Wufuding Shen Decoction for treating Parkinson's disease and preparation method thereof

Abstract

The application discloses a wufutudingshen decoction for treating Parkinson's disease and a preparation method thereof. The wufutudingshen decoction is composed of main materials and auxiliary materials. The main materials comprise kadsura longipedunculata, fig wisteria, clematis, kadsura pinnata and Uncaria, and the auxiliary materials comprise radix paeoniae alba and licorice. Compared with the prior art, the application is based on the Zhuang medical etiology and pathogenesis of Parkinson's disease, and through rigorous compatibility and network pharmacology, the main drugs of clematis, Uncaria, kadsura longipedunculata, fig wisteria and kadsura pinnata, the auxiliary drug of radix paeoniae alba and the drug carrier of licorice are screened out. Seven drugs are used together to achieve the functions of 'dispelling wind, nourishing yin, balancing qi and blood, regulating the Xiqiao and dredging the two channels'. The Zhuang medicine formula has the functions of improving the symptoms of dyskinesia, reducing the loss of substantia nigra-striatal dopaminergic neurons, adjusting the intestinal flora and metabolites of the PD model and promoting the proliferation of regulatory T cells.

Description

Technical Field

[0001] The present invention relates to the technical field of traditional Chinese medicines, and in particular to a Wuteng Dingshen Decoction for treating Parkinson's disease and a preparation method thereof. Background Art

[0002] Parkinson's disease (PD) is a very common neurodegenerative disease. Approximately 3 in every 1,000 people worldwide suffer from PD. The disease is characterized by the degeneration and death of dopamine (DA) neurons in the substantia nigra of the midbrain and a significant decrease in DA content in the striatum. It can cause patients to have a variety of motor and non-motor symptoms, seriously affecting their ability to care for themselves and placing a heavy burden on their families and society. Currently, dopa replacement therapy is the preferred treatment for PD, but studies have found that long-term use of the drug will still lead to many limitations, such as reduced efficacy and symptom fluctuations. In-depth research on the pathophysiological mechanisms of PD, finding new, appropriate and effective intervention measures, and developing new therapeutic targets are key issues that need to be addressed.

[0003] Zhuang medicine is an important component of traditional Chinese medicine, boasting a long history. Long-standing clinical practice has yielded a wealth of effective single, compound, secret, and proven prescriptions. Zhuang medicine does not have a specific name for Parkinson's disease. In ancient times, the condition was often categorized as "tremor" or "cramp," but modern Chinese medicine has designated it "tremor and cramp disease." With the increasing understanding of the disease, modern Zhuang people often describe it as "fengsen (hand tremors)," "disease (foot tremors)," and "dangsen (body tremors)," hence the Zhuang name for Parkinson's disease, "disease (cramps)."

[0004] Historically, there have been very few records of Zhuang medicine prescriptions for PD. Based on the basic theories and treatment principles of Zhuang medicine, combined with the characteristics of PD and years of clinical observation, the inventors proposed a core pathogenesis hypothesis of "yin deficiency and blood deficiency, endogenous wind toxins, apraxia of the cerebral arteries, and obstruction of the two pathways (dragon and fire pathways)." Treatment should focus on "dispelling wind and nourishing yin, balancing qi and blood, and regulating the cerebral arteries and the two pathways." Through rigorous compatibility and network pharmacological screening, they developed a Zhuang medicine prescription, "Wuteng Dingshen Decoction," to alleviate symptoms. This prescription primarily combines the Guangxi Zhuang Autonomous Region's specialty herbs Millettia reticulata, Fufang Teng, Jiaojiao Teng, Kuanjin Teng, and Uncaria rhynchophylla, along with white peony root and licorice. This Zhuang medicine compound boasts significant efficacy and minimal toxicity, improving a range of motor and non-motor symptoms in PD patients. Furthermore, the medicinal ingredients are readily available at traditional Chinese medicine stores, making them inexpensive, and the preparation process simple, making it convenient to use. Moreover, based on Wuteng Dingshen Decoction, it was further improved by adding auxiliary medicinal ingredients to enhance the efficacy of Wuteng Dingshen Decoction. Summary of the Invention

[0005] The present application aims to provide a Wutudingshen decoction for treating Parkinson's disease, aiming to provide a Zang medicine compound with significant curative effect on improving a series of motor and non-motor symptoms of Parkinson's disease patients and small toxic and side effects according to the basic theory of Zang medicine.

[0006] The present application also aims to provide a preparation method of the Wutudingshen decoction for treating Parkinson's disease, aiming to improve the preparation method of the Wutudingshen decoction.

[0007] To achieve the above-mentioned purpose, the present application provides a Wutudingshen decoction for treating Parkinson's disease, 1, the components of the Wutudingshen decoction are as follows:

[0008] The components of the main material include: caudate vitex, cissus japonica, semen momordicae, kadsura longipedunculata and Uncaria.

[0009] The components of the auxiliary material include: white peony root and licorice.

[0010] Preferably, in the above technical solution, the components of the main material are proportioned by weight as follows: 5-15g caudate vitex, 10-20g cissus japonica, 5-15g semen momordicae, 5-15g kadsura longipedunculata and 10-20g Uncaria, and the components of the auxiliary material are proportioned by weight as follows: 5-15g white peony root and 2-8g licorice.

[0011] Preferably, in the above technical solution, the components of the main material are proportioned by weight as follows: 8-12g caudate vitex, 13-17g cissus japonica, 8-12g semen momordicae, 8-12g kadsura longipedunculata and 13-17g Uncaria, and the components of the auxiliary material are proportioned by weight as follows: 13-17g white peony root and 3-6g licorice.

[0012] Preferably, in the above technical solution, the components of the main material are proportioned by weight as follows: 10g caudate vitex, 15g cissus japonica, 10g semen momordicae, 10g kadsura longipedunculata and 15g Uncaria, and the components of the auxiliary material are proportioned by weight as follows: 15g white peony root and 5g licorice.

[0013] Preferably, in the above technical solution, the components of the auxiliary material further include eucommia and dragon bone.

[0014] Preferably, in the above technical solution, the components of the main material are proportioned by weight as follows: 8-12g caudate vitex, 13-17g cissus japonica, 8-12g semen momordicae, 8-12g kadsura longipedunculata and 13-17g Uncaria, and the components of the auxiliary material are proportioned by weight as follows: 5-10g eucommia, 5-10g dragon bone, 13-17g white peony root and 3-6g licorice.

[0015] Preferably, in the above technical solution, the components of the auxiliary material further include Millettia pinnata and Sabia schumanniana.

[0016] Preferably, in the above technical solution, the components of the main material are proportioned by weight as 8-12 g of Spatholobus suberectus Dunn, 13-17 g of Euonymus fortunei (Turcz.) Hand.-Mazz, 8-12 g of Menispermum dauricum DC, 8-12 g of Sinomenium acutum Rehd et Wils, 13-17 g of Uncaria rhynchophylla (Miq.) Juss, and the components of the auxiliary material are proportioned by weight as 5-10 g of Eucommia ulmoides Oliv, 5-10 g of Cornus officinalis Sieb et Zucc, 5-10 g of Millettia pinnata (L.) Panzer, 8-12 g of Sargentodoxa cuneata Oliv, 13-17 g of Paeonia lactiflora Pall and 3-6 g of Glycyrrhiza uralensis Fisch.

[0017] According to the above technical solution, the components of the Wufuding Decoction are mainly composed of Spatholobus suberectus Dunn, Euonymus fortunei (Turcz.) Hand.-Mazz, Menispermum dauricum DC, Sinomenium acutum Rehd et Wils, Uncaria rhynchophylla (Miq.) Juss, Paeonia lactiflora Pall and Glycyrrhiza uralensis Fisch, which are characteristic Zhuang medicines in Guangxi Zhuang Autonomous Region, and the properties of each raw medicinal material are as follows:

[0018] Spatholobus suberectus Dunn, also known as Xuefengteng, Xuefeng, Xuefeng, Sanye Xuefengteng, Jiucengfeng, is the stem of Spatholobus suberectus Dunn, Baimahuang Youmahuang or Liangyeyuanhuangdeng. It is bitter and sweet, warm in nature, and belongs to the liver and kidney channels. Spatholobus suberectus Dunn has the effects of invigorating blood, tonifying blood, relaxing sinews and activating collaterals, regulating menstruation and relieving pain, and is mainly used to treat lumbago and knee pain, numbness and paralysis, rheumatic arthralgia, blood deficiency and emaciation, and irregular menstruation.

[0019] Euonymus fortunei (Turcz.) Hand.-Mazz, also known as Jinxianfeng, Jiuniuzao, Kuaifangfeng, Luoshengteng, Paowangcao, is a evergreen shrub of Euonymus fortunei (Turcz.) Hand.-Mazz. It is bitter and slightly warm in nature. It has the effects of relaxing sinews and activating collaterals, and stopping bleeding and removing stasis, and is mainly used to treat lumbar muscle strain, rheumatic arthralgia, hemoptysis, metrorrhagia, irregular menstruation, fractures and hemorrhage from trauma.

[0020] Tuber Fleeceflower Stem, also known as Qiteng, Shouwuteng, is the stem or leafy stem of Polygonum multiflorum Thunb. It is sweet and slightly bitter in taste, and is neutral in nature, and belongs to the heart and liver channels. Tuber Fleeceflower Stem has the effects of nourishing the heart and calming the mind, expelling wind, and freeing the collaterals, and is mainly used to treat insomnia, dreaminess, blood deficiency and body pain, numbness of the skin, rheumatic arthralgia, and pruritus of wind rashes.

[0021] Sinomenium acutum Rehd et Wils, also known as Songenteng, Shujinteng, Damoshshen, is the stem of Sinomenium acutum Rehd et Wils. It is bitter in taste and cool in nature, and belongs to the liver channel. Sinomenium acutum Rehd et Wils has the effects of relaxing sinews and activating collaterals, and expelling wind and relieving pain, and is mainly used to treat rheumatic arthralgia, sciatica, lumbar muscle strain, and injuries from falls.

[0022] Uncaria rhynchophylla (Miq.) Miq. ex Havil.), also known as Da Gouding, Shuang Gouding, is the dry stem branches of Uncaria rhynchophylla, Uncaria macrophylla, Uncaria sinensis, Uncaria sinensis or Uncaria sinensis. It tastes sweet and slightly cold; it belongs to the liver and pericardium channels. Uncaria has the effects of calming wind and calming, stopping convulsions, clearing liver and liver, and dispersing wind and heat. It is mainly used to treat internal liver wind, convulsions, high fever, convulsions, infantile convulsions, headache and dizziness, cold with convulsions, infantile convulsions, headache and dizziness, and other diseases.

[0023] Os Draconis, also known as Lu Hu Yisheng, Nagabone, Shenglonggu, Calcined Dragon Bone, Five-Flower Dragon Bone, Qinghua Longgu, Flower Dragon Bone, and White Dragon Bone, is a fossil bone of ancient large mammals such as elephants, rhinoceroses, and three-toed horses. It tastes bitter and astringent, and is cold in nature; it belongs to the heart and liver channels. Dragon bone has the effects of calming the liver and calming the liver, calming the spirit, and astringing and astringing. It is mainly used to treat restlessness, insomnia, convulsions, dizziness, and other diseases.

[0024] Sinomenium acutum (Thunb.) Rehd. et Wils., also known as Dafengteng, Fufengsan, He Fangji, Paifengteng, Qingfangji, is the dry tuber of Sinomenium acutum and Sinomenium acutum. It tastes bitter and pungent, and is flat; it belongs to the liver and spleen channels. Qingfengteng has the effects of expelling wind and dampness, unblocking meridians and collaterals, and promoting urination; it is mainly used to treat arthralgia, rheumatoid arthritis, edema, foot disease, heart and stomach pain, and other diseases.

[0025] Millettia speciosa Champ., also known as Meilvyaoduoteng, Shantaluo, Dalishu. It tastes sweet and is flat; it belongs to the lung and kidney channels. Millettia speciosa has the effects of tonifying deficiency and moistening the lungs, strengthening the tendons and activating the collaterals; it is mainly used to treat muscle strain, rheumatoid arthritis, lung heat, lung deficiency cough, pulmonary tuberculosis, chronic bronchitis, chronic hepatitis, and other diseases.

[0026] Eucommia ulmoides Oliv., also known as Tiaoxipei, Sizhong, Silk Cotton Skin, Jade Silk Skin. It tastes sweet and slightly bitter, and is warm; it belongs to the liver and kidney channels. Eucommia ulmoides has the effects of tonifying liver and kidney and strengthening tendons and bones; it is mainly used to treat kidney deficiency, rheumatism, muscle weakness, stiffness, and other diseases.

[0027] A preparation method of Wutengding Shen Decoction for treating Parkinson's disease, as described above, characterized in that the method comprises:

[0028] (1) accurately weigh the dry medicinal materials of Millettia speciosa, Euonymus fortunei, Caulis Polygoni Multiflori, Millettia speciosa, Uncaria rhynchophylla, Paeonia lactiflora and Glycyrrhiza uralensis, respectively;

[0029] (2) Place the dried medicinal materials in a clay pot, add 500-700 mL of water, boil over high heat, and simmer over low heat until the medicinal liquid is 200-300 mL. Divide the mixture into two portions, each portion containing 100-150 mL of Wuteng Dingshen Decoction.

[0030] Preferably, in the above technical solution, the Wuteng Dingshen Decoction is an oral medicine, and the method of taking it is to take it warm once in the morning and once in the evening after meals.

[0031] Compared with the prior art, the present invention has the following beneficial effects:

[0032] (1) The pharmacology of the Wuteng Dingshen Decoction of the present invention in treating Parkinson's disease is as follows: Uncaria rhynchophylla is cool in nature, sweet and bitter in taste, and Polygonum multiflorum is flat in nature and sweet in taste. Both can dispel wind poison, regulate Qi and blood, and clear the fire path; Millettia reticulata can nourish blood and dispel wind, and is good at clearing the fire path; Radix Achyranthis Bidentatae is good at clearing the dragon path; Rhizoma Cibotii mainly replenishes Qi and blood, and clears the two paths; Wuteng is used in the prescription as the main drug; White Peony Root nourishes blood, softens the liver, and restrains Yin, and is a supporting drug with the effect of strengthening the main drug; Licorice harmonizes the other drugs and is a supporting drug. The seven drugs are used together to achieve the effect of "dispelling wind and nourishing Yin, balancing Qi and blood, regulating Qi and blood, and clearing the two paths". This Wuteng Dingshen Decoction can improve the symptoms of movement disorders, reduce the loss of dopamine neurons in the substantia nigra-striatum, and has the effect of adjusting the intestinal flora and its metabolites in PD models and promoting the proliferation of regulatory T cells.

[0033] (2) Wuteng Dingshen Decoction has achieved positive intervention effects in both animal experiments and clinical practice. Network pharmacology analysis and animal experiments have also revealed the effective mechanism of action of this Zhuang medicine prescription in treating Parkinson's disease, clinically improving the clinical symptoms of Parkinson's patients and improving their quality of life.

[0034] (3) The pharmacological mechanism of the Wuteng Dingshen Decoction of the present invention in treating Parkinson's disease is as follows: the five vines are used as the main ingredients in the prescription; white peony root nourishes blood, softens the liver, and restrains yin; dragon bone calms the liver, calms the mind, and eucommia bark nourishes the liver, nourishes the kidney, replenishes blood, and improves intelligence; oxalis strengthens qi, transports blood, and activates collaterals; and Qingfengteng dispels wind, dredges the collaterals, and relieves pain. These five ingredients serve as auxiliary ingredients to enhance the efficacy of the main ingredient and assist in treating secondary symptoms; liquorice harmonizes the other ingredients and serves as a supporting ingredient. Behavioral evaluation and striatal DA results in animal experiments of the improved Wuteng Dingshen Decoction showed that the therapeutic effect was significantly enhanced. BRIEF DESCRIPTION OF THE DRAWINGS

[0035] The purpose, features and advantages of the present invention will be further described with reference to the accompanying drawings and in conjunction with the embodiments.

[0036] Figure 1 The graphs show the distance and speed of the mine test after drug administration, as well as the DA content in the striatum of the Control group, MPTP group, MPTP+WTlow group, MPTP+WTmid group, and MPTP+WThig group;

[0037] Figure 2 is a graph of the abundance metabolism of bifidobacterium in the control group, the MPTP group and the MPTP+WThig group;

[0038] Figure 3 is a graph of the abundance metabolism of bifidobacterium in the control group, the MPTP group and the MPTP+WThig group;

[0039] Figure 4 is a LDA difference contribution graph of the peripheral blood metabolites of mice treated with Wutuding Shen Decoction;

[0040] Figure 5 is a graph of discriminant analysis and cluster analysis of Wutuding Shen Decoction;

[0041] Figure 6 is a graph of the Tregs in the spleen of the control group, the MPTP group and the MPTP+WThig group;

[0042] Figure 7 is a graph of the logic gate range of flow cytometry of the method;

[0043] Figure 8 is a column chart of the Foxp3 transcription factor level in the spleen of the control group, the MPTP group and the MPTP+WThig group;

[0044] Figure 9 is a column chart of the scores of patients before and after treatment in the clinical experiment;

[0045] Figure 10 is a column chart of the open period improvement rate of patients before and after treatment in the clinical experiment and a bar chart of the defecation frequency of patients per week;

[0046] Figure 11 is a graph of predicting the potential target and action mechanism of Wutuding Shen Decoction in treating Parkinson's disease. DETAILED DESCRIPTION

[0047] The test mouse conditions and medication conditions of the animal experiment involved in the present application are as follows:

[0048] (1) Animals used in the experiment

[0049] The animals were obtained from the Scientific Experimental Center of Guangxi University of Traditional Chinese Medicine. The C57BL / 6 mice used in the experiment were SPF grade and about 8-10 weeks old. The animals were raised under specific pathogen-free (SPF) safety conditions. The breeding environment was maintained at 22±2°C, the relative humidity was maintained at 50%-60%, artificial light was used to simulate natural conditions with 12 hours of light and dark each, and sufficient food and water were provided during breeding. All experimental operators have the "Certificate of Qualification for Professional Technical Examination of Laboratory Animals". All animal experiments comply with the relevant provisions of the "Ethics Certificate for Laboratory Animals". Laboratory Animal Production License No. SCXK (Xiang)

[0050] 2019-0004, approved by the Ethics Committee of Guangxi University of Traditional Chinese Medicine (Ethics No. DW20230411-048).

[0051] (2) Medication and experimental conditions

[0052] Drug preparation: All the medicinal materials involved in the present invention are single-ingredient Chinese herbal medicine slices, purchased uniformly by the Pharmacy Department of the International Zhuang Medicine Hospital Affiliated to Guangxi Zhuang University of Traditional Chinese Medicine and prepared according to requirements, and the liquid is prepared at 0.1g / ml;

[0053] Dosage conversion for mouse experiments: According to the animal dosage in "Laboratory Animals and Animal Experimental Techniques", the daily dose for mice (mg / kg) = 9.01 × human dose (mg / kg); the standard human body weight is 60kg, and the daily dose of Wuteng Dingshen Decoction for adults is 80g. Therefore, the daily dose for mice = 9.01 ×

[0054] (80 / 60) = 12013 mg / kg. According to the above method, the daily dose for mice is calculated to be 240.26 mg (the mice are administered at a weight of 20 g each), which is approximately equal to 240 mg. The dosage volume for mice is 0.2 mL / 10 g, which corresponds to 0.5 g / mL of Wuteng Dingshen Decoction. This dose is used as the medium dose for animal administration, the high dose is 1.0 g / mL, and the low dose is 0.25 g / mL.

[0055] Example 1

[0056] A Wuteng Dingshen Decoction for treating Parkinson's disease is composed of a main ingredient and auxiliary ingredients, wherein the main ingredient components are 10g of Millettia reticulata, 15g of Thunbergia fortunei, 10g of Polygonum multiflorum, 10g of Caulis Achyranthis Bidentatae and 15g of Uncaria rhynchophylla; and the auxiliary ingredients are 15g of White Peony Root and 5g of Licorice.

[0057] A method for preparing Wuteng Dingshen Decoction for treating Parkinson's disease, the method comprising:

[0058] (1) Respectively, accurately take 10g of the dry medicinal materials of Broussonetia kaempferi, 15g of Taxillus delavayi, 10g of Tetrastigma hemsleyanum, 10g of Tetrastigma obtectum, 15g of Uncaria rhynchophylla, 15g of Paeonia lactiflora and 5g of Glycyrrhiza uralensis, etc.

[0059] (2) Put the dry medicinal materials in a pot, add 500mL of water, boil with high heat, and slowly simmer with small heat until the medicinal liquid is 200mL, and divide it into two portions, each of which is five-teng Dingshen decoction of 100mL.

[0060] The use method of the above five-teng Dingshen decoction is as follows: internal use, taking twice a day, specifically, 100mL of each of the patient's morning and evening after meals, 3 weeks for a course of treatment.

[0061] Example 2

[0062] A five-teng Dingshen decoction for treating Parkinson's disease, which is composed of main materials and auxiliary materials, the main material components are 5g of Broussonetia kaempferi, 10g of Taxillus delavayi, 5g of Tetrastigma hemsleyanum, 5g of Tetrastigma obtectum and 10g of Uncaria rhynchophylla; the auxiliary material components are 10g of Paeonia lactiflora and 2g of Glycyrrhiza uralensis.

[0063] The preparation method and the taking method are the same as those of example 1.

[0064] Example 3

[0065] A five-teng Dingshen decoction for treating Parkinson's disease, which is composed of main materials and auxiliary materials, the main material components are 15g of Broussonetia kaempferi, 20g of Taxillus delavayi, 15g of Tetrastigma hemsleyanum, 15g of Tetrastigma obtectum and 20g of Uncaria rhynchophylla; the auxiliary material components are 20g of Paeonia lactiflora and 8g of Glycyrrhiza uralensis.

[0066] The preparation method and the taking method are the same as those of example 1.

[0067] Comparative example 1

[0068] It is basically the same as example 1, the difference lies in that the main materials are different, and the main materials lack Tetrastigma hemsleyanum, Taxillus delavayi and Tetrastigma obtectum. The Dingshen decoction is composed of main materials and auxiliary materials, the main material components are 10g of Broussonetia kaempferi and 15g of Uncaria rhynchophylla; the auxiliary material components are 15g of Paeonia lactiflora and 5g of Glycyrrhiza uralensis.

[0069] Comparative example 2

[0070] It is basically the same as example 1, the difference lies in that the main materials are different, and the main materials lack Taxillus delavayi and Tetrastigma obtectum. The Dingshen decoction is composed of main materials and auxiliary materials, the main material components are 10g of Broussonetia kaempferi, 10g of Tetrastigma hemsleyanum and 15g of Uncaria rhynchophylla; the auxiliary material components are 15g of Paeonia lactiflora and 5g of Glycyrrhiza uralensis.

[0071] Comparative example 3

[0072] The difference between the embodiment 1 and the embodiment 2 is that the main material is different, and the main material is replaced by Tripterygium instead of Taxillus vinceotii. The Ding Shen decoction is composed of the main material and the auxiliary material. The main material is composed of 10 g of Millettia pinnata, 15 g of Tripterygium, 10 g of Cissus jamaicensis, 10 g of Entada phaseoloides, and 15 g of Uncaria tomentosa. The auxiliary material is composed of 15 g of Paeonia lactiflora and 5 g of Glycyrrhiza uralensis.

[0073] I. Animal experiment 1

[0074] The purpose of the experiment is to detect the influence of Ding Shen decoction with different components on the behavior, metabolites, and other indicators of MPTP model mice.

[0075] The C57BL / 6 mice were randomly divided into 5 groups, and the specific treatment of each group of mice is as follows:

[0076] ① Control group (Control): equal amount of physiological saline intraperitoneal injection.

[0077] ② Model group (MPTP): continuous intraperitoneal injection of MPTP (1-methyl-4-phenyl-1, 2, 3, 6-tetrahydropyridine) for 5 days to establish a Parkinson's disease model.

[0078] ③ Example 1: continuous intraperitoneal injection of MPTP for 5 days, and 0.50 g / mL dose of Ding Shen decoction prepared in example 1 was administered orally for 14 days;

[0079] ④ Comparative example 1: continuous intraperitoneal injection of MPTP for 5 days, and 0.50 g / mL dose of Ding Shen decoction prepared in comparative example 1 was administered orally for 14 days;

[0080] ⑤ Comparative example 2: continuous intraperitoneal injection of MPTP for 5 days, and 0.50 g / mL dose of Ding Shen decoction prepared in comparative example 2 was administered orally for 14 days;

[0081] ⑥ Comparative example 3: continuous intraperitoneal injection of MPTP for 5 days, and 0.50 g / mL dose of Ding Shen decoction prepared in comparative example 3 was administered orally for 14 days.

[0082] (1) Open field test

[0083] The behavior of the experimental mice was evaluated by the open field test, and the evaluation results were used to represent the movement ability of the mice. The open field test results are shown in the following table 1-2.

[0084] Table 1: Open field test distance baseline and distance after the experiment

[0085]

[0086] As shown in Table 1, the MPTP model mice administered with the Wufuding Shen Tang of Example 1 were significantly higher than the MPTP group mice, and similar to the blank group mice, as compared with the MPTP group; while the mice of Comparative Examples 1-3 were higher than the MPTP group mice but significantly lower than the mice of Example 1.

[0087] Table 2: Speed baseline and speed after the experiment of the open field test

[0088]

[0089]

[0090] As shown in Table 2, the speed of the mice of Comparative Examples 1-3 was lower than that of the mice of Example 1.

[0091] (2) Striatal DA determination

[0092] Striatal sampling: After the open field test, the experimental mice were intraperitoneally injected with 1% sodium pentobarbital sodium at 35 mg / kg of body weight for anesthesia, and the brain was removed after anesthesia, and the bilateral striatum was taken for determination of striatal dopamine. The content of dopamine in the striatum was detected by high performance liquid chromatography, and the detection steps were as follows: ① sample pretreatment: after the behavior test, the mice were sacrificed by cervical dislocation method, and the striatal tissue was taken quickly. The striatal tissue was weighed and added to the tissue homogenate solution (0.4M perchloric acid solution containing 0.1mM disodium ethylenediaminetetraacetate) at m / v=1:10, and homogenized. The homogenate was centrifuged at 15000 rpm at 4°C for 20 min, and the supernatant was detected by high performance liquid chromatography-mass spectrometry. ② Standard preparation: accurately weigh the standard into a 100 mL volumetric flask, dissolve and dilute with 1% formic acid, shake well, and prepare a 500 mg / L standard stock solution, which is stored in a 4°C refrigerator. When used, dilute with acetic acid (pH=5.7) to the required concentration. ③ Analysis conditions: chromatographic column: Shim-pack GIST C18 column (75x2.1mm, 2μm); automatic sample injector sample tray temperature control: 4°C; column temperature: room temperature; mobile phase: acetic acid-ethyl acetate (20mmol / L, pH=5.7): acetonitrile=95:5 (v / v); flow rate: 0.2mL / min; injection volume: 5μL; sampling time: positive ion mode 5min, negative ion mode 10min. ④ Qualitative and quantitative analysis of samples. The results of striatal DA determination are shown in Table 3.

[0093] Table 3: Striatal DA determination table

[0094] Group Striatal DA value (ng / g) Difference from MPTP group Blank group —— —— MPTP group 95.86 —— Example 1 144.81 48.95 Comparative Example 1 99.55 3.69 Comparative Example 2 105.38 9.52 Comparative Example 3 126.46 30.60

[0095] II. Animal experiment 2

[0096] The experimental purpose is to detect the behavior, metabolites, etc. of MPTP model mice when the dose of the Wutuding Shen decoction (main ingredient: 10 g of Broussonetia kaempferi, 15 g of Ficus microcarpa, 10 g of Periploca sepium, 10 g of Tetrastigma obtectum, and 15 g of Uncaria rhynchophylla; auxiliary ingredient: 15 g of Radix Paeoniae Alba and 5 g of Radix Glycyrrhizae) is different.

[0097] The C57BL / 6 mice were randomly divided into 5 groups, and the specific treatments of the mice in each group were as follows:

[0098] ① Control group (Control): equal amount of normal saline was injected intraperitoneally.

[0099] ② Model group (MPTP): MPTP (1-methyl-4-phenyl-1, 2, 3, 6-tetrahydropyridine) was continuously injected intraperitoneally for 5 days to establish a Parkinson's disease model.

[0100] ③ Low-dose Wutuding Shen decoction group (MPTP+WTlow): MPTP was continuously injected intraperitoneally for 5 days, and the mice were given 0.25 g / mL dose of Wutuding Shen decoction by gavage for 14 days;

[0101] ④ Medium-dose Wutuding Shen decoction group (MPTP+WTmid): MPTP was continuously injected intraperitoneally for 5 days, and the mice were given 0.50 g / mL dose of Wutuding Shen decoction by gavage for 14 days;

[0102] ⑤ High-dose Wutuding Shen decoction group (MPTP+WThig): MPTP was continuously injected intraperitoneally for 5 days, and the mice were given 0.75 g / mL dose of Wutuding Shen decoction by gavage for 14 days;

[0103] The Wutuding Shen decoction was prepared according to Example 1.

[0104] (1) Behavior detection method and results thereof

[0105] The behavior of the experimental mice was evaluated by using the open field test, and the evaluation results were used to represent the movement ability of the mice. The open field test results are shown in Figure 1 -a, Figure 1 -b, Figure 1 -c.

[0106] From Figure 1 -a, it can be seen that the movement path of the open field test is used as behavior evaluation, and it can be seen that the movement path of the MPTP model mice is less, indicating that the MPTP model has obvious movement disorders; at the same time Figure 1 -b and Table 1-2, it can be seen that after treatment with the Wutuding Shen decoction, the total distance of the MPTP model mice in the MPTP+WTlow, MPTP+WTmid, and MPTP+WThig groups in the open field test increased significantly, and even the total distance of the MPTP model mice in the MPTP+WTmid group was similar to that of the mice in the blank group; from Figure 1-c and Table 1-2 can be seen, after the treatment of Wuding Shen decoction, MPTP + WTlow, MPTP + WTmid and MPTP + WThig group of MPTP model mice in the open field test speed significantly faster, close to the speed of the blank group mice.

[0107] (2) striatal DA and substantia nigra content determination

[0108] Striatum and substantia nigra sampling: after the behavioral test, the experimental mice were intraperitoneally injected with 1% sodium pentobarbital, anesthetized at 35 mg / kg body weight, and then the brain was separated and the bilateral striatum was taken for determination of striatal dopamine. The results of striatal DA content are shown in Table 5 and Figure 1 -d; the remaining brain was fixed in 4% formaldehyde in 0.1 mol / L PB (pH 7.3) for 4 h. Then it was transferred into 20% and 30% sugar solution in turn, and after the tissue block sank, coronal frozen serial sections were made, with a thickness of 25 μm. Three sets of sections (10 sections each) were collected from 2.80 mm-3.80 mm (substantia nigra site) behind the fontanel, and then floated in 0.01 mol / L PBS. The results of the substantia nigra in the methionine hydroxylase are shown in Figure 2 .

[0109] Striatum detection method and operation steps in animal experiment 1 are the same.

[0110] From Figure 1 -d can be seen, the high dose group improved obviously High performance liquid chromatography-mass spectrometry analysis, after the treatment of Wuding Shen decoction, striatal dopamine (DA) content was significantly higher than that of the model group, with the best effect in the high dose group, the difference from MPTP was 174.9, P = 0.0178.

[0111] The method for detecting the tyrosine hydroxylase in the substantia nigra of the midbrain: the number of tyrosine hydroxylase positive neurons in the substantia nigra compacta of the midbrain is calculated by immunofluorescence staining, and the steps are as follows: after the substantia nigra part of the midbrain is taken out, it is rinsed in 0.01 mol / L PBS buffer (pH 7.3) for 3 times, and then it is placed in a blocking solution containing 0.3% Triton X-100 and 10% goat serum for 30 min at room temperature. The blocked section is transferred to a 1:300 dilution of mouse anti-TH monoclonal antibody and incubated in a refrigerator at 4°C for 48 h. The section is taken out and rinsed in 0.01 mol / L PBS for 10 min each time for 3 times, and then it is transferred to a 1:200 dilution of FITC-labeled goat anti-mouse IgG antibody and incubated at room temperature for 4-6 h. After being washed with 0.01 M PBS, the brain slice is mounted on a glass slide, glycerol phosphate buffer is used for mounting, and the fluorescence upright microscope is used for observation and photography. The excitation wavelength of FITC is 488 nm, and the TH immunopositive neurons show green color. The number of TH positive neurons in the SNpc of 10 brain sections (1 set) of each mouse is counted respectively.

[0112] From Figure 2 It can be seen that the tyrosine hydroxylase (TH) staining positive cells in the substantia nigra compacta of the midbrain are significantly reduced, and the TH positive cells are increased after the intervention of Wutuding decoction, and the effect of the medium and high dose groups is better.

[0113] (3) Intestinal flora

[0114] Collection of fecal samples: on the second day after the last gavage of each group of mice, each mouse was placed in a high-pressure sterilized cage box without bedding to freely defecate, and the formed feces were quickly collected in a sterilized EP tube and stored at -80°C.

[0115] Intestinal flora detection method: 16S rDNA amplification and sequencing were used to detect the composition of the fecal flora. The specific steps are as follows: Take an appropriate amount of feces (>50 mg), add ultrapure water at 10 μL / g, shake thoroughly to homogenize, let it stand for 30 minutes, centrifuge at 12,000 rpm at 4°C for 20 minutes, aspirate the supernatant, filter it through a 0.22 μm water filter, and transfer it to a gas phase bottle for measurement. Sequences of the V4-V5 region of 16S rDNA were selected for high-throughput sequencing analysis. The measured raw data were sequence optimized and bioinformatics analyzed. OTU (operational taxonomic unit) clustering was performed, and based on taxonomic information, statistical analysis of community structure was performed at the phylum, class, order, family, genus, and species levels. The spliced ​​optimized sequences were clustered into OTUs using the software USEARCH and annotated with species information. On the basis of the above analysis, the Alpha diversity analysis (Chao and Ace species richness statistics, Shannon and Simpson species diversity statistics) was performed using mothurrua software, and rarefaction curves and Venn diagrams were produced.

[0116] The intestinal flora of MPTP model mice is as follows Figures 3-5 As shown, K represents the blank group, M represents the MPTP model mice, and G represents the high-dose Wuteng Dingshen Decoction group. Figure 3 It can be seen that the intervention of Wuteng Dingshen Decoction increased the abundance metabolomics of Bifidobacterium.

[0117] (4) Peripheral blood metabolite content

[0118] Peripheral blood metabolite content detection method: collect peripheral blood, and detect peripheral blood metabolite content by using high performance liquid chromatography tandem high resolution mass spectrometer, steps are as follows: ① sample pretreatment: 100 μL of serum is taken in a 1.5 mL centrifuge tube, 10 μL of 100 ppb d9-TMAO solution is added, vortex mixing is uniformly carried out for 30 s; 300 μL of methanol is added, vortex mixing is uniformly carried out for 3 min, centrifugation is carried out at 4 DEG C for 10 min, 100 ul of supernatant is taken in a centrifuge tube, the above process is repeated for secondary centrifugation for 3 min, the supernatant is filtered through a 0.22 μm filter into a sample bottle, and 10 ul is taken for analysis. Standard preparation: an appropriate amount of standard is weighed in a 100 mL volumetric flask, methanol is used for constant volume, shaking is uniformly carried out, a standard stock solution is configured, and the standard stock solution is stored in a refrigerator at-20 DEG C. When used, the standard stock solution is diluted to the required concentration with ethyl acetate / water (1:1, v / v) containing 20 mM ascorbic acid. ② Chromatographic column analysis condition: Shim-pack GIST C18 column (75x2.1 mm, 2 μm); automatic sample disc temperature control: 4 DEG C; column temperature: room temperature; mobile phase: acetic acid-acetic acid (20 mmol / L, pH = 5.7): ethyl acetate = 95:5 (v / v); flow rate: 0.2 mL / min; sample amount: 5 μL; sampling time: positive ion mode 5 min, negative ion mode 10 min. ③ Mass spectrometry analysis condition, ionization mode: electrospray ionization negative ion mode (ESI-); capillary voltage 3.0 kV; cone hole voltage 40 V; ion source temperature 100 DEG C; desolvation gas temperature 300 DEG C; desolvation gas flow rate 600 L / h; collector flow rate 50 L / h; collection mode: MSe Continuum mode, negative ion mode; mass number collection range: 100-1200 Da, scanning time: 0.2 sec, high-energy channel voltage: 10-65 V. The data collected are subjected to data pretreatment, and finally the final peak number obtained after QC sample screening is subjected to quality control.

[0119] From Figure 4 and Figure 5 It can be seen that Wutuding Shen Decoction significantly up-regulates 104 metabolites in the peripheral blood of the MPTP model, among which, the products of the amino acid metabolic pathway are the most (n = 9, Figure 5 ).

[0120] (5) Flow cytometry

[0121] Flow cytometry is used to detect the Tregs in the spleen (as shown in Table 5 and Figure 6 Table 6) and the Foxp3 transcription factor level in the spleen (as shown in Table 5 and Figure 8 Table 6), and the flow cytometry logical gate range is shown in Figure 7 .

[0122] Table 5 Tregs in the spleen of the blank group, MPTP and MPTP+WThig

[0123]

[0124] From Table 5 and Figures 6-7 It can be seen that the proportion of Tregs in the spleen of the blank control group, MPTP model group and Wutuding Shen Decoction high-dose treatment group was significantly different between groups (F = 5.102, P = 0.020) by flow cytometry. The proportion of Tregs in the Wutuding Shen Decoction treatment group was significantly higher than that in the MPTP model group (Mean diff = 4.213, P = 0.012).

[0125] Table 6: Spleen Foxp3 Transcription Factor Level Table

[0126]

[0127] From Table 6 and Figure 8 It can be seen that the results of RT-qPCR detection of spleen Foxp3 transcription factor level suggest that there is a significant difference in spleen Foxp3 transcription factor level between the three groups (F = 34.77, P < 0.001). The Foxp3 level in the Wutuding Shen Decoction treatment group was significantly higher than that in the blank control group (Mean diff = 1.754, P < 0.001) and the MPTP model group (Mean diff = 2.182, P < 0.001).

[0128] III. Clinical Cases

[0129] (1) Study Objectives

[0130] Case Source: This study will select 16 cases of PD patients who meet the inclusion criteria, have obtained the approval of the ethics committee (approval number: 2024-102-1) and signed the informed consent form from July 2024 to December 2024 in the inpatient and outpatient departments of the Brain Disease Department of the International Zhuang Medicine Hospital Affiliated to the Guangxi Zhuang Autonomous Region University of Chinese Medicine. This study included 16 subjects, of which 7 were male and 9 were female. The age range of the subjects was 40-74 years, with an average age of (59.50 ± 9.359) years. The disease duration ranged from 0.5 to 7.0 years, with an average disease duration of (3.78 ± 2.359) years.

[0131] The 16 patients met the Western diagnostic criteria and Chinese diagnostic criteria:

[0132] ① Western diagnostic criteria for Parkinson's disease: According to the "Chinese Parkinson's Disease Diagnostic Criteria (2016 Edition)" formulated by the Parkinson's Disease and Movement Disorder Group of the Chinese Medical Association and the Neurology Division of the Chinese Medical Association.

[0133] ②Diagnosis criteria of Parkinson's disease in TCM: According to the diagnostic criteria in the "TCM Old Age Tremor Diagnosis and Efficacy Evaluation Standard" formulated by the National Chinese Medicine Association in 1992, two deputy chief physicians or above independently diagnose the patient, and the results are consistent. If the results are inconsistent, a third deputy chief physician or above will be involved in the diagnosis, and the same results of the two diagnoses will be used to determine the diagnostic classification.

[0134] (2) Treatment method

[0135] On the basis of the original anti-Parkinsonian drugs, the patients were treated with Wutuding Shen Decoction prepared in Example 1, and the administration method was to take one dose after each meal twice a day, with a course of 3 weeks.

[0136] (3) Experimental evaluation

[0137] The evaluation index of efficacy observation includes general clinical data and efficacy observation index:

[0138] 1. General clinical data: Collect and record the patient's gender, age, disease duration, and severity of clinical manifestations to evaluate their baseline data.

[0139] 2. Efficacy observation index: ① The main efficacy index is evaluated by UPDRS III part score; ② Secondary efficacy index: Unified Parkinson's Disease Rating Scale (UPDRS I, II, IV); each efficacy observation index is evaluated once before treatment and 3 weeks after treatment.

[0140] Efficacy evaluation criteria: The efficacy is judged by the UPDRS-III score and improvement rate before and after treatment, and the judgment method is as follows: ① The higher the score, the worse the efficacy; ② The higher the improvement rate, the better the efficacy.

[0141] Safety index, including the following four aspects:

[0142] ① Vital signs: body temperature, heart rate, respiration, blood pressure (after 10 minutes of quiet rest); physical examination, blood oxygen, etc.

[0143] ② Complete three routines, liver and kidney function, blood clotting function, blood lipid, blood sugar, etc. Safety index examination;

[0144] ③ Complete ECG, head CT or MRI examination, etc.

[0145] ④ Record the time of adverse reactions, related symptoms and corresponding treatment measures of patients in a timely manner after entering the group.

[0146] (5) Statistical analysis

[0147] SPSS 26.0 software was used for statistical processing, and the results were as followsFigure 9 The two groups of normal distribution measurement data are expressed as mean ± standard deviation (x ± s), the paired t test is used for the comparison of the two groups before and after treatment, the independent sample t test is used for the comparison between the two groups, the non-parametric test of rank sum test is used for the measurement data not meeting the normal distribution, the x 2 test is used for the count data, the rank sum test is used for the rank data, and the test results are taken as the level of 0.05, P<0.05 indicates that the difference is statistically significant.

[0148] From Figure 9 -a can be seen that there is no significant difference in the left rotation dopa equivalent treatment dose (L-dopa Equivalent Dose, LED) of the patient before and after treatment (t = 2.055, P = 0.058); from Figure 9 -b can be seen that the H-Y stage after treatment has a downward trend (t = 2.070, P = 0.056); from Figure 9 -c can be seen that the UPDRS-III off stage score after treatment is significantly decreased (t = 2.305, P = 0.036); from Figure 9 -d can be seen that the on stage score after treatment is also decreased (t = 3.591, P = 0.003).

[0149] From Figure 10 -a can be seen that the on stage improvement rate after treatment has improved (t =-2.026, P = 0.061); from Figure 10 -b can be seen that in terms of non-motor symptoms, the number of defecations per week of the patient after treatment is significantly increased (z =-2.333, P = 0.020).

[0150] (6) Specific cases before and after clinical treatment

[0151] Zhang Qiong, female, 58 years old, a resident of Nanning City, was admitted to the hospital due to "slowness of movement, weakness of limbs for 5 years". The patient was diagnosed with Parkinson's disease in the past and treated with oral administration of Madopar, entacapone and other drugs, but still had repeated episodes of limb rigidity after drug efficacy, accompanied by difficulty falling asleep, dry and hard stool and other conditions. The patient now seeks treatment from a traditional Chinese medicine doctor. Physical examination: slight rigidity of limbs, decreased flexibility. Pale red tongue, yellow fur, slippery pulse. After 1 course of treatment with the drug of the embodiment, the patient's limb flexibility and sleep improved, and the average defecation frequency was 1 time per 2 days.

[0152] Ge Feng, male, 74 years old, a resident of Nanning City, Binxiang County, the patient began to have limb rigidity, slowness of movement, dry and hard stool in 2022, and was clinically diagnosed with Parkinson's disease. The patient has been treated with oral administration of Madopar, piribedil and entacapone for a long time, and has experienced a reduction in drug efficacy and a high incidence of nightmares in the past 2 months. After 1 course of treatment with the drug of the embodiment, the patient's oral drug efficacy was prolonged, the patient's subjective limb strength was improved, the patient's sleep at night was improved, and the patient's constipation was relieved.

[0153] Luo Meng, male, 68 years old, a resident of Guilin City, has had head shaking, limb tremor and limb stiffness for nearly 20 years. He has been taking Madopar and Pramipexole tablets for a long time. The above symptoms have been repeated. In the past year, he has had involuntary limb twisting after taking medicine. He has also had frequent urination, 3 days of defecation, 1 solution, many dreams, poor sleep, shouting and limb dancing in sleep. Physical examination: bilateral upper limb tremor, increased muscle tone of the four limbs, dark red tongue, yellowish and slightly greasy fur, and stringy pulse. After 2 courses of treatment with the medicine of the present application, the patient's drug efficacy was enhanced, the degree of limb twisting after taking medicine was reduced, the frequency of urination was improved, the sleep time at night was prolonged, the shouting in dreams was reduced, and the constipation was relieved.

[0154] Bai De, male, 69 years old, a resident of Nanning Liangqing District, has had involuntary limb shaking and small step walking since 2021. In the past 3 months, his mobility has worsened, he has difficulty getting up at night, he has occasional hallucinations, he has poor sleep at night, he often talks in his sleep, he has fallen out of bed, he has frequent urination, he gets up 3 times a night, and he has constipation. Physical examination: left upper limb tremor, agitated gait, difficulty in starting, pale tongue, white fur, and thin pulse. After 1 course of treatment with the medicine of the present application, the patient's sleep at night was improved, the number of times he got up at night was reduced to 1 time per night, his limb movement was more flexible than before, the small step walking and leg difficulty in starting were reduced, and he no longer had hallucinations.

[0155] III. Network pharmacology analysis

[0156] Based on network pharmacology analysis, the potential target and mechanism of action of Wutudingshen Decoction in the treatment of Parkinson's disease were predicted. From Figure 11 -a can be seen, Wutudingshen Decoction has a total of 93 drug-disease intersection protein targets, a PPI network is constructed therefrom, and the top 10 genes are screened according to the Degree value and node size; from Figure 11 -b can be seen, the top 10 genes include AKT1, CAT, IL-6, TNF, IL-1B, MAPK3, CASP3, etc.; from Figure 11 -c can be seen, KEGG pathway analysis is performed on the 93 intersection targets, and the results show that there is significant enrichment in the MAPK signaling pathway, the PI3K-AKT signaling pathway, and the IL-17 signaling pathway. The results suggest that Wutudingshen Decoction may play a therapeutic role in improving Parkinson's disease through the above targets and pathways.

[0157] Example 4

[0158] A Wutuding Shen Decoction for treating Parkinson's disease, the Wutuding Shen Decoction is composed of main ingredients and auxiliary ingredients, the main ingredients are 10g of Millettia nitida, 15g of Ficus repens, 10g of Periploca sepium, 10g of Tetrastigma obtectum and 15g of Uncaria rhynchophylla, and the auxiliary ingredients are 8g of Eucommia ulmoides, 8g of Cornus officinalis, 8g of Millettia pachycarpa, 10g of Schefflera arboricola, 15g of Paeonia lactiflora and 5g of Glycyrrhiza uralensis.

[0159] The preparation method and the taking method are the same as those in Embodiment 1.

[0160] Embodiment 5

[0161] A Wutuding Shen Decoction for treating Parkinson's disease, the Wutuding Shen Decoction is composed of main ingredients and auxiliary ingredients, the main ingredients are 8g of Millettia nitida, 13g of Ficus repens, 8g of Periploca sepium, 8g of Tetrastigma obtectum and 13g of Uncaria rhynchophylla, and the auxiliary ingredients are 5g of Eucommia ulmoides, 5g of Cornus officinalis, 5g of Millettia pachycarpa, 8g of Schefflera arboricola, 13g of Paeonia lactiflora and 3g of Glycyrrhiza uralensis.

[0162] The preparation method and the taking method are the same as those in Embodiment 1.

[0163] Embodiment 6

[0164] A Wutuding Shen Decoction for treating Parkinson's disease, the Wutuding Shen Decoction is composed of main ingredients and auxiliary ingredients, the main ingredients are 12g of Millettia nitida, 17g of Ficus repens, 12g of Periploca sepium, 12g of Tetrastigma obtectum and 17g of Uncaria rhynchophylla, and the auxiliary ingredients are 10g of Eucommia ulmoides, 10g of Cornus officinalis, 10g of Millettia pachycarpa, 12g of Schefflera arboricola, 17g of Paeonia lactiflora and 6g of Glycyrrhiza uralensis.

[0165] The preparation method and the taking method are the same as those in Embodiment 1.

[0166] Comparative Example 4

[0167] The same as Embodiment 4, the difference lies in that the auxiliary ingredients are different, and the auxiliary ingredients lack Millettia pachycarpa and Schefflera arboricola.

[0168] The main ingredients are 10g of Millettia nitida, 15g of Ficus repens, 10g of Periploca sepium, 10g of Tetrastigma obtectum and 15g of Uncaria rhynchophylla, and the auxiliary ingredients are 8g of Eucommia ulmoides, 8g of Cornus officinalis, 15g of Paeonia lactiflora and 5g of Glycyrrhiza uralensis.

[0169] Comparative Example 5

[0170] The same as Embodiment 4, the difference lies in that the auxiliary ingredients are different, and the auxiliary ingredients lack Cornus officinalis and Schefflera arboricola.

[0171] A Wutudingshen decoction for treating Parkinson's disease, the composition of the Wutudingshen decoction is composed of main materials and auxiliary materials, the main material components are 10g of kadsura longipedunculata, 15g of ficus repens, 10g of periploca forrestii, 10g of periploca forrestii and 15g of Uncaria, and the auxiliary material components are 8g of eucommia ulmoides, 8g of Millettia speciosa, 15g of paeonia lactiflora and 5g of licorice.

[0172] Four, animal experiment 3

[0173] The purpose of the experiment is to detect the effect of the modified auxiliary component of Dingshen decoction on the behavior, metabolite and other indicators of MPTP model mice.

[0174] The C57BL / 6 mice were randomly divided into 5 groups, and the specific treatment of each group of mice was as follows:

[0175] ① Control group (Control): equal amount of physiological saline intraperitoneal injection.

[0176] ② Model group (MPTP): continuous intraperitoneal injection of MPTP (1-methyl-4-phenyl-1, 2, 3, 6-tetrahydropyridine) for 5 days, and Parkinson's disease model was established.

[0177] ③ Example 4: continuous intraperitoneal injection of MPTP for 5 days, 0.50g / mL dose of Wutudingshen decoction prepared in example 1 was administered by gavage for 14 days;

[0178] ④ Comparative example 4: continuous intraperitoneal injection of MPTP for 5 days, 0.50g / mL dose of Dingshen decoction prepared in comparative example 4 was administered by gavage for 14 days;

[0179] ⑤ Comparative example 5: continuous intraperitoneal injection of MPTP for 5 days, 0.50g / mL dose of Dingshen decoction prepared in comparative example 5 was administered by gavage for 14 days.

[0180] (1) Open field test

[0181] The behavior of the experimental mice was evaluated by open field test, and the evaluation results were used to represent the movement ability of the mice, and the open field test results are shown in the following table 7-8.

[0182] Table 7: Open field test distance baseline and distance after the experiment

[0183]

[0184] As can be seen from table 7, compared with the MPTP group, the MPTP model mice administered with the Wutudingshen decoction of example 4 were significantly higher than the MPTP group mice, and were similar to the blank group mice; while the mice of comparative examples 4-5 were lower than the mice of example 4, and comparative example 5 was slightly lower than comparative example 4.

[0185] Table 8: Open field test speed baseline and speed after the experiment

[0186]

[0187] As shown in Table 8, the mice of Comparative Examples 4-5 had lower speed than the mice of Example 4 group.

[0188] (2) Striatal DA assay

[0189] Striatal sampling: The sampling and detection method was the same as described above and will not be repeated here. The results of the striatal DA assay are shown in Table 9.

[0190] Table 9. Striatal DA assay

[0191]

[0192]

[0193] The present application can be implemented in various different ways and is not limited to the described embodiments, and it will be understood by those of ordinary skill in the art that the present application can be implemented by other specific forms without changing the technical idea or essential characteristics of the present application. Therefore, it should be understood that the above-described embodiments are exemplary and are not intended to limit the present application.

Claims

1. A Wuteng Dingshen Decoction for treating Parkinson's disease, characterized in that: The Wuteng Dingshen Decoction is composed of main ingredients and auxiliary ingredients. The components of the main ingredient are 5-15g of Millettia repens, 10-20g of Euonymus fortunei, 5-15g of Polygonum multiflorum, 5-15g of Rhizoma Cynanchum and 10-20g of Uncaria rhynchophylla in a weight ratio; The auxiliary material comprises 5-15g of white peony root and 2-8g of liquorice in a weight ratio.

2. The Wuteng Dingshen Decoction for treating Parkinson's disease according to claim 1, characterized in that: The components of the main ingredient are 8-12g of Millettia reticulata, 13-17g of Euonymus fortunei, 8-12g of Polygonum multiflorum, 8-12g of Caulis Trichosanthis, and 13-17g of Uncaria rhynchophylla in a weight ratio, and the components of the auxiliary material are 13-17g of Paeonia lactiflora and 3-6g of Licorice in a weight ratio.

3. The Wuteng Dingshen Decoction for treating Parkinson's disease according to claim 2, characterized in that: The components of the main material are 10g of Millettia reticulata, 15g of Thunbergia fortunei, 10g of Polygonum multiflorum, 10g of Caulis Achyranthis Bidentatae, and 15g of Uncaria rhynchophylla in a weight ratio, and the components of the auxiliary material are 15g of Paeonia lactiflora and 5g of Licorice in a weight ratio.

4. A Wuteng Dingshen Decoction for treating Parkinson's disease, characterized in that: The Wuteng Dingshen Decoction is composed of main ingredients and auxiliary ingredients. The components of the main ingredients are 8-12g of Millettia reticulata, 13-17g of Eucommia ulmoides, 8-12g of Polygonum multiflorum, 8-12g of Cynanchum wilfordii, and 13-17g of Uncaria rhynchophylla in a weight ratio. The components of the auxiliary ingredients are 5-10g of Eucommia ulmoides, 5-10g of Dragon Bone, 13-17g of White Peony Root, and 3-6g of Licorice in a weight ratio.

5. A Wuteng Dingshen Decoction for treating Parkinson's disease, characterized in that: The Wuteng Dingshen Decoction is composed of main ingredients and auxiliary ingredients. The components of the main ingredients are 8-12g of Millettia reticulata, 13-17g of Eucommia ulmoides, 8-12g of Polygonum multiflorum, 8-12g of Cynanchum wilfordii, and 13-17g of Uncaria rhynchophylla in a weight ratio. The components of the auxiliary ingredients are 5-10g of Eucommia ulmoides, 5-10g of Dragon Bone, 5-10g of Oxyphylla strychnifolia, 8-12g of Caulis Achyranthis Bidentatae, 13-17g of White Peony Root and 3-6g of Licorice in a weight ratio.

6. A method for preparing the Wuteng Dingshen Decoction for treating Parkinson's disease according to any one of claims 1 to 5, characterized in that: The method comprises: (1) Accurately weigh the dry medicinal materials of the main ingredient and the auxiliary ingredient respectively; (2) The dried medicinal materials are mixed with 500-700 mL of water and boiled over high heat, then simmered over low heat until the medicinal liquid is 200-300 mL. The medicinal liquid is divided into two parts to prepare Wuteng Dingshen Decoction.

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