Traditional Chinese medicine composition as well as preparation method and application thereof

Abstract

The invention provides a traditional Chinese medicine composition. Compared with the prior art, the traditional Chinese medicine composition has the advantages that the traditional Chinese medicines are reasonably matched, so that all the components have the effect of synergistically improving insomnia, and uncomfortable symptoms such as exhaustion, cognitive competence impairment and irritability of insomnia patients are improved; meanwhile, a transcriptome sequencing technology is adopted, the effect of the traditional Chinese medicine composition on signal channels of patients with insomnia and mild and moderate senile dementia is analyzed and studied, and it is found that the traditional Chinese medicine composition can remarkably reverse disease-related channels on disease-related cell models, remarkably reverse insomnia disorder channels in neuronal cells and microglia cells, and remarkably reverse insomnia disorder channels in neuronal cells and microglia cells. Disorder pathways of mild and moderate senile dementia diseases are obviously reversed in neuronal cells and vascular endothelial cells; therefore, the traditional Chinese medicine composition can be used for improving insomnia and preventing and treating mild and moderate senile dementia, and the traditional Chinese medicine composition is prepared from medicinal and edible medicinal materials and is safe and reliable.

Description

Technical Field

[0001] The present invention belongs to the technical field of traditional Chinese medicine, and in particular relates to a traditional Chinese medicine composition, a preparation method and application thereof. Background Art

[0002] Insomnia usually refers to a subjective experience in which patients are unable to fall asleep or stay asleep despite adequate sleep opportunities and environment, resulting in low rest recovery time and quality, thus affecting normal daytime activities. The main clinical manifestations are difficulty falling asleep, difficulty maintaining sleep, or dreaming and waking up early, as well as fatigue, impaired cognitive ability, irritability and other daytime functional disorders. Many studies have shown that long-term insomnia is an independent risk factor for cardiovascular and cerebrovascular diseases, depression, anxiety and diabetes. The increasing pace of life and the influx of various pressures have led to a continuous increase in the incidence of insomnia. According to statistics, the incidence of insomnia among adults in my country is 38.2%, and more than 300 million Chinese have sleep disorders, which is higher than the level in developed countries. Insomnia has become a major disease that affects people's lives and physical health and needs to be solved urgently. In addition, insomnia is closely related to mental disorders and cognitive decline. Studies have shown that insomnia may increase the risk of Alzheimer's disease, and insomnia is also one of the clinical symptoms of Alzheimer's disease. Among patients with cognitive impairment, the incidence of sleep disorders is much higher than that of people without cognitive impairment. A study on patients with Alzheimer's disease and those without it found that the incidence of difficulty falling asleep, fragmented sleep, restlessness at night, sleep-wake rhythm disorders, and obstructive sleep apnea syndrome in patients with Alzheimer's disease was significantly higher than that in patients without it. Moreover, patients with different types of cognitive impairment have different types of sleep disorders. For example, patients with mild cognitive impairment mainly have long sleep latency, less total sleep time, and low sleep efficiency; patients with Alzheimer's disease mainly have circadian rhythm disorders and changes in sleep structure.

[0003] Sleep is essential for brain health and function. It promotes a variety of physiological processes, including homeostatic regulation of neuronal activity, synaptic strength, and clearance of metabolic waste. Microglia are the main immune cells in the brain and play a key role in brain homeostasis by regulating neuronal activity, pruning synapses, and clearing cell debris and harmful aggregates. Alzheimer's disease (AD) is a degenerative disease of the central nervous system that occurs in the elderly and pre-elderly, characterized by progressive cognitive dysfunction and behavioral impairment. It is the main type of senile dementia. The main pathological characteristics of AD are β-amyloid deposition to form neuritic plaques, neurofibrillary tangles formed by abnormal phosphorylation of tau protein, neuronal loss and glial cell proliferation. In addition, changes and dysfunction of the cerebrovascular system are an important part of the pathophysiology of AD. This process promotes neurodegeneration, inflammation, β-amyloid accumulation and tau protein phosphorylation, and promotes the occurrence and progression of AD.

[0004] Therefore, the treatment of insomnia has become a prominent medical and social problem that needs to be solved urgently. Insomnia is often accompanied by other health problems, and sometimes it is difficult to determine the causal relationship between the two. Whether it is "primary" or "secondary", independent clinical intervention is required for insomnia itself to prevent the symptoms from being prolonged or recurring. Currently, in the treatment of insomnia, drugs are usually selected to intervene for patients. However, conventional Western medicines are not ideal during the medication period and have high side effects, which requires a more reasonable medication plan for patients.

[0005] Traditional Chinese medicine believes that insomnia belongs to the category of "insomnia". The Yellow Emperor's Classic of Internal Medicine once recorded insomnia as "cannot sleep", "cannot sleep eyes", and "cannot lie down". Traditional Chinese medicine believes that insomnia is essentially a syndrome of deficiency and excess, which is closely related to dysfunction of organs such as the heart, liver, kidney, and spleen, and mental malnutrition or restlessness. Since insomnia patients have a long course of illness, are prone to recurrent attacks, and are difficult to recover quickly, their treatment has always been one of the thorny problems faced by clinicians. In recent years, a large number of clinical practices have confirmed that traditional Chinese medicine has good clinical efficacy in the treatment of insomnia, and has unique advantages that Western medicine does not have, which can enable patients to obtain "twice the result with half the effort" clinical benefits. Therefore, it is very necessary to provide a Chinese medicine composition for improving insomnia for insomnia patients. Summary of the invention

[0006] In view of this, the technical problem to be solved by the present invention is to provide a Chinese medicine composition for improving insomnia, cognitive impairment or dementia, and a preparation method and application thereof.

[0007] The present invention provides a traditional Chinese medicine composition, comprising the following components:

[0008] 2-10 parts by weight of dried tangerine peel, 3-10 parts by weight of loblolly leaves, 0.3-1 parts by weight of cloves, 3-10 parts by weight of poria, 5-15 parts by weight of liquorice, 0.3-1 parts by weight of galangal, 0.1-0.5 parts by weight of cinnamon, 5-15 parts by weight of amomum, 3-10 parts by weight of hawthorn, 1-3 parts by weight of hovenia dulcis, and 0-3 parts by weight of citron.

[0009] Preferably, the following components are included:

[0010] 2-10 parts by weight of dried tangerine peel, 3-10 parts by weight of lophatherum gracile, 0.3-1 parts by weight of clove, 3-10 parts by weight of tuckahoe, 5-15 parts by weight of licorice, 0.3-1 parts by weight of galangal, 0.1-0.5 parts by weight of cinnamon, 5-15 parts by weight of amomum, 3-10 parts by weight of hawthorn, 1-3 parts by weight of hovenia dulcis, and 1-3 parts by weight of citron.

[0011] Preferably, the following components are included:

[0012] 5 parts by weight of dried tangerine peel, 5 parts by weight of lophatherum gracile, 0.5 parts by weight of clove, 5 parts by weight of tuckahoe, 10 parts by weight of licorice, 0.5 parts by weight of galangal, 0.3 parts by weight of cinnamon, 10 parts by weight of amomum, 5 parts by weight of hawthorn, 2 parts by weight of hovenia dulcis and 1 part by weight of citron.

[0013] Preferably, the following components are included:

[0014] 3 parts by weight of dried tangerine peel, 10 parts by weight of loblolly leaf, 0.3 parts by weight of clove, 10 parts by weight of tuckahoe, 5 parts by weight of liquorice, 1 part by weight of galangal, 0.1 parts by weight of cinnamon bark, 15 parts by weight of amomum, 3 parts by weight of hawthorn, 1 part by weight of hovenia dulcis and 3 parts by weight of citron.

[0015] Preferably, the following components are included:

[0016] 10 parts by weight of dried orange peel, 3 parts by weight of lophatherum gracile, 1 part by weight of clove, 3 parts by weight of tuckahoe, 15 parts by weight of liquorice, 0.3 parts by weight of galangal, 0.5 parts by weight of cinnamon bark, 5 parts by weight of amomum villosum, 10 parts by weight of hawthorn and 3 parts by weight of Hovenia dulcis;

[0017] Or it includes the following components: 2 parts by weight of dried tangerine peel, 5 parts by weight of lophatherum gracile, 0.5 parts by weight of clove, 5 parts by weight of tuckahoe, 9 parts by weight of liquorice, 0.9 parts by weight of galangal, 0.3 parts by weight of cinnamon bark, 6 parts by weight of amomum villosum, 5 parts by weight of hawthorn and 2 parts by weight of Hovenia dulcis.

[0018] The present invention also provides a method for preparing the above-mentioned Chinese medicine composition, comprising the following steps:

[0019] The Chinese medicine composition is obtained after the dried tangerine peel, loblolly leaf, clove, poria, liquorice, galangal, cinnamon bark, amomum villosum, hawthorn, hovenia dulcis fruit and citron are mixed and extracted.

[0020] The present invention also provides a use of the above-mentioned traditional Chinese medicine composition in the preparation of medicines and / or preparations for preventing, improving or treating insomnia;

[0021] And / or, use of the above-mentioned Chinese medicine composition in the preparation of drugs and / or preparations for preventing, improving or treating cognitive impairment or dementia.

[0022] Preferably, the medicine and / or preparation is selected from one or more of wine, granules, liquid beverage, tea substitute, powder, oral liquid, tablet, pill, paste and capsule.

[0023] The present invention also provides a use of the above-mentioned traditional Chinese medicine composition in the preparation of a drug and / or preparation for reversing and / or inhibiting a disorder in a disease signal pathway;

[0024] The disease signaling pathways include insomnia signaling pathways and mild to moderate Alzheimer's disease signaling pathways.

[0025] Preferably, the insomnia signaling pathways include autophagy-animal, viral carcinogenesis, neutrophil extracellular trap formation, osteoclast differentiation, FcγR-mediated phagocytosis, Th1 and Th2 cell differentiation, neurotrophin signaling pathway, endocrine resistance, Toll-like receptor signaling pathway, T cell receptor signaling pathway, phosphatidylinositol signaling system, phosphoinositide metabolism, and NOD-like receptor signaling pathway;

[0026] The mild to moderate Alzheimer's disease signaling pathways include mitochondrial autophagy-animals, synaptic vesicle circulation, thermogenesis, Alzheimer's disease, neurodegenerative pathways-multiple diseases, oxidative phosphorylation, myocardial contraction, circadian entrainment, dopaminergic synapses, retrograde endocannabinoid signaling, mineral absorption, and amyotrophic lateral sclerosis.

[0027] The invention provides a traditional Chinese medicine composition, comprising the following components: 2-10 parts by weight of dried tangerine peel, 3-10 parts by weight of lophatherum gracile, 0.3-1 part by weight of clove, 3-10 parts by weight of tuckahoe, 5-15 parts by weight of liquorice, 0.3-1 part by weight of galangal, 0.1-0.5 part by weight of cinnamon bark, 5-15 parts by weight of amomum villosum, 3-10 parts by weight of hawthorn, 1-3 parts by weight of hovenia dulcis and 0-3 parts by weight of citron. Compared with the prior art, the present invention rationally combines the Chinese medicines so that the components can synergistically improve insomnia and improve the uncomfortable symptoms of insomnia patients such as fatigue, impaired cognitive ability, irritability, etc.; at the same time, the present invention adopts transcriptome sequencing technology to analyze and study the effects of the Chinese medicine combination on the signal pathways of insomnia and mild to moderate Alzheimer's patients in disease cell models, and finds that the Chinese medicine combination can significantly reverse disease-related pathways in disease cell models, especially can significantly reverse insomnia and mild to moderate Alzheimer's disease disorder pathways in neuronal cells, microglia and vascular endothelial cells; therefore, the Chinese medicine combination can be used to improve insomnia, prevent and treat mild to moderate Alzheimer's disease, cognitive decline, etc., and the Chinese medicine combination is all medicinal and edible medicinal materials, safe and reliable. BRIEF DESCRIPTION OF THE DRAWINGS

[0028] Figure 1 This is a heat map of pathways related to the regulation of insomnia and mild to moderate Alzheimer's disease by single-drug in Examples 1 to 4 of the present invention and Comparative Examples 1 to 4;

[0029] Figure 2 Heat map of pathways for regulating insomnia and mild to moderate Alzheimer's disease by the Chinese medicinal compositions prepared in Examples 1 to 4 of the present invention and Comparative Examples 1 to 4;

[0030] Figure 3 The invention relates to the regulation of the expression of dopamine D2 receptor (DRD2) gene and beta-amyloid precursor protein (APP) gene in SH-SY5Y cells induced by retinoic acid by a traditional Chinese medicine composition. DETAILED DESCRIPTION

[0031] The following will be combined with the embodiments of the present invention to clearly and completely describe the technical solutions in the embodiments of the present invention. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without making creative work are within the scope of protection of the present invention.

[0032] The invention provides a traditional Chinese medicine composition, comprising the following components: 2-10 parts by weight of dried tangerine peel, 3-10 parts by weight of lophatherum gracile, 0.3-1 part by weight of clove, 3-10 parts by weight of tuckahoe, 5-15 parts by weight of liquorice, 0.3-1 part by weight of galangal, 0.1-0.5 part by weight of cinnamon bark, 5-15 parts by weight of amomum villosum, 3-10 parts by weight of hawthorn, 1-3 parts by weight of hovenia dulcis and 0-3 parts by weight of citron.

[0033] According to the present invention, the content of tangerine peel in the traditional Chinese medicine composition is preferably 4 to 8 parts by weight, more preferably 5 to 6 parts by weight, and even more preferably 5 parts by weight; in some embodiments provided by the present invention, the content of tangerine peel in the traditional Chinese medicine composition is specifically 5 parts by weight, 3 parts by weight, 10 parts by weight or 2 parts by weight; tangerine peel is the dried mature peel of the Rutaceae plant Citrus reticulata Blanco and its cultivated varieties, which has the effects of regulating qi and strengthening the spleen, drying dampness and resolving phlegm, and can be used for abdominal distension, loss of appetite, vomiting and diarrhea, and cough with excessive phlegm.

[0034] According to the present invention, the content of the Lophatherum gracile in the traditional Chinese medicine composition is preferably 4 to 8 parts by weight, more preferably 5 to 6 parts by weight, and even more preferably 5 parts by weight; in some embodiments provided by the present invention, the content of the Lophatherum gracile in the traditional Chinese medicine composition is specifically 5 parts by weight, 10 parts by weight or 3 parts by weight; the Lophatherum gracile is the dried stems and leaves of the Poaceae plant Lophatherum gracile Brongn., which has the effects of clearing away heat and purging fire, relieving restlessness and quenching thirst, and promoting diuresis and relieving stranguria, and can be used for fever, thirst, short, red and painful urination, and sores in the mouth and tongue.

[0035] According to the present invention, the content of the clove in the traditional Chinese medicine composition is preferably 0.4-0.8 parts by weight, more preferably 0.4-0.6 parts by weight, and even more preferably 0.5 parts by weight; in some embodiments provided by the present invention, the content of the clove in the traditional Chinese medicine composition is specifically 0.5 parts by weight, 0.3 parts by weight or 1 part by weight; the clove is the dried flower bud of Eugeniacaryophyllata Thunb., a plant of the Myrtaceae family, which has the effects of warming the middle and relieving adverse reactions, tonifying the kidney and assisting yang, and can be used for spleen and stomach deficiency and cold, hiccups and vomiting, anorexia, vomiting and diarrhea, cold pain in the heart and abdomen, and impotence due to kidney deficiency.

[0036] According to the present invention, the content of the Poria cocos in the traditional Chinese medicine composition is preferably 4 to 8 parts by weight, more preferably 5 to 6 parts by weight, and even more preferably 5 parts by weight; in some embodiments provided by the present invention, the content of the Poria cocos in the traditional Chinese medicine composition is specifically 5 parts by weight, 10 parts by weight or 3 parts by weight; Poria cocos is the dried sclerotium of the Polyporaceae fungus Poria cocos (Schw.) Wolf, which has the effects of promoting diuresis and eliminating dampness, strengthening the spleen and calming the mind, and can be used for edema, oliguria, phlegm and fluid, dizziness and palpitations, spleen deficiency and poor appetite, loose stools and diarrhea, restlessness, palpitations and insomnia.

[0037] According to the present invention, the content of the licorice in the traditional Chinese medicine composition is preferably 8 to 12 parts by weight, more preferably 9 to 11 parts by weight, and even more preferably 10 parts by weight; in some embodiments provided by the present invention, the content of the licorice in the traditional Chinese medicine composition is specifically 10 parts by weight, 5 parts by weight, 15 parts by weight or 9 parts by weight; licorice is the dried root and rhizome of the leguminous plants Glycyrrhiza uralensis Fisch., Glycyrrhiza inflata Bat. or Glycyrrhiza glabra L., which has the effects of tonifying the spleen and replenishing qi, clearing away heat and detoxifying, removing phlegm and relieving cough, relieving acute pain, and harmonizing various medicines, and can be used for spleen and stomach weakness, fatigue, palpitations and shortness of breath, cough with sputum, acute pain in the abdomen and limbs, carbuncle, sore and poison, and relieve drug toxicity and potency.

[0038] According to the present invention, the content of the galangal in the traditional Chinese medicine composition is preferably 0.4-0.8 weight parts, more preferably 0.4-0.6 weight parts, and even more preferably 0.5 weight parts; in some embodiments provided by the present invention, the content of the galangal in the traditional Chinese medicine composition is specifically 0.5 weight parts, 1 weight part, 0.3 weight parts or 0.9 weight parts; galangal is the dried rhizome of Alpinia officinarum Hance, a plant of the ginger family, which has the effects of warming the stomach and stopping vomiting, dispersing cold and relieving pain, and can be used for cold pain in the abdomen, vomiting due to stomach cold, belching and acid regurgitation.

[0039] According to the present invention, the content of cinnamon in the traditional Chinese medicine composition is preferably 0.2-0.4 parts by weight, more preferably 0.3 parts by weight; in some embodiments provided by the present invention, the content of cinnamon in the traditional Chinese medicine composition is specifically 0.3 parts by weight, 0.1 parts by weight or 0.5 parts by weight; cinnamon is the dried bark of Cinnamomum cassia Presl, a plant of the Lauraceae family, and has the effects of replenishing fire and assisting yang, guiding fire back to the origin, dispersing cold and relieving pain, and warming and unblocking meridians, and can be used for impotence, cold uterus, cold pain in the waist and knees, asthma due to kidney deficiency, floating of virtual yang, dizziness and red eyes, cold pain in the heart and abdomen, vomiting and diarrhea due to deficiency and cold, abdominal pain due to cold hernia, dysmenorrhea and amenorrhea.

[0040] According to the present invention, the content of Amomum villosum in the traditional Chinese medicine composition is preferably 8 to 12 parts by weight, more preferably 9 to 11 parts by weight, and even more preferably 10 parts by weight; in some embodiments provided by the present invention, the content of Amomum villosum in the traditional Chinese medicine composition is specifically 10 parts by weight, 15 parts by weight, 5 parts by weight or 6 parts by weight; Amomum villosum is the dried mature fruit of the ginger family plants Amomum villosum Lour., Amomum villosum Lour. var. xanthioides TLWu et Senjen or Amomum longiligulare TLWu, which has the effects of removing dampness and stimulating appetite, warming the spleen and stopping diarrhea, regulating qi and stabilizing the fetus, and can be used for dampness and turbidity blocking the middle part of the body, abdominal distension without hunger, spleen and stomach deficiency and cold, vomiting and diarrhea, nausea during pregnancy, and fetal movement disorder.

[0041] According to the present invention, the content of hawthorn in the traditional Chinese medicine composition is preferably 4 to 8 parts by weight, more preferably 5 to 6 parts by weight, and even more preferably 5 parts by weight; in some embodiments provided by the present invention, the content of hawthorn in the traditional Chinese medicine composition is specifically 5 parts by weight, 3 parts by weight or 10 parts by weight; hawthorn is the dried mature fruit of the Rosaceae plant Cralaegus pinnatifida. Bge. var. major NEBr. or Crataegus pinnatifida Bge., which has the effects of digesting food and strengthening the stomach, promoting qi and dispersing blood stasis, clearing turbidity and lowering lipids, and can be used for meat stagnation, stomach distension, diarrhea and abdominal pain, blood stasis amenorrhea, postpartum blood stasis, heart and abdomen stabbing pain, chest pain, hernia pain, and hyperlipidemia.

[0042] According to the present invention, the content of the Hovenia dulcis in the traditional Chinese medicine composition is preferably 1 to 2.5 parts by weight, more preferably 1 to 2 parts by weight, and even more preferably 2 parts by weight; in some embodiments provided by the present invention, the content of the Hovenia dulcis in the traditional Chinese medicine composition is specifically 2 parts by weight, 1 part by weight or 3 parts by weight; Hovenia dulcis is the mature seed of the Rhamnaceae plants Hovenia dulcis, Hovenia dulcis and Hovenia dulcis, which has the effects of detoxifying alcohol, quenching thirst, relieving restlessness, stopping vomiting, and promoting urination, and can be used for drunkenness, thirst, vomiting, and difficulty in urination.

[0043] According to the present invention, the content of citron in the traditional Chinese medicine composition is preferably 1 to 3 parts by weight, more preferably 1 to 2.5 parts by weight, more preferably 1 to 2 parts by weight, and most preferably 1 part by weight. Citron is the dried mature fruit of Citrusmedica L. or Citrus wilsoniiTanaka of the Rutaceae family, and has the effects of soothing the liver and regulating qi, relieving fullness, and resolving phlegm, and can be used for liver and stomach qi stagnation, chest and flank distension and pain, fullness in the epigastrium, vomiting and belching, and cough with excessive phlegm.

[0044] In a specific embodiment provided by the present invention, the traditional Chinese medicine composition preferably includes 2-10 weight parts of dried tangerine peel, 3-10 weight parts of lophatherum gracile, 0.3-1 weight parts of cloves, 3-10 weight parts of Poria cocos, 5-15 weight parts of licorice, 0.3-1 weight parts of galangal, 0.1-0.5 weight parts of cinnamon, 5-15 weight parts of Amomum villosum, 3-10 weight parts of hawthorn, 1-3 weight parts of Hovenia dulcis and 1-3 weight parts of citron.

[0045] In another specific embodiment provided by the present invention, the Chinese medicine composition comprises the following components: 2-10 parts by weight of dried tangerine peel, 3-10 parts by weight of loofah leaves, 0.3-1 parts by weight of cloves, 3-10 parts by weight of Poria cocos, 5-15 parts by weight of licorice, 0.3-1 parts by weight of galangal, 0.1-0.5 parts by weight of cinnamon, 5-15 parts by weight of Amomum villosum, 3-10 parts by weight of hawthorn and 1-3 parts by weight of Hovenia dulcis.

[0046] In a specific embodiment provided by the present invention, the Chinese medicine composition comprises the following components: 5 parts by weight of dried tangerine peel, 5 parts by weight of lophatherum gracile, 0.5 parts by weight of cloves, 5 parts by weight of Poria cocos, 10 parts by weight of licorice, 0.5 parts by weight of galangal, 0.3 parts by weight of cinnamon, 10 parts by weight of Amomum villosum, 5 parts by weight of hawthorn, 2 parts by weight of Hovenia dulcis and 1 part by weight of citron.

[0047] In a specific embodiment provided by the present invention, the Chinese medicine composition comprises the following components: 3 parts by weight of dried tangerine peel, 10 parts by weight of loblolly leaves, 0.3 parts by weight of cloves, 10 parts by weight of Poria cocos, 5 parts by weight of licorice, 1 part by weight of galangal, 0.1 parts by weight of cinnamon, 15 parts by weight of Amomum villosum, 3 parts by weight of hawthorn, 1 part by weight of Hovenia dulcis and 3 parts by weight of citron.

[0048] In a specific embodiment provided by the present invention, the Chinese medicine composition comprises the following components: 10 parts by weight of dried tangerine peel, 3 parts by weight of lophatherum gracile, 1 part by weight of clove, 3 parts by weight of Poria cocos, 15 parts by weight of licorice, 0.3 parts by weight of galangal, 0.5 parts by weight of cinnamon, 5 parts by weight of Amomum villosum, 10 parts by weight of hawthorn and 3 parts by weight of Hovenia dulcis.

[0049] In another specific embodiment provided by the present invention, the Chinese medicine composition comprises the following components: 2 parts by weight of dried tangerine peel, 5 parts by weight of lophatherum gracile, 0.5 parts by weight of cloves, 5 parts by weight of Poria cocos, 9 parts by weight of licorice, 0.9 parts by weight of galangal, 0.3 parts by weight of cinnamon, 6 parts by weight of Amomum villosum, 5 parts by weight of hawthorn and 2 parts by weight of Hovenia dulcis.

[0050] The present invention rationally combines the Chinese medicines so that the components can synergistically improve insomnia, improve the uncomfortable symptoms of insomnia patients such as fatigue, impaired cognitive ability, irritability, etc., and at the same time, the present invention adopts cell transcriptome sequencing technology to analyze and study the signal pathways of the Chinese medicine combination on insomnia and mild to moderate Alzheimer's patients, and finds that the Chinese medicine combination can significantly reverse the disease-related pathways in disease-related cell lines, especially significantly reverse the disordered pathways of insomnia disease in neuronal cells and microglial cells, and significantly reverse the disordered pathways of mild to moderate Alzheimer's disease in neuronal cells and vascular endothelial cells. Therefore, the Chinese medicine composition can be used to improve insomnia and prevent and treat mild to moderate Alzheimer's disease; and the Chinese medicine composition is all medicinal and edible medicinal materials, which is safe and reliable.

[0051] The Chinese medicine composition provided by the present invention can be prepared by a preparation method well known to those skilled in the art, and the above components can be directly mixed to obtain the composition, or the above components can be mixed, extracted and then dried to obtain the composition, without any special restrictions; the extraction method can be a method well known to those skilled in the art, without any special restrictions, and in the present invention, preferably, ultrasonic and Soxhlet methods are used for water extraction; the drying method can be a method well known to those skilled in the art, without any special restrictions, and freeze drying is preferred in the present invention.

[0052] The present invention also provides an application of the above-mentioned traditional Chinese medicine composition in preparing medicines and / or preparations for preventing, improving or treating insomnia.

[0053] The present invention also provides a use of the above-mentioned traditional Chinese medicine composition in the preparation of medicines and / or preparations for preventing, improving or treating cognitive impairment or dementia.

[0054] According to the present invention, the medicine and / or preparation includes but is not limited to one or more of wine, granules, liquid beverages, tea substitutes, powders, oral liquids, tablets, pills, ointments and capsules.

[0055] The present invention also provides an application of the above-mentioned Chinese medicine composition in the preparation of a drug and / or preparation for reversing and / or inhibiting the disorder of a disease signal pathway; the disease signal pathway includes an insomnia signal pathway and a mild to moderate Alzheimer's disease signal pathway.

[0056] According to the present invention, the insomnia signaling pathway includes one or more of autophagy-animal, viral carcinogenesis, neutrophil extracellular trapformation, osteoclast differentiation, FcγR-mediated phagocytosis, Th1 and Th2 cell differentiation, neurotrophin signaling pathway, endocrine resistance, Toll-like receptor signaling pathway, T cell receptor signaling pathway, phosphatidylinositol signaling system, inositol phosphate metabolism and NOD-like receptor signaling pathway.

[0057] According to the present invention, the mild to moderate Alzheimer's disease signaling pathways include one or more of mitochondrial autophagy-animal, synaptic vesicle cycle, thermogenesis, Alzheimer's disease, pathways of neurodegeneration-multiple diseases, oxidative phosphorylation, cardiac muscle contraction, circadian entrainment, dopaminergic synapse, retrograde endocannabinoid signaling, mineral absorption and amyotrophic lateral sclerosis.

[0058] In order to further illustrate the present invention, the following describes in detail a Chinese medicine composition, a preparation method and application thereof in combination with embodiments.

[0059] The reagents used in the following examples are all commercially available.

[0060] Examples 1 to 4 and Comparative Examples 1 to 4

[0061] The weight proportions of the components of the Chinese medicine composition are shown in Table 1.

[0062] Table 1 Chinese medicine composition examples and comparative examples by weight

[0063]

[0064] High-throughput transcriptome screening technology was used to detect the effects of the Chinese medicine combination on insomnia and mild to moderate Alzheimer's disease-related pathways, including neural, immune, metabolic, and inflammatory pathways in neurons, microglia, and vascular endothelial cells.

[0065] The specific experimental process is:

[0066] 1) Extraction of Chinese medicine: The Chinese medicine compositions of Examples 1 to 4 and Comparative Examples 1 to 4 contain the following 14 single medicines: dried tangerine peel, light bamboo leaf, clove, Poria cocos, liquorice, galangal, cinnamon, amomum, hawthorn, citron, Hovenia dulcis, bergamot, wolfberry, patchouli. The medicinal materials were extracted with water by ultrasonic extraction or Soxhlet extraction, and the extracted medicinal solution was prepared into freeze-dried powder using a freeze dryer. An appropriate amount of freeze-dried powder was weighed and dissolved in PBS buffer to prepare an extract mother solution with a concentration of 50 mg / mL.

[0067] 2) The above 14 extracts were used to treat neuronal cells (SH-SY5Y cells differentiated by retinoic acid for 72 hours) in DMEM medium at a concentration of 200ug / mL for 24 hours; interleukin-1β (IL-1β)-induced HMC3 microglia were treated with the above 14 extracts at a concentration of 200ug / mL in HMC3 special medium for 24 hours; IL-1β-induced HUVEC-T1 vascular endothelial cells were treated with the above 14 extracts at a concentration of 100ug / mL in DMEM medium for 24 hours.

[0068] 3) The drug screen cells were lysed, and the cell lysate was directly reverse transcribed and amplified into double-stranded cDNA, and then adapters were added by Tn5 transposase, and the purified product was used for library construction and sequencing.

[0069] 4) Data processing: The solvent control PBS treatment group was used as the control group, and the drug treatment group was compared with the control group to calculate the change fold of the relevant pathway genes and background genes.

[0070] 5) Gene set enrichment analysis technology was used to conduct enrichment analysis of signal pathways related to insomnia and mild to moderate Alzheimer's disease. When FDR < 0.25, the pathway was considered to be significantly regulated.

[0071] Figure 1 The heat map of pathways related to insomnia and mild to moderate Alzheimer's disease regulated by single drug in the embodiments and comparative examples, wherein Figure 1 A check mark in the middle indicates that the drug can effectively regulate this pathway. Figure 1 The results showed that 14 kinds of Chinese medicines regulate 25 pathways related to insomnia and mild to moderate Alzheimer's disease in neurons, microglia and vascular endothelial cells: Insomnia signaling pathways include autophagy-animal, viral carcinogenesis, neutrophil extracellular trap formation, osteoclast differentiation, FcγR-mediated phagocytosis, Th1 and Th2 cell differentiation, neurotrophin signaling pathway, endocrine resistance, Toll-like receptor signaling pathway, T cell receptor signaling pathway, phosphatidylinositol signaling system, phosphoinositide metabolism, and NOD-like receptor signaling pathway;

[0072] Mild to moderate Alzheimer's disease signaling pathways include mitochondrial autophagy-animals, synaptic vesicle circulation, heat production, Alzheimer's disease, neurodegenerative pathways-multiple diseases, oxidative phosphorylation, myocardial contraction, circadian entrainment, dopaminergic synapses, retrograde endocannabinoid signaling, mineral absorption, and amyotrophic lateral sclerosis. Red indicates that the pathway is significantly upregulated, blue indicates that the pathway is significantly downregulated, and white indicates no significant effect. The results show that any single Chinese medicine in the composition can act on at least one of the signaling pathways related to insomnia and mild to moderate Alzheimer's disease.

[0073] Figure 2 Heat map of pathways related to insomnia and mild to moderate Alzheimer's disease regulated by the Chinese medicine compositions prepared in Examples 1 to 4 and Comparative Examples 1 to 4, Figure 2The cross in the middle indicates that the Chinese medicine composition is ineffective in regulating the pathway. The results showed that the Chinese medicine composition prepared in Examples 1 to 4 completely regulated 25 abnormal pathways related to insomnia and mild to moderate Alzheimer's disease in neurons, microglia and vascular endothelial cells: it had a positive regulatory effect on insomnia signaling pathways in neurons, including autophagy-animal, viral carcinogenesis, neutrophil extracellular trap formation, osteoclast differentiation, and FcγR-mediated phagocytosis; it had a positive regulatory effect on insomnia signaling pathways in microglia, including Th1 and Th2 cell differentiation, neurotrophin signaling pathway, endocrine resistance, Toll-like receptor signaling pathway, It has a positive regulatory effect on T cell receptor signaling pathway, phosphatidylinositol signaling system, phosphoinositide metabolism, and NOD-like receptor signaling pathway; it has a positive regulatory effect on mild to moderate Alzheimer's disease pathways in neuronal cells, including mitochondrial autophagy-animals, synaptic vesicle circulation, heat production, Alzheimer's disease, neurodegenerative pathways-multiple diseases, and oxidative phosphorylation; it has a positive regulatory effect on mild to moderate Alzheimer's disease pathways in vascular endothelial cells, including myocardial contraction, circadian entrainment, dopaminergic synapses, retrograde endocannabinoid signals, mineral absorption, and amyotrophic lateral sclerosis. The Chinese medicine composition prepared in comparative examples 1 to 4 has no significant effect on individual pathways of 25 abnormal pathways related to insomnia and mild to moderate Alzheimer's disease in neuronal cells, microglia, and vascular endothelial cells, or has a reverse regulatory effect. The Chinese medicine composition prepared in comparative example 1 has no regulatory effect on the insomnia pathway in neuronal cells, including FcγR-mediated phagocytosis and neutrophil extracellular trap formation, and has no regulatory effect on endocrine resistance in microglia; it has a negative regulatory effect on the mild to moderate Alzheimer's disease pathway in neuronal cells, including oxidative phosphorylation, and has no regulatory effect on mineral absorption in vascular endothelial cells. The Chinese medicine composition prepared in comparative example 2 has no regulatory effect on the insomnia pathway in neuronal cells, including FcγR-mediated phagocytosis, and has a negative regulatory effect on autophagy-animal and viral carcinogenesis; it has no regulatory effect on endocrine resistance in microglia; it has a negative regulatory effect on thermogenesis and oxidative phosphorylation in neuronal cells for the mild to moderate Alzheimer's disease pathway; and has no regulatory effect on mineral absorption in vascular endothelial cells. The Chinese medicine composition prepared in comparative example 3 has no regulatory effect on the insomnia pathway in neuronal cells, including osteoclast differentiation, and has no regulatory effect on the T cell receptor signaling pathway in microglia; has no regulatory effect on the synaptic vesicle cycle of the mild to moderate Alzheimer's disease pathway in neuronal cells, and has a negative regulatory effect on mitochondrial autophagy-animals; has no regulatory effect on circadian entrainment and dopaminergic synapses in vascular endothelial cells, and has a negative regulatory effect on retrograde endocannabinoid signals. The Chinese medicine composition prepared in comparative example 4 has a negative regulatory effect on the formation of neutrophil extracellular traps in neuronal cells and a negative regulatory effect on endocrine resistance in microglia; the mild to moderate Alzheimer's disease pathway has a negative regulatory effect on oxidative phosphorylation in neuronal cells, and has a negative regulatory effect on amyotrophic lateral sclerosis in vascular endothelial cells.The results showed that the Chinese medicine composition composed of 10 to 11 kinds of medicinal materials in Examples 1 to 4 can act on 25 abnormal signal pathways related to insomnia and mild to moderate Alzheimer's disease, and the components cooperate with each other to produce a synergistic effect, thereby playing a role in improving insomnia and mild to moderate Alzheimer's disease.

[0074] Example 5

[0075] qPCR was used to detect the effect of the Chinese medicine combination on the expression of dopamine D2 receptor (DRD2) gene and amyloid beta precursor protein (APP) gene in neurons formed by SH-SY5Y cells induced by retinoic acid (RA) for 72 hours and treated for 24 hours.

[0076] Treat neuronal cells (SH-SY5Y cells differentiated by retinoic acid induction for 72 hours) in DMEM medium with the extract of the Chinese herbal medicine composition at a concentration of 200 ug / mL for 24 hours;

[0077] 1. Dilute the extract mother solutions prepared in Examples 1 to 4 and Comparative Examples 1 to 4 to extracts with a final concentration of 200 μg / mL;

[0078] 2. SH-SY5Y cells were plated at 80,000 cells / well in a 12-well plate and induced with retinoic acid for 72 h;

[0079] 3. Add the extract of the Chinese herbal medicine composition at a final concentration of 200 μg / mL, and set up a solvent control group, and treat the cells for a total of 24 hours;

[0080] 4. Wash the cells twice with PBS, extract RNA using a kit and convert it into cDNA.

[0081] 5. qPCR reaction, program: 95°C, 3 min; (95°C, 3 s; 58°C, 30 s) 40 cycles.

[0082] 6. Data processing: The gene expression level of the blank control group is 1, and the gene expression level of the drug-treated group is greater than 1, which means the gene expression is up-regulated, and less than 1, which means it is down-regulated. Figure 3 .

[0083] Depend on Figure 3As a result, the Chinese medicine composition provided in Examples 1 to 4 can upregulate DRD2 gene expression and inhibit APP gene expression in retinoic acid-induced human neuronal cells SH-SY5Y. DRD2 plays an important role in regulating dopamine signal transduction, and dopamine plays a key role in sleep-wake regulation. APP is closely related to Alzheimer's disease. APP can be decomposed by α-proteinase, β-proteinase and γ-proteinase; the continuous action of β-proteinase and γ-proteinase can decompose APP to produce β-amyloid protein. β-amyloid protein can lead to the formation of senile plaques in the brain and apoptosis of nerve cells, and is an important factor leading to Alzheimer's disease. Therefore, the Chinese medicine composition can play an anti-insomnia and anti-senile dementia role by upregulating DRD2 gene expression and inhibiting APP gene.

[0084] Example 6: Application of Chinese medicine composition

[0085] According to the transcriptome analysis, the Chinese medicine composition was prepared as a tea substitute according to the ratio of Example 1: 5 parts of dried tangerine peel, 5 parts of light bamboo leaves, 0.5 parts of cloves, 5 parts of Poria cocos, 10 parts of licorice, 0.5 parts of galangal, 0.3 parts of cinnamon, 10 parts of Amomum villosum, 5 parts of hawthorn, 2 parts of Hovenia dulcis and 1 part of citron.

[0086] Ten volunteers with insomnia, cognitive or memory impairment were recruited to try the tea substitute. They used it once a night, 10g each time, for 3 consecutive days. They filled out questionnaires based on their feelings before and after use. The questionnaires included whether there were adverse reactions, whether the symptoms related to insomnia, cognitive or memory impairment (difficulty falling asleep, night awakening, sleep time, sleep quality, daytime sleepiness and emotions, cognitive and memory functions) were improved, etc. The results are shown in Table 2. As shown in Table 2, none of the 10 volunteers had adverse reactions after drinking the tea substitute; among them, 7 volunteers had significant improvements in their symptoms related to insomnia, cognitive or memory impairment.

[0087] Table 2 Generation tea drink tasting results

[0088]

[0089] The preferred embodiments of the present invention disclosed above are only used to help explain the present invention, but the present invention is not limited thereto. Those skilled in the art can understand that within the technical concept of the present invention, the technical solution of the present invention can be modified, or some technical features can be combined in any other way. These modifications or combinations do not make the essence of the corresponding technical solution deviate from the spirit and scope of each implementation of the technical solution of the present invention, and should be regarded as the contents disclosed by the present invention, which belong to the protection scope of the present invention.

Claims

1. A Chinese medicine composition, characterized in that: The components include: 2-10 parts by weight of dried tangerine peel, 3-10 parts by weight of loblolly leaves, 0.3-1 parts by weight of cloves, 3-10 parts by weight of poria, 5-15 parts by weight of liquorice, 0.3-1 parts by weight of galangal, 0.1-0.5 parts by weight of cinnamon, 5-15 parts by weight of amomum, 3-10 parts by weight of hawthorn, 1-3 parts by weight of hovenia dulcis, and 0-3 parts by weight of citron.

2. The Chinese medicine composition according to claim 1, characterized in that: The components include: 2-10 parts by weight of dried tangerine peel, 3-10 parts by weight of lophatherum gracile, 0.3-1 parts by weight of clove, 3-10 parts by weight of tuckahoe, 5-15 parts by weight of liquorice, 0.3-1 parts by weight of galangal, 0.1-0.5 parts by weight of cinnamon bark, 5-15 parts by weight of amomum villosum, 3-10 parts by weight of hawthorn, 1-3 parts by weight of hovenia dulcis, and 1-3 parts by weight of citron.

3. The Chinese medicine composition according to claim 1, characterized in that The components include: 5 parts by weight of dried tangerine peel, 5 parts by weight of lophatherum gracile, 0.5 parts by weight of clove, 5 parts by weight of tuckahoe, 10 parts by weight of licorice, 0.5 parts by weight of galangal, 0.3 parts by weight of cinnamon, 10 parts by weight of amomum, 5 parts by weight of hawthorn, 2 parts by weight of hovenia dulcis and 1 part by weight of citron.

4. The Chinese medicine composition according to claim 1, characterized in that: The invention comprises the following components: 3 parts by weight of dried tangerine peel, 10 parts by weight of loblolly leaves, 0.3 parts by weight of cloves, 10 parts by weight of tuckahoe, 5 parts by weight of liquorice, 1 part by weight of galangal, 0.1 parts by weight of cinnamon bark, 15 parts by weight of amomum, 3 parts by weight of hawthorn, 1 part by weight of hovenia dulcis and 3 parts by weight of citron.

5. The Chinese medicine composition according to claim 1, characterized in that: The components include: 10 parts by weight of dried orange peel, 3 parts by weight of lophatherum gracile, 1 part by weight of clove, 3 parts by weight of tuckahoe, 15 parts by weight of liquorice, 0.3 parts by weight of galangal, 0.5 parts by weight of cinnamon bark, 5 parts by weight of amomum villosum, 10 parts by weight of hawthorn and 3 parts by weight of Hovenia dulcis; Or it includes the following components: 2 parts by weight of dried tangerine peel, 5 parts by weight of lophatherum gracile, 0.5 parts by weight of clove, 5 parts by weight of tuckahoe, 9 parts by weight of liquorice, 0.9 parts by weight of galangal, 0.3 parts by weight of cinnamon bark, 6 parts by weight of amomum villosum, 5 parts by weight of hawthorn and 2 parts by weight of Hovenia dulcis.

6. A method for preparing the Chinese medicine composition according to claim 1, characterized in that: The following steps are involved: The Chinese medicine composition is obtained after the dried tangerine peel, loblolly leaf, clove, poria, liquorice, galangal, cinnamon bark, amomum villosum, hawthorn, hovenia dulcis fruit and citron are mixed and extracted.

7. Use of the Chinese medicine composition according to any one of claims 1 to 5 in the preparation of medicines and / or preparations for preventing, improving or treating insomnia; And / or, use of the Chinese medicine composition according to any one of claims 1 to 5 in the preparation of drugs and / or preparations for preventing, improving or treating cognitive impairment or dementia.

8. The use according to claim 7, characterized in that: The medicine and / or preparation is selected from one or more of wine, granules, liquid beverage, tea substitute, powder, oral liquid, tablet, pill, paste and capsule.

9. Use of the Chinese medicine composition according to any one of claims 1 to 5 in the preparation of drugs and / or preparations for reversing and / or inhibiting disorder of disease signal pathways; The disease signaling pathways include insomnia signaling pathways and mild to moderate Alzheimer's disease signaling pathways.

10. The use according to claim 9, characterized in that: The insomnia signaling pathways include autophagy-animal, viral carcinogenesis, neutrophil extracellular trap formation, osteoclast differentiation, FcγR-mediated phagocytosis, Th1 and Th2 cell differentiation, neurotrophin signaling pathway, endocrine resistance, Toll-like receptor signaling pathway, T cell receptor signaling pathway, phosphatidylinositol signaling system, phosphoinositide metabolism, and NOD-like receptor signaling pathway; The mild to moderate Alzheimer's disease signaling pathways include mitochondrial autophagy-animals, synaptic vesicle circulation, thermogenesis, Alzheimer's disease, neurodegenerative pathways-multiple diseases, oxidative phosphorylation, myocardial contraction, circadian entrainment, dopaminergic synapses, retrograde endocannabinoid signaling, mineral absorption, and amyotrophic lateral sclerosis.

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