Traditional Chinese medicine composition, preparation method thereof and application of traditional Chinese medicine composition in preparation of medicine for preventing and treating stress insomnia

Abstract

The invention relates to the technical field of traditional Chinese medicines, in particular to a traditional Chinese medicine composition, a preparation method thereof and application of the traditional Chinese medicine composition in preparation of medicines for preventing and treating stress insomnia. The composition provided by the invention is composed of vinegar-processed rhizoma cyperi, green tangerine peel, pinellia ternate, poria with hostwood, salvia miltiorrhiza, ginseng, rhizoma acori graminei, nacre mother of pearl, crude dragon bone and ginger, and the comprehensive advantages of the composition in treatment of stress insomnia are shown through a multi-target and multi-channel regulation mechanism in combination with significant regulation effects of GO function enrichment and KEGG signal channels. The multi-layer action mechanism provides a unique treatment potential for the formula, and particularly has an important clinical application prospect in the aspect of coping with complex stress response and nerve dysfunction.

Description

Technical Field

[0001] The present invention relates to the technical field of traditional Chinese medicines, and in particular to a traditional Chinese medicine composition and a preparation method thereof, and application of the traditional Chinese medicine composition in preparing medicines for preventing and treating stress-induced insomnia. Background Art

[0002] Stress insomnia is a common sleep disorder caused by sudden or continuous stress events. Its clinical manifestations mainly include difficulty falling asleep, frequent night awakenings, early awakenings, and decreased sleep quality. Epidemiological studies have shown that the incidence of stress insomnia in modern society is on the rise, especially in high-pressure working environments and fast-paced lives. Its social impact is mainly manifested in the long-term negative effects on individual health, including memory loss, emotional instability, reduced work efficiency, and other health problems that may be induced, such as cardiovascular disease and diabetes.

[0003] At present, the methods for treating stress-induced insomnia are mainly divided into two categories: drug therapy and non-drug therapy. In terms of drug therapy, benzodiazepines and non-benzodiazepines are commonly used hypnotic agents. Although they have rapid onset and significant efficacy, long-term use may lead to adverse reactions such as drug dependence, drug tolerance, and daytime sleepiness. In addition, certain antidepressants and antianxiety drugs are also used to improve sleep, but these drugs may cause side effects such as decreased cognitive function and emotional disorder, especially in the elderly. In traditional Chinese medicine theory, stress-induced insomnia is often related to liver depression and spleen deficiency, and deficiency of both heart and spleen. Commonly used Chinese patent medicines for stress insomnia, especially liver depression and spleen deficiency type, commonly used Chinese patent medicines are: Tianwang Buxin Dan, which has the effect of nourishing the heart and calming the mind, suitable for insomnia caused by insufficient qi and blood and restlessness; Guipi Pills, suitable for palpitations and insomnia caused by spleen deficiency, can nourish the spleen and calm the mind; Suanzaoren Decoction, nourishing yin and calming the mind, often used for insomnia of yin deficiency and hyperactivity of fire type; Chaihu Shugan Powder, specially used to regulate liver qi and relieve liver depression, can relieve insomnia caused by depression. Although the above Chinese patent medicines can relieve symptoms to a certain extent, they also have some limitations: 1. Stress insomnia often belongs to liver depression and spleen deficiency type, heart and spleen deficiency type, patients often have bad mood, anxiety and other manifestations, even if the Chinese patent medicine for calming the mind is used, it may not be effectively dredged due to emotional factors, resulting in poor efficacy. 2. The ingredients of many Chinese patent medicines are relatively simple, and they may not be able to provide comprehensive conditioning for liver depression and spleen deficiency combined with heart and spleen deficiency type insomnia, resulting in no significant improvement in symptoms. 3. Individual differences in drug efficacy: Different patients have different responses to Chinese patent medicines. Some patients may be sensitive to certain Chinese patent medicines, but not to others, resulting in uneven treatment effects. In summary, Chinese patent medicines can play a certain role in regulating and relieving symptoms of stress-induced insomnia, but their effects are affected by many factors, and the clinical effects are always unstable.

[0004] It can be seen that the limitations of traditional Chinese medicine treatment are that its mechanism of action is complex, its effect is slow, and it is difficult to standardize, which makes clinical application and efficacy evaluation challenging. In addition, the ingredients of traditional Chinese medicine are diverse and may interact with other drugs, so special attention should be paid when using it. Non-drug treatments such as cognitive behavioral therapy (CBT-I) and sleep hygiene education, although they avoid the problem of drug dependence, their efficacy is slow to show and require long-term persistence of patients to achieve the desired effect.

[0005] In summary, the existing means of treating stress-induced insomnia have significant limitations. It is difficult to avoid the side effects and dependence of drugs, and it is impossible to achieve good therapeutic effects through non-drug treatment in the short term. Therefore, it is particularly important to develop a safe, effective and long-term treatment method, which is also the unique advantage of traditional Chinese medicine formula in the treatment of stress-induced insomnia. 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 and a preparation method thereof and application of the composition in preparing a medicine for preventing and treating stress-induced insomnia.

[0007] The raw materials of the Chinese medicine composition provided by the invention include: vinegar-cooked cyperus rotundus, green peel, pinellia tuber, poria, salvia miltiorrhiza, ginseng, calamus, mother-of-pearl, raw dragon bone and ginger.

[0008] According to long-term clinical practice, the present invention optimizes the drug components and obtains a Chinese medicine composition capable of treating insomnia, particularly stress-induced insomnia. In the medicine, vinegar-flavored cyperus and calamus are used as the main drugs, and act together on the pathological state of liver depression and spleen deficiency. Green peel, pinellia tuber and tuckahoe are used as the assistant drugs, which further enhance the effects of soothing the liver and relieving depression and strengthening the spleen and nourishing the heart, and are particularly effective for treating insomnia symptoms caused by mood swings and indigestion. Salvia miltiorrhiza, ginseng, mother-of-pearl and dragon bone are used as adjuvant drugs, which jointly enhance the tranquilizing effect of the prescription. Ginger is used as the guiding drug to harmonize the drugs, reduce the side effects of the drugs and enhance the efficacy.

[0009] Experiments show that the composition of the composition of the present invention is reasonable, and replacing some of the components will lead to a decrease in the drug effect, for example, the decrease in the expression level of related proteins is inhibited. In other words, the drug flavor compatibility of the composition of the present invention is more reasonable, so that the drug combination of the present invention can effectively alleviate the symptoms of stress-induced insomnia and achieve the purpose of treatment.

[0010] Furthermore, the present invention optimizes the dosage of each component in the Chinese medicine composition. Compared with other dosage ratios, the following ratio can bring better effects in improving insomnia.

[0011] In the present invention, the Chinese medicine composition is prepared from the following raw materials in parts by weight: 12-18 parts of vinegar cyperus, 12-16 parts of green peel, 12-20 parts of pinellia, 12-18 parts of poria, 12-19 parts of salvia miltiorrhiza, 8-12 parts of ginseng, 10-18 parts of calamus, 20-30 parts of mother-of-pearl, 25-35 parts of raw dragon bones and 3-6 parts of ginger.

[0012] In some embodiments, the Chinese medicine composition is made of the following raw materials in parts by weight: 12 parts of vinegar-soaked cyperus, 15 parts of green peel, 12 parts of pinellia, 15 parts of poria, 15 parts of salvia miltiorrhiza, 9 parts of ginseng, 15 parts of calamus, 30 parts of mother-of-pearl, 30 parts of dragon bones and 5 parts of ginger.

[0013] In other embodiments, the Chinese medicine composition is made of the following raw materials in parts by weight: 18 parts of vinegar-soaked cyperus, 12 parts of green peel, 20 parts of pinellia, 12 parts of poria, 19 parts of salvia miltiorrhiza, 8 parts of ginseng, 18 parts of calamus, 20 parts of mother-of-pearl, 35 parts of dragon bones and 3 parts of ginger.

[0014] In other embodiments, the Chinese medicine composition is made of the following raw materials in parts by weight: 15 parts of vinegar-soaked cyperus, 16 parts of green peel, 15 parts of pinellia, 18 parts of poria, 12 parts of salvia miltiorrhiza, 12 parts of ginseng, 10 parts of calamus, 30 parts of mother-of-pearl, 25 parts of dragon bones and 6 parts of ginger.

[0015] Furthermore, the present invention also provides an extract, which is obtained by extracting the above-mentioned traditional Chinese medicine composition.

[0016] The extraction methods used in the extract of the present invention include but are not limited to percolation, decoction, heating reflux extraction, immersion extraction, ultrasound-assisted extraction, supercritical fluid extraction, microwave-assisted extraction, sublimation extraction, steam distillation extraction or enzyme-assisted extraction.

[0017] The solvent used in extracting the extract of the present invention includes at least one of water, ethanol or its aqueous solution, propylene glycol or its aqueous solution, glycerol or its aqueous solution, acetic acid or its aqueous solution.

[0018] Furthermore, the present invention also provides a method for preparing the extract as described above, which comprises soaking the Chinese medicine composition as described above in water and then decocting it, and concentrating and drying the decoction to obtain the extract.

[0019] Preferably, the water decoction extraction is performed twice, each time for 1.5 to 2 hours, with 8 to 15 times the weight of the medicinal material added in water for the first time and 6 to 12 times the weight of the medicinal material added in water for the second time.

[0020] Preferably, the concentration is performed to a relative density of 1.25 to 1.40 (50-60°C). The drying is performed under reduced pressure at a temperature of 60 to 80°C.

[0021] Furthermore, the present invention also provides the use of the above-mentioned Chinese medicine composition, the above-mentioned extract or the extract prepared by the above-mentioned preparation method in the preparation of drugs for preventing and treating stress-induced insomnia.

[0022] In the present invention, the treatment includes regulating the expression level of proteins and / or targets, prolonging sleep latency, and prolonging sleep duration.

[0023] In some embodiments, the protein includes at least one of FOS, PLA2G4A, ELK1, and IL1B.

[0024] In some embodiments, the targets include: MAPK signaling pathway, IL-17 signaling pathway, NF-kappaB signaling pathway, Th17 cell differentiation, VEGF signaling pathway, calcium signaling pathway, regulation of TRP channels by inflammatory mediators, cholinergic synapses, relaxin signaling pathway, apoptosis, p53 signaling pathway, necroptosis, B cell receptor signaling pathway, cGMP-PKG signaling pathway, antifolate resistance, Th1 and Th2 cell differentiation, GnRH signaling pathway, NOD-like receptor signaling pathway, Toll-like receptor signaling pathway or at least one of the T cell receptor signaling pathway.

[0025] The present invention also provides a drug for preventing and treating stress-induced insomnia, which comprises a pharmaceutically acceptable adjuvant and the aforementioned Chinese medicine composition, the aforementioned extract or the extract prepared by the aforementioned preparation method.

[0026] The dosage forms of the drug described in the present invention include but are not limited to granules, ointments, oral liquids, micropills, dripping pills, honey pills, water pills, tablets, decoctions, granules, capsules, tinctures, syrups, suppositories, gels and / or sprays.

[0027] The pharmaceutically acceptable excipients include fillers, diluents, lubricants, suspending agents, emulsifiers, disintegrants, plasticizers, preservatives, flavoring agents, edible flavors, sweeteners, acidulants, edible colorings, and a mixture of one or more of the above.

[0028] The drugs described in the present invention also include other drugs for treating insomnia, and the other drugs for treating insomnia include: at least one of diazepam, estazolam, alprazolam, triazolam, clonazepam, lorazepam, oxazepam, zopiclone, eszopiclone, zaleplon, eszopiclone, melatonin, ramelteon, agomelatine or doxepin.

[0029] In the present invention, in the medicine, the Chinese medicine composition and other therapeutic agents may exist independently of each other or may exist in combination, and the present invention does not limit this.

[0030] Furthermore, the present invention also provides a method for preventing and treating insomnia, which comprises administering the above-mentioned drug.

[0031] The administration methods of the drug include, but are not limited to, oral administration, chewing, swallowing, sublingual administration, oral mucosal administration, nasogastric administration, gastric tube administration, duodenal administration, jejunostomy administration, colostomy administration, rectal administration, enema, suppository insertion, anal drip, rectal foam administration, and rectal gel administration. Preferably, the administration method is oral administration.

[0032] In the present invention, the object of prevention and treatment is a human or a mammal, and the mammal includes equine, bovine, porcine, ovine, canine, feline, primate or rodent.

[0033] The present invention provides a composition composed of vinegar cyperus rotundus, green peel, pinellia tuber, poria, salvia miltiorrhiza, ginseng, calamus, mother-of-pearl, raw dragon bone and ginger, which has the effect of preventing and treating stress-induced insomnia. Experiments show that this Chinese medicine formula demonstrates its comprehensive advantages in the treatment of stress-induced insomnia through multi-target and multi-pathway regulatory mechanisms, combined with the significant regulatory effects of GO functional enrichment and KEGG signaling pathways. This multi-level mechanism of action provides the formula with unique therapeutic potential, especially in coping with complex stress reactions and neurological disorders, and has important clinical application prospects. BRIEF DESCRIPTION OF THE DRAWINGS

[0034] Figure 1 a shows differentially expressed genes in insomnia, and different columns in b represent the gene expression of tissues in different samples;

[0035] Figure 2 This indicates the potential target of Chinese medicine combination for the treatment of stress-induced insomnia;

[0036] Figure 3 A "protein-protein interaction" network diagram showing potential targets for Chinese medicine combination in the treatment of insomnia;

[0037] Figure 4 The functional enrichment cluster diagram of potential targets of Chinese medicine combination for the treatment of insomnia is shown;

[0038] Figure 5 The effects of each prescription on the animals' sleep latency and sleep duration are shown;

[0039] Figure 6 The band diagrams of FOS, PLA2G4A, ELK1, and IL1B proteins in the hippocampus of mice in each group are shown, among which A is the blank group; B is the model group; C is the Chinese medicine group;

[0040] Figure 7The band diagrams of FOS, PLA2G4A, ELK1 and IL1B proteins in the hippocampus of mice in each group are shown, among which: A is the blank group; B is the model group; C is the prescription 2 group; and D is the prescription 3 group. DETAILED DESCRIPTION

[0041] The present invention provides a Chinese medicine composition and a preparation method thereof and an application thereof in the preparation of a drug for preventing and treating stress-induced insomnia. Those skilled in the art can refer to the content of this article and appropriately improve the process parameters to achieve the above. It should be particularly noted that all similar substitutions and modifications are obvious to those skilled in the art and are considered to be included in the present invention. The method and application of the present invention have been described through preferred embodiments, and relevant personnel can obviously modify or appropriately change and combine the method and application of this article without departing from the content, spirit and scope of the present invention to implement and apply the technology of the present invention.

[0042] The test materials used in the present invention are all common commercial products and can be purchased in the market.

[0043] The medicinal materials involved in the present invention include:

[0044] Vinegar-cured Cyperus rotundus: Cyperus rotundus is the dried rhizome of the Cyperaceae plant Cyperus rotundus L. The processed product obtained by roasting or boiling with vinegar is called Vinegar-cured Cyperus rotundus.

[0045] Green peel is the peel of the dried young or immature fruit of Citrus reticulata Blanco and its cultivated varieties of the Rutaceae family.

[0046] Pinellia is the dried tuber of Pinellia ternata (Thunb.) Breit., a plant of the Araceae family.

[0047] Poria is the part in the middle of the Poria sclerotium (white Poria) that contains pine roots or fine pine wood heart.

[0048] Salvia miltiorrhiza is the dried root and rhizome of Salvia miltiorrhiza Bunge., a plant of the Lamiaceae family.

[0049] Ginseng is the root of Panax ginseng CA Mey. [P. schin-sengNees], a plant of the Araliaceae family.

[0050] Acorus tatarinowii Schott is the dried rhizome of Acorus tatarinowii Schott, a plant of the Araceae family.

[0051] Mother of pearl is the nacreous layer of the pearl oyster, the pearl oyster of the family Pinctadae, or the shells of several river clams of the family Cercidiidae.

[0052] Dragon bones are skeletal fossils of ancient mammals such as elephants, rhinos, and three-toed horses.

[0053] Ginger is the fresh rhizome of Zingiber officinale Rosc., a plant of the ginger family.

[0054] TCM pathogenesis of stress-induced insomnia:

[0055] Stress insomnia refers to insomnia caused by factors such as psychological pressure and mood swings, which is often manifested as symptoms such as difficulty falling asleep, easy awakening, and dreaminess. In traditional Chinese medicine theory, liver depression and spleen deficiency combined with deficiency of both heart and spleen are the main pathological mechanisms leading to stress insomnia. The liver governs the release of qi and regulates emotions. If the liver qi is stagnant, the mood is easily volatile and the mind is restless, leading to insomnia. The spleen governs transportation and transformation, which is the source of qi and blood biochemical transformation. If the spleen is weak, the qi and blood biochemical transformation is insufficient, and the mind is not nourished, which can also cause insomnia. The heart governs the mind. If the heart and spleen are both deficient, the mind is not nourished, and the spirit is not at home, thereby causing symptoms such as difficulty falling asleep, easy awakening, and dreaminess. Therefore, the present invention aims to relieve liver depression, strengthen the spleen and nourish the heart through reasonable compatibility of Chinese medicines, thereby achieving the purpose of treating stress insomnia.

[0056] Pharmacological action: The present invention is derived from the empirical prescription of the inventor's long-term clinical practice. Vinegar-scented Cyperus has the effects of soothing the liver and relieving depression, regulating menstruation and relieving pain, while Acorus calamus can open the mind and calm the heart, eliminate dampness and soothe the stomach. The two are the main drugs and work together on the pathological state of liver depression and spleen deficiency. Qingpi, Pinellia, and Poria are the ministerial drugs. Qingpi is warm in nature, bitter and pungent in taste, and enters the liver and stomach meridians. It is good at soothing the liver and regulating qi, eliminating accumulation and removing stagnation; Pinellia is warm in nature, pungent in taste, and enters the spleen and stomach meridians. It has the effects of drying dampness and resolving phlegm, and descending adverse vomiting; Poria is flat in nature, sweet and light in taste, and enters the heart, spleen, and kidney meridians, and can strengthen the spleen and eliminate dampness, calm the heart and tranquilize the mind. The combination of these three drugs further strengthens the effects of soothing the liver and relieving depression and strengthening the spleen and nourishing the heart, and is particularly effective for treating insomnia symptoms caused by mood swings and indigestion. The liver-soothing effect of the green peel is combined with the phlegm-removing and adverse-relief effect of the pinellia tuber, which can effectively relieve the discomfort symptoms such as chest tightness and nausea, while the spleen-strengthening and dampness-removing effect of Poria cocos helps to improve the function of the digestive system, thereby providing support for improving sleep quality. The three complement each other and strengthen the effects of soothing the liver and relieving depression and strengthening the spleen and nourishing the heart. Salvia miltiorrhiza, ginseng, mother-of-pearl, and dragon bone are used as adjuvants. Salvia miltiorrhiza promotes blood circulation and removes blood stasis, calms the nerves and calms the mind, ginseng greatly replenishes vital energy, invigorates the spleen and benefits the lungs, mother-of-pearl calms the nerves, clears the liver and improves eyesight, and dragon bone calms the nerves and calms the liver and suppresses yang, and they jointly enhance the calming effect of the prescription. Ginger is used as a drug to coordinate various drugs, alleviate the side effects of drugs, and enhance drug efficacy. Through this compatibility, the drug combination of the present invention can effectively alleviate the symptoms of stress-induced insomnia and achieve therapeutic purposes.

[0057] Drug pair analysis:

[0058] ① The compatibility of vinegar-flavored cyperus and Acorus tatarinowii is based on the principle of "liver is responsible for dispersing and spleen is responsible for transportation and transformation" in traditional Chinese medicine theory. Vinegar-flavored cyperus is warm in nature, pungent and bitter in taste, and enters the liver and spleen meridians. It has the effects of soothing the liver and relieving depression, regulating menstruation and relieving pain, while Acorus tatarinowii is pungent and warm in nature, and enters the heart and stomach meridians. It can open the mind and calm the heart, eliminate dampness and harmonize the stomach. The combination of the two can play a synergistic role in the pathological state of liver depression and spleen deficiency, thereby effectively alleviating the symptoms of stress-induced insomnia. The combination of the liver-soothing effect of vinegar-flavored cyperus and the heart-calming effect of Acorus tatarinowii can not only improve mood, but also regulate spleen and stomach function and enhance overall anti-stress ability.

[0059] ② Pinellia and Acorus calamus: Pinellia is warm in nature, pungent and bitter in taste, and enters the spleen, stomach, and lung meridians. It has the effects of drying dampness and resolving phlegm, relieving adverse reactions and stopping vomiting, and eliminating lumps and dispersing nodules. Acorus calamus is pungent and warm in nature, and enters the heart and stomach meridians. It can open the mind and calm the heart, remove dampness and harmonize the stomach. The combination of the two can play a synergistic role in the pathological state of internal obstruction of phlegm and dampness and restlessness, thereby effectively relieving the symptoms of stress-induced insomnia. The combination of the phlegm-resolving effect of Pinellia and the calming effect of Acorus calamus can not only improve sleep quality, but also regulate spleen and stomach functions and enhance overall anti-stress ability.

[0060] ③ Ginseng and mother-of-pearl: Ginseng tastes sweet and slightly bitter, is slightly warm in nature, and enters the spleen, lung, and heart meridians. It has the effects of greatly replenishing vital energy, replenishing the spleen and lungs, promoting fluid production and quenching thirst, and calming the nerves and improving intelligence. Mother-of-pearl is salty and cold in nature, and enters the liver and heart meridians. It has the effects of calming the liver and suppressing yang, clearing the liver and improving eyesight, and calming the nerves. The combination of the two can play a synergistic role in the pathological state of deficiency of both heart and spleen and hyperactivity of liver yang, thereby effectively alleviating the symptoms of stress-induced insomnia. The combination of the qi-replenishing effect of ginseng and the calming effect of mother-of-pearl can not only improve sleep quality, but also regulate heart and spleen functions and enhance overall anti-stress ability.

[0061] ④ Poria and Salvia miltiorrhiza: Poria is sweet in taste, neutral in nature, and enters the heart and spleen meridians. It has the effects of calming the mind and tranquilizing the nerves, strengthening the spleen and removing dampness. Salvia miltiorrhiza is bitter in taste, slightly cold in nature, and enters the heart and liver meridians. It has the effects of promoting blood circulation and removing blood stasis, clearing the heart and relieving restlessness. The combination of the two can play a synergistic role in the pathological state of deficiency of both heart and spleen and internal blood stasis, thereby effectively relieving the symptoms of stress-induced insomnia. The combination of the calming effect of Poria and the promoting blood circulation and removing blood stasis effect of Salvia miltiorrhiza can not only improve sleep quality, but also regulate the function of the heart and spleen and enhance the overall anti-stress ability.

[0062] Through the analysis of drug pairs, it can be seen that the different medicinal flavors in the traditional Chinese medicine composition provided by the present invention are irreplaceable, and the interaction between them produces a good therapeutic effect on stress-induced insomnia.

[0063] The numerical ranges and parameters involved in the present invention have been presented in the specific embodiments as accurately as possible. However, any numerical value inherently inevitably contains standard deviations due to individual test methods. Therefore, unless otherwise explicitly stated, it should be understood that all numerical ranges or specific data used in the present disclosure may have certain reasonable deviations within a certain range, for example: within ±10%, ±5%, ±1% or ±0.5%. The present invention is further described below in conjunction with the embodiments:

[0064] Example 1

[0065] Prescription 1: 12g of vinegar-soaked Cyperus rotundus, 15g of Citrus aurantium, 12g of Pinellia ternata, 15g of Poria, 15g of Salvia miltiorrhiza, 9g of Ginseng, 15g of Acorus calamus, 30g of mother of pearl, 30g of Dragon Bone, and 5g of Ginger.

[0066] Take the above 10 herbs and soak them in water for 0.5-2h, decoct them twice, each time for 1.5-2 hours, add 8-15 times of water for the first time, and 6-12 times of water for the second time, filter, combine the two decoctions, concentrate under reduced pressure to a thick paste (50-60℃) with a relative density of 1.25-1.4, dry under reduced pressure (60-80℃), and obtain a dry paste for later use. Mix the above dry paste, appropriate amount of dextrin and drug powder, crush, and pass through a 60-mesh sieve for later use.

[0067] Example 2

[0068] Prescription 2: 12g of vinegar-soaked Cyperus rotundus, 15g of dried tangerine peel, 12g of Pinellia ternata, 15g of Poria cocos, 15g of Salvia miltiorrhiza, 9g of Ginseng, 15g of Atractylodes macrocephala, 30g of mother of pearl, 30g of raw dragon bone, and 5g of ginger.

[0069] The preparation method is the same as Example 1.

[0070] Example 3

[0071] Prescription 3: 12g of costus root, 15g of green peel, 12g of pinellia, 15g of poria, 15g of salvia miltiorrhiza, 9g of codonopsis pilosula, 15g of calamus, 30g of mother of pearl, 30g of raw dragon bone, and 5g of ginger.

[0072] The preparation method is the same as Example 1.

[0073] Efficacy verification:

[0074] 1. Network pharmacology to select targets and predict pathways:

[0075] (1) The multi-target and multi-pathway therapeutic advantages of traditional Chinese medicine formulas

[0076] This Chinese medicine formula has a total of 226 targets on the human body. Through the transcriptome detection data of stress-induced insomnia collected clinically, it was found that there are 3655 related expression genes in stress-induced insomnia ( Figure 1). A Venn diagram was used to draw a map of drug and disease targets, and it was found that the Chinese medicine formula can act on 40 disease targets ( Figure 2 ), indicating that this Chinese medicine formula has multi-target action characteristics.

[0077] (2) Network diagram of protein-protein interaction of targets of traditional Chinese medicine formula for the treatment of stress-induced insomnia

[0078] In the STRING database, we first set the species to "Homo sapiens", then input 40 targets of this traditional Chinese medicine formula for the treatment of stress-induced insomnia, and obtained the "Protein-Protein Interaction" (PPI) network diagram ( Figure 3 (a), involving 40 nodes and 85 edges. The TSV files obtained from the STRING online analysis tool were imported into Cytoscape 3.7.1 for visualization and analysis ( Figure 3 The results showed that the 40 targets interacted well with each other and had the potential to treat stress-induced insomnia.

[0079] (3) Functional enrichment of targets in traditional Chinese medicine formulas for the treatment of stress-induced insomnia

[0080] GO and KEGG are important tools in enrichment analysis, mainly used to explore the biological processes and signaling pathways involved in the treatment of diseases with clinical drugs, as well as the key genes in the process. Using the ClusterProfiler package, we performed GO analysis on 40 targets of traditional Chinese medicine formulas for the treatment of stress-induced insomnia, and obtained a total of 831 items (adjusted P < 0.05), including 795 biological processes (BP), 35 molecular functions (MF) and 78 KEGG. We combined the up-regulated and down-regulated expression of the 40 target genes in the clinical transcriptome and mapped the top 18 BPs ( Figure 4 a), MF ( Figure 4 b) and KEGG ( Figure 4 The circular diagram in (c) is used to intuitively understand the biological processes and signaling pathways of traditional Chinese medicine formulas for the treatment of stress-induced insomnia. The top 20 KEGG signaling pathways of traditional Chinese medicine formulas for the treatment of stress-induced insomnia are shown in Table 1.

[0081] Table 1 Top 20 KEGG signaling pathways

[0082]

[0083] 2. Regulatory effect of Chinese herbal medicine formula on FOS / PLA2G4A / ELK1 signaling pathway in stress-induced insomnia model mice

[0084] (1) Verification path selection

[0085] The top 20 KEGG signaling pathways obtained through enrichment analysis showed that the MAPK signaling pathway was significantly involved in the regulation of traditional Chinese medicine formulas in the treatment of stress-induced insomnia. Therefore, we selected the downstream protein target FOS / PLA2G4A / ELK1 signaling pathway of this signaling pathway to carry out animal experimental verification based on stress-induced insomnia model mice.

[0086] Animal grouping and modeling method:

[0087] A. Animal Grouping:

[0088] Thirty SPF C57BL / 6J healthy male mice, 6-8 weeks old, weighing 18-22 g, were purchased from Beijing Huafukang Biotechnology Co., Ltd. During the experiment, mice were free to eat and drink, the room temperature was 18-26°C, the relative humidity was 50%-70%, and the light-dark cycle was 12h / 12h.

[0089] After one week of adaptive feeding of the mice, 30 mice were divided into five groups according to their spontaneous activity and body weight, namely, blank group, model group, prescription group 1, prescription group 2, and prescription group 3, with 6 mice in each group.

[0090] B. Preparation of stress-induced insomnia mouse model (using PCPA intraperitoneal injection + modified multi-platform water environment method): According to relevant literature, the stress-induced insomnia mouse model was prepared, and PCPA was intraperitoneally injected at 350 mg / kg at 8:00-9:00 every day, once a day, for 3 consecutive days. At the same time, the stress-induced insomnia mouse model was prepared by the modified multi-platform water environment method: a water tank (60cm×35cm×20cm) was prepared, and the water tank was filled with water about 0.2cm deep under the platform. Six large platforms (12cm in diameter, 5cm in height, and 15cm in diameter between platforms) were placed in the water tank for the blank group mice, and wire mesh was placed on the platform to allow the mice to sleep without falling into the water. 18 circular platforms (3cm in diameter, 5cm in bottom height, and about 6.5cm in diameter between platforms) were placed in the water tank for the other groups of mice. Mice can drink water and exercise freely on the platform. During REM sleep, the decrease in the muscle tension of the mouse's abdominal muscles will cause the mouse to fall into the water or wake up by touching the water surface, causing sleep deprivation in the mouse. During the modeling period, mice had free access to food pellets and water bottles on the top of the water tank. Housing environment: 12h / 12h light-dark cycle, light on from 8:00, constant temperature (22±2)℃, water in the water tank was changed twice a day during the entire sleep deprivation period. In order to reduce stress, each mouse was familiar with the environment for 3 days before the experiment. Mice were deprived of sleep for 24 hours a day for 3 days.

[0091] C. Verification of modeling success:

[0092] 24 hours after the modeling, the sleep latency and sleep duration of the righting test were used to evaluate the model. The disappearance of the righting reflex of the animal for more than 1 min was used as the standard for falling asleep, and the sleep time was recorded; the turning over 3 times within 1 min was used as the standard for falling asleep, and the sleep end time was recorded. Compared with the blank group, the sleep latency of the model group and prescription 1, 2, and 3 groups was prolonged (P < 0.05) and the sleep duration was shortened (P < 0.05), indicating that the stress-induced insomnia model in mice was successfully established. Figure 5 .

[0093] (2) Western blotting experiment steps

[0094] The hippocampus tissues of the brains of the stress-induced insomnia model mice (blank group, model group, prescription 1 group, prescription 2 group, and prescription 3 group) were taken and quickly frozen with liquid nitrogen, and the hippocampus regions were cut for subsequent protein immunoblotting experiments. In order to avoid sample degradation, the frozen samples needed to be quickly frozen in liquid nitrogen and then stored in a -80°C refrigerator.

[0095] The hippocampus of each group of mice was taken out from the refrigerator and homogenized using RIPA lysis buffer. After homogenization, the samples were placed on ice for 30 minutes to ensure that the cells were fully lysed and the proteins were released. Cell debris was removed by centrifugation (12000 rpm, 4°C, 10 minutes), and the supernatant was collected as a total protein sample. Subsequently, the protein concentration was determined by the BCA method to ensure that the protein content in each group of samples was consistent. According to the molecular weight of the target protein, an appropriate SDS-PAGE separation gel was selected. The protein sample (20 μg) was mixed with the loading buffer and boiled for 10 minutes to denature the protein. The sample was then loaded into the gel and the protein was separated by electrophoresis (120V, 2 hours). After the electrophoresis was completed, the protein was transferred from the gel to the PVDF membrane by wet transfer, and the transfer time was 2 hours, and the voltage was set to 100V.

[0096] After transfer, the membrane was blocked with 5% non-fat milk powder solution for 1 hour at room temperature. After blocking, the membrane was incubated with diluted primary antibodies (FOS, PLA2G4A, ELK1 antibodies) at 4°C overnight. The next day, the membrane was washed 3 times with TBST for 5 minutes each time, and the membrane was incubated with secondary antibodies for 1 hour. After incubation, the membrane was washed again with TBST 3 times, and finally developed with ECL chemiluminescent substrate.

[0097] After obtaining the image of the protein bands through the imaging system, the optical density of each band was quantitatively analyzed using ImageJ analysis software. The optical density value of the band was determined by the software and normalized with the optical density of the internal reference protein β-actin. Finally, the relative changes in protein expression levels between groups were calculated, and statistical analysis was performed to compare the differences between groups.

[0098] (3) Western blotting results

[0099] The results of immunoblotting experiments showed that (Table 2, Figure 6 ), compared with the blank group, the expression of FOS, PLA2G4A, ELK1, and IL1B proteins in the stress-induced insomnia model mice were significantly increased (P < 0.01); compared with the model group, the expression of FOS, PLA2G4A, ELK1, and IL1B proteins in the Chinese medicine formula group were significantly decreased (P < 0.01). This suggests that this Chinese medicine formula can treat stress-induced insomnia by participating in the regulation of the FOS / PLA2G4A / ELK1 signaling pathway.

[0100] Table 2 Comparison of FOS, PLA2G4A, ELK1, and IL1B protein expression levels in the hippocampus of mice in each group (n=6, ±s)

[0101]

[0102] Note: a compared with the blank group, P < 0.01; b compared with the model group, P < 0.01.

[0103] 3. Drug ingredient conversion experiment:

[0104] At the same time, the Chinese medicine group in Example 1 was set as Prescription Group 1: 12g of vinegar cyperus, 15g of green peel, 12g of pinellia, 15g of poria, 15g of salvia miltiorrhiza, 9g of ginseng, 15g of calamus, 30g of mother-of-pearl, 30g of dragon bone, and 5g of ginger. Prescription Group 2 and Prescription Group 3 in Examples 2-3 were set as parallel controls. The formulas of Prescription 2 and Prescription 3 are as follows (the preparation methods of the granules of Prescription 2 and 3 are the same as those of Prescription Group 1):

[0105] Prescription 2: Vinegar-soaked Cyperus rotundus 12g, dried tangerine peel 15g, Pinellia 12g, Poria 15g, Salvia miltiorrhiza 15g, Ginseng 9g, Atractylodes macrocephala 15g, Mother of Pearl 30g, Raw Dragon Bone 30g, Ginger 5g (Green peel and Acorus calamus are replaced with dried tangerine peel and Atractylodes macrocephala)

[0106] Prescription 3: 12g of costus root, 15g of green peel, 12g of pinellia tuber, 15g of poria, 15g of salvia miltiorrhiza, 9g of codonopsis pilosula, 15g of calamus, 30g of mother of pearl, 30g of raw dragon bone, 5g of ginger (replace costus root and codonopsis root with vinegar cyperus and ginseng)

[0107] The results of immunoblotting experiments showed that (Table 3, Table 4, Figure 7Compared with prescription group 1, the protein expressions of FOS, PLA2G4A, ELK1, and IL1B in prescription group 2 were significantly increased (P < 0.01), indicating that the efficacy of prescription group 1 was better than that of prescription group 2; compared with prescription group 1, the protein expressions of FOS, PLA2G4A, ELK1, and IL1B in prescription group 3 were significantly increased (P < 0.01), indicating that the efficacy of prescription group 1 was better than that of prescription group 3.

[0108] Table 3 Comparison of FOS, PLA2G4A, ELK1 and IL1B protein expression levels in the hippocampus of mice in each group

[0109] (n=6, ±s)

[0110]

[0111] Note: aCompared with prescription 1 group, P < 0.01.

[0112] Table 4 Comparison of FOS, PLA2G4A, ELK1 and IL1B protein expression levels in the hippocampus of mice in each group

[0113] (n=6, ±s)

[0114]

[0115] Note: aCompared with prescription 1 group, P < 0.01.

[0116] The above results show that the Chinese medicine formula (Example 1, i.e., Prescription Group 1) can treat stress-induced insomnia by participating in the regulation of the FOS / PLA2G4A / ELK1 signaling pathway, and its efficacy is better than that of Prescription Group 1 and Prescription Group 2.

[0117] Conclusion: The Chinese medicine formula of Example 1 demonstrates its comprehensive advantages in the treatment of stress-induced insomnia through multi-target and multi-pathway regulatory mechanisms, combined with the significant regulatory effects of GO functional enrichment and KEGG signaling pathways. This multi-level mechanism of action provides the formula with unique therapeutic potential, especially in coping with complex stress reactions and neurological disorders, and has important clinical application prospects.

[0118] 4. The efficacy of Chinese medicine formula on patients with electromagnetic radiation intolerance and stress-induced insomnia

[0119] 4.1. Research subjects:

[0120] A total of 120 patients with electromagnetic radiation intolerance and stress insomnia who met the diagnostic criteria were selected, and a unified random number table was generated using SPSS statistical software. The patients were divided into treatment group 1, treatment group 2, and western medicine group according to the order of their visit.

[0121] 4.2. Diagnostic criteria:

[0122] Refer to the diagnostic criteria of (Chinese Classification and Diagnostic Criteria for Mental Disorders CCMD-3).

[0123] 4.3. Inclusion criteria:

[0124] (1) Those who meet the above diagnostic criteria;

[0125] (2) Aged 18 to 55 years old;

[0126] (3) PSQI range of 11-15 points;

[0127] (4) Long-term exposure to complex electromagnetic environment, total electromagnetic radiation time ≥ 3 months

[0128] (5) Volunteer to participate in this study and sign the informed consent.

[0129] 4.4. Exclusion criteria:

[0130] (1) Those who have participated in other clinical trials in the past three months;

[0131] (2) Those who are unwilling to cooperate.

[0132] 4.5. Elimination criteria:

[0133] (1) Subjects who do not meet the inclusion criteria after enrollment;

[0134] (2) Subjects who did not take medication according to the study protocol after enrollment.

[0135] 4.6. Termination criteria:

[0136] (1) Those who have serious adverse events and are not suitable to continue the study plan;

[0137] (2) Those who have some sudden illness and are not suitable to continue this study plan;

[0138] (3) The subjects have poor compliance and fail to complete the study according to the study protocol.

[0139] 4.7. Sample size estimation and randomization method:

[0140] (1) Sample size estimation: With reference to literature reports, previous clinical experience, and research basis, and in consultation with statistical experts, SPSS statistical software was used to set the test level α = 0.05 (one-sided), the test power 1-β = 0.90, the first overall rate (estimate) π1 = 60%, and the second overall rate (estimate) π2 = 85%. The resulting sample size of each group was n = 36, plus 10% dropout cases, and the total sample size of the two groups was N = 80.

[0141] (2) Randomization method: SPSS statistical software was used to generate a unified random number table, and patients were divided into the Chinese medicine group and the Western medicine group according to the order of their visit.

[0142] 4.8 Treatment options

[0143] Treatment group 1: Drug composition: 10 g of Pinellia ternata, 20 g of Coix seeds, 15 g of stir-fried Semen Ziziphi Spinosae, 15 g of Acorus calamus, 15 g of Atractylodes macrocephala, 20 g of Poria, 15 g of roasted Polygala tenuifolia, 20 g of Platycladus orientalis, 6 g of Licorice. One dose of the drug was made into 2 bags of granules (produced by Tianjin Xinjingtang Chinese Medicine Clinic Co., Ltd., batch number: 20230801), each bag was 7 g. One bag each time, twice a day, 30 minutes after meals in the morning and evening.

[0144] Treatment group 2: Drug composition: vinegar cyperus rotundus 12g, green peel 15g, pinellia 12g, poria 15g, salvia miltiorrhiza 15g, ginseng 9g, calamus 15g, mother of pearl 30g, dragon bone 30g and ginger 5g. One dose of the drug was made into 2 bags of granules (produced by Tianjin Xinjingtang Chinese Medicine Clinic Co., Ltd., batch number: 202301003), each bag of 7g. Take 1 bag each time, twice a day, 30 minutes after meals in the morning and evening.

[0145] Positive control group: oral Zolpidem tartrate tablets [Sanofi (Hangzhou) Pharmaceutical Co., Ltd., imported drug registration number: H20181237, 10 mg per tablet], 10 mg each time, once a day, before bedtime. The treatment cycle for both groups was 4 weeks, and no other sedative hypnotic drugs were taken during the treatment period.

[0146] 4.9. Observation items:

[0147] (1) General observation items: including name, gender, age, marital status, position, contact information, etc. This information is confidential and cannot be disclosed. It is only used for this study.

[0148] (2) Quantitative items: total sleep time before and after treatment

[0149] (3) Scale measurements: sleep parameters (sleep efficiency, sleep structure, wakefulness index), sleep quality (PSQI score) and quality of life (SGRQ score).

[0150] 4.10. Evaluation of efficacy:

[0151] (1) Improvement in sleep duration: Compare the sleep duration recorded before and after treatment.

[0152] (2) Improvement of sleep parameters: Comparison of the levels of quantitative components of sleep efficiency, sleep structure, and wakefulness index before and after treatment.

[0153] (3) Improvement in sleep quality: Compare the total scores of the PSQI scale before and after treatment.

[0154] (4) Improvement in quality of life: Compare the total score of the SGRQ scale before and after treatment.

[0155] 4.11. Efficacy determination:

[0156] The clinical efficacy was determined by the PSQI total score reduction rate, PSQI total score reduction rate = (PSQI total score before treatment - PSQI total score after treatment) / PSQI total score before treatment × 100%. Cured: Insomnia completely disappeared, PSQI total score reduction rate ≥ 80%; markedly effective: Insomnia almost completely disappeared, PSQI total score reduction rate ≥ 50%, but < 80%; improved: Insomnia significantly alleviated, PSQI total score reduction rate ≥ 30% but < 50%; ineffective: Insomnia did not improve, PSQI total score reduction rate < 30%. Total effective = cured + markedly effective + improved.

[0157] The TCM syndrome score reduction rate was used to determine the efficacy of TCM syndrome. TCM syndrome score reduction rate = (TCM syndrome score before treatment - TCM syndrome score after treatment) / TCM syndrome score before treatment × 100%. Clinical cure: clinical symptoms and signs disappeared or basically disappeared, TCM syndrome score reduction rate ≥ 95%; marked effect: clinical symptoms and signs were significantly improved, TCM syndrome score reduction rate ≥ 70% but < 95%; improvement: clinical symptoms and signs were improved, TCM syndrome score reduction rate ≥ 30% but < 70%; ineffective: symptoms and signs were not significantly improved or aggravated, TCM syndrome score reduction rate < 30%. Total effectiveness = clinical cure + marked effect + improvement.

[0158] 4.12. Statistical methods: SPSS 22.0 software was used to process the data. The measurement data were expressed as (x±s) and the t test was used. The enumeration data were expressed as percentage (%) and the chi-square test was used. P < 0.05 was considered statistically significant.

[0159] Experimental results

[0160] 4.13.1 Treatment outcomes

[0161] 4.13.1.1 The PSQI scores and efficacy of the two groups of patients before and after treatment were compared. The results are shown in Table 5:

[0162] Table 5 Comparison of PSQI scores before and after treatment in the treatment group and the control group (x±s)

[0163]

[0164] Note: Compared with before treatment, *: P<0.05, compared with the control group, #: P<0.05.

[0165] in conclusion:

[0166] By comparing the various indicators of the treatment group and the control group, it can be clearly seen that the treatment group has significant improvements in sleep quality, sleep time, sleep time, sleep efficiency, sleep disorders and daytime dysfunction. Specifically, the indicators of the treatment group were significantly lower after treatment than before treatment, and the difference was statistically significant compared with the control group (P<0.05). This shows that this treatment method has a significant effect on improving the sleep condition of patients with stress insomnia. In addition, although the control group also improved to a certain extent, the effect was not as obvious as that of the treatment group.

[0167] 4.13.2 Comparison of TCM syndrome efficacy between the two groups of patients

[0168] The total effective rate of TCM syndrome in the treatment group was 96%, while that in the control group was 70%. The treatment group was superior to the control group (χ 2 =12.43, P <0.01).

[0169] Table 6 Comparison of TCM syndrome efficacy between two groups of male patients with stress-induced insomnia [cases (%)]

[0170]

[0171] 4.13.3 Comparison of TCM syndromes between the two groups of patients with stress-induced insomnia

[0172]

[0173] By comparing the changes in various TCM syndrome indicators between the treatment group and the control group, it can be seen that the treatment group had significantly higher improvements in abdominal distension, irritability, fatigue, and mental fatigue than the control group. Specifically, the median of each indicator decreased significantly after treatment, and statistical analysis showed that the difference was extremely significant (p<0.001). This shows that the Chinese medicine formula has a significant effect in improving the TCM syndrome of patients with stress-induced insomnia, and the overall symptom improvement in the treatment group is relatively large, suggesting that this treatment plan has a high application value in clinical practice.

[0174] The above are only preferred embodiments of the present invention. It should be pointed out that, for ordinary technicians in this technical field, several improvements and modifications can be made without departing from the principles of the present invention. These improvements and modifications should also be regarded as the scope of protection of the present invention.

Claims

1. A Chinese medicine composition comprising: Vinegar-scented Cyperus rotundus, Citrus aurantium, Pinellia ternata, Poria, Salvia miltiorrhiza, Ginseng, Acorus calamus, mother of pearl, raw dragon bone and ginger.

2. The Chinese medicine composition according to claim 1, characterized in that: Made from the following raw materials in parts by weight: 12-18 parts of vinegar-soaked cyperus, 12-16 parts of green peel, 12-20 parts of pinellia, 12-18 parts of poria, 12-19 parts of salvia miltiorrhiza, 8-12 parts of ginseng, 10-18 parts of calamus, 20-30 parts of mother of pearl, 25-35 parts of raw dragon bone and 3-6 parts of ginger.

3. The Chinese medicine composition according to claim 1, characterized in that Made from the following raw materials in parts by weight: 12 parts of vinegar-soaked cyperus, 15 parts of green peel, 12 parts of pinellia, 15 parts of poria, 15 parts of salvia miltiorrhiza, 9 parts of ginseng, 15 parts of calamus, 30 parts of mother-of-pearl, 30 parts of dragon bones and 5 parts of ginger.

4. An extract obtained by extracting the Chinese medicine composition according to any one of claims 1 to 3.

5. The method for preparing the extract according to claim 4, comprising: soaking the Chinese medicine composition according to any one of claims 1 to 3 in water and then decocting it in water, and concentrating and drying the decoction to obtain the extract.

6. The preparation method according to claim 5, characterized in that: The water decoction extraction is performed twice, each time for 1.5 to 2 hours, with 8 to 15 times the weight of the medicinal material added in water for the first time and 6 to 12 times the weight of the medicinal material added in water for the second time.

7. The preparation method according to claim 5, characterized in that: The concentration is to a relative density of 1.25 to 1.

40.

8. Use of the Chinese medicine composition according to any one of claims 1 to 3, the extract according to claim 4 or the extract prepared by the preparation method according to any one of claims 5 to 7 in the preparation of a drug for preventing and treating stress-induced insomnia.

9. The use according to claim 8, characterized in that: The treatment includes regulating the expression level of proteins and / or targets, prolonging sleep latency, and prolonging sleep duration; The protein includes at least one of FOS, PLA2G4A, ELK1, and IL1B.

10. A drug for preventing and treating stress-induced insomnia, comprising a pharmaceutically acceptable excipient and the Chinese medicine composition according to any one of claims 1 to 3, the extract according to claim 4, or the extract prepared by the preparation method according to any one of claims 5 to 7.

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