline) and an example 4 group (maximum concentration liquid medicine prepared by the compound extract of the Chinese herbal medicine according to example 4). Fasting was 12h (without water deprivation) before the experiment, and gavage was performed in both groups, 1mL/20g body weight, 1 time/day, and 15 days. And recording and observing the death condition, the daily weight increase condition and the daily water intake of the mouse, observing and recording the appearance signs, the behavior and the activity, the mental state, the appetite, the fur, the complexion, the respiration, the excrement and the color of the mouse, the existence of abnormal secretions such as the genitals of the mouth, the nose and the eyes and the like, and continuously observing for 15 days. After 15 days, the patient is killed after neck removal, dissected, and visually observed for main organs such as heart, liver, spleen, lung, kidney, gastrointestinal tract and the like, and has obvious pathological morphological changes.

The test results show that the administration dosage is 55 times of the clinical dosage of the adult calculated by the adult weight of 60 kg. After the mice are perfused with the extract with the maximum concentration and the maximum volume, the appearance signs, the behavior activity, the mental state, the appetite, the urine color, the fur, the skin color and the breath of the mice in the group of example 4 have normal breathing, and the genitals of the nose, the eyes, the mouth and the mouth have no abnormal secretion; the color of the stool particles is dark, the hardness of the particles is moderate, and the constipation phenomenon is avoided. After dissection, the internal organs of both groups of mice were normal. The mice in example 4 and normal groups showed no significant difference in body weight and food intake (P > 0.05) as detailed in Table 8, table 9 and Table 10.

TABLE 8 weight change of mice during dosing (g, mean + -SD)

TABLE 9 change in water intake of mice during dosing (mL, mean + -SD)

TABLE 10 mouse abnormalities, deaths, and anatomical changes

The basis for selecting total glucosides of paeony as a positive control drug is as follows:

the specific implementation scheme is as follows: SPF grade Balb/C mice 32, female mice, weight 18-22g. In addition to the normal group, the other mice replicated the sjogren syndrome mouse model using the scopolamine hydrochloride method, and after 10 days the model replication was completed, which were randomly grouped (n = 8). Mice were divided into normal group, total glucosides of paeony group, hydroxychloroquine group, and example 2 group. The total glucosides of paeony liquid (converted into mouse dose of 0.182g/kg according to clinical dose) is administered to the total glucosides of paeony group except normal group mice; hydroxychloroquine group is given hydroxychloroquine sulfate liquid medicine (converted into mouse dose of 0.03g/kg according to clinical dose); the group of example 2 was administered with the corresponding test drug solution. The gavage volume of the mice is 1mL/20g of body weight, 1 time per day, and the gavage is continuously carried out for 15 days. The weight change and water intake of the mice were measured every 2 days during the period. After the experiment was completed, mouse spleen weight and spleen coefficient were measured. The test results are shown in tables 11, 12 and 13.

Through the investigation of the weight, the water intake and the spleen coefficient of the mice, the intervention effect of the two positive drugs on the SS mice is compared. The results show that the treatment effect of the total glucosides of paeony group is obviously better than that of the hydroxychloroquine group, and the treatment effect of the group in example 2 is equivalent to that of the hydroxychloroquine group. Therefore, the total glucosides of paeony is preferably used as a positive control drug in the test.

TABLE 11 weight change (g, mean + -SD) of mice during dosing

Note: the normal group of the cells is set to v, ^* P<0.05or ^** P<0.01; vs. total glucosides of paeony group, ^# P<0.05or ^## P<0.01。

TABLE 12 variation in mouse Water intake during dosing (mL, mean + -SD)

TABLE 13 mouse spleen weight and spleen coefficient variation (Mean + -SD)

Claims

1. a traditional Chinese medicine composition for preventing or treating sjogren syndrome is characterized by comprising 2g-30g of selfheal, 2g-30g of desmodium, 2g-30g of spreading hedyotis herb, 2g-30g of chrysanthemum and 2g-30g of honeysuckle.

2. The preparation method of the traditional Chinese medicine composition for preventing or treating sjogren's syndrome according to claim 1 is characterized by comprising the following steps of decocting and mixing 2g-30g of selfheal, 2g-30g of desmodium, 2g-30g of spreading hedyotis herb, 2g-30g of chrysanthemum and 2g-30g of honeysuckle flower with water to obtain a finished product.

3. The use of a Chinese medicinal composition according to claim 1 or 2 in the preparation of a medicament for the prevention or treatment of sjogren's syndrome.