Composition for treating mitochondrial dysfunction in Parkinson's disease and preparation method thereof

Abstract

A composition for treating mitochondrial dysfunction in Parkinson's disease, comprising an extract; the extract is extracted from a raw material drug; the raw material drug comprises Rehmannia glutinosa, Fructus Corni, Lycium barbarum, Cistanche deserticola, Morinda officinalis, Polygala tenuifolia, and Acorus tatarinowii. In the raw material drug, the mass ratio of Rehmannia glutinosa, Fructus Corni, Lycium barbarum, Cistanche deserticola, Morinda officinalis, Polygala tenuifolia, and Acorus tatarinowii is 1:1:1:1:1:0.5:0.5. Also included: the raw material drug further comprises Gastrodia elata: Uncaria rhynchophylla. In the raw material drug, the mass ratio of Rehmannia glutinosa, Fructus Corni, Lycium barbarum, Cistanche deserticola, Morinda officinalis, Polygala tenuifolia, Acorus tatarinowii, Gastrodia elata, and Uncaria rhynchophylla is equal to 1:1:1:1:1:0.5:0.5:0.5:1.

Description

Technical Field

[0001] The present application belongs to the technical field of Chinese medicine compositions and preparation methods thereof, and in particular to a Chinese medicine composition for treating mitochondrial dysfunction in Parkinson's disease and a preparation method thereof. Background Art

[0002] Parkinson's disease (PD) is a degenerative disease with resting tremor, muscle rigidity and bradykinesia as the main symptoms. At present, the pathogenesis of PD has not been fully elucidated. Clinically, drugs such as levodopa and dopamine receptor agonists are commonly used for treatment, but they can only control symptoms, but cannot effectively delay or prevent the progression of the disease, nor can they achieve the purpose of cure. In addition, serious side effects and complications will occur as the medication time increases.

[0003] Parkinson's disease mitochondrial dysfunction refers to a state in which the fasting triglyceride and / or total cholesterol levels in the blood are abnormally elevated. Its types include hypertriglyceridemia, hypercholesterolemia, mixed Parkinson's disease mitochondrial dysfunction, and high low-density lipoprotein levels. In terms of etiology, Parkinson's disease mitochondrial dysfunction can be divided into genetic factors and non-genetic factors.

[0004] As a common metabolic disorder, mitochondrial dysfunction in Parkinson's disease is closely related to lipid metabolism imbalance and has been shown to significantly increase the risk of cardiovascular disease. Previous studies have shown that ischemic cardiomyopathy, hypertensive cardiomyopathy, and diabetic cardiomyopathy are common causes of myocardial damage and necrosis. Myocardial cell damage can also be found in conditions such as hyperthyroidism, renal failure, and viral myocarditis. The emergence and development of these diseases are often related to abnormal lipid metabolism. For example, patients with coronary heart disease and diabetes often have dyslipidemia.

[0005] Histopathological studies further revealed that mitochondrial dysfunction in Parkinson's disease may lead to myocardial structural disorder, myocardial cell fibrosis and even interstitial hemorrhage.

[0006] A high-fat diet can induce cell apoptosis, death and autophagy, thus participating in a variety of disease processes. From a biochemical point of view, a high-fat diet can stimulate inflammatory responses in the body, promote the generation of reactive oxygen species, inhibit the expression of nuclear factor E2-related factor 2 (Nrf2) in the antioxidant system, and affect the structure and function of mitochondria, ultimately causing damage to cardiomyocytes.

[0007] According to statistics, about one-third of heart disease cases worldwide can be attributed to mitochondrial dysfunction in Parkinson's disease, highlighting its severity as an important public health issue.

[0008] Parkinson's disease mitochondrial dysfunction not only has a high prevalence, but also has a trend of being diagnosed at a younger age. In the United States, the proportion of people with high total cholesterol levels is about 17%, and among adults aged 20 and above, the prevalence of Parkinson's disease mitochondrial dysfunction is as high as 10%. In China, the overall prevalence of dyslipidemia in adults is as high as 40.4%. The total number of deaths caused by Parkinson's disease mitochondrial dysfunction ranks second among all metabolic diseases.

[0009] Statins are the first choice for lowering serum total cholesterol. They effectively regulate blood lipid levels in plasma by inhibiting methylhydroxyglutaryl coenzyme A (HM-CoA) reductase, a key enzyme in the synthesis of cholesterol in the body. However, it is worth noting that lipid-lowering drugs usually require patients to take them for a long time or even for life, which may be accompanied by adverse reactions such as loss of appetite, indigestion, headache, dizziness, joint pain and abnormal liver function, all of which may affect the patient's quality of life and treatment compliance.

[0010] Therefore, developing new drugs with independent intellectual property rights to treat mitochondrial dysfunction in Parkinson's disease not only has important clinical and economic value, but also has far-reaching significance for scientific research practice. Summary of the invention

[0011] In order to provide a traditional Chinese medicine composition for treating mitochondrial dysfunction in Parkinson's disease and a preparation method thereof, a convenient form of medication can be provided, while ensuring the effectiveness of the drug ingredients, while reducing the toxic and side effects of the drugs, and improving the user experience and ensuring the drug effect.

[0012] The technical solution of the present application to solve the above technical problems is a composition for treating mitochondrial dysfunction in Parkinson's disease, comprising: an extract; the extract is extracted from raw materials; the raw materials include Rehmannia glutinosa, Fructus Corni, Lycium barbarum, Cistanche deserticola, Morinda officinalis, Polygala tenuifolia, and Acorus tatarinowii.

[0013] It can be that in the raw material medicine, the mass ratio of Rehmannia glutinosa, Fructus Corni, Lycium barbarum, Cistanche deserticola, Morinda officinalis, Polygala tenuifolia and Acorus calamus is 1:1:1:1:1:0.5:0.5.

[0014] Yes, it can also include: the raw materials also include Gastrodia elata and Uncaria rhynchophylla.

[0015] It can be that in the raw materials, the mass ratio of Rehmannia glutinosa, Cornus officinalis, Lycium barbarum, Cistanche deserticola, Morinda officinalis, Polygala tenuifolia, Acorus calamus, Gastrodia elata and Uncaria rhynchophylla is equal to 1:1:1:1:1:0.5:0.5:0.5:1.

[0016] The technical solution of the present application to solve the above technical problem can also be a Chinese patent medicine for treating mitochondrial dysfunction in Parkinson's disease, wherein the Chinese patent medicine is a tablet, a pill, a granule or an oral liquid, and is characterized in that it includes the composition.

[0017] The technical solution of the present application to solve the above technical problem can also be a medicine for treating mitochondrial dysfunction in Parkinson's disease, wherein the medicine is a tablet, a pill, a granule or an oral liquid, characterized in that it includes the composition. The medicine can include western medicine.

[0018] The technical solution of the present application to solve the above technical problems can also be a composition for treating mitochondrial dysfunction in Parkinson's disease, characterized in that it includes a combination of raw materials, and the raw materials include Rehmannia glutinosa, Cornus officinalis, Lycium barbarum, Cistanche deserticola, Morinda officinalis, Polygala tenuifolia, and Acorus tatarinowii.

[0019] It can be that in the raw material drug combination, the mass ratio of Rehmannia glutinosa, Cornus officinalis, Lycium barbarum, Cistanche deserticola, Morinda officinalis, Polygala tenuifolia, and Acorus calamus is equal to 1:1:1:1:1:0.5:0.5, and the mass of one portion is 30 g.

[0020] Yes, the raw material combination includes Gastrodia elata: Uncaria rhynchophylla.

[0021] The mass ratio of Rehmannia glutinosa, Cornus officinalis, Lycium barbarum, Cistanche deserticola, Morinda officinalis, Polygala tenuifolia, Acorus calamus, Gastrodia elata and Uncaria rhynchophylla is 1:1:1:1:1:0.5:0.5:0.5:1.

[0022] The technical solution of the present application to solve the above technical problems can also be a method for preparing a decoction for treating mitochondrial dysfunction in Parkinson's disease, comprising: step S1: weighing the composition described in any one of claims 6 to 9 according to the formula, putting it into a decoction device, and adding 7-9 times the mass of water; step S2: heating it to boiling with fire, and keeping it boiling for more than 1 hour; separating the composition and the medicinal liquid to obtain medicinal liquid A; step S3: using the separated composition obtained in step S2, adding 4-6 times the mass of water again; heating it to boiling with fire, and keeping it boiling for more than 1 hour; separating the composition and the medicinal liquid; Step S4: combine the medicinal liquid A and the medicinal liquid B, and concentrate them to 60-90 ml to form the medicinal liquid C; Step S5: cool the medicinal liquid C, add ethanol to make the volume of ethanol account for more than 60% to obtain the medicinal liquid D; Step S5: let the medicinal liquid D stand for more than 12 hours, and take the supernatant to obtain the medicinal liquid F; Step S6: remove the ethanol from the medicinal liquid F, and volatilize it to a state without alcohol taste to obtain the medicinal liquid G; Step S7: add water to the medicinal liquid G to a volume of 70-110 ml to obtain the medicinal liquid H; Step S8: boil the medicinal liquid H and concentrate it to half of the volume to obtain the target medicinal liquid I.

[0023] One of the technical effects of the above technical scheme is that Rehmannia glutinosa, Fructus Corni, Lycium barbarum, Cistanche deserticola, and Morinda officinalis nourish the liver and kidney; Polygala tenuifolia and Acorus calamus dissipate phlegm and open the mind, improve intelligence and calm the mind; Gastrodia elata and Uncaria rhynchophylla calm the liver and stop wind, stop tremors and stop spasms. The whole formula is mainly used to nourish the liver and kidney, stop wind and stop spasms, and is supplemented by dissolving phlegm and nourishing qi and blood, which together play the effects of nourishing the liver and kidney, calming the liver and extinguishing wind, stopping tremors and stopping spasms, so as to improve symptoms such as tremors, rigidity, and hypokinesia.

[0024] One of the technical effects of the above technical scheme is that cooked rehmannia root is made by steaming raw rehmannia root with rice wine. It has a strong smell, is sweet but not bitter, is slightly warm but not cold, is moist and nourishing, and enters the kidney meridian. Its function is also changed from clearing to nourishing. It is good at nourishing yin and blood, replenishing the kidney and essence, and increasing marrow and improving intelligence.

[0025] One of the technical effects of the above technical solution is that the flavor of cornus is sweet, sour, slightly warm, and enters the liver and kidney meridians. It has a mild tonic effect, strengthens yang without adding fire, nourishes yin without greasy diaphragm, astringes without retaining evil, and has the effects of nourishing liver and kidney, astringing and consolidating. Rehmannia glutinosa and cornus can jointly play multiple effects of nourishing liver and kidney, nourishing blood and consolidating essence, and supplementing liver yang.

[0026] One of the technical effects of the above technical solution is that Cistanche deserticola tastes sweet and salty, is warm in nature, and enters the kidney and large intestine meridians, and has the effects of tonifying kidney yang, benefiting essence and blood, and moistening the intestines and relieving constipation. Studies have shown that Cistanche deserticola has neuroprotective effects such as anti-apoptosis, anti-oxidation, anti-aging, and anti-cerebral ischemia-reperfusion injury. The main component of Cistanche deserticola, phenylethanoid glycoside, can inhibit the activity of Caspase-3 and counteract the apoptosis of dopaminergic neurons caused by MPP+ (full English name + Chinese).

[0027] One of the technical effects of the above technical solution is that Morinda officinalis is pungent and sweet in taste and slightly warm in nature. It enters the kidney and liver meridians. It nourishes the kidney and boosts yang, dispels wind and dampness, and strengthens tendons and bones.

[0028] One of the technical effects of the above technical solution is that Uncaria rhynchophylla is sweet in taste and cool in nature, enters the liver and heart meridians, can calm liver wind, clear away liver heat, and calm wind and tremors to treat symptoms of liver heat and wind.

[0029] One of the technical effects of the above technical solution is that Polygala tenuifolia has a warm nature and a bitter taste, and its warmth can nourish the kidney. Li Zhongzi believes that Polygala tenuifolia can "replenish deficiency, remove evil spirits, improve wisdom and benefit the nine orifices, and cure forgetfulness.

[0030] One of the technical effects of the above technical solution is that both Acorus calamus and Polygala tenuifolia enter the heart and open the mind. Acorus calamus removes phlegm and opens the mind, removes dampness and stimulates appetite, and is often used for coma and forgetfulness caused by phlegm and turbidity blocking the mind; Polygala tenuifolia connects the heart and kidneys and has the effects of calming the mind, improving intelligence, removing phlegm and opening the mind, and is often used for insomnia, dreaminess, palpitations, and forgetfulness. Polygala tenuifolia connects the kidneys and connects with the heart, dispersing phlegm and resolving stagnation; Acorus calamus is pungent, dispersing, warming, and dispersing filth and removing dampness. The two medicines are used together to complement each other, and the power of removing phlegm and opening the mind, improving intelligence and calming the mind, and connecting the heart and kidneys is enhanced.

[0031] One of the technical effects of the above technical scheme: Gastrodia elata is sweet in taste and mild in nature, enters the liver meridian, nourishes fluid and calms the liver, calms wind and suppresses yang, and is also good at treating wind, and is mainly used to treat "wind deficiency, dizziness and headache" and various symptoms caused by wind and phlegm disturbance.

[0032] One of the technical effects of the above technical solution is that Uncaria rhynchophylla is sweet and cool in nature, enters the liver and heart meridians, can calm liver wind, clear liver heat, and calm wind and tremor to treat symptoms of liver heat and wind.

[0033] In this application, the composition and compatibility of the Chinese medicine prescription were specially designed and optimized. A new Chinese medicine formula was developed to effectively treat mitochondrial dysfunction in Parkinson's disease. BRIEF DESCRIPTION OF THE DRAWINGS

[0034] Figure 1 is a schematic diagram of the mouse treatment timeline;

[0035] Figure 2 This is a schematic diagram of the experimental results of behavioral changes in mice;

[0036] Figure 3 This is a schematic diagram of immunohistochemical staining of the substantia nigra pars compacta (SNPc) in the midbrain of mice;

[0037] Figure 4 This is a schematic diagram of TH-positive cells in the substantia nigra compacta of mice;

[0038] Figure 5 This is a schematic diagram of the changes in mitochondrial complexes I to IV in the brain tissue of each group of mice

[0039] Figure 6 is a schematic diagram of flow cytometry detection of mitochondria;

[0040] Figure 7 is a schematic diagram of mitochondrial membrane potential;

[0041] Figure 8 It is a schematic diagram of the expression results of proteins related to mitochondrial dynamics;

[0042] Fig. 9 It is a schematic diagram of a process for preparing a decoction for treating mitochondrial dysfunction in Parkinson's disease;

[0043] Fig.10 It is a schematic diagram of a process for preparing a decoction for treating mitochondrial dysfunction in Parkinson's disease. DETAILED DESCRIPTION

[0044] The content of this application is further described in detail below in conjunction with the accompanying drawings. It should be noted that the following is a description of the preferred embodiments of the present invention and does not constitute any limitation to the present invention. The description of the preferred embodiments of the present invention is only an illustration of the general principles of the present invention. The numbers such as "first", "second" and "A" and "B" involved in the present invention are only for the convenience of explanation and do not represent a temporal or spatial order relationship.

[0045] Traditional Chinese medicine is characterized by low cost and few side effects. It also has its own regulatory characteristics and advantages such as comprehensiveness, integrity, and bidirectionality. Compared with extremely expensive Western medicine, it is a low-cost solution.

[0046] Some formulas in traditional Chinese medicine also have some side effects, and traditional methods cannot accurately identify the effective ingredients and toxic ingredients. How to simplify the formula, improve the safety of medication, and reduce the complexity and cost of medication is a technical problem to be solved.

[0047] The applicant has been exploring the treatment of Parkinson's disease through Chinese medicine compound. How to combine various Chinese medicine formulas to form a compound, and develop a high-tech Chinese medicine modern compound with independent intellectual property rights, concentrated effective components, clear target, significant efficacy, and less toxic and side effects is the direction that has been explored.

[0048] In the present application, a composition for treating mitochondrial dysfunction in Parkinson's disease is designed, comprising: an extract; the extract is extracted from a raw material drug; the raw material drug comprises Rehmannia glutinosa, Fructus Corni, Lycium barbarum, Cistanche deserticola, Morinda officinalis, Polygala tenuifolia, and Acorus tatarinowii. In the raw material drug, the mass ratio of Rehmannia glutinosa, Fructus Corni, Lycium barbarum, Cistanche deserticola, Morinda officinalis, Polygala tenuifolia, and Acorus tatarinowii is 1:1:1:1:1:0.5:0.5.

[0049] The composition for treating mitochondrial dysfunction in Parkinson's disease also includes: the raw material medicine also includes Gastrodia elata: Uncaria rhynchophylla. In the raw material medicine, the mass ratio of Rehmannia glutinosa, Fructus Corni, Lycium barbarum, Cistanche deserticola, Morinda officinalis, Polygala tenuifolia, Acorus calamus, Gastrodia elata and Uncaria rhynchophylla is 1:1:1:1:1:0.5:0.5:0.5:1.

[0050] In the present application, a Chinese patent medicine for treating mitochondrial dysfunction in Parkinson's disease is designed. The Chinese patent medicine is in the form of tablets, pills, granules or oral liquid, and is characterized in that it comprises the above-mentioned composition.

[0051] In the present application, a medicine for treating mitochondrial dysfunction in Parkinson's disease is designed, and the medicine is in the form of tablets, pills, granules or oral liquid, including the above-mentioned combination.

[0052] In the present application, a composition for treating mitochondrial dysfunction in Parkinson's disease is designed, comprising a combination of raw materials, wherein the raw materials include Rehmannia glutinosa, Fructus Corni, Lycium barbarum, Cistanche deserticola, Morinda officinalis, Polygala tenuifolia, and Acorus tatarinowii.

[0053] In the raw material drug combination, the mass ratio of Rehmannia glutinosa, Cornus officinalis, Lycium barbarum, Cistanche deserticola, Morinda officinalis, Polygala tenuifolia, and Acorus calamus is 1:1:1:1:1:0.5:0.5, and the mass of one portion is 30 g.

[0054] The raw material combination also includes Gastrodia elata: Uncaria rhynchophylla. The mass ratio of Rehmannia glutinosa, Fructus Corni, Lycium barbarum, Cistanche deserticola, Morinda officinalis, Polygala tenuifolia, Acorus calamus, Gastrodia elata and Uncaria rhynchophylla is 1:1:1:1:1:0.5:0.5:0.5:1.

[0055] A Chinese medicine formula for improving Parkinson's disease mitochondrial dysfunction includes: cooked rehmannia: cornus: wolfberry: Cistanche deserticola: Morinda officinalis: Gastrodia elata: Uncaria rhynchophylla: Polygala tenuifolia: Acorus calamus = 30g; 30g; 30g; 30g: 30g; 15g; 30g; 15g; 15g. Among them, cooked rehmannia, cornus, wolfberry, Cistanche deserticola, Morinda officinalis nourish the liver and kidney; Polygala tenuifolia and Acorus calamus resolve phlegm, open the mind, improve intelligence and calm the mind; Gastrodia elata and Uncaria rhynchophylla calm the liver and stop wind, stop tremors and stop spasms. The whole formula is mainly used to nourish the liver and kidney, stop wind and stop spasms, and is supplemented with resolving phlegm and nourishing qi and blood, which together play the effects of nourishing the liver and kidney, calming the liver and extinguishing wind, stopping tremors and stopping spasms, so as to improve symptoms such as tremors, rigidity, and hypokinesia.

[0056] Rehmannia glutinosa is made from raw Rehmannia glutinosa steamed with rice wine. It has a strong flavor, sweet but not bitter, slightly warm but not cold, moist and nourishing. It also enters the kidney meridian, and its function changes from clearing to tonic. It is good at nourishing blood and yin, replenishing essence and benefiting marrow.

[0057] Fructus Corni is sweet, sour, slightly warm in nature, and enters the liver and kidney meridians. It has a mild tonic effect, strengthens yang without adding fire, nourishes yin without greasy diaphragm, and astringes without retaining evil. It has the effects of nourishing liver and kidney, astringing and consolidating. Rehmannia glutinosa and Fructus Corni have multiple effects of nourishing liver and kidney, nourishing blood and consolidating essence, and supplementing liver yang.

[0058] Cistanche deserticola tastes sweet and salty, is warm in nature, and enters the kidney and large intestine meridians. It has the effects of tonifying kidney yang, benefiting essence and blood, and moistening the intestines and relieving constipation. Studies have shown that Cistanche deserticola has anti-apoptosis, anti-oxidation, anti-aging, and anti-cerebral ischemia-reperfusion injury neuroprotective effects. The main component of Cistanche deserticola, phenylethanoid glycoside, can inhibit the activity of Caspase-3 and counteract the apoptosis of dopaminergic neurons caused by MPP+ (full English name + Chinese).

[0059] Morinda officinalis, pungent, sweet, slightly warm in nature. Enters the kidney and liver meridians. It nourishes the kidney and boosts yang, dispels wind and dampness, and strengthens tendons and bones.

[0060] Uncaria rhynchophylla is sweet in taste and cool in nature. It enters the liver and heart meridians. It can calm liver wind, clear liver heat, and calm wind and tremors to treat symptoms of liver heat and wind.

[0061] Acorus calamus has a strong aroma and smell, and it has the woody aroma of spring and harmony. It tastes bitter and warm in nature, and has the effects of opening the mind and calming the spirit, removing dampness and soothing the stomach. Acorus calamus removes the foul air of the four seasons and can be used to treat phlegm and dampness that obstruct the mind. The "Compendium of Materia Medica" says: "Taking it for a long time will make you feel light, not forgetful, not confused, prolong your life, improve your mind, raise your ambitions, and prevent you from aging."

[0062] Polygala has a warm nature and a bitter taste. Its warmth can nourish the kidneys. Li Zhongzi believes that Polygala can: "replenish deficiency, remove evil spirits, improve wisdom and benefit the nine orifices, and cure forgetfulness."

[0063] Both Acorus calamus and Polygala tenuifolia enter the heart and open the orifices. Acorus calamus removes phlegm and opens the orifices, removes dampness and stimulates appetite, and is often used for coma and forgetfulness caused by phlegm and turbidity blocking the orifices; Polygala tenuifolia connects the heart and kidneys and has the effects of calming the mind and improving intelligence, dispersing depression and resolving phlegm, and is often used for insomnia, dreaminess, palpitations, and forgetfulness. Polygala tenuifolia connects the kidneys and connects with the heart, dispersing phlegm and resolving knots; Acorus calamus is pungent, dispersing, warming, and dispersing filth and removing dampness. The two herbs are used together to complement each other, and the power of removing phlegm and opening the orifices, improving intelligence and calming the mind, and connecting the heart and kidneys is enhanced.

[0064] Gastrodia elata is sweet and mild in nature. It enters the liver meridian, nourishes fluid and calms the liver, calms wind and suppresses yang. It is also good at treating wind. It is mainly used to treat "wind-deficiency, dizziness and headache" and various symptoms caused by wind and phlegm.

[0065] The combination of Gastrodia elata and Uncaria rhynchophylla can be traced back to the "Compendium of Medical Prescriptions" by Sun Yunxian in the Yuan Dynasty. Both medicines are commonly used to treat liver wind movement, epilepsy and convulsions. The combination of the two medicines complements each other to enhance the effects of clearing heat and calming the liver, calming wind and stopping spasms. Many prescriptions based on the Gastrodia elata-Uncaria rhynchophylla pair have an improvement effect on bradykinesia, resting tremor, muscle rigidity, postural instability and sleep disorders in PD; they can also reduce the dosage of levodopa, thereby reducing its adverse reactions.

[0066] like Fig. 9 As shown, in a method for preparing a decoction for treating mitochondrial dysfunction in Parkinson's disease, the components of the above-mentioned composition are weighed according to proportion, decocted with water twice, each time for 1 hour, the decoctions are combined, filtered, and the filtrate is concentrated to 70ml, cooled, ethanol is added to make the alcohol content reach 60%, and allowed to stand overnight. The supernatant is taken, the ethanol is recovered, evaporated until there is no alcohol taste, water is added to 90ml, boiled, and made into a 200% aqueous solution, sterilized at 0.1mPa for 15min, and stored at 4°C for use.

[0067] In a method for preparing a decoction for treating mitochondrial dysfunction in Parkinson's disease, each component is first soaked in warm water for 30 minutes; Gastrodia elata is first decocted for 15 minutes, and then the other components except Uncaria rhynchophylla are added and boiled for 30 minutes using a normal medicine decoction device, and finally Uncaria rhynchophylla is added and boiled for 15 minutes. After boiling, the residue is removed, filtered with gauze, and the decoction is concentrated at 37 degrees to obtain the target medicine solution.

[0068] Specifically, Fig.10 , a method for preparing a decoction for treating mitochondrial dysfunction in Parkinson's disease, comprising:

[0069] Step S1: After weighing the composition according to the proportion of each component, put it into a decoction device, and add 8 times the mass of water; the amount of water should at least cover the composition by more than 2 cm;

[0070] Step S2: heating to boiling, and keeping boiling for more than 1 hour; separating the composition and the liquid medicine to obtain liquid medicine A;

[0071] Step S3: Add 5 times the mass of water to the separated composition obtained in step S2, with the amount of water at least covering the composition by more than 2 cm; heat to boiling, and keep boiling for more than 1 hour; separate the composition and the liquid medicine to obtain liquid medicine B;

[0072] Step S4: combining the drug solution A and the drug solution B and concentrating them to 70 ml to form drug solution C;

[0073] Step S5: cooling the drug solution C, and adding ethanol so that the volume ratio of ethanol is more than 60% to obtain drug solution D;

[0074] Step S5: the drug solution D is allowed to stand for more than 12 hours, and the supernatant is taken to obtain drug solution F;

[0075] Step S6: The drug solution F is subjected to a deethanol treatment to evaporate to a state without alcohol taste, thereby obtaining the drug solution G;

[0076] Step S7: Add water to the drug solution G to a volume of 90 ml to obtain drug solution H.

[0077] Step S8: The drug solution H is boiled, concentrated to half its volume, and sterilized at 0.1 mPa for 15 min to obtain the target drug solution I.

[0078] The above formula, through a large number of clinical observations, has been found to not only relieve the clinical symptoms of PD and delay the progression of the disease, but also has less toxic side effects. Early animal experiments have shown that it can significantly improve the rotational behavior of PD model rats, increase the content of striatal dopamine (DA) and its metabolites, increase the expression of tyrosine hydroxylase (TH) and its THmRNA, and has anti-oxidative stress and certain anti-apoptotic effects. It can be used to treat mitochondrial dysfunction in Parkinson's disease. Diseases can be treated through multiple pathways and at multiple links or targets.

[0079] The applicant conducted a series of drug efficacy verification experiments. According to the drug efficacy, it was divided into multiple groups, and the drug-containing serum was extracted for high-throughput screening experiments. The high-throughput screening used an in vitro cell model of Parkinson's disease, including PC12 cell line and SH-SY5Y cell line. The PD in vitro cell model was used for high-throughput screening, the prescription compatibility was optimized, and the target formula group was obtained. The SH-SY5Y cell line is a human neuroblastoma cell; the PC12 cell line is a cell derived from the adrenal medulla pheochromocytoma of adult rats; the SH-SY5Y PD model and the PC12 PD model mean that the above cells are used as PD models for experimental verification.

[0080] The multiple groups are:

[0081] Group A: Rehmannia glutinosa: Fructus Corni: Lycium barbarum: Cistanche deserticola: Morinda officinalis; mass ratio is 1:1:1:1:1=30g:30g:30g:30g:30g:30g

[0082] Group B: Gastrodia elata: Uncaria rhynchophylla; mass ratio is 1:2 = 15g:30g

[0083] Group C: Polygala tenuifolia: Acorus calamus; mass ratio is 1:1 = 15g:15g

[0084] Group D: Scorpion: Bombyx batryticatus; mass ratio is 1:2 = 5g:10g

[0085] Group E: Pueraria root; 30g

[0086] Group F: Rehmannia glutinosa: Cornus officinalis: Lycium barbarum: Cistanche deserticola: Morinda officinalis: Gastrodia elata: Uncaria rhynchophylla: Polygala tenuifolia: Acorus calamus: Pueraria lobata: Bombyx batryticatus: Scorpio.

[0087] The mass ratio of each component is: 6:6:6:6:6:3:6:3:3:6:2:1.

[0088] The mass ratio of each component can also be:

[0089] 30g:30g:30g30g:30g:15g:30g:15g:15g:30g:10g:5g.

[0090] Group G: blank control group NC.

[0091] The efficacy of the target formula group was verified by observing the effects of the target formula group on the ultrastructure of substantia nigra dopaminergic neurons in PD model mice, and the content of DA and its metabolites; the mechanism of action was deeply explored from the aspects of autophagy, brain mitochondrial function, mitochondrial dynamics, and the dynamic balance regulation of mitochondrial fusion and fission by observing the effects of the target formula group on mitochondrial complex enzyme activity, mitochondrial transmembrane potential, mitochondrial respiratory function, ATPase synthesis activity, ROS generation rate, mitochondrial dynamics-related proteins, and autophagy-related proteins and their mRNA expression under the intervention of autophagy inducers and blockers.

[0092] Effect and mechanism verification:

[0093] The experimental mice were divided into a normal group (Sham), a PD model group (Model), a positive drug control group (Madopar), and a Chinese medicine control group ( XFDCT ), the target formula group (Op Rx), MPTP-induced (1-methyl-4-phenyl-1,2.3,6-tetrahydropyridine-induced) 30 mg / kg·d intraperitoneal injection was used to establish a subacute PD mouse model, and samples were collected after 14 days of intervention in each group. The positive drug control group (Madopar) used the imported Western medicine "Madopar". The Chinese medicine control group ( XFDCT) is the formulation of the above-mentioned group F. The target formulation group is the formulation of the above-mentioned group A + group C + group B.

[0094] Compare the behavioral effects of the above groups on PD mice:

[0095] like Figure 1 First, MPTP was used for induction. Five days after intraperitoneal injection, the model mice showed significant motor dysfunction, and the climbing time was significantly increased compared with that before modeling (P<0.01), indicating that the PD model was successfully established. After 7 and 14 days of treatment with the target formula group, the motor ability of the mice was greatly restored, and the climbing time of the target formula group was significantly shorter than that of the PD model group (P<0.01). The results are shown in Figure 2 As shown in the pole test picture on the left.

[0096] like Figure 2 , as shown in the middle Rotarod test, after 5 consecutive days of MPTP intraperitoneal injection modeling, the latency of the mouse rotarod movement (first fall time) was significantly reduced compared with that before modeling (P<0.01), indicating that the motor function of the model mice was significantly reduced; the latency of the Op Rx group was higher than that of the model group after 7 days of treatment (P<0.01), indicating that Chinese medicine treatment played a significant role in the recovery of motor function in PD mice;

[0097] like Figure 2 As shown in the Tremor score on the right, 5 days after the intraperitoneal injection of MPTP, the mice developed obvious symptoms of tremor paralysis (P<0.01), but the tremor paralysis symptoms did not persist for a long time. Within a few hours to a few days after the cessation of MPTP injection, the tremor paralysis symptoms of mice in each group gradually decreased and became difficult to observe. Therefore, there was no difference between the groups after subsequent intervention treatment.

[0098] Effects on TH expression in substantia nigra pars compacta of PD mice:

[0099] After 14 days of Chinese medicine gavage treatment, immunohistochemical staining was performed on the substantia nigra pars compacta (SNPc) of the midbrain slices of mice. Figure 3 As shown in the figure, the number of neurons in the substantia nigra pars compacta in the model group decreased and their arrangement was scattered. Figure 4 The number of TH-positive cells in the substantia nigra of the Op Rx group was significantly reduced compared with that in the model group after 14 days of treatment with traditional Chinese medicine (P<0.01). This indicates that the optimized prescription treatment induces enhanced TH expression in the substantia nigra of mice and has a neuroprotective effect on PD mice.

[0100] Effects on the activity of mitochondrial complexes I to IV in the substantia nigra of PD mice:

[0101] Effects of Chinese medicine on the activities of mitochondrial respiratory chain complex enzymes I-IV in the substantia nigra of subacute PD model mice Figure 5 The results in the table show that 14 days after modeling and drug administration, the activity of mitochondrial complex I in the model group was significantly reduced compared with the normal control group (P<0.01). After 14 days of treatment with the optimized prescription, the activity of mitochondrial complex I in the Op Rx group was significantly increased compared with the model group (P<0.01). In addition, there was no statistical difference in the activities of mitochondrial complex II, III, and IV among the groups, indicating that Chinese medicine treatment has a restorative effect on the activity of mitochondrial complex I in PD model mice.

[0102] Effects on mitochondrial membrane potential in the substantia nigra of PD mice:

[0103] To further observe the protective effect of the optimized Chinese medicine prescription on mitochondria in PD mice, the JC-1 dye method was used to detect the mitochondrial membrane potential, the ratio of Green / Red (i.e., monomer / aggregate / monomer) in the cells. When the ratio increased, the mitochondrial membrane potential level of the cells decreased, and when the ratio decreased, the mitochondrial membrane potential level of the cells increased. Figure 6 and Figure 7 As shown in the data, the green / red fluorescence ratio of the model group was significantly increased (P<0.001), indicating that the mitochondrial membrane potential level of MPTP-induced PD mice was significantly decreased, mitochondrial permeability was increased, channel pores were open, and a large number of neurons were in the early stages of apoptosis; the green / red fluorescence ratio of the target formula group after 14 days of treatment was significantly lower than that of the model group (P<0.01), indicating that the optimized formula treatment can improve the mitochondrial membrane potential of MPTP-induced mice, reduce mitochondrial damage, and thus play a protective role in DA neurons.

[0104] Effects on mitochondrial dynamics-related proteins in the substantia nigra of PD mice:

[0105] Western blot was used to detect the expression of mitochondrial dynamics-related proteins in the substantia nigra of each group of mice. The results are shown in Figure 8 Obviously, compared with the normal control group, the expression of mitochondrial fission proteins Fis1 and Drp1 in MPTP-induced PD model mice was significantly increased (P<0.01), and the expression of mitochondrial fusion protein Mfn2 was significantly inhibited (P<0.01). After 14 days of treatment with the optimized prescription, the expression of fission proteins Fis1 and Drp1 decreased to varying degrees (P<0.01), and the content of mitochondrial fusion protein Mfn2 was significantly increased (P<0.01), indicating that the increase of MPTP-induced mitochondrial fission and the inhibition of mitochondrial fusion can be reversed by traditional Chinese medicine treatment.

[0106] The compound formula in this application can be further applied in clinical practice to promote the recovery of motor and non-motor symptoms of PD patients, delay disease progression, improve patients' quality of life, reduce medical expenses, and alleviate the burden on families, society and medical insurance. It has very important academic value and social significance.

[0107] Although the present invention is illustrated and described according to the preferred embodiment and several alternatives, the invention is not limited by the specific description in this specification. Other additional replacement or equivalent components can also be used to practice the present invention.

Claims

1. A composition for treating mitochondrial dysfunction in Parkinson's disease, characterized in that: include: Extracts; Extracts are made from raw materials; The raw materials include Rehmannia glutinosa, Cornus officinalis, Lycium barbarum, Cistanche deserticola, Morinda officinalis, Polygala tenuifolia, and Acorus calamus.

2. The composition for treating mitochondrial dysfunction in Parkinson's disease according to claim 1, characterized in that: Among the raw materials, the mass ratio of Rehmannia glutinosa, Fructus Corni, Lycium barbarum, Cistanche deserticola, Morinda officinalis, Polygala tenuifolia and Acorus calamus is 1:1:1:1:1:0.5:0.

5.

3. The composition for treating mitochondrial dysfunction in Parkinson's disease according to claim 1, characterized in that: Also includes: The raw materials also include Gastrodia elata and Uncaria rhynchophylla.

4. The composition for treating mitochondrial dysfunction in Parkinson's disease according to claim 3, characterized in that: Among the raw materials, the mass ratio of Rehmannia glutinosa, Cornus officinalis, Lycium barbarum, Cistanche deserticola, Morinda officinalis, Polygala tenuifolia, Acorus calamus, Gastrodia elata, and Uncaria rhynchophylla is 1:1:1:1:1:0.5:0.5:0.5:

1.

5. A drug for treating mitochondrial dysfunction in Parkinson's disease, characterized in that: include: The composition according to claims 1 to 4; Include any of the following technical features: TA10: The drug is a Chinese patent medicine, which is in the form of tablets, pills, granules or oral liquid; TA20: The drug is a finished medicine, which is in the form of tablets, pills, granules or oral liquid.

6. A composition for treating mitochondrial dysfunction in Parkinson's disease, characterized in that: It includes a combination of raw materials, and the raw materials include Rehmannia glutinosa, Cornus officinalis, Lycium barbarum, Cistanche deserticola, Morinda officinalis, Polygala tenuifolia, and Acorus calamus.

7. The composition for treating mitochondrial dysfunction in Parkinson's disease according to claim 6, characterized in that: In the raw material drug combination, the mass ratio of Rehmannia glutinosa, Cornus officinalis, Lycium barbarum, Cistanche deserticola, Morinda officinalis, Polygala tenuifolia, and Acorus calamus is 1:1:1:1:1:0.5:0.5, and the mass of one portion is 30 g.

8. The composition for treating mitochondrial dysfunction in Parkinson's disease according to claim 6, characterized in that: The raw material combination includes Gastrodia elata and Uncaria rhynchophylla.

9. The composition for treating mitochondrial dysfunction in Parkinson's disease according to claim 8, characterized in that: The mass ratio of Rehmannia glutinosa, Cornus officinalis, Lycium barbarum, Cistanche deserticola, Morinda officinalis, Polygala tenuifolia, Acorus calamus, Gastrodia elata and Uncaria rhynchophylla is 1:1:1:1:1:0.5:0.5:0.5:

1.

10. A method for preparing a decoction for treating mitochondrial dysfunction in Parkinson's disease, characterized in that: include: Step S1: weigh the composition according to any one of claims 6 to 9 according to the formula, put it into a decoction device, and add 7-9 times the weight of water; Step S2: heating to boiling, and keeping boiling for more than 1 hour; separating the composition and the liquid medicine to obtain liquid medicine A; Step S3: Add 4-6 times the mass of water to the separated composition obtained in step S2; heat it to boiling and keep boiling for more than 1 hour; separate the composition and the liquid medicine to obtain liquid medicine B; Step S4: combining the drug solution A and the drug solution B, and concentrating them to 60-90 ml to form drug solution C; Step S5: cooling the drug solution C, and adding ethanol so that the volume ratio of ethanol is more than 60% to obtain drug solution D; Step S5: the drug solution D is allowed to stand for more than 12 hours, and the supernatant is taken to obtain drug solution F; Step S6: The drug solution F is subjected to a deethanol treatment to evaporate to a state without alcohol taste, thereby obtaining the drug solution G; Step S7: adding water to the drug solution G to a volume of 70-110 ml to obtain drug solution H; Step S8: The drug solution H is boiled and concentrated to half its volume to obtain the target drug solution I.

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