APPLICATION OF TRADITIONAL CHINESE MEDICINE COMPOSITION IN PREPARATION OF DRUGS FOR TREATING NEURITIS

Abstract

A traditional Chinese medicine composition for treating neuritis. The traditional Chinese medicine composition is prepared mainly from Notopterygii Rhizoma et Radix, Angelicae Pubescentis Radix, Poria, Saposhnikoviae Radix, Schizonepetae Herba, Chuanxiong Rhizoma, Platycodonis Radix, Bupleuri Radix, Peucedani Radix, Aurantii Fructus, and Glycyrrhizae Radix et Rhizoma. The traditional Chinese medicine composition can significantly relieve facial paralysis, movement disorders, and other related symptoms of facial neuritis in a rat, remarkably improve expression of VEGFb, VEGFR1, and VEGFR2 proteins in postoperative side facial nerves of the rat, and promote recovery and regeneration of damaged rear axons.

Claims

1. A method for treating facial neuritis in a subject, comprising administrating a therapeutically effective amount of a traditional Chinese medicine composition to the subject, wherein the traditional Chinese medicinal composition comprises consists of the following traditional Chinese medicine components: Notopterygii Rhizoma et Radix 50-100 parts by weight, Angelicae Pubescentis Radix 50-100 parts by weight, Poria 50-100 parts by weight, Saposhnikoviae Radix 50-100 parts by weight, Schizonepetae Herba 50-100 parts by weight, Chuanxiong Rhizoma 50-100 parts by weight, Platycodonis Radix 50-100 parts by weight, Bupleuri Radix 50-100 parts by weight, Peucedani Radix 50-100 parts by weight, Aurantii Fructus 50-100 parts by weight and Glycyrrhizae Radix et Rhizoma 5-50 parts by weight.

2. (canceled)

3. The method according to claim 1, characterized in that the traditional Chinese medicine composition consists of the following traditional Chinese medicine components: Schizonepetae Herba 75 parts by weight, Saposhnikoviae Radix 75 parts by weight, Notopterygii Rhizoma et Radix 75 parts by weight Angelicae Pubescentis Radix 75 parts by weight, Bupleuri Radix 75 parts by weight, Peucedani Radix 75 parts by weight Chuanxiong Rhizoma 75 parts by weight, Aurantii Fructus 75 parts by weight, Poria 75 parts by weight Platycodonis Radix 75 parts by weight, and Glycyrrhizae Radix et Rhizoma 25 parts by weight.

4. (canceled)

5. The method according to claim 1, characterized in that the neuritis is caused by one or more of infection, poisoning, genetic defect, nutritional disorder, immune damage, metabolic disorder, endocrine disorder, congenital malformation, blood circulation disorder, abnormal hyperplasia, wind-cold and trauma.

6. The method according to claim 1, characterized in that the traditional Chinese medicine composition can improve the symptoms of neuritis, and the symptoms include irritation symptoms and/or destruction symptoms, wherein the irritation symptoms show pain and numbness, and the destruction symptoms show weakness and paralysis.

7. The method according to claim 1, characterized in that the traditional Chinese medicine composition can improve the symptoms of facial neuritis, including shallow nasolabial groove, distortion of commissure, air leakage and salivation in speech, loss of facial random movements, large palpebral fissure, inability to successfully complete frowning, closing eyes, whistling and other movements, and loss of one or more facial expressions.

8. The method according to claim 1, characterized in that the traditional Chinese medicine composition can be prepared into a preparation with pharmaceutically acceptable excipients; and the preparation comprises granules, mixtures, tablets, capsules, pills, dustpowders, powders, syrups, microcapsules and ointments.

9. (canceled)

10. (canceled)

11. The method according to claim 1, characterized in that the traditional Chinese medicine composition is JINGFANG Preparation, wherein the JINGFANG Preparation is JINGFANG KELI or JINGFANG HEJI.

Description

BRIEF DESCRIPTION OF THE DRAWINGS

[0036] FIG. 1-2: Study on Nerve Conduction Velocity of Lateral Nerve Potential of Rats in Each Group after Administration for 14 Days

[0037] FIG. 3-4: Expression of Beclin1 Protein in Facial Nerve Tissue of Rats in Each Group after Administration for 14 Days

[0038] FIG. 5-6: Expression of P62 Protein in Facial Nerve Tissue of Rats in Each Group after Administration for 14 Days

DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0039] The summary of the present invention is described in further detail by way of examples in order to make the purse and the technical proposal of the present invention clearer, but it should not be understood that the scope of the above subject area of the present invention is limited to the following examples. Corresponding substitutions or modifications made in accordance with ordinary knowledge and conventional means in the art without departing from the above-mentioned techniques of the present invention are included in the present invention.

Example 1 Preparation of Granule (Commercially Available JINGFANG KELI)

Prescription:

[0040] Schizonepetae Herba 75 g, Saposhnikoviae Radix 75 g, Notopterygii Rhizoma et Radix 75 g, Angelicae Pubescentis Radix 75 g, Peucedani Radix 75 g [0041] Chuanxiong Rhizoma 75 g, Aurantii Fructus 75 g, Poria 75 g, Platycodonis Radix 75 g, and Glycyrrhizae Radix et Rhizoma 25 g;

Preparation Method:

[0042] Among the above eleven herbs, extract volatile oils of Schizonepetae Herba, Saposhnikoviae Radix, Notopterygii Rhizoma et Radix, Angelicae Pubescentis Radix, Peucedani Radix, Chuanxiong Rhizoma and Aurantii Fructus respectively, and collect the distilled aqueous solution of Chuanxiong Rhizoma and Aurantii Fructus separately; percolate the residues of Chuanxiong Rhizoma, Aurantii Fructus and Poria with 25% ethanol solution prepared from the above aqueous solution as solvent according to the percolation method under the terms of fluid extract and extract; decoct the residues of Schizonepetae Herba, Saposhnikoviae Radix, Notopterygii Rhizoma et Radix, Angelicae Pubescentis Radix and Peucedani Radix together with the rest of Bupleuri Radix, Platycodonis Radix and Glycyrrhizae Radix et Rhizoma in water for 1.5 hours twice, combine decoction, filter, and concentrate filtrate into thick paste; combine the percolated liquid and the thick paste, mix evenly, stand, filter, concentrate the filtrate into clear paste with a relative density of 1.30 (80-85 C.), take 1 part of the clear paste, add 6 parts of sucrose, mix evenly to make granules, dry, add the volatile oils such as Schizonepetae Herba, and mix evenly.

Example 2 Preparation of Granule

Prescription:

[0043] Schizonepetae Herba 50 g, Saposhnikoviae Radix 50 g, Notopterygii Rhizoma et Radix 50 g, Angelicae Pubescentis Radix 50 g, Peucedani Radix 50 g [0044] Chuanxiong Rhizoma 50 g, Aurantii Fructus 50 g, Poria 50 g, Platycodonis Radix 50 g, and Glycyrrhizae Radix et Rhizoma 5 g.

Preparation Method: Same as Example 1.

Example 3 Preparation of Granule

Prescription:

[0045] Schizonepetae Herba 100 g, Saposhnikoviae Radix 100 g, Notopterygii Rhizoma et Radix 100 g, Angelicae Pubescentis Radix 100 g, Peucedani Radix 100 g [0046] Chuanxiong Rhizoma 100 g, Aurantii Fructus 100 g, Poria 100 g, Platycodonis Radix 100 g, and Glycyrrhizae Radix et Rhizoma 50 g.

Preparation Method: Same as Example 1.

Example 4 Preparation of Mixture (Commercially Available JINGFANG HEJI)

Prescription:

[0047] Schizonepetae Herba 97 g, Saposhnikoviae Radix 97 g, Notopterygii Rhizoma et Radix 97 g, Angelicae Pubescentis Radix 97 g, Peucedani Radix 97 g [0048] Chuanxiong Rhizoma 97 g, Aurantii Fructus 97 g, Poria 97 g, Platycodonis Radix 97 g, and Glycyrrhizae Radix et Rhizoma 32.4 g;

Preparation Method:

[0049] Distill Schizonepetae Herba, Saposhnikoviae Radix, Notopterygii Rhizoma et Radix, Angelicae Pubescentis Radix, Aurantii Fructus, Chuanxiong Rhizoma and Peucedani Radix and extract volatile oils respectively, and collect the distilled aqueous solution separately; percolate the residues of Chuanxiong Rhizoma, Aurantii Fructus and Poria with 25% ethanol solution prepared from the above aqueous solution as solvent according to the percolation method under the terms of fluid extract and extract, collect percolate and recover ethanol under reduced pressure. Decoct the remaining five kinds of residues with Bupleuri Radix, Platycodonis Radix and Glycyrrhizae Radix et Rhizoma in water for three times, filter, combine filtrate, concentrate to about 1300 mL, combine with percolate, stand still, filter, concentrate to about 1000 mL, add 3 g of sodium benzoate and the above volatile oil, stir evenly, and add water to make it 1000 mL.

Comparative Example 1: Decoction

[0050] Bupleuri Radix 30 g, Scutellaria baicalensis 17 g, Schizonepetae Herba 14 g, Saposhnikoviae Radix 15 g, Rhizoma typhonii 15 g, Gastrodia elata 15 g [0051] Scorpion 15 g, Glycyrrhizae Radix et Rhizoma 10 g, Garden balsam stem 15 g, Notopterygii Rhizoma et Radix 20 g, Earthworm 20 g

Preparation Method:

[0052] Prepare decoction to 500 mL by conventional decoction.

Comparative Example 2: Decoction

[0053] Schizonepetae Herba 10 g, Saposhnikoviae Radix 10 g, Bupleuri Radix 12 g, Scutellaria baicalensis 10 g, Notopterygii Rhizoma et Radix 10 g, Angelicae Pubescentis Radix 10 g, Chuanxiong Rhizoma 12 g [0054] Mint 10 g, Honeysuckle 10 g, Forsythia suspensa 10 g, Angelica dahurica 10 g, Pueraria lobata 20 g, Hedyotis diffusa 15 g [0055] Red paeony root 10 g, Great burdock achene 10 g and Glycyrrhizae Radix et Rhizoma 9 g

[0056] Preparation method: Prepare decoction to 500 mL by conventional decoction.

Comparative Example 3: Decoction

[0057] Schizonepetae Herba 10 g, Saposhnikoviae Radix 10 g, Bupleuri Radix 12 g, Scutellaria baicalensis 10 g, Notopterygii Rhizoma et Radix 10 g, Chuanxiong Rhizoma 12 g, Angelica sinensis 10 g [0058] Mint 10 g, Honeysuckle 10 g, Forsythia suspensa 10 g, Pueraria lobata 20 g, Hedyotis diffusa 15 g, Cicada slough 6 g [0059] Red paeony root 10 g, Great burdock achene 10 g, Glycyrrhizae Radix et Rhizoma 9 g, Rhizoma cimicifugae 10 g, Great burdock achene 10 g, and Acorus gramineus 10 g Preparation method: Prepare decoction to 500 mL by conventional decoction.

Pharmacological Experiment

[0060] The inventor carried out related pharmacodynamic experimental research to prove the efficacy of JINGFANG Preparation in the present invention in treating facial neuritis. It should be noted that the following experimental studies are carried out on the basis of proving drug safety by acute toxicity test and long-term toxicity test, and the dosage in the experimental studies is within the safe dose range. The medicines selected in the following pharmacodynamic tests are prepared from the representative formula of the present invention; pharmacodynamic experiments have also been carried out by the inventor of the medicines prepared from other formulations contained in the present invention, and the experimental results have the same or similar effects, but due to space limitations, they are not listed here.

[0061] In addition, the following pharmacodynamic experiments only take some animal models as examples to verify the efficacy of the present invention. Here, only the pharmacodynamic experimental results of the traditional Chinese medicine composition for treating facial neuritis are displayed. The inventor has also done relevant pharmacodynamic experiments for neuritis caused by other types and other pathogenic factors mentioned in the present invention, and the experimental results show the same or similar effects, so the pharmacodynamic experimental results will not be listed one by one.

1 Materials

1.1 Experimental Drugs and Reagents

1.1.1 Drugs

[0062] Granules obtained in Examples 1, 2 and 3 of the present invention; [0063] Example 4 Mixture; [0064] Decoctions obtained in Comparative examples 1-3.

1.1.2 Dose

[0065] Example 1 Granule: 2.02 g/kg (low dose), 4.05 g/kg (medium dose), 8.10 g/kg (high dose); [0066] Example 2 Granule: 4.05 g/kg; [0067] Example 3 Granule: 4.05 g/kg; [0068] Example 4 Mixture: 4.05 ml/kg; [0069] Decoction in Comparative example 1: 18 mL/kg; [0070] Decoction in Comparative example 2: 18 mL/kg; [0071] Decoction in Comparative example 3: 18 mL/kg;

1.2 Experimental Animals

[0072] SD rats, SPF grade, 180-220 g, experimental animal license number: SYXK (Lu) 20180008, provided by Lunan Pharmaceutical Group Co., Ltd., were fed adaptively under standard conditions for one week before the experiment.

2. Methods

2.1 Molding Method and Grouping

[0073] A number of rats, half male and half female, were taken. An appropriate amount of ether was poured into the small beaker to completely infiltrate the cotton ball, and the small beaker was put into the rat anesthesia chamber after the ether container was tightly covered. Then, the rat is quickly put into the anesthesia chamber before the anesthesia chamber is closed tightly. When the rats fell spontaneously and fast breathing slowed down, they were taken out of the anesthesia chamber to check their corneal reflexes and pain reflexes. If the rat did not close their eyes and dodge when touching their cornea with the tip of the hemostat, it was proved that the corneal reflexes disappeared; if the rat did not struggle or dodge when clamping the toe of the rat with appropriate force with a hemostat, it was proved that the pain reflexes disappeared and the rats' general anesthesia was effective. Then, the right side of the rat was fixed upwards on the surgical table in a lateral position because the right side of the face is the surgical area to be sterilized with 1% iodophor solution. A cortex of about 1 cm long was cut at the midpoint of the line connecting the right tragus and the pupil of the right eye of the rat, with the incision perpendicular to the connecting line. After the subcutaneous tissue of the incision was separated to the muscular surface, the facial nerve was separated along the exposed buccal branch of the rat facial nerve towards the right auricle of the rat until the right facial nerve stem of the rat was exposed. The facial nerve was separated and clamped with the front end of the needle holder for 90 s. After the clamping damage was formed, the wound was sutured with 3-0 silk suture, and applied with erythromycin ointment to prevent infection. The modeled rats were taken to form the model group, three dose groups of Example 1 (high, medium, and low), the groups of Examples 2-4, and the groups of Comparative examples 1-3, with 10 rats in each group, half male and half female. The operation of 10 rats in the blank group (half male and half female) was basically the same as above, but the right facial nerve stem was only exposed without any injury treatment.

2.2 Administration

[0074] Rats in each administration group are given corresponding drugs by intragastric administration, while rats in the blank group and model group are given the same amount of normal saline by intragastric administration once a day for 14 days.

3. Statistical Processing

[0075] SPSS 22.0 software is used for statistical analysis of the data. The measurement data are expressed by (x+s). One-way ANOVA is used for comparison among multiple groups, and independent sample T test is used for analysis between two groups. The difference is statistically significant with P<0.05.

4 Observation Indexes and Experimental Results

4.1 Rat Behavioral Score

4.1.1 Scoring Criteria

TABLE-US-00001 TABLE 1 Scoring Table of Tentacle Movement Score Tentacle movement Position of tentacle 1 No tentacle movement Backward 2 Mild tremor Backward 3 Large tremor Backward 4 Normal tentacle movement Backward 5 Normal Forward

TABLE-US-00002 TABLE 2 Observation Table of Eye Closure and Blink Reflex Score Blink reflex and eye closure 1 No blink reflex or eye closure 2 Ocular muscle tension with no blink reflex 3 Blink reflex with 50% eye closure 4 Blink reflex with 75% eye closure 5 Complete closure, positive blink reflex

4.1.2 Statistical Results

TABLE-US-00003 TABLE 3 Behavioral Scores of Rats after Administration on 0 d (x s, n = 10) Behavioral score Group 0 d Blank group 9.79 0.51 Model group 1.71 0.54* Example 1 High dose group 1.68 0.44* Medium dose group 1.73 0.49* Low dose group 1.72 0.36* Example 2 group 1.75 0.46* Example 3 group 1.69 0.48* Example 4 group 1.68 0.58* Comparative example 1 group 1.71 0.52* Comparative example 2 group 1.73 0.50* Comparative example 3 group 1.70 0.41* Note: Compared with the blank group, *P < 0.01.

TABLE-US-00004 TABLE 4 Behavioral Scores of Rats after Administration for 14 d (x s, n = 10) Behavioral score Group 14 d Blank group 9.80 0.42.sup. Model group 2.20 0.82*.sup. Example 1 High dose group 6.68 0.97*.sup.# Medium dose group 6.65 0.88*.sup.# Low dose group 5.10 1.06*.sup.# Example 2 group 5.40 1.03*.sup.# Example 3 group 5.90 0.97*.sup.# Example 4 group 6.80 1.05*.sup.# Comparative example 1 group 3.30 1.42*.sup.# Comparative example 2 group 4.20 1.40*.sup.# Comparative example 3 group 3.70 1.34*.sup.# Note: Compared with the blank group, *P < 0.01; Compared with the model group, .sup.#P < 0.01.

[0076] The results of behavioral scores of facial neuritis showed that among the comparison between the blank group and model group, three dose groups (high, medium, low) in Example 1, Example 2-4 groups, and Comparative example 1-4 groups after administration on Od, the difference was statistically significant (P<0.01), which proved that the model was successful; on the 14th day of administration, among the model group and blank group, three dose groups (high, medium and low) in Example 1, Example 2-4 groups, and Comparative example 1-4 groups respectively, the difference was statistically significant (P<0.01).

4.2 Expression of VEGFb, VEGFR1 and VEGFR2 Protein in Lateral Nerves of Rats after Administration for 14 Days

TABLE-US-00005 TABLE 5 Expression of VEGFb Protein in Lateral Nerves of Rats after Administration for 14 Days (x s, n = 10) Group VEGFb/Actin Blank group 1.72 0.07.sup. Model group 0.76 0.06.sup. Example 1 High dose group 1.59 0.15*.sup.# Medium dose group 1.58 0.14*.sup.# Low dose group 1.19 0.13*.sup.# Example 2 group 1.25 0.09*.sup.# Example 3 group 1.46 0.11*.sup.# Example 4 group 1.62 0.11*.sup.# Comparative example 1 group 0.92 0.12*.sup.% Comparative example 2 group 1.06 0.14*.sup.# Comparative example 3 group 0.98 0.11*.sup.# Note: Compared with the blank group, *P < 0.01; Compared with the model group, .sup.#P < 0.01, .sup.%P < 0.05.

TABLE-US-00006 TABLE 6 Expression of VEGFR1 Protein in Lateral Nerves of Rats after Administration for 14 Days (x s, n = 10) Group VEGFR1/Actin Blank group 1.58 0.05.sup. Model group 0.68 0.04.sup. Example 1 High dose group 1.36 0.14*.sup.# Medium dose group 1.35 0.10*.sup.# Low dose group 1.05 0.05*.sup.# Example 2 group 1.13 0.15*.sup.# Example 3 group 1.27 0.15*.sup.# Example 4 group 1.38 0.09*.sup.# Comparative example 1 group 0.86 0.15*.sup.# Comparative example 2 group 0.95 0.12*.sup.# Comparative example 3 group 0.89 0.13*.sup.# Note: Compared with the blank group, *P < 0.01; Compared with the model group, .sup.#P < 0.01.

TABLE-US-00007 TABLE 7 Expression of VEGFR2 Protein in Lateral Nerves of Rats after Administration for 14 Days (x s, n = 10) Group VEGFR2/Actin Blank group 1.56 0.03.sup. Model group 0.61 0.05.sup. Example 1 High dose group 1.53 0.15*.sup.# Medium dose group 1.54 0.13*.sup.# Low dose group 1.27 0.12*.sup.# Example 2 group 1.36 0.14*.sup.# Example 3 group 1.43 0.13*.sup.# Example 4 group 1.55 0.08*.sup.# Comparative example 1 group 0.99 0.08*.sup.# Comparative example 2 group 1.09 0.05*.sup.# Comparative example 3 group 1.04 0.08*.sup.# Note: Compared with the blank group, *P < 0.01; Compared with the model group, .sup.#P < 0.01.

[0077] To sum up, according to the pharmacodynamic experimental results, the traditional Chinese medicine composition of the present invention can obviously relieve the facial paralysis related symptoms of facial neuritis, significantly improve the expression of VEGFb, VEGFR1 and VEGFR2 proteins in the lateral nerves of rats after operation, promote the recovery and regeneration of axons after injury, and effectively treat facial neuritis.

4.3 Study on Nerve Conduction Velocity of Lateral Nerve Potential of Rats after Administration for 14 Days

[0078] After administration for 14 days, rats were anesthetized by intraperitoneal injection, and facial nerves were separated. Stimulating electrodes and recording electrodes were placed along the separated facial nerves from proximal to distal. Nerve conduction velocity CV (m.Math.s.sup.1)=distance between stimulating electrodes and recording electrodes/conduction time. The mean value was calculated by repeated measurement for 10 times.

[0079] As shown in FIG. 1, after administration for 14 days, the rats in each group were compared in potential nerve conduction velocity test, which showed that among the comparison between the model group and blank group, three dose groups (high, medium and low) in Example 1, and Example 2-4 groups respectively, the difference was statistically significant (P<0.01).

[0080] FIG. 2 is a comparison of facial nerve conduction velocity of rats in Comparative examples 1-3, model group and blank group. Comprehensive comparison shows that the traditional Chinese medicine composition of the examples of the present invention can stimulate electrical conduction of facial nerve of rats and effectively treat facial neuritis.

4.4 Expression of Beclin1 Protein and P62 Protein in Facial Nerve Tissue of Rats

[0081] After administration for 14 days, the expression of Beclin1 protein in facial nerve tissue was detected by immunohistochemistry and P62 protein in facial nerve tissue was detected by RT-PCR.

[0082] FIG. 3, FIG. 4, FIG. 5 and FIG. 6 show the expression levels of Beclin1 protein and P62 protein in facial nerve tissue of rats after administration for 14 days. The results showed that the traditional Chinese medicine composition of the present invention can reduce the expression of Beclin1 protein in the facial nerve tissue of rats, increase the level of P62 protein, participate in the regulation mechanism of autophagy, and promote the recovery of facial nerve function. Among the comparison between the model group and blank group, three dose groups (high, medium and low) in Example 1, and Example 2-4 groups respectively, the difference was statistically significant (P<0.01).

[0083] To sum up, through the behavior score of rats, it can be seen that the traditional Chinese medicine composition of the present invention can obviously relieve facial paralysis, dyskinesia and other related symptoms of facial neuritis of rats; can significantly increase the expression of VEGFb, VEGFR1 and VEGFR2 proteins in the lateral nerve of rats after operation, promote the recovery and regeneration of axons after injury, stimulate the electrical conduction of facial nerve in rats, improve the electrical conduction velocity of facial nerve, reduce the expression of Beclin1 protein in facial nerve tissue of rats, increase the level of P62 protein, and promote the recovery of facial nerve function. Therefore, the traditional Chinese medicine composition of the present invention has the effect of treating facial neuritis and can improve the related symptoms of facial neuritis.