Application of traditional Chinese medicine composition in preparation of medicine for treating nerve deafness

Abstract

The invention relates to application of a traditional Chinese medicine composition in preparation of a medicine for treating nerve deafness. The traditional Chinese medicine composition is prepared from the following raw material medicines in parts by weight: 5-25 parts of ligusticum wallichii and 1.5-8 parts of gastrodia elata. The traditional Chinese medicine composition is simple in compatibility and simple and convenient in preparation method, and has an obvious effect on treating nerve deafness.

Description

Technical Field

[0001] The invention belongs to the technical field of traditional Chinese medicines, relates to a new use of a traditional Chinese medicine composition, and specifically relates to the use of the traditional Chinese medicine composition in preparing a medicine for treating neurological deafness. Background Art

[0002] Neural hearing loss refers to hearing loss or complete deafness caused by damage to the auditory nervous system due to various factors, mostly sensorineural hearing loss. The lesions are located in the hair cells of the auditory organ, the auditory nerve or the auditory centers at all levels, which manifests as a disorder in the perception of sound and the conduction of nerve impulses, causing hearing loss or even disappearance, often accompanied by tinnitus.

[0003] The goal of treating sensorineural hearing loss is to restore or partially restore the lost hearing first, and then try to preserve and utilize the remaining hearing. The best treatment time is within 1 week of the onset of symptoms, and generally treatment will be effective within 3 months. The main treatment methods include medication, surgery, and hearing aids. Summary of the invention

[0004] The purpose of the present invention is to provide an application of a Chinese medicine composition in the preparation of a drug for treating sensorineural deafness, characterized in that the Chinese medicine composition is prepared from the following raw materials in the following weight proportions: 5-25 parts of Chuanxiong and 1.5-8 parts of Gastrodia. Wherein, Chuanxiong is the dried rhizome of the plant Ligusticumchuanxiong Hort. of the Umbelliferae family; and Gastrodia is the dried tuber of the plant Gastrodia elata Bl. of the Orchidaceae family.

[0005] Furthermore, the Chinese medicine composition is made of raw materials in the following weight ratios: 8-15 parts of Chuanxiong and 2-6 parts of Gastrodia elata.

[0006] Furthermore, the Chinese medicine composition is made of raw materials in the following weight ratios: 8-10 parts of Chuanxiong and 2.5-5 parts of Gastrodia elata.

[0007] Furthermore, the Chinese medicine composition is made of raw materials in the following weight ratios: 9-10 parts of Chuanxiong and 2.5-3 parts of Gastrodia elata.

[0008] Furthermore, the Chinese medicine composition is made of raw materials in the following weight ratio: 10 parts of Chuanxiong and 2.5 parts of Gastrodia elata.

[0009] Furthermore, the Chinese medicine composition is made of raw materials in the following weight ratio: 9 parts of Chuanxiong and 5 parts of Gastrodia elata.

[0010] Specifically, the drug is selected from an oral dosage form, an injection dosage form or an external administration preparation.

[0011] Specifically, the medicine is selected from decoctions, tablets, capsules, granules, pills, injections, powders, decoctions, suspensions, dispersants, syrups, suppositories, gels, aerosols, patches, and oral liquids.

[0012] The composition of the present invention described above can be prepared by directly grinding several medicinal materials into powder according to traditional methods, or by obtaining extracts through conventional means so as to be prepared into different dosage forms, thereby obtaining better therapeutic effects and being more conducive to the preparation of modern pharmaceutical dosage forms.

[0013] Furthermore, the method for preparing the Chinese medicine composition comprises the following steps:

[0014] (1) taking Chuanxiong and Gastrodia elata in a weight ratio of 5-25 of Chuanxiong and 1.5-8 of Gastrodia elata, drying, crushing and mixing them to obtain a mixture;

[0015] (2) extracting the mixture by reflux with 70%-90% ethanol for 2-3 times, each time for 1-3 hours, to obtain an extract and a medicinal residue;

[0016] (3) combining the extracts and filtering to obtain a filtrate;

[0017] (4) The filtrate is concentrated to a clear paste having a relative density of 1.21-1.30 at 55-65°C.

[0018] Furthermore, the preparation method of the Chinese medicine composition also includes:

[0019] (5) decocting the medicinal residue with water for 1-2 hours, combining the decoction, filtering, and concentrating the filtrate after the decoction to a clear paste with a relative density of 1.21-1.30 at 55-65° C., preferably decocting for 1 hour;

[0020] (6) Combine the clear pastes obtained in step (4) and step (5).

[0021] Specifically, step (2) is to extract the mixture by reflux using 90% ethanol twice to obtain an extract, each time for 2 hours.

[0022] Furthermore, the drug is selected from capsules, and the preparation method, in addition to the above steps, further comprises: adding drug-acceptable excipients, such as an appropriate amount of dextrin, to the clear paste obtained in step (4) and / or step (5), vacuum drying, mixing evenly, sieving, granulating, granulating, and encapsulating.

[0023] Experimental studies have shown that the Chinese medicine composition of the present invention can upregulate the cAMP content in the cochlear tissue in a mouse model of sensorineural deafness, and significantly improve the PKA mRNA expression level, and has a significant improvement effect on mouse deafness damage; in a rat model of sensorineural deafness, it can significantly reduce the ABR response threshold and significantly reduce the CAT content in the cochlear tissue, thereby mediating the oxidative stress response and improving the cochlear microcirculation disorder, indicating that the Chinese medicine composition will play an important role in the treatment or prevention of sensorineural deafness. DETAILED DESCRIPTION

[0024] The present invention discloses an application of a Chinese medicine composition, and those skilled in the art can refer to the content of this article and appropriately improve the process parameters to achieve it. It is particularly important to point out that all similar substitutions and modifications are obvious to those skilled in the art, and they are all considered to be included in the present invention. The method and application of the present invention have been described through preferred embodiments, and relevant personnel can obviously modify or appropriately change and combine the methods and applications described herein without departing from the content, spirit and scope of the present invention to implement and apply the technology of the present invention.

[0025] As used herein, the term "treat" refers to any indicator of success in treating or ameliorating an injury, condition, disorder or symptom (e.g., pain), including any objective or subjective parameter, such as elimination; improvement; reduction of symptoms or making symptoms, injuries, conditions or disorders more tolerable to the patient; reduction of symptom or disorder frequency or duration; or, in some cases, prevention of the onset of symptoms or disorders. Treatment or amelioration of symptoms can be based on any objective or subjective parameter, including, for example, the results of a physical examination.

[0026] The raw materials and reagents used in the following examples can all be purchased from the market.

[0027] Example 1 Preparation of Chinese medicine composition capsules

[0028] Take 784g of Chuanxiong and 196g of Gastrodia elata, dry, crush and mix them; then use 90% ethanol to reflux and extract twice to obtain an extract and medicinal residues, each time for 2 hours, combine the extracts, and filter to obtain a filtrate; the filtrate is concentrated to a clear paste with a relative density of 1.27 at 55-65°C; the medicinal residues are decocted with water twice, each time for 1 hour, the decoctions are combined, filtered, and the filtrate after the decoction is concentrated to a clear paste with a relative density of 1.27 at 55-65°C, and the clear paste is combined with the above-mentioned clear paste to obtain a traditional Chinese medicine composition; add drug-acceptable excipients to the clear paste of the traditional Chinese medicine composition, vacuum dry, mix evenly, and sieve; granulate, granulate, and encapsulate to make 1000 capsules for standby use, with each capsule containing 0.34g, equivalent to 0.98g of the original medicinal material.

[0029] Experimental Example 2 Therapeutic Effect of Traditional Chinese Medicine Composition on D-galactose-induced Neurological Deafness Model in Mice 1. Experimental Materials

[0030] 1.1 Animals

[0031] ICR mice, male, SPF grade, 60 mice, weighing 16-18 g, were purchased from Yangzhou University, animal certificate number: A202402280049.

[0032] Rearing environment: room temperature 20-26℃, relative humidity controlled at 40-70%. Rearing in separate cages, 5 per group.

[0033] 1.2 Drugs

[0034] The Chinese medicine composition prepared in Example 1, Jiangsu Kangyuan Pharmaceutical Co., Ltd.; prednisone (Tianjin Tianyao Pharmaceutical Co., Ltd., batch number H12020201); D-galactose (Sigma, batch number 59-23-4); chloral hydrate (Sinopharm Chemical Reagent Co., Ltd., batch number 20200320).

[0035] 1.3 Instruments

[0036] Electronic balance (Sartorius Scientific Instruments, model BS224S); low-temperature centrifuge (eppendorf, Germany, model 5804R); purified water instrument (Millipore, Germany, model: Milli-Q Plus); animal weight scale (Shanghai Xiangchuan Electronic Scale Co., Ltd., model: CX-SC-DA); microplate reader (MeiGu Molecular, USA, model Flexstation3); AB step one plus real-time fluorescence quantitative PCR instrument (Applied biosystems, model: 4376592).

[0037] 2. Dosage design

[0038] The daily dosage of the Chinese medicine composition is 4.08g of crude drug / day. According to the body surface area conversion method, the equivalent dose for mice is 4.08g / 60kg*12.3≈0.84g of crude drug / kg. According to the 1 / 4:1 / 2:1 dosing method, the low dose is 0.84g of crude drug / kg, the medium dose is 1.68g of crude drug / kg, and the high dose is 3.36g of crude drug / kg. Oral administration, 0.2mL / 10g, once a day.

[0039] The daily dose of prednisone is 10 mg / day. According to the body surface area conversion method, the equivalent dose for mice is 10 mg / 60 kg*12.3=2.05 mg / kg.

[0040] 3. Experimental Methods

[0041] ICR mice were adaptively fed for 3 days and were allowed to eat feed and water freely. They were randomly divided into normal control group, model control group, amoxicillin capsule control group, and high, medium and low dose groups of the Chinese medicine composition according to their body weight, with 10 mice in each group.

[0042] A neurological deafness mouse model was induced with a dose of 150 mg / kg of D-galactose. After weighing the required drug, dissolve it in physiological saline to obtain the D-galactose solution of the required concentration, seal it, protect it from light, and store it for later use. Except for the normal control group, the remaining mice were intraperitoneally injected with 150 mg / kg of D-galactose (0.1 mL / 10 g) once a day for 60 consecutive days; the normal control group was intraperitoneally injected with 0.9% physiological saline under the same conditions. After the model was successfully prepared, it was administered by gavage. Blank control group and model group: physiological saline, once a day, for 14 consecutive days. Positive drug group, low-, medium-, and high-dose groups of Chinese medicine composition: the corresponding drugs were given once a day for 14 consecutive days.

[0043] At the end of the experiment, in accordance with the ethical principles of experimental animal welfare, mice were anesthetized by intraperitoneal injection of 10% chloral hydrate at a rate of 0.3 mL / 100 g of mouse body weight, and then killed by cervical dislocation, and bilateral cochleae were removed. The cAMP content in the homogenate of mouse cochlear tissue was detected by ELISA. The expression level of PKA mRNA in mouse cochlear tissue was detected by QPCR.

[0044] 4. Statistical methods

[0045] The results were statistically analyzed using t-test.

[0046] 5. Experimental Results

[0047] 5.1 Effect of D-galactose on cAMP content in cochlear tissue homogenate of mouse model of sensorineural deafness

[0048] The results of cAMP content in the cochlear tissue homogenate of mice in each group showed that compared with the normal control group, the cAMP content in the cochlear tissue homogenate of mice in the model group was significantly reduced (P < 0.001), indicating that the model was successfully established; compared with the model group, the cAMP content in the positive drug, medium-dose group and high-dose group of Chinese medicine combination was significantly increased (P < 0.001, P < 0.01), indicating that drug treatment can increase the cAMP content in cochlear tissue. The results are shown in Table 1.

[0049] Table 1: cAMP content in cochlear tissue of mice in each group

[0050]

[0051] Note: Compared with the normal control group, ### P<0.001; compared with the model control group, ***P<0.001, **P<0.01.

[0052] 5.2 Effects on PKA mRNA expression in cochlear tissue of D-galactose-induced neurological deafness mouse model

[0053] The results of the expression level of PKA mRNA in the cochlear tissue of mice in each group showed that compared with the normal control group, the expression level of PKA mRNA in the cochlear tissue of mice in the model group was significantly reduced (P < 0.001), indicating that the model was successfully established; compared with the model group, the expression level of PKA mRNA in the cochlear tissue of the medium-dose group was increased (P < 0.05), and the expression level of PKA mRNA in the positive drug group and the high-dose group was significantly increased (P < 0.001). The results are shown in Table 2.

[0054] Table 2: PKA mRNA expression levels in cochlear tissues of mice in each group

[0055]

[0056] Note: Compared with the normal control group, ### P<0.001; compared with the model control group, ***P<0.001, *P<0.05.

[0057] 6. Experimental conclusion

[0058] The Chinese herbal composition at 3.36 and 1.68 g / kg can significantly increase the cAMP content in cochlear tissue and significantly improve the expression level of PKA mRNA, indicating that the Chinese herbal composition has a certain therapeutic effect on D-galactose-induced sensorineural deafness in mice.

[0059] Experimental Example 3 Therapeutic effect of Chinese medicine composition on the rat model of neurological deafness induced by cochlear ischemia-reperfusion

[0060] 1. Experimental Materials

[0061] 1.1 Animals

[0062] SD rats, male, SPF grade, 80, weighing 240-260 g, were purchased from Sibeifu (Suzhou) Biotechnology Co., Ltd., animal certificate number: NO.202367043.

[0063] Rearing environment: room temperature 20-26℃, relative humidity controlled at 40-70%. Rearing in separate cages, 5 per group.

[0064] 1.2 Drugs and reagents

[0065] The Chinese medicine composition was prepared according to the method of Example 1 and provided by Jiangsu Kangyuan Pharmaceutical Co., Ltd.; dexamethasone sodium phosphate injection (Guizhou Guangzheng Pharmaceutical Co., Ltd., batch number 19032034); chloral hydrate (Sinopharm Chemical Reagent Co., Ltd., batch number 20200320); catalase (CAT) kit (Nanjing Jiancheng, batch number 20190413).

[0066] 1.3 Instrument

[0067] Electronic balance (Sartorius Scientific Instruments, model BS224S); electronic balance (G&G, model TC3K); low-temperature centrifuge (eppendorf, Germany, model 5804R); microplate reader (Megu Molecular, USA, model Flexstation 3); brainstem auditory evoked potential instrument (MADSEN, Denmark, model: Octavus).

[0068] 2. Dosage design

[0069] The daily dosage of the Chinese medicine composition is 4.08 g of crude drug / day. According to the body surface area conversion method, the equivalent dose for rats is 4.08 g / 60 kg*6.2=0.42 g of crude drug / kg. According to the 1 / 4:1 / 2:1 dosing method, the medium dose is 0.84 g of crude drug / kg and the high dose is 1.68 g of crude drug. Intragastric administration, 0.1 mL / 10 g, once a day.

[0070] The dosage of dexamethasone sodium phosphate injection is 14.5 mg / day. According to the body surface area conversion method, the equivalent dose for rats is 14.5 mg / 60 kg*6.2=1.5 mg / kg.

[0071] 3. Experimental Methods

[0072] 60 rats with appropriate body weight were selected from 80 rats, and the rats were randomly divided into a normal control group, a model group, a low-, medium-, and high-dose group of the Chinese medicine composition, and a dexamethasone group, with 10 rats in each group, and the CIRI model was prepared by vascular occlusion. Conventional anesthesia, fixation, disinfection and skin preparation were performed, and an incision was made in the middle of the front neck of the rat, and the subclavian artery and branch vertebral artery were exposed. The common carotid artery was clamped with a sterile arteriolar clamp to form local ischemia for 1 hour and then released to restore blood supply and form reperfusion. The control group only made an incision, and the incision was sutured after exposing the relevant artery. The low-, medium-, and high-dose groups of the Chinese medicine composition and the dexamethasone group were given the corresponding drugs after model preparation, and the normal control group and the model group were given equal doses of normal saline for gavage, and the intervention was continuous for 7 days.

[0073] ABR threshold test: After the drug administration, the ABR threshold was measured in a dark room. Except for the normal control group, which measured the ABR threshold before the experiment and before being killed, the ABR thresholds of the animals in the other groups were measured at two time points: 24 hours after modeling and before drug administration, and 7 days after modeling. At the end of the experiment, all rats were anesthetized and cochlear tissue was taken for detection and CAT activity level.

[0074] 4. Statistical methods

[0075] The results were statistically analyzed using t-test.

[0076] 5. Experimental Results

[0077] 5.1 Effects on ABR response threshold of rat model of neurological deafness induced by cochlear ischemia-reperfusion

[0078] Compared with the normal control group, the ABR response threshold of the model group animals increased significantly (P < 0.001); compared with the model group, the ABR response threshold of the positive drug and the medium and high dose groups of the Chinese medicine combination were significantly reduced (P < 0.05, P < 0.001). The results are shown in Table 3.

[0079] Table 3: Effects on ABR response thresholds in rat neurological deafness model of cochlear ischemia-reperfusion

[0080]

[0081] Note: Compared with the normal control group, ### P<0.001; compared with the model control group, ***P<0.01, *P<0.05.

[0082] 5.2 Effects of CAT levels in cochlear tissue of rats with neurological deafness induced by cochlear ischemia-reperfusion

[0083] Compared with the normal control group, the CAT content in the cochlear tissue of the model group animals was significantly reduced (P < 0.001); compared with the model group, the CAT activity of the positive drug and the medium and high dose groups of the Chinese medicine combination was significantly increased (P < 0.01, P < 0.001). The results are shown in Table 4.

[0084] Table 4: Effects of cochlear ischemia-reperfusion on CAT levels in rat neural deafness model

[0085]

[0086] Note: Compared with the normal control group, ### P<0.001; compared with the model control group, ***P<0.01, **P<0.01.

[0087] 6. Experimental conclusion

[0088] The Chinese medicine composition can significantly reduce the ABR response threshold, improve auditory nerve function damage, and significantly reduce the CAT content in cochlear tissue, indicating that the Chinese medicine composition has a certain therapeutic effect on the rat model of sensorineural deafness with cochlear ischemia-reperfusion.

[0089] The above embodiments of the present invention are merely examples for clearly illustrating the present invention, and are not intended to limit the embodiments of the present invention. For those skilled in the art, other different forms of changes or modifications can be made based on the above description. It is not necessary and impossible to list all the embodiments here. Any modifications, equivalent substitutions and improvements made within the spirit and principles of the present invention should be included in the protection scope of the claims of the present invention.

Claims

1. Use of a Chinese medicine composition in the preparation of a drug for treating neurological deafness, characterized in that: The traditional Chinese medicine composition is prepared from raw materials in the following weight proportions: 5-25 parts of Chuanxiong and 1.5-8 parts of Gastrodia elata.

2. The use according to claim 1, characterized in that: The traditional Chinese medicine composition is prepared from raw materials in the following weight proportions: 8-15 parts of Chuanxiong and 2-6 parts of Gastrodia elata.

3. The use according to claim 1, characterized in that: The traditional Chinese medicine composition is prepared from raw materials in the following weight proportions: 8-10 parts of Chuanxiong and 2.5-5 parts of Gastrodia elata.

4. The use according to claim 1, characterized in that: The traditional Chinese medicine composition is prepared from raw materials in the following weight proportions: 10 parts of Chuanxiong and 2.5 parts of Gastrodia elata.

5. The use according to any one of claims 1 to 4, characterized in that: The drug is selected from oral dosage forms, injection dosage forms or external administration preparations.

6. The use according to claim 5, characterized in that: The medicine is selected from decoctions, tablets, capsules, granules, pills, injections, powders, decoctions, suspensions, dispersants, syrups, suppositories, gels, aerosols, patches, and oral liquids.

7. The use according to claim 1, characterized in that: The method for preparing the Chinese medicine composition comprises the following steps: (1) Take two components, Chuanxiong and Gastrodia elata, in a weight ratio of 5-25 of Chuanxiong and 1.5-8 of Gastrodia elata. drying, crushing and mixing the obtained mixture; (2) extracting the mixture with 70%-90% ethanol reflux for 2-3 times to obtain an extract and medicinal residues, 1-3 hours each time; (3) combining the extracts and filtering to obtain a filtrate; (4) The filtrate is concentrated to a clear paste having a relative density of 1.21-1.30 at 55-65°C.

8. The use according to claim 7, characterized in that: The preparation method of the Chinese medicine composition also includes: (5) decocting the medicinal residue with water for 1-2 hours, combining the decoction, filtering, and concentrating the filtrate after the decoction to a clear paste with a relative density of 1.21-1.30 at 55-65° C., preferably decocting for 1 hour; (6) Combine the clear pastes obtained in step (4) and step (5).

9. The use according to claim 6 or 7, characterized in that: Step (2) is to extract the mixture by reflux with 90% ethanol twice to obtain an extract, each time for 2 hours.

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