Five-vine tranquilizing decoction for treating Parkinson's disease and preparation method of five-vine tranquilizing decoction

Abstract

The invention discloses a five-vine tranquilizing decoction for treating Parkinson's disease and a preparation method thereof, the five-vine tranquilizing decoction is composed of main materials and auxiliary materials, the main materials comprise suberect spatholobus stem, wintercreeper, tuber fleeceflower stem, tinospora sinensis merr and uncaria rhynchophylla; the auxiliary materials comprise the following components: radix paeoniae alba and liquorice. Compared with the prior art, according to the strong medicine etiology and pathogenesis of Parkinson's disease, the traditional Chinese medicine composition takes tuber fleeceflower stem, uncaria, suberect spatholobus stem, wintercreeper and tinospora sinensis merr as main medicines, takes white paeony root as an auxiliary medicine and takes liquorice as an auxiliary medicine through strict compatibility and network pharmacology screening, and the seven medicines are combined to achieve the effects of dispelling wind and nourishing yin, balancing qi and blood, regulating qi and blood and dredging two ways. The Zhuang prescription has the effects of improving the dyskinesia symptom, relieving the loss of black matter-striatum dopaminergic neurons, regulating intestinal flora and metabolites of a PD model and promoting the proliferation of regulatory T cells.

Description

Technical Field

[0001] The 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. About 3 in 1,000 people worldwide suffer from PD. The disease is mainly characterized by the degeneration and death of dopamine (DA) neurons in the substantia nigra of the midbrain and a significant decrease in the DA content in the striatum. It can cause patients to have a variety of motor and non-motor disorder symptoms, seriously affecting their self-care ability and placing a heavy burden on their families and society. Currently, dopa replacement therapy is used as the first choice for the treatment of PD, but studies have found that long-term use of the drug will still lead to many limitations such as decreased efficacy and symptom fluctuations. In-depth research on the pathophysiological mechanism 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 part of traditional Chinese medicine with a long history. Long-term clinical practice has accumulated a large number of effective single prescriptions, compound prescriptions, secret prescriptions and proven prescriptions. There is no disease name "Parkinson's disease" in Zhuang medicine. In ancient times, the disease was mostly classified as "tremor" and "contraction". Modern Chinese medicine named it "tremor and constriction disease". With the popularization of disease knowledge, modern Zhuang people often describe the disease as "Fengsen (hand tremor)", "Dingsen (foot tremor)", and "Dangsen (body tremor)", so Zhuang medicine named Parkinson's disease "Dingfengsen".

[0004] For a long time, there are very few records of Zhuang medicine prescriptions for the treatment of PD. Based on the basic theories and treatment principles of Zhuang medicine, combined with the characteristics of PD and many years of clinical observation, the inventor proposed that "yin deficiency and blood deficiency, endogenous wind poison, apraxia of the brain, and blockage of the two routes (dragon route and fire route)" are the core pathogenesis hypothesis of the disease. The treatment should be based on "dispelling wind and nourishing yin, balancing qi and blood, and regulating the two routes". Through rigorous compatibility and network pharmacological screening, the self-made Zhuang medicine prescription "Wuteng Dingshen decoction" is used to improve the symptoms of the disease. The prescription is mainly composed of the characteristic Zhuang medicines of Guangxi Zhuang Autonomous Region, Millettia repens, Thunb., Polygonum multiflorum, Caulis gentiana, Uncaria rhynchophylla, white peony root and licorice. The Zhuang medicine compound has significant efficacy and small toxic and side effects, and can improve a series of motor and non-motor symptoms of PD patients. At the same time, the medicinal materials can be purchased in Chinese medicine stores on the market, the medicinal materials are cheap, the preparation process is simple, and it is easy to use. And on the basis of Wu Teng Ding Shen Decoction, it was further improved by adding auxiliary medicinal ingredients to enhance the efficacy of Wu Teng Ding Shen Decoction. Summary of the invention

[0005] The purpose of the present invention is to provide a Wuteng Dingshen Decoction for treating Parkinson's disease, aiming to provide a Zhuang medicine compound which has significant therapeutic effect on improving a series of motor and non-motor symptoms of patients with Parkinson's disease and has small toxic and side effects based on the basic theory of Zhuang medicine.

[0006] The present invention also aims to provide a method for preparing Wuteng Dingshen Decoction for treating Parkinson's disease, aiming at improving the efficacy of Wuteng Dingshen Decoction.

[0007] To achieve the above object, the present invention provides a Wuteng Dingshen Decoction for treating Parkinson's disease. 1. The ingredients of the Wuteng Dingshen Decoction are as follows:

[0008] The main ingredients include: Millettia spatholobi, Euonymus fortunei, Caulis ternatae, Caulis gentiana and Uncaria rhynchophylla;

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

[0010] Preferably, in the above technical scheme, the components of the main ingredient are matched by weight in the following ratio: 5-15g Millettia spatholobi, 10-20g Thunbergia fortunei, 5-15g Polygonum multiflorum, 5-15g Caulis Trichosanthis and 10-20g Uncaria rhynchophylla, and the components of the auxiliary material are matched by weight in the following ratio: 5-15g White Peony Root and 2-8g Licorice.

[0011] Preferably, in the above technical scheme, the components of the main ingredient are matched by weight to form 8-12g of Millettia spatholobi, 13-17g of Thunbergia fortunei, 8-12g of Polygonum multiflorum, 8-12g of Caulis Trichosanthis, and 13-17g of Uncaria rhynchophylla, and the components of the auxiliary material are matched by weight to form 13-17g of Paeonia lactiflora and 3-6g of Licorice.

[0012] Preferably, in the above technical scheme, the components of the main ingredient are matched by weight with 10g Millettia spatholobi, 15g Thunbergia fortunei, 10g Caulis Polygoni Multiflori, 10g Caulis Achyranthis Bidentatae, and 15g Uncaria rhynchophylla, and the components of the auxiliary material are matched by weight with 15g Paeonia lactiflora and 5g Licorice.

[0013] Preferably, in the above technical solution, the auxiliary material components also include Eucommia ulmoides and Dragon Bone.

[0014] Preferably, in the above technical scheme, the components of the main ingredient are matched by weight to form 8-12g of Millettia spatholobi, 13-17g of Thunbergia fortunei, 8-12g of Polygonum multiflorum, 8-12g of Caulis Trichosanthis, and 13-17g of Uncaria rhynchophylla, and the components of the auxiliary materials are matched by weight to form 5-10g of Eucommia ulmoides, 5-10g of Ossobucus ursinus, 13-17g of Paeonia lactiflora and 3-6g of Licorice.

[0015] Preferably, in the above technical solution, the auxiliary material components also include Herba Cynanchifoliae and Herba Polygoni Multiflori.

[0016] Preferably, in the above technical scheme, the components of the main ingredient are matched by weight to form 8-12g of Millettia reticulata, 13-17g of Caulis ophiopogonis, 8-12g of Caulis oxyphyllae, 8-12g of Caulis gentiana, and 13-17g of Uncaria rhynchophylla, and the components of the auxiliary materials are matched by weight to form 5-10g of Eucommia ulmoides, 5-10g of Ossobucus ursinica, 5-10g of Radix Achyranthis Bidentatae, 8-12g of Caulis quinatae, 13-17g of Paeonia lactiflora and 3-6g of Licorice.

[0017] According to the above technical scheme, the formula of Wuteng Dingshen Decoction is mainly composed of the Guangxi Zhuang Autonomous Region's characteristic Zhuang medicine Millettia spatholobi, Thunb. Ficus microcarpa, Caulis ophiopogonis, Caulis hyssop, Uncaria rhynchophylla, Paeonia lactiflora and Licorice, and the characteristics of each raw material are as follows:

[0018] Spatholobus suberectus Dunn: (scientific name Spatholobus suberectus Dunn), also known as blood wind vine, blood wind, blood vine, three-leaf Spatholobus, nine-layer wind, is the vine stem of leguminous plants such as dense flower bean, white flower oil vine, fragrant flower rock bean vine or bright leaf rock bean vine. It tastes bitter and sweet, and is warm in nature; it enters the liver and kidney meridians. Spatholobus suberectus has the effects of promoting blood circulation, replenishing blood, relaxing muscles and tendons, regulating menstruation and relieving pain. It is mainly used to treat low back and knee pain, numbness and paralysis, rheumatism, blood deficiency and chlorosis, irregular menstruation and other symptoms.

[0019] Euonymus fortunei (Turcz.) Hand.-Mazz is an evergreen vine shrub of the genus Euonymus in the family Celastraceae. Also known as golden thread wind, nine-ox-made, wall-keeping wind, vine stone, and wall-climbing grass. ① "Compendium of Materia Medica": bitter, slightly warm, non-toxic. ② "Guizhou Folk Medicine": pungent, neutral. Euonymus fortunei has the effects of relaxing muscles and tendons, stopping bleeding and removing blood stasis. It is mainly used to treat lumbar muscle strain, rheumatic arthralgia, hemoptysis, metrorrhagia, irregular menstruation, fractures from falls, and traumatic bleeding.

[0020] Tuber Fleeceflower Stem: (scientific name Tuber Fleceflower Stem), also known as Chess Vine and Shouwu Vine, is the stem or leaf stem of Polygonum multiflorum. It tastes sweet and slightly bitter, and is neutral in nature. It enters the heart and liver meridians. Tuber Fleceflower Stem has the effects of nourishing the heart and calming the mind, dispelling wind, and unblocking meridians. It is mainly used to treat insomnia, dreaminess, body pain due to blood deficiency, skin numbness, rheumatic pain, urticaria and itching.

[0021] Kuanjinteng: Also known as Songgenteng, Shujinteng, Dasongshen, it is the stem of the Chinese blue ox gall of the Menispermaceae family. It tastes bitter and is cool in nature; it enters the liver meridian. Kuanjinteng has the effects of relaxing muscles and tendons, dispelling wind and relieving pain. It is mainly used to treat rheumatic pain, sciatica, lumbar muscle strain, traumatic injuries and other diseases.

[0022] Uncaria rhynchophylla: (scientific name: Uncariarhynchophylla (Miq.) Miq. ex Havil.), also known as big hooked ding and double hooked twigs, is the dried hooked stems and branches of the Rubiaceae plants Uncaria rhynchophylla, big-leafed Uncaria rhynchophylla, hairy Uncaria rhynchophylla, Chinese Uncaria rhynchophylla or sessile fruit Uncaria rhynchophylla. It tastes sweet and is slightly cold in nature; it enters the liver and pericardium meridians. Uncaria rhynchophylla has the effects of calming wind and relieving convulsions, stopping spasms, clearing the liver and calming the liver, dispersing wind and dissipating heat. It is mainly used to treat symptoms such as internal liver wind movement, epilepsy and convulsions, high fever convulsions, acute convulsions in children, headache and dizziness, cold and convulsions, crying in children, headache and red eyes, and poor rash.

[0023] Dragon bone: (scientific name Os Draconis), also known as Lu Hu Yisheng, Naga bone, raw dragon bone, calcined dragon bone, five-flowered dragon bone, green dragon bone, flower dragon bone, white dragon bone, is a bone fossil of ancient large mammals such as elephants, rhinoceroses, and three-toed horses. It tastes sweet and astringent, and is slightly cold in nature; it enters the heart and liver meridian. Dragon bone has the effects of calming the liver and suppressing yang, calming the mind, and astringing and consolidating; it is mainly used to treat restlessness, palpitations and insomnia, epilepsy, madness, liver yang vertigo, slippery symptoms, wet sores, itchy rashes, and sores that do not heal for a long time.

[0024] Sinomenium acutum (Thunb.) Rehd. et Wils., also known as Dafengteng, Chuifengsan, Heifangji, Paifengteng, Qingfangji, is the dried tuber of Sinomenium acutum and Maoqingteng of Menispermaceae. It tastes bitter and pungent, and is neutral in nature. It enters the liver and spleen meridians. Sinomenium acutum has the effects of removing rheumatism, unblocking meridians, and promoting urination. It is mainly used to treat rheumatism, rheumatism, edema, beriberi, heart and stomach pain, carbuncle, malignant ulcer and other diseases.

[0025] Millettia speciosa Champ., also known as beautiful millettia, mountain lotus root, and powerful potato. It tastes sweet and is neutral in nature. It enters the lung and kidney meridians. Millettia speciosa Champ. has the effects of nourishing the lungs, strengthening the tendons and activating the collaterals. It is mainly used to treat lumbar muscle strain, rheumatoid arthritis, lung heat, lung deficiency cough, tuberculosis, chronic bronchitis, chronic hepatitis, spermatorrhea and other diseases.

[0026] Eucommia ulmoides Oliv., also known as shredded skin, Sizhong, silk cotton skin, jade skin. It tastes sweet and slightly spicy, warm in nature; it enters the liver and kidney meridians. Eucommia ulmoides has the effect of nourishing the liver and kidneys and strengthening the tendons and bones; it is mainly used to treat kidney deficiency and low back pain, rheumatic arthralgia, weak tendons and bones, poor flexion and extension, waist and knee pain, kidney yang deficiency, blood stasis and other diseases.

[0027] A method for preparing the Wuteng Dingshen Decoction for treating Parkinson's disease as described above, characterized in that the method comprises:

[0028] (1) Accurately weigh the dry medicinal materials such as Millettia spatholobi, Euonymus fortunei, Polygonum multiflorum, Caulis gentiana, 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 Wu Teng Ding Shen 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 for the treatment of Parkinson's disease is as follows: Uncaria rhynchophylla is cool in nature, sweet and bitter in taste, Polygonum multiflorum is flat in nature, sweet in taste, and both can dispel wind poison, regulate the intestinal tract, and clear the fire path; Millettia spatholobi can nourish blood and dispel wind, and is good at clearing the fire path; Caulis lappa is good at clearing the dragon path; Caulis clematis mainly invigorates qi and blood, and clears the two paths; Wuteng is used in the prescription as the main drug; White Peony nourishes blood, softens the liver, and restrains yin, and is a supporting drug, which has the effect of strengthening the main drug; Licorice harmonizes all the 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 the intestinal tract, 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 the PD model and promoting the proliferation of regulatory T cells.

[0033] (2) Wuteng Dingshen Decoction has achieved positive intervention effects in animal experiments and clinical practice, and 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 disease patients and improving the quality of life of patients;

[0034] (3) The pharmacology of the Wuteng Dingshen Decoction of the present invention in treating Parkinson's disease is as follows: Wuteng is used in the prescription as the main drug; white peony root nourishes blood, softens the liver and restrains yin, dragon bone calms the liver, calms the mind, eucommia nourishes the liver, nourishes the kidney, replenishes blood and improves intelligence, radix strychnifolia replenishes qi, transports blood and activates collaterals, and qingfengteng dispels wind, dredges collaterals and relieves pain. The five are all auxiliary drugs to enhance the efficacy of the main drug and assist in the treatment of secondary and concurrent symptoms; licorice harmonizes the various drugs and is a supporting drug. The behavioral evaluation and striatal DA results of the animal experiment of the improved Wuteng Dingshen Decoction showed that the therapeutic effect was significantly enhanced. BRIEF DESCRIPTION OF THE DRAWINGS

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

[0036] Figure 1 It is a graph of the distance and speed of the mine experiment 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 the immunofluorescence staining of tyrosine hydroxylase in the substantia nigra of the brain in the Control group, MPTP group, MPTP+WTlow group, MPTP+WTmid group, and MPTP+WThig group;

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

[0039] Figure 4 This is the LDA difference contribution diagram of Wuteng Dingshen Decoction on peripheral blood metabolites of mice;

[0040] Figure 5 This is a diagram of the discriminant analysis and cluster analysis of Wuteng Dingshen Decoction;

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

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

[0043] Figure 8 is a bar graph of the levels of Foxp3 transcription factor in spleen of the Control group, MPTP group, and MPTP+WThig group;

[0044] Fig. 9 It is a bar chart of the scores of the patients in the clinical trial before and after treatment;

[0045] Fig.10 It is a bar graph of the improvement rate of the opening period of clinical trial patients before and after treatment and a bar graph of the number of bowel movements per week of patients;

[0046] Fig.11 It is a diagram predicting the potential targets and mechanisms of action of Wuteng Dingshen Decoction in the treatment of Parkinson's disease. DETAILED DESCRIPTION

[0047] The experimental mice and medication conditions of the animal experiments involved in the present invention are as follows:

[0048] (1) Experimental animals

[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℃, the relative humidity was maintained at 50%-60%, the artificial light simulated the natural conditions with 12 hours of light and dark respectively, and sufficient food and drinking water were given during breeding. All experimental operators have the "Certificate of Qualification of Professional Technical Examination for 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 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-flavor Chinese herbal medicine slices, which are purchased uniformly by the Pharmacy Department of the International Zhuang Medicine Hospital Affiliated to Guangxi Zhuang Autonomous Region University of Traditional Chinese Medicine and prepared as required, and the liquid medicine is prepared at 0.1g / ml;

[0053] Dose conversion for mouse experiments: According to the animal dosage calculation in "Experimental Animals and Animal Experimental Techniques": 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, so the daily dose for mice = 9.01 ×

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

[0055] Example 1

[0056] A Wuteng Dingshen Decoction for treating Parkinson's disease, the Wuteng Dingshen Decoction consisting of a main ingredient and auxiliary ingredients, the main ingredient comprising 10g of Millettia spatholobi, 15g of Thunbergia fargesii, 10g of Polygonum multiflorum, 10g of Caulis Achyranthis Bidentatae and 15g of Uncaria rhynchophylla; the auxiliary ingredients comprising 15g of Paeonia lactiflora and 5g of Licorice.

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

[0058] (1) Accurately weigh 10 g of Millettia spatholobi, 15 g of Euonymus fortunei, 10 g of Polygonum multiflorum, 10 g of Rhizoma Cynoglossi, 15 g of Uncaria rhynchophylla, 15 g of Paeonia lactiflora, and 5 g of Glycyrrhiza uralensis.

[0059] (2) Place the dried medicinal materials in a clay pot, add 500 mL of water, boil over high heat, and simmer over low heat until the medicinal liquid is 200 mL. Divide the mixture into two portions, each portion containing 100 mL of Wu Teng Ding Shen Decoction.

[0060] The method of using the above-mentioned Wu Teng Ding Shen Decoction is as follows: for oral administration, take twice a day, specifically the patient takes 100 mL each time after meals in the morning and evening, and one course of treatment is 3 weeks.

[0061] Example 2

[0062] A Wuteng Dingshen Decoction for treating Parkinson's disease, the Wuteng Dingshen Decoction consisting of a main ingredient and auxiliary ingredients, the main ingredient comprising 5g of Millettia spatholobi, 10g of Thunbergia fortunei, 5g of Polygonum multiflorum, 5g of Caulis Achyranthis Bidentatae and 10g of Uncaria rhynchophylla; the auxiliary ingredient comprising 10g of Paeonia lactiflora and 2g of Licorice.

[0063] The preparation method and administration method are the same as those in Example 1.

[0064] Example 3

[0065] A Wuteng Dingshen Decoction for treating Parkinson's disease, the Wuteng Dingshen Decoction is composed of a main ingredient and auxiliary ingredients, the main ingredient components are 15g of Millettia reticulata, 20g of Thunbergia fortunei, 15g of Polygonum multiflorum, 15g of Caulis Achyranthis Bidentatae and 20g of Uncaria rhynchophylla; the auxiliary ingredients are 20g of Paeonia lactiflora and 8g of Licorice.

[0066] The preparation method and administration method are the same as those in Example 1.

[0067] Comparative Example 1

[0068] The method is basically the same as Example 1, except that the main ingredients are different, and the main ingredients lack Caulis Polygoni Multiflori, Caulis Tetrapanacis Fangos and Caulis Tetrapanacis Kusnezoffii. The Dingshen Decoction is composed of main ingredients and auxiliary ingredients, the main ingredient components are 10g Millettia Spatholobi and 15g Uncaria Rhynchophylla; the auxiliary ingredients are 15g Paeonia lactiflora and 5g Licorice.

[0069] Comparative Example 2

[0070] The method is basically the same as Example 1, except that the main ingredients are different, and the main ingredients lack euphorbia fargesii and radix scutellariae. The dingshen decoction is composed of main ingredients and auxiliary ingredients, the main ingredients are 10g of caulis spatholobi, 10g of caulis spatholobi and 15g of uncaria; the auxiliary ingredients are 15g of white peony root and 5g of liquorice.

[0071] Comparative Example 3

[0072] The method is basically the same as Example 1, except that the main ingredients are different, and Tripterygium wilfordii is added to replace Euphorbia fortunei. The Dingshen Decoction is composed of main ingredients and auxiliary ingredients, the main ingredients are 10g Millettia spatholobi, 15g Tripterygium wilfordii, 10g Polygonum multiflorum, 10g Caulis gentiana and 15g Uncaria rhynchophylla; the auxiliary ingredients are 15g White Peony Root and 5g Licorice.

[0073] 1. Animal Experiments 1

[0074] The purpose of the experiment is to detect the effects of different components of Dingshen Decoction on the behavior, metabolites and other indicators of MPTP model mice.

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

[0076] ①Blank group (Control): an equal amount of normal saline was injected intraperitoneally.

[0077] ② Model group (MPTP): MPTP (1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine) was injected intraperitoneally for 5 consecutive days to establish a Parkinson's disease model.

[0078] ③ Example 1: MPTP was intraperitoneally injected continuously for 5 days, and Wuteng Dingshen Decoction prepared in Example 1 was gavaged intragastricly at a dose of 0.50 g / mL for 14 days;

[0079] ④Comparative Example 1: MPTP was intraperitoneally injected continuously for 5 days, and the Dingshen Decoction prepared in Comparative Example 1 at a dose of 0.50 g / mL was intragastrically administered for 14 days;

[0080] ⑤Comparative Example 2: MPTP was intraperitoneally injected continuously for 5 days, and the Dingshen Decoction prepared in Comparative Example 2 at a dose of 0.50 g / mL was intragastrically administered for 14 days;

[0081] ⑥Comparative Example 3: MPTP was intraperitoneally injected continuously for 5 days, and the Dingshen Decoction prepared in Comparative Example 3 at a dose of 0.50 g / mL was intragastrically administered for 14 days.

[0082] (1) Open field test

[0083] The open field test was used to conduct behavioral evaluation on the experimental mice. The evaluation results were used to indicate the motor ability of the mice. The results of the open field test are shown in Table 1-2 below.

[0084] Table 1. Baseline distance of open field test and distance after the test

[0085]

[0086] As can be seen from Table 1, compared with the MPTP group, the MPTP model mice administered with Wuteng Dingshen Decoction of Example 1 had significantly higher mileage than the MPTP group mice and were similar to the blank group mice; while the mileage of the mice in Comparative Examples 1-3 was higher than that of the MPTP group mice but significantly lower than that of the Example 1 group mice.

[0087] Table 2 Baseline speed and speed after the open field test

[0088]

[0089]

[0090] As can be seen from Table 2, the speed of the mice in Comparative Examples 1-3 is lower than that of the mice in Example 1.

[0091] (2) Striatal DA measurement

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

[0093] Table 3 Striatal DA measurement table

[0094] Group Striatal DA value (ng / g) Difference with 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] 2. Animal Experiments 2

[0096] The purpose of the experiment is to detect the behavior, metabolites, etc. of MPTP model mice when the Wuteng Dingshen Decoction (the main ingredients are 10g Millettia reticulata, 15g Thunb. 10g Polygonum multiflorum, 10g Caulis Achyranthes bidentata and 15g Uncaria rhynchophylla; the auxiliary ingredients are 15g Paeonia lactiflora and 5g Licorice) is administered at different doses.

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

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

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

[0100] ③ Wuteng Dingshen Decoction low-dose group (MPTP+WTlow): MPTP was intraperitoneally injected for 5 consecutive days, and 0.25 g / mL Wuteng Dingshen Decoction was intragastrically administered for 14 days;

[0101] ④Wuteng Dingshen Decoction medium-dose group (MPTP+WTmid): continuous intraperitoneal injection of MPTP for 5 days, and oral gavage of 0.50 g / mL Wuteng Dingshen Decoction for 14 days;

[0102] ⑤ Wuteng Dingshen Decoction high-dose group (MPTP+WThig): continuous intraperitoneal injection of MPTP for 5 days, and oral gavage of 0.75g / mL Wuteng Dingshen Decoction for 14 days;

[0103] The Wuteng Dingshen Decoction was prepared according to Example 1.

[0104] (1) Behavioral testing methods and results

[0105] The open field test was used to evaluate the behavior of the experimental mice. The evaluation results were used to indicate the motor ability of the mice. The results of the open field test are as follows: Figure 1 -a, Figure 1 -b, Figure 1 -c.

[0106] from Figure 1 -a shows that the open field test movement path is used as a behavioral assessment, and it can be seen that the MPTP model mice have fewer movement paths, indicating that the MPTP model has obvious movement disorders; Figure 1 -b and Table 1-2 show that after treatment with Wuteng Dingshen Decoction, the total distance traveled in the open field test of the MPTP model mice in the MPTP+WTlow, MPTP+WTmid and MPTP+WThig groups increased significantly, and even the total distance traveled in the open field test of the MPTP model mice in the MPTP+WTmid group was similar to that of the mice in the blank group; Figure 1-c and Table 1-2, it can be seen that after treatment with Wuteng Dingshen Decoction, the open field test speed of the MPTP model mice in the MPTP+WTlow, MPTP+WTmid and MPTP+WThig groups was significantly accelerated, approaching the speed of the blank group mice.

[0107] (2) Determination of DA content in the striatum and substantia nigra

[0108] Sampling of striatum and substantia nigra: After behavioral testing, the experimental mice were intraperitoneally injected with 1% sodium pentobarbital at 35 mg / kg body weight for anesthesia. After anesthesia, 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 placed in 0.1mol / LPB (pH 7.3) containing 4% formaldehyde and fixed for 4 hours. Then it was transferred to 20% and 30% sugar solutions. After the tissue block sank, coronal frozen continuous sections were made, with a thickness of 25μm. Three sets of sections (10 slices per set) were collected from 2.80mm-3.80mm behind the anterior fontanelle (substantia nigra), and then floated in 0.01mol / L PBS. The results of tyrosine hydroxylase in the substantia nigra of the midbrain were as follows Figure 2 shown.

[0109] The striatum detection method was the same as the operating steps in animal experiment 1.

[0110] from Figure 1 -dIt can be seen that the improvement in the medium and high-dose groups was more obvious. HPLC-MS analysis showed that the content of dopamine (DA) in the striatum was significantly increased after treatment with Wuteng Dingshen Decoction compared with the model group, and the high-dose group had the best effect, with a difference of 174.9 with MPTP, P=0.0178.

[0111] Method for detecting tyrosine hydroxylase in the substantia nigra of the midbrain: Immunofluorescence staining was used to calculate the number of neurons positive for tyrosine hydroxylase in the compact part of the substantia nigra of the midbrain. The steps are as follows: Take a set of slices from the substantia nigra of the midbrain and rinse them three times in 0.01mol / LPBS buffer (pH 7.3), then place them in a blocking solution containing 0.3% TritonX-100 and 10% goat serum for blocking at room temperature for 30 minutes, transfer the blocked slices to a 1:300 diluted mouse anti-TH monoclonal antibody, and incubate them in a 4°C refrigerator for 48 hours. Take out the slices and rinse them three times with 0.01mol / LPBS, 10 minutes each time, then transfer the slices to a 1:200 diluted FITC-labeled goat anti-mouse IgG antibody, incubate them at room temperature for 4-6 hours, and after sufficient rinsing with 0.01MPBS, mount the brain slices on a slide, seal them with glycerol phosphate buffer, and observe and photograph them under a fluorescent upright microscope. The excitation wavelength of FITC is 488 nm, and TH-immunopositive neurons appear green. The number of TH-positive neurons in SNpc was counted in 10 brain sections (1 set) for each mouse.

[0112] from Figure 2 It can be seen that the number of cells positive for tyrosine hydroxylase (TH) staining in the compact part of the midbrain substantia nigra was significantly reduced, and the number of TH-positive cells increased after intervention with Wuteng Dingshen Decoction, with the best effect in the medium and high dose groups.

[0113] (3) Intestinal flora

[0114] Collection of fecal samples: On the second day after the last oral gavage of each group of mice, each mouse was placed individually in an autoclaved cage without bedding to allow free defecation. The feces were quickly collected in a sterilized EP tube and stored at -80°C for later use.

[0115] Intestinal flora detection method: 16SrDNA amplification and sequencing were used to detect the composition of fecal flora. The specific steps are as follows: Take an appropriate amount of feces (>50mg), add ultrapure water at 10μL / g, fully shake to homogenize, let stand for 30min, centrifuge at 4℃, 12000rpm for 20min, aspirate the supernatant, filter with 0.22μm water membrane, and transfer to a gas phase bottle for measurement. The V4-V5 region sequence of 16SrDNA was selected for high-throughput sequencing analysis. The measured raw data was sequence optimized and bioinformatics analysis was performed. OTU (operational taxonomic unit) was used for clustering, and based on taxonomic information, the community structure was statistically analyzed at the classification levels of phylum, class, order, family, genus, and species. The above spliced ​​optimized sequences were clustered into OTUs using the software USEARCH and species information was annotated. On the basis of the above analysis, the mothurrua software was used to conduct Alpha diversity analysis (Chao and Ace species richness statistics, Shannon and Simpson species diversity statistics), and to produce rarefaction curves and Venn diagrams.

[0116] The intestinal flora of MPTP model mice is as follows Figure 3-Figure 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 levels

[0118] Detection method of peripheral blood metabolite content: Peripheral blood was collected and the metabolite content of peripheral blood was detected by high performance liquid chromatography tandem high resolution mass spectrometry. The steps are as follows: ① Sample pretreatment: 100 μL of serum was extracted and placed in a 1.5 mL centrifuge tube, 10 μL of 100 ppb d9-TMAO solution was added, and vortexed for 30 seconds; then 300 μL of methanol was added, vortexed for 3 minutes, centrifuged at 4 ° C for 10 minutes, 100 ul of supernatant was placed in a centrifuge tube, and the above process was repeated for a second centrifugation for 3 minutes. The supernatant was filtered through a 0.22 μm filter into a sample injection bottle, and 10 ul was taken for analysis. Standard preparation: Weigh an appropriate amount of standard in a 100 mL volumetric flask, dilute to volume with methanol, shake well, and prepare a standard stock solution, which was stored in a refrigerator at -20 ° C. When used, it was diluted to the required concentration with acetonitrile / water (1:1, v / v) containing 20 mM ascorbic acid. ②Chromatographic column analysis conditions: Shim-pack GIST C18 column (75x2.1mm, 2μm); automatic sampler sample tray temperature control: 4℃; column temperature: room temperature; mobile phase: acetic acid-acetic acid (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. ③Mass spectrometry analysis conditions, ionization mode: electrospray ionization negative ion mode (ESI-); capillary voltage 3.0kV; cone voltage 40V; ion source temperature 100℃; desolvation gas temperature 300℃; desolvation gas flow rate 600L / h; collector flow rate 50L / h; acquisition mode: MSe Continuum mode, negative ion mode; mass number acquisition range: 100-1200Da, scanning time: 0.2sec, high energy channel voltage: 10-65V. The collected data are preprocessed and finally the final peak number is obtained after quality control (QC sample) screening.

[0119] from Figure 4 and Figure 5 It can be seen that Wuteng Dingshen Decoction significantly upregulated 104 metabolites in the peripheral blood of the MPTP model, among which the products of the amino acid metabolic pathway were the most (n=9, Figure 5 ).

[0120] (5) Flow cytometry

[0121] Flow cytometry was used to detect Tregs in the spleen (see Table 5 and Figure 6 ) and spleen Foxp3 transcription factor levels (as shown in Tables 6 and Figure 8 ), the range of flow cytometry logic gates is as follows Figure 7 shown.

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

[0123]

[0124] From Table 5 and Figure 6-7 It can be seen that there were significant differences in the proportion of spleen Tregs in the blank control group, MPTP model group, and Wuteng Dingshen Decoction high-dose treatment group (F=5.102, P=0.020) using flow cytometry. The proportion of Tregs in the Wuteng Dingshen Decoction treatment group was significantly higher than that in the MPTP model group (Mean diff=4.213, P=0.012).

[0125] Table 6. Foxp3 transcription factor levels in spleen

[0126]

[0127] From Table 6 and Figure 8 It can be seen that the results of RT-qPCR detection of spleen Foxp3 transcription factor levels indicated that there were significant differences in spleen Foxp3 transcription factor levels among the three groups (F=34.77, P<0.001), and the Foxp3 level in the Wuteng Dingshen 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] 3. Clinical Cases

[0129] (1) Research subjects

[0130] Case source: This study will select 16 PD patients who were admitted to the inpatient and outpatient departments of the Department of Encephalopathy of the International Zhuang Medicine Hospital Affiliated to Guangxi University of Traditional Chinese Medicine from July 2024 to December 2024 or from previous cases who met the inclusion criteria, obtained approval from the Ethics Committee (approval number: 2024-102-1) and signed informed consent as research subjects. A total of 16 subjects were included in this study, including 7 males and 9 females. The age range of the subjects was 40-74 years old, with an average age of (59.50±9.359) years old. The course of disease ranged from 0.5-7.0 years, with an average course of disease of (3.78±2.359) years.

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

[0132] ① Western medicine diagnostic criteria for Parkinson's disease: in accordance with the "Chinese Parkinson's Disease Diagnostic Criteria (2016 Edition)" formulated in 2016 by the Parkinson's Disease and Movement Disorders Group of the Neurology Branch of the Chinese Medical Association and the Parkinson's Disease and Movement Disorders experts of the Neurology Branch of the Chinese Medical Association.

[0133] ② TCM diagnostic criteria for Parkinson's disease: Based on the diagnostic criteria in the TCM Diagnosis and Therapeutic Effect Evaluation Criteria for Geriatric Tremor Syndrome formulated by the Geriatrics Society of the All-China Association of Traditional Chinese Medicine in 1992, two associate chief physicians or above independently perform dialectical classification on the patients, and the dialectical conclusion is taken if the results are consistent; if the dialectical results are inconsistent, a third TCM physician with the title of associate chief or above will perform dialectical classification, and the dialectical classification is drawn based on the same results of the two dialectics.

[0134] (2) Treatment methods

[0135] The patient was treated with the original anti-Parkinson's western medicine and Wuteng Dingshen Decoction prepared in Example 1 was added. The method of administration was to take the decoction once in the morning and once in the evening after each meal. The course of treatment was 3 weeks.

[0136] (3) Experimental evaluation

[0137] Efficacy observation and evaluation indicators, including general clinical data and efficacy observation indicators:

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

[0139] 2. Efficacy observation indicators: ① The main efficacy indicator is evaluated by the unified comprehensive Parkinson's disease rating scale (UPDRSIII) in the off-stage period; ② Secondary efficacy indicators: unified comprehensive Parkinson's disease rating scale (UPDRSSI, II, IV); each efficacy observation indicator is evaluated once before treatment and 3 weeks after treatment.

[0140] Efficacy evaluation criteria: based on the UPDRS-III scores and improvement rate before and after treatment, 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 indicators include 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 Improve the inspection of three major routine, liver and kidney function, coagulation function, blood lipids, blood sugar and other safety indicators;

[0144] ③Complete electrocardiogram, head CT or MRI and other examinations;

[0145] ④ After enrollment, timely record the time of adverse reactions, related symptoms and corresponding treatment measures of the patients.

[0146] (5) Statistical analysis

[0147] SPSS 26.0 software was used for statistical analysis. Fig. 9 As shown in the figure, the measurement data of the two groups that conformed to the normal distribution were expressed as mean ± standard deviation (x ± s). The paired t test was used for the intra-group comparison before and after treatment, and the independent sample t test was used for the comparison between the two groups. The non-parametric rank sum test was used for the measurement data that did not conform to the non-normal distribution; the counting data were expressed as x 2 The rank sum test was used for the rank data, and the test results were based on the 0.05 level, with P < 0.05 indicating that the difference was statistically significant.

[0148] from Fig. 9 -a shows that there is no significant difference in the L-dopa equivalent therapeutic dose (LED) of patients before and after treatment (t=2.055, P=0.058); Fig. 9 -b shows that the HY stage after treatment has a downward trend (t=2.070, P=0.056); Fig. 9 -c shows that the UPDRS-III off-period score decreased significantly after treatment (t=2.305, P=0.036); Fig. 9 -dIt can be seen that the opening score was also reduced after treatment (t=3.591, P=0.003).

[0149] from Fig.10 -a shows that the improvement rate of the opening period after treatment has increased (t = -2.026, P = 0.061); Fig.10 -b It can be seen that in terms of non-motor symptoms, the number of bowel movements per week of patients increased significantly after treatment (z=-2.333, P=0.020).

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

[0151] Zhang Mouqiong, female, 58 years old, from Nanning, was diagnosed with Parkinson's disease in other hospitals and treated with oral drugs such as Madopar and Entacapone, but she still had repeated limb stiffness after the drug effect, accompanied by difficulty falling asleep, dry and hard stools, etc. She is now seeking treatment from Chinese and Zhuang medicine. Physical examination: slightly stiff limbs and decreased flexibility. Pale red tongue, yellow fur, slippery pulse, after one course of treatment according to the embodiment of the present invention, limb flexibility and sleep improved, and the average bowel movement was once every two days.

[0152] Ge Moufeng, male, 74 years old, from Binyang, Nanning, began to experience limb stiffness, slow movement, and dry and hard stools in 2022. He was clinically diagnosed with Parkinson's disease and was treated with long-term oral administration of Madopar, Piribedil, and Entacapone. In the past two months, he experienced drug efficacy reduction and frequent nighttime dreams and easy awakening. After one course of treatment according to the present invention, the patient's oral drug efficacy was prolonged, his limb strength was improved, his sleep at night was improved, and his constipation was relieved.

[0153] Luo Mouning, male, 68 years old, from Guilin, has had head shaking, limb tremors, and limb stiffness for nearly 20 years. He has been taking Madopar and Pramipexole tablets for a long time. The above symptoms are repeated from time to time. In the past year, he has had involuntary twisting of limbs after taking the medicine. When he went to the doctor, he also had frequent urination, bowel movements once every three days, many dreams, poor sleep, and shouting and limb dancing during sleep. Physical examination: tremor in both upper limbs, increased muscle tension in the limbs, dark red tongue, slightly greasy yellow fur, and stringy pulse. After two courses of treatment according to the embodiment of the present invention, the patient's drug efficacy was enhanced, the degree of limb twisting after taking the medicine was alleviated, frequent urination was improved, the night sleep time was prolonged, shouting in dreams was reduced compared with before, and constipation was relieved.

[0154] Bai Moude, male, 69 years old, from Liangqing District, Nanning City, began to gradually experience involuntary tremors of the limbs and small steps in 2021. In the past three months, the difficulty in movement has increased, it is difficult to get up and move at night, there are occasional hallucinations, poor sleep at night, easy to talk in sleep, and there have been falls from the bed, frequent urination, getting up 3 times / night, and constipation. Physical examination: tremor in the left upper limb, panic gait, difficulty starting, pale tongue, white fur, and thin pulse. After one course of treatment according to the embodiment of the present invention, the patient's sleep at night improved, the number of getting up at night was reduced to 1 time / night, the limbs were more flexible when getting up at night, and the small steps and difficulty in starting the legs during the day were reduced, and no hallucinations recurred.

[0155] 3. Network Pharmacology Analysis

[0156] Based on network pharmacology analysis, the potential targets and mechanisms of Wuteng Dingshen Decoction in the treatment of Parkinson's disease were predicted. Fig.11 -a As can be seen, Wuteng Dingshen Decoction obtained a total of 93 drug-disease intersection protein targets, and a PPI network was constructed for them. The top 10 genes were selected according to the degree value and node size; Fig.11 -b shows that the top 10 genes include AKT1, CAT, IL-6, TNF, IL-1B, MAPK3, CASP3, etc.; Fig.11 -c It can be seen that the KEGG pathway analysis of 93 intersection targets showed that the MAPK signaling pathway, PI3K-AKT signaling pathway, and IL-17 signaling pathway were significantly enriched. The results suggest that Wuteng Dingshen Decoction may play a therapeutic role in improving Parkinson's disease through the above targets and pathways.

[0157] Example 4

[0158] A Wuteng Dingshen Decoction for treating Parkinson's disease. The Wuteng Dingshen Decoction is composed of a main ingredient and an auxiliary ingredient. The main ingredient is 10g of Millettia reticulata, 15g of Eleutherodactyla fortunei, 10g of Polygonum multiflorum, 10g of Caulis Achyranthis Bidentatae and 15g of Uncaria rhynchophylla. The auxiliary ingredient is 8g of Eucommia ulmoides, 8g of Ossobucus urinariae, 8g of Radix Achyranthis Bidentatae, 10g of Caulis Achyranthis Bidentatae, 15g of Radix Paeoniae Alba and 5g of Radix Glycyrrhizae.

[0159] The preparation method and administration method are the same as those in Example 1.

[0160] Example 5

[0161] A Wuteng Dingshen Decoction for treating Parkinson's disease. The Wuteng Dingshen Decoction is composed of a main ingredient and an auxiliary ingredient. The main ingredient is 8g of Millettia spatholobi, 13g of Eleutherodactyla fortunei, 8g of Polygonum multiflorum, 8g of Caulis Achyranthis Bidentatae and 13g of Uncaria rhynchophylla. The auxiliary ingredient is 5g of Eucommia ulmoides, 5g of Ossobucus urinariae, 5g of Rhizoma Cynoglossi, 8g of Caulis Achyranthis Bidentatae, 13g of Paeonia lactiflora and 3g of Licorice.

[0162] The preparation method and administration method are the same as those in Example 1.

[0163] Example 6

[0164] A Wuteng Dingshen Decoction for treating Parkinson's disease. The Wuteng Dingshen Decoction is composed of a main ingredient and an auxiliary ingredient. The main ingredient is 12g of Millettia spatholobi, 17g of Eleutherodactyla fortunei, 12g of Polygonum multiflorum, 12g of Caulis Achyranthis Bidentatae and 17g of Uncaria rhynchophylla. The auxiliary ingredient is 10g of Eucommia ulmoides, 10g of Ossobucus urinariae, 10g of Radix Achyranthis Bidentatae, 12g of Caulis Achyranthis Bidentatae, 17g of Paeonia lactiflora and 6g of Licorice.

[0165] The preparation method and administration method are the same as those in Example 1.

[0166] Comparative Example 4

[0167] It is basically the same as Example 4, except that the auxiliary materials are different and lack Rhizoma Cynoglossi and Rhizoma Cynoglossi.

[0168] The components of the main material are 10g of Millettia spatholobi, 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 materials are 8g of Eucommia ulmoides, 8g of Ossobucus urinariae, 15g of Paeonia lactiflora and 5g of Licorice.

[0169] Comparative Example 5

[0170] It is basically the same as Example 4, except that the auxiliary materials are different and lack dragon bone and Qingfengteng.

[0171] A Wuteng Dingshen Decoction for treating Parkinson's disease. The Wuteng Dingshen Decoction is composed of a main ingredient and an auxiliary ingredient. The main ingredient is 10g of Millettia spatholobi, 15g of Eucommia fortunei, 10g of Polygonum multiflorum, 10g of Caulis Achyranthis Bidentatae and 15g of Uncaria rhynchophylla, and the auxiliary ingredient is 8g of Eucommia ulmoides, 8g of Radix Achyranthis Bidentatae, 15g of Paeonia lactiflora and 5g of Licorice.

[0172] IV. Animal Experiments 3

[0173] The purpose of the experiment is to detect the effects of the auxiliary components in the improved Dingshen Decoction on the behavior, metabolites and other indicators of MPTP model mice.

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

[0175] ①Blank group (Control): an equal amount of normal saline was injected intraperitoneally.

[0176] ② Model group (MPTP): MPTP (1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine) was injected intraperitoneally for 5 consecutive days to establish a Parkinson's disease model.

[0177] ③ Example 4: MPTP was intraperitoneally injected continuously for 5 days, and Wuteng Dingshen Decoction prepared in Example 1 was gavaged intragastricly at a dose of 0.50 g / mL for 14 days;

[0178] ④Comparative Example 4: MPTP was intraperitoneally injected continuously for 5 days, and the Dingshen Decoction prepared in Comparative Example 4 at a dose of 0.50 g / mL was intragastrically administered for 14 days;

[0179] ⑤Comparative Example 5: MPTP was intraperitoneally injected continuously for 5 days, and the Dingshen Decoction prepared in Comparative Example 5 at a dose of 0.50 g / mL was gavage for 14 days.

[0180] (1) Open field test

[0181] The open field test was used to conduct behavioral evaluation on the experimental mice. The evaluation results were used to indicate the motor ability of the mice. The results of the open field test are shown in Tables 7-8 below.

[0182] Table 7 Baseline distance of open field test and distance after the test

[0183]

[0184] As can be seen from Table 7, compared with the MPTP group, the MPTP model mice administered with Wuteng Dingshen Decoction of Example 4 were significantly higher than the mice in the MPTP group and were similar to the mice in the blank group; while the mice in Comparative Examples 4-5 were lower than the mice in the Example 4 group, and Comparative Example 5 was slightly lower than the mice in Comparative Example 4.

[0185] Table 8 Baseline speed of open field test and speed after the test

[0186]

[0187] It can be seen from Table 8 that the speed of the mice in Comparative Example 4-5 is lower than that of the mice in Example 4 group.

[0188] (2) Striatal DA measurement

[0189] Striatal sampling: The sampling and detection methods are consistent with those described above and will not be described in detail. The results of striatal DA determination are shown in Table 9.

[0190] Table 9 Striatal DA measurement table

[0191]

[0192]

[0193] The present invention can be implemented in various ways and is not limited to the embodiments described. A person skilled in the art can understand that the present invention can be implemented in other specific ways without changing the technical concept or essential features of the present invention. Therefore, it should be understood that the embodiments described above are exemplary and are not intended to limit the present invention.

Claims

1. A Wuteng Dingshen Decoction for treating Parkinson's disease, characterized in that: The ingredients of the Wuteng Dingshen Decoction are as follows: The main ingredients include: Millettia spatholobi, Euonymus fortunei, Caulis ternatae, Caulis gentiana and Uncaria rhynchophylla; The components of the auxiliary material include: white peony root and liquorice.

2. The Wuteng Dingshen Decoction for treating Parkinson's disease according to claim 1, characterized in that: The components of the main material are 5-15g of Millettia spatholobi, 10-20g of Thunbergia fortunei, 5-15g of Polygonum multiflorum, 5-15g of Caulis Achyranthis Bidentatae and 10-20g of Uncaria rhynchophylla in a weight ratio, and the components of the auxiliary material are 5-15g of Paeonia lactiflora and 2-8g 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 ingredient are 8-12g of Millettia spatholobi, 13-17g of Thunbergia fortunei, 8-12g of Polygonum multiflorum, 8-12g of Caulis Achyranthis Bidentatae, 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.

4. The Wuteng Dingshen Decoction for treating Parkinson's disease according to claim 3, characterized in that: The components of the main material are 10g of Millettia spatholobi, 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.

5. The Wuteng Dingshen Decoction for treating Parkinson's disease according to claim 1, characterized in that: The auxiliary materials also include Eucommia ulmoides and Dragon Bone.

6. The Wuteng Dingshen Decoction for treating Parkinson's disease according to claim 5, 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 materials are 5-10g of Eucommia ulmoides, 5-10g of Ossobucus ursinus, 13-17g of Paeonia lactiflora and 3-6g of Licorice in a weight ratio.

7. The Wuteng Dingshen Decoction for treating Parkinson's disease according to claim 5, characterized in that: By weight, the auxiliary materials also include Radix Codonopsis pilosulae and Rhizoma Cynoglossi.

8. The Wuteng Dingshen Decoction for treating Parkinson's disease according to claim 7, characterized in that: The components of the main ingredient are 8-12g of Millettia reticulata, 13-17g of Eleutherodactyla fortunei, 8-12g of Polygonum multiflorum, 8-12g of Caulis Achyranthes bidentata, and 13-17g of Uncaria rhynchophylla, and the components of the auxiliary materials are 5-10g of Eucommia ulmoides, 5-10g of Osmanthus fragrans, 5-10g of Dioscorea macrophylla, 8-12g of Caulis Achyranthes bidentata, 13-17g of Paeonia lactiflora and 3-6g of Licorice.

9. A method for preparing Wuteng Dingshen Decoction for treating Parkinson's disease according to any one of claims 1 to 8, 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, and the medicinal liquid is divided into two portions to prepare Wu Teng Ding Shen Decoction.

10. The method for preparing Wuteng Dingshen Decoction for treating Parkinson's disease according to claim 9, characterized in that: The Wuteng Dingshen Decoction is an internal medicine, and the method of taking it is to take it warm once in the morning and once in the evening after meals.

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