Traditional Chinese medicine formula essential oil for improving or treating insomnia

Abstract

The invention provides traditional Chinese medicine formula essential oil for improving or treating insomnia, and relates to the technical field of traditional Chinese medicine essential oil. The traditional Chinese medicine formula essential oil is prepared from radix bupleuri essential oil, longan aril essential oil, tuber fleeceflower stem essential oil, allium macrostemon essential oil, angelica sinensis essential oil and borneol. The traditional Chinese medicine formula essential oil can significantly improve the sleep quality of drosophila melanogaster; through sniffing administration on an insomnia model mouse, the insomnia state of the insomnia model mouse can be obviously improved, the behavior activity of the model mouse can be recovered, the sleep rate of the mouse can be obviously increased, the sleep latency period can be obviously shortened, the sleep time can be obviously prolonged, the levels of hypothalamic homogenate 5-HT, GABA and Glu of the mouse can be obviously increased, and the level of hypothalamic homogenate DA can be obviously reduced; the traditional Chinese medicine composition has a remarkable treatment effect on insomnia model mice. The traditional Chinese medicine formula essential oil provided by the invention has a relatively good application prospect in preparation of aromatherapy medicines for treating insomnia.

Description

Technical Field

[0001] The invention relates to the technical field of traditional Chinese medicine essential oils, and in particular to a traditional Chinese medicine formula essential oil for improving or treating insomnia. Background Art

[0002] Insomnia refers to a sleep disorder syndrome in which the initiation and maintenance of sleep are impaired, resulting in the inability of sleep quality to meet individual physiological needs, and has a significant impact on the patient's daily activities, such as slow reaction, fatigue, and difficulty concentrating. Long-term sleep disorders can lead to metabolic disorders, reduced immunity, anxiety, persistent headaches, depression, forgetfulness and many other problems, seriously affecting normal work and life, and accelerating the aging process of the human body, seriously threatening people's quality of life and physical and mental health, while also increasing the risk of obesity, diabetes, hypertension, heart disease and stroke. It is very difficult to treat insomnia. At present, the methods for treating insomnia mainly include drug therapy, cognitive behavioral therapy, psychotherapy, etc. Drug therapy is mainly based on chemical synthetic sedatives, such as benzodiazepines, tricyclic antidepressants and melatonin. Frequent use can lead to adverse effects such as renal damage, respiratory depression, withdrawal reactions and rebound insomnia. Psychotherapy and cognitive behavioral therapy are the first treatments for insomnia, but due to the limitation of relevant resources, they cannot be promoted on a large scale.

[0003] Chinese herbal essential oil is a volatile oily liquid extracted from Chinese herbal medicine. It can be extracted from the roots, stems, flowers, leaves, seeds, fruits and other parts of Chinese herbal medicine. Chinese herbal essential oil comes directly from nature and has complex ingredients. It mainly includes terpenoid compounds, aromatic compounds, aliphatic compounds and nitrogen-containing and sulfur-containing compounds. Compared with chemical products synthesized by humans, it has the characteristics of low toxicity and high safety. There are many non-drug intervention measures for sleep regulation. Aromatherapy, as a unique means, has a long history of clinical practice. Aromatherapy, as a traditional Chinese medicine nursing technology, is one of the commonly used non-drug therapies for insomnia. Aromatherapy has the characteristics of simple operation, significant efficacy, high safety, few side effects, and no dependence. Some scholars have confirmed that aromatherapy can effectively improve the symptoms of patients with insomnia. Chinese herbal essential oil is the most representative product of aromatherapy. While calming the nerves, it has been proven to have pharmacological activities such as anti-anxiety, anti-depression, and neuroprotection. At present, aromatherapy for insomnia mainly uses single plant essential oils with high fragrance, such as lavender, bergamot, and citrus, which have general effects, slow onset, inability to fully regulate body functions, and incomplete treatment. Therefore, it is very necessary to develop a Chinese medicine formula essential oil that has a precise efficacy, quick effect, and regulates body functions for improving or treating insomnia. Summary of the invention

[0004] 1. Technical issues to be solved

[0005] In view of the deficiencies of the prior art, the present invention provides a traditional Chinese medicine essential oil for improving or treating insomnia.

[0006] (II) Technical solution

[0007] To achieve the above objectives, the present invention is implemented through the following technical solutions:

[0008] The invention provides a traditional Chinese medicine essential oil for improving or treating insomnia. The traditional Chinese medicine essential oil consists of the following components in parts by weight: 1-10 parts of bupleurum essential oil, 1-10 parts of longan pulp essential oil, 1-10 parts of tuberose essential oil, 1-10 parts of Allium macrostemon essential oil, 1-10 parts of angelica essential oil and 1-6 parts of borneol.

[0009] Furthermore, the traditional Chinese medicine formula essential oil is composed of the following ingredients in parts by weight: 4-8 parts of bupleurum essential oil, 4-8 parts of longan pulp essential oil, 4-8 parts of tuberose essential oil, 3-7 parts of Allium macrostemon essential oil, 4-8 parts of angelica essential oil, and 2-5 parts of borneol.

[0010] Furthermore, the traditional Chinese medicine formula essential oil is composed of the following ingredients in parts by weight: 6 parts of bupleurum essential oil, 6 parts of longan pulp essential oil, 6 parts of tuberose essential oil, 5 parts of Allium macrostemon essential oil, 6 parts of angelica essential oil, and 3 parts of borneol.

[0011] Specifically, bupleurum essential oil, longan meat essential oil, flos cholerae essential oil, allium macrostemon essential oil, and angelica essential oil are extracted from corresponding Chinese medicinal plants using conventional essential oil extraction methods, or purchased from the market.

[0012] Specifically, the extraction methods include pressing extraction, supercritical carbon dioxide extraction, steam distillation extraction, immersion extraction, chemical solvent extraction, and oil separation.

[0013] Specifically, the distillation concentration of the above-mentioned single essential oil is controlled to be above 99%.

[0014] The present invention also provides the use of the traditional Chinese medicine formula essential oil in the preparation of a medicine for improving or causing insomnia. Specifically, the traditional Chinese medicine formula essential oil is added with medically acceptable excipients to prepare an inhalation dosage form for administration by inhalation.

[0015] Furthermore, the effective content of the essential oil in the Chinese medicine formula in the medicine for improving or insomnia is 5-20%.

[0016] Bupleurum has a slightly fragrant smell, slightly bitter taste, and slightly cold nature. It enters the liver, gallbladder, and heart meridians. It has the functions of dispelling wind and heat, relieving liver depression, and raising Yang Qi. It can be used to treat various diseases caused by liver depression and qi stagnation. Modern research has found that Bupleurum extracts contain chemical components such as bupleurosaponin, brass, and volatile oils, which have specific anti-inflammatory, antioxidant, liver-protective, anti-tumor, and anti-depression effects. Bupleurum volatile oil contains a variety of active ingredients, mainly including linalool and limonene. Modern medical research has proved that Bupleurum volatile oil has anti-inflammatory and antibacterial properties, but there is no report on the use of diesel volatile oil in the treatment of insomnia.

[0017] Longan flesh is sweet, warm in nature, and enters the heart and spleen meridians. It has the effects of nourishing the heart and spleen, nourishing blood and calming the mind. It is often used for insufficient qi and blood, palpitations, forgetfulness and insomnia, and blood deficiency and sallow complexion. Longan flesh is rich in nutrients, and its main chemical components are sugars, lipids, saponins, polypeptides, polyphenols, volatile components, amino acids and trace elements. It has many physiological effects and health care mechanisms such as anti-aging, enhancing immunity, anti-tumor, regulating endocrine, and inhibiting bacteria.

[0018] The taste of Polygonum multiflorum is sweet and slightly bitter, and it is neutral in nature. It is in the heart and liver meridians. It has the effects of nourishing the heart and calming the mind, dispelling wind, and unblocking the meridians. It is used for insomnia, dreaminess, body pain due to blood deficiency, numbness of the skin, rheumatic pain, and urticaria pruritus. The chemical composition of Polygonum multiflorum is complex, mainly including quinones, flavonoids, diphenylethylene and its derivatives. In addition, Polygonum multiflorum also contains a large number of steroids, benzene derivatives, anthocyanins and volatile components. Modern pharmacological studies have found that Polygonum multiflorum has a good effect in anti-inflammatory and antioxidant, lowering blood sugar and lipids, regulating the immune system and nervous system, and absorbing moisture and relieving itching. Studies have shown that among the active ingredients of Polygonum multiflorum, Polygonum multiflorum glycosides, total flavonoids, and total anthraquinones are the main components for improving sleep. There is no report on the application of Polygonum multiflorum volatile components for improving insomnia.

[0019] Xiebai is pungent, bitter and warm; it enters the heart, lung, stomach and large intestine meridians; it can promote yang and disperse nodules, promote qi and remove stagnation; it is used for chest pain, heart pain, fullness and pain in the abdomen, and heaviness after diarrhea; Xiebai mainly contains steroidal saponins, essential oils, polysaccharides, fatty acids, volatile oils and other biologically active ingredients, which have pharmacological effects such as lipid-lowering, anti-oxidation, antibacterial, anti-tumor, and protection of myocardial cells, and is used to treat hyperlipidemia, angina pectoris, coronary heart disease and other diseases.

[0020] Angelica sinensis is sweet, pungent, and warm in nature. It enters the liver, heart, and spleen meridians. It has the effects of nourishing blood and activating blood circulation, regulating menstruation and relieving pain, and moistening the intestines and relieving constipation. It is often used for blood deficiency and sallow complexion, dizziness and palpitations, irregular menstruation, amenorrhea and dysmenorrhea, abdominal pain due to deficiency and cold, rheumatic arthralgia, traumatic injuries, carbuncle ulcers, and constipation due to dry intestines. Angelica sinensis contains a variety of ingredients such as lactones, alkaloids, organic acids or phenols, amino acids, trace elements, vitamins, and neutral volatile oils. In modern pharmacological research, Angelica sinensis polysaccharides, ferulic acid, and Angelica sinensis volatile oils are the main compounds that play the role of Angelica sinensis in hematopoiesis, immunomodulation, anti-tumor, liver protection, and neuroprotection.

[0021] Borneol tastes pungent and bitter, and is cool in nature. It enters the heart, spleen, and lung meridians. It has the effects of refreshing the mind, clearing away heat and relieving pain. It is often used for fever, convulsions, stroke, phlegm, qi stagnation, coma, chest pain, red eyes, mouth sores, sore throat, and pus in the ear canal. Its main ingredient is borneol, which has multiple pharmacological effects such as anti-inflammatory, analgesic, antibacterial, and cardiovascular protection.

[0022] (III) Beneficial effects

[0023] The present invention is based on the principle of TCM syndrome differentiation and treatment, and is reasonably compatible for insomnia of deficiency of both heart and spleen. Longan pulp with sweet, warm and moist texture is used as the main medicine, which can not only tonify spleen qi but also nourish heart blood. Angelica sinensis for nourishing blood and nourishing heart and Polygonum multiflorum for calming nerves and settling mind are used as the ministerial medicine, and the main medicine is used to nourish heart blood and achieve the effect of calming nerves at the same time; Bupleurum and Allium macrostemon are used as adjuvants, which can soothe liver and relieve depression, promote qi circulation and remove stagnation, and are combined with the tonic medicines in the main and ministerial medicines, so that they can be replenished without stagnation, and can be sparse without dispersing, which can not only tonify heart and spleen but also regulate qi mechanism; finally, Borneol is used as the adjuvant, which can clear away heat and relieve restlessness, and is assisted by the adjuvant to soothe liver and regulate qi; all medicines play the effects of tonifying heart and spleen, nourishing blood and calming nerves, and soothing liver and regulating qi. The present invention is based on the compatibility theory, and the volatile components of each medicinal material in the prescription are used as the main active ingredients to prepare the Chinese medicine formula essential oil composed of Bupleurum essential oil, Longan pulp essential oil, Polygonum multiflorum essential oil, Allium macrostemon essential oil, Angelica sinensis essential oil, and Borneol.

[0024] The results of the study showed that the Chinese medicine formula essential oil of the present invention can significantly improve the sleep quality of fruit flies; administering the drug to insomnia model mice by sniffing can significantly improve the insomnia state of insomnia model mice, restore the behavioral activities of model mice, significantly increase the sleep rate of mice, significantly shorten the sleep latency, and significantly prolong the sleep time, and significantly increase the levels of 5-HT, GABA, and Glu in the hypothalamus homogenate of mice, and significantly reduce the level of DA in the hypothalamus homogenate, which has a significant therapeutic effect on insomnia model mice. The Chinese medicine formula essential oil provided by the present invention has a good application prospect in the preparation of aromatherapy drugs for the treatment of insomnia. DETAILED DESCRIPTION

[0025] In order to make the purpose, technical solution and advantages of the embodiments of the present invention clearer, the technical solution in the embodiments of the present invention will be clearly and completely described below in combination with the embodiments of the present invention. Obviously, the described embodiments are 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 creative work are within the scope of protection of the present invention.

[0026] Example 1

[0027] A traditional Chinese medicine essential oil for improving or treating insomnia. The traditional Chinese medicine essential oil consists of the following ingredients in parts by weight: 3 parts of bupleurum essential oil, 3 parts of longan pulp essential oil, 3 parts of tuberose essential oil, 2 parts of Allium macrostemon essential oil, 3 parts of angelica essential oil and 1 part of borneol.

[0028] The above-mentioned bupleurum essential oil, longan pulp essential oil, smilax china essential oil, allium macrostemon essential oil and angelica essential oil are all commercially available, and the distillation concentration of the single essential oil is above 99%.

[0029] The Chinese medicine formula essential oil is added with medically acceptable auxiliary materials to prepare an inhalation dosage form for administration by sniffing; wherein the effective content of the Chinese medicine formula essential oil is 20%.

[0030] Example 2

[0031] A traditional Chinese medicine formula essential oil for improving or treating insomnia. The traditional Chinese medicine formula essential oil consists of the following components in parts by weight: 7 parts of bupleurum essential oil, 8 parts of longan pulp essential oil, 7 parts of smilax glabra essential oil, 6 parts of allium macrostemon essential oil, 8 parts of angelica essential oil and 5 parts of borneol.

[0032] Example 3

[0033] A traditional Chinese medicine formula essential oil for improving or treating insomnia. The traditional Chinese medicine formula essential oil consists of the following components in parts by weight: 10 parts of bupleurum essential oil, 10 parts of longan pulp essential oil, 10 parts of tuberose essential oil, 10 parts of Allium macrostemon essential oil, 10 parts of angelica essential oil, and 6 parts of borneol.

[0034] Example 4

[0035] A traditional Chinese medicine essential oil for improving or treating insomnia. The traditional Chinese medicine essential oil consists of the following components in parts by weight: 4 parts of bupleurum essential oil, 4 parts of longan pulp essential oil, 4 parts of tuberose essential oil, 3 parts of Allium macrostemon essential oil, 4 parts of angelica essential oil, and 2 parts of borneol.

[0036] Example 5

[0037] A traditional Chinese medicine formula essential oil for improving or treating insomnia. The traditional Chinese medicine formula essential oil consists of the following components in parts by weight: 8 parts of bupleurum essential oil, 8 parts of longan pulp essential oil, 8 parts of tuberose essential oil, 7 parts of Allium macrostemon essential oil, 8 parts of angelica essential oil, and 5 parts of borneol.

[0038] Example 6

[0039] A traditional Chinese medicine essential oil for improving or treating insomnia. The traditional Chinese medicine essential oil consists of the following components in parts by weight: 5 parts of bupleurum essential oil, 7 parts of longan pulp essential oil, 7 parts of tuberose essential oil, 4 parts of Allium macrostemon essential oil, 6 parts of angelica essential oil and 4 parts of borneol.

[0040] Example 7

[0041] A traditional Chinese medicine essential oil for improving or treating insomnia. The traditional Chinese medicine essential oil consists of the following components in parts by weight: 6 parts of bupleurum essential oil, 6 parts of longan pulp essential oil, 6 parts of tuberose essential oil, 5 parts of Allium macrostemon essential oil, 6 parts of angelica essential oil, and 3 parts of borneol.

[0042] Test Example 1

[0043] 1 Experimental animals and groups

[0044] 120 wild-type Drosophila Melanieogaster Canton Special strains with similar health status were selected, half of which were male and half were female. After culturing in basal medium for 3 days before the experiment, they were randomly divided into 6 groups, namely blank group and test groups 1-5, each group had 20 flies, half of which were male and half were female.

[0045] 2 Preparation of basal culture medium and drug-containing culture medium

[0046] Basic culture medium: Take 6.5g corn flour, 0.5g agar, 4.4g sucrose, add 70mL distilled water, heat and stir until slightly viscous, cool slightly, add 0.45g yeast powder and 0.45mL propionic acid, mix well, pour into the fruit fly tube, the culture medium thickness is about 2.5-3cm, leave it for 12 hours, and set aside.

[0047] Drug-containing culture medium: Add Chinese herbal formula essential oils 1-5 to the above-mentioned basic culture medium to prepare drug-containing culture medium containing Chinese herbal formula essential oils 1-5 at a concentration of 1%; the specific ingredients of Chinese herbal formula essential oils 1-5 are as follows:

[0048] Traditional Chinese medicine formula essential oil 1: 6 parts of bupleurum essential oil, 6 parts of longan pulp essential oil, 6 parts of polyanthus essential oil, 5 parts of Allium macrostemon essential oil, 6 parts of angelica essential oil, 3 parts of borneol;

[0049] Traditional Chinese medicine formula essential oil 2: 6 parts of bupleurum essential oil, 6 parts of smilax glabra essential oil, 5 parts of allium macrostemon essential oil, 6 parts of angelica essential oil, and 3 parts of borneol;

[0050] Traditional Chinese medicine formula essential oil 3: 6 parts of bupleurum essential oil, 6 parts of longan meat essential oil, 5 parts of Xiebai essential oil, and 3 parts of borneol;

[0051] Chinese medicine formula essential oil 4: 6 parts of longan pulp essential oil, 6 parts of Polygonum multiflorum essential oil, 6 parts of Chinese angelica essential oil, 3 parts of borneol;

[0052] Chinese medicine formula essential oil 5: 6 parts of bupleurum essential oil, 6 parts of longan meat essential oil, 6 parts of tuberose essential oil, 5 parts of Allium macrostemon essential oil, and 6 parts of angelica essential oil.

[0053] 3 Experimental methods

[0054] The blank group and the experimental group 1-5 fruit flies were respectively transferred into the blank culture medium and the corresponding Chinese medicine formula essential oil 1-5 drug-containing culture medium, and the drug was administered for 3 consecutive days. After the drug administration, the sleep of each group of fruit flies was monitored for 24 hours. The specific method is: a small amount of fruit fly culture medium is placed at the lower end of the glass tube, and the fruit flies to be observed are moved into the glass tube. A fruit fly is placed in each glass tube, and the opening is sealed with cotton. Then it is placed for 6 hours. After the fruit flies in the glass tube are fully adapted to the new environment, the sleep of the experimental fruit flies is monitored. The monitoring time is from 7:00 am to 7:00 am the next day. The sleep of the fruit flies is continuously recorded within 24 hours. The monitoring standard is that the fruit fly is stationary for more than 5 minutes, indicating that the fruit fly is in a sleeping state. The sleep time and sleep frequency of the fruit flies are recorded.

[0055] 4 Results

[0056] The results of the sleep time and sleep frequency of each group of fruit flies in 24 hours are shown in Table 1. Compared with the blank group, the sleep time and sleep frequency of fruit flies in test groups 1-5 increased significantly within 24 hours, with significant differences (P<0.05). The average sleep time of fruit flies in test groups 1-5 also increased compared with the blank group, with no significant difference (P>0.05). Among the test groups 1-5, the sleep time of fruit flies in test group 1 in 24 hours was significantly higher than that in test groups 2-5, with significant differences (P<0.05); the sleep frequency of fruit flies in test group 1 in 24 hours was also significantly higher than that in test groups 2-4, with significant differences (P<0.05). The above results show that Chinese medicine formula essential oils 1-5 can significantly increase the sleep time and sleep frequency of fruit flies throughout the day, among which Chinese medicine formula essential oil 1 (Chinese medicine formula essential oil of the present invention) has the most significant effect on improving the sleep of fruit flies.

[0057] The present invention is based on the principle of TCM syndrome differentiation and treatment, and is reasonably compatible for insomnia of deficiency of both heart and spleen. Longan pulp with sweet, warm and moist texture is used as the main medicine, which can not only tonify spleen qi but also nourish heart blood. Angelica sinensis for nourishing blood and nourishing heart and Polygonum multiflorum for calming nerves and settling mind are used as the ministerial medicine, and the main medicine is used to nourish heart blood and achieve the effect of calming nerves at the same time; Bupleurum and Allium macrostemon are used as adjuvants, which can soothe liver and relieve depression, promote qi circulation and remove stagnation, and are combined with the tonic medicines in the main and ministerial medicines, so that they can be replenished without stagnation, and can be sparse without dispersing, which can not only tonify heart and spleen but also regulate qi mechanism; finally, Borneol is used as the adjuvant, which can clear away heat and relieve restlessness, and is assisted by the adjuvant to soothe liver and regulate qi; all medicines play the effects of tonifying heart and spleen, nourishing blood and calming nerves, and soothing liver and regulating qi. The present invention is based on the compatibility theory, and the volatile components of each medicinal material in the prescription are used as the main active ingredients to prepare the Chinese medicine formula essential oil composed of Bupleurum essential oil, Longan pulp essential oil, Polygonum multiflorum essential oil, Allium macrostemon essential oil, Angelica sinensis essential oil, and Borneol. The above test results show that the various drugs in the traditional Chinese medicine formula essential oil of the present invention work synergistically and each component is indispensable. This combination is a key technical means to improve sleep quality.

[0058] Table 1 24h sleep time and sleep frequency of each group of fruit flies

[0059] Grouping Sleep time (min) Sleep times (times) Average sleep time per session (min) Blank Group <![CDATA[567.47±36.79 a ]]> <![CDATA[113.47±13.94 a ]]> 5.00±0.17 Experimental Group 1 <![CDATA[764.19±54.01 b ]]> <![CDATA[141.52±20.19 bc ]]> 5.40±0.37 Experimental Group 2 <![CDATA[677.39±44.28 c ]]> <![CDATA[132.82±14.78 c ]]> 5.10±0.57 Experimental Group 3 <![CDATA[691.77±60.31 ce ]]> <![CDATA[134.58±17.09 c ]]> 5.14±0.39 Experimental Group 4 <![CDATA[709.68±37.49 ed ]]> <![CDATA[136.74±21.04 c ]]> 5.19±0.66 Experimental Group 5 <![CDATA[729.39±30.74 d ]]> <![CDATA[139.46±16.97 bc ]]> 5.23±0.44

[0060] Note: In the same column of data, the same lowercase letter shoulders or no letter means P>0.05; different lowercase letter shoulders mean P<0.05.

[0061] Test Example 2

[0062] 1 Experimental Materials

[0063] 1.1 Experimental animals

[0064] KM mice, half male and half female, SPF grade, weighing 18-22 g, were purchased from Jinan Pengyue Experimental Animal Breeding Co., Ltd., animal certificate number: 370726201100902753. Laboratory use license number: SYXK (Yu) 2020-0004, production license number: SCXK (Lu) 20190003. The experiment began after one week of adaptive feeding, and the feeding conditions were: temperature (26±2)℃, relative humidity (49±5)%, and free access to food and water.

[0065] 1.2 Instruments

[0066] Electronic balance (AX324ZH), Ohaus Instrument (Changzhou) Co., Ltd.; precision balance (JA1103N), Henan Zhongliang Scientific Instrument Co., Ltd.; FA(N) / JA(N) series electronic balance, Shanghai Minqiao Precision Instrument Co., Ltd.; grinder (JXFSTPRP-48), Shanghai Jingxin Industrial Development Co., Ltd.; electronic constant temperature water bath (DZKW-4), Beijing Zhongxing Weiye Instrument Co., Ltd.; water bath (XMTD-7000), Gongyi Yuhua Instrument Co., Ltd.; KDC-160HR high-speed refrigerated centrifuge, Zhongjia Branch of Keda Innovation Co., Ltd.; 680 microplate reader, BIO-RAD, USA; adjustable pipette, Shanghai Leibo Analytical Instrument Co., Ltd.; nebulizer; surgical instruments.

[0067] 1.3 Reagents and drugs

[0068] p-Chlorophenylalanine (PCPA), Aladdin Technology Co., Ltd., batch number: #K1912044; Compound diazepam tablets, Jining Ankang Pharmaceutical Co., Ltd., approval number: National Medicine Standard H10970219, production batch number: 20200404' Pentobarbital sodium salt: Ruibio, Germany, batch number: 2020 / 12; Normal saline, Henan Kelun Pharmaceutical Co., Ltd., batch number: C1200719C2; NaHCO3: Tianjin Hengxing Chemical Reagent Manufacturing Co., Ltd.; Mouse 5-hydroxytryptamine (5-HT) ELIS A detection kit, Suzhou Calvin Biotechnology Co., Ltd., batch number, E20210101A; mouse dopamine (DA) ELISA detection kit, Suzhou Calvin Biotechnology Co., Ltd., batch number, E20210101A; mouse γ-aminobutyric acid (GABA) ELISA detection kit, Suzhou Calvin Biotechnology Co., Ltd., batch number, E20210101A; mouse glutamate (Glu) ELISA detection kit, Suzhou Calvin Biotechnology Co., Ltd., batch number, E20210101A.

[0069] The traditional Chinese medicine formula essential oil of the present invention comprises: 6 mL of bupleurum, 6 mL of longan pulp, 6 mL of polyphylla, 5 mL of Allium macrostemon, 6 mL of angelica sinensis and 3 g of borneol. The above single essential oils are all obtained by taking original plants as raw materials and adopting supercritical carbon dioxide extraction. The distillation concentration of the above single essential oils is above 99%.

[0070] 2 Experimental content

[0071] 2.1 Grouping

[0072] A blank group was set up, and the model mice were randomly divided into model, diazepam tablet group, and high-dose and low-dose groups of Aconitum carmichaelii essential oil, with a total of 5 groups, 12 mice in each group.

[0073] 2.2 Modeling

[0074] According to the weight of the mice, 400 mg / kg PCPA was intraperitoneally injected, with a dosage of 0.1 ml / 10 g, for 3 consecutive days. PCPA was prepared before use.

[0075] Preparation of PCPA: Dissolve 0.1 g NaHCO3 powder in 2 mL distilled water to make a 5% NaHCO3 aqueous solution, mix with 0.9% NaCl solution, adjust the pH to 7-8, add p-chlorophenylalanine (PCPA) powder, and add a few drops of Tween-80 to promote dissolution, so that the concentration of PCPA is 40 mg / mL.

[0076] When the model mice showed loss of circadian rhythm, strong aggression, messy hair, and weight loss, it indicated that the model was successfully established.

[0077] 2.3 Drug administration

[0078] The successfully modeled mice were divided into a model group, a diazepam tablet group (0.625 mg / kg, administered by gavage), and a high-dose and low-dose group of Chinese medicine essential oil (10uL of essential oil was used per mouse for high-dose and 5uL for low-dose, administered by inhalation), and related experiments were carried out three days after administration.

[0079] When sniffing essential oils, first place the mice in a homemade closed sniffing box (45×30×25cm), then put distilled water or different doses of Chinese medicine essential oil solutions into the nebulizer cup, start the air compressor pump, and atomize the solution into fine mist particles. After it diffuses throughout the sniffing box, start timing. The sniffing administration lasts for 40 minutes, once a day, for three consecutive days.

[0080] 2.4 General observation

[0081] On the third day after the successful establishment of the insomnia model, the general activities of the mice were observed three days after the administration. The specific evaluation indicators of the mouse activity are as follows: "++" mice have lost their circadian rhythm, are highly aggressive, have messy hair, and have significantly reduced weight; "+" mice have circadian rhythm disorders, are aggressive, have messy hair, and have reduced weight; "normal" mice have normal activities and reactions; "--" mice have less activity and reduced reaction; "-" mice have very little activity and significantly reduced reaction.

[0082] 2.5 Behavioral assay

[0083] At the end of sniffing on day 3, the rats were immediately injected intraperitoneally with sodium pentobarbital for the righting test (40 mg / kg (0.1 mL / 10 g) to detect the sleep latency and sleep time. The disappearance of the righting reflex of the mice for more than 1 min was considered a sign of falling asleep. The number of sleeping rats, sleep latency and sleep duration of each group were recorded.

[0084] 2.6 Determination of biochemical indicators

[0085] After completing the behavioral test, the mouse hypothalamus was taken, homogenate was prepared, centrifuged (3000rm, 4°C, 10min), the supernatant was taken, and the contents of 5-HT, DA, GABA, NE, and Glu were determined according to the instructions of the ELISA kit.

[0086] 2.7 Statistical analysis

[0087] Graphpad Prism 8.2.0 statistical software was used for statistical analysis. The comparison between the two groups of data was performed using the t test, P < 0.05 indicated a statistical difference, P < 0.01 indicated a significant statistical difference, and the data were expressed as mean ± standard deviation (SD). The rank data were tested using the Radit test.

[0088] 3 Results

[0089] 3.1 Effects of Chinese herbal essential oils on the general state of PCPA-induced insomnia model in mice

[0090] As shown in Table 2, the mice in the modeling group showed loss of circadian rhythm, strong aggression, messy hair, and weight loss. As shown in Table 3, 3 days after administration, the mice in each medication group recovered to normal, or showed less activity and low response compared with the model group, indicating that the insomnia state of the mice in each medication group was significantly improved.

[0091] Table 2 Activity of mice in each group when insomnia model was successfully established

[0092] Grouping ++ + normal －－ － blank 0 0 0 0 0 Model 8 4 0 0 0 Compound diazepam group 7 5 0 0 0 High-dose group of Chinese medicine essential oil 9 3 0 0 0 Low-dose group of Chinese medicine essential oil 9 3 0 0 0

[0093] Table 3 Activity of mice in each group after 3 days of administration

[0094] Grouping ++ + normal －－ － blank 0 0 0 0 0 Model 7 2 3 0 0 Compound diazepam group 0 0 2 2 8 High-dose group of Chinese medicine essential oil 0 0 2 2 8 Low-dose group of Chinese medicine essential oil 0 0 2 3 7

[0095] 3.2 Effects of Chinese herbal essential oils on sleep in a mouse insomnia model induced by PCPA

[0096] As shown in Table 4, the sleep rate of the blank group was 100%, the sleep rate of the model group was only 41.66%, and the sleep rates of the high-dose group and low-dose group of Chinese medicine essential oil were 91.67% and 83.33%, respectively. The sleep rate of the high-dose group was consistent with that of the compound diazepam group. Compared with the blank group, the sleep latency of the model group mice was significantly prolonged (P<0.01) and the sleep time was significantly shortened (P<0.01); compared with the model group, the high-dose group, low-dose group and compound diazepam group of Chinese medicine essential oil significantly shortened the sleep latency of mice (P<0.01) and significantly prolonged the sleep time (P<0.01).

[0097] Table 4 Effects of Chinese herbal essential oils on sleep in the PCPA-induced insomnia model in mice

[0098]

[0099]

[0100]

[0101] Note: Compared with the blank group: # P<0.05, ## P<0.01; compared with the model group: *P<0.05, **P<0.01

[0102] 3.3 Effects of Chinese herbal essential oils on biochemical parameters of PCPA-induced insomnia model in mice

[0103] As shown in Table 5, compared with the blank group, the levels of 5-HT, GABA, and Glu in the hypothalamic homogenate of the mice in the model group were significantly decreased (P<0.01), and the level of DA in the hypothalamic homogenate was significantly increased (P<0.01); compared with the model group, the high-dose group, low-dose group and compound diazepam group of Chinese herbal formula essential oil could significantly increase the levels of 5-HT and GABA in the hypothalamic homogenate of the mice (P<0.01), and the level of DA in the hypothalamic homogenate was significantly decreased (P<0.01); the high-dose group and compound diazepam group of Chinese herbal formula essential oil could significantly increase the level of Glu in the hypothalamic homogenate of the mice (P<0.01), and the low-dose group could significantly increase the level of Glu in the hypothalamic homogenate of the mice (P<0.05).

[0104] Table 5 Effects of Chinese herbal essential oils on 5-HT, DA, GABA, and Glu levels in hypothalamic homogenate of mice with insomnia induced by PCPA ( n=12)

[0105]

[0106] Note: Compared with the blank group: # P<0.05, ## P<0.01; compared with the model group: *P<0.05, **P<0.01

[0107] 4 Conclusion

[0108] The present invention adopts the method of intraperitoneal injection of PCPA to successfully construct a mouse insomnia model, and the mouse insomnia model is used to further verify the therapeutic effect of the Chinese medicine formula essential oil of the present invention on insomnia. The above test results show that the Chinese medicine formula essential oil of the present invention is administered to insomnia model mice by sniffing, which can significantly improve the insomnia state of the model mice, restore the behavioral activities of the model mice, significantly increase the sleep rate of mice, significantly shorten the sleep latency, and significantly prolong the sleep time. It can significantly increase the levels of 5-HT, GABA, and Glu in the hypothalamic homogenate of mice, and significantly reduce the level of DA in the hypothalamic homogenate. The Chinese medicine formula essential oil provided by the present invention has a significant therapeutic effect on insomnia model mice.

[0109] The above embodiments are only used to illustrate the technical solutions of the present invention, rather than to limit the same. Although the present invention has been described in detail with reference to the aforementioned embodiments, those skilled in the art should understand that the technical solutions described in the aforementioned embodiments may still be modified, or some of the technical features may be replaced by equivalents. However, these modifications or replacements do not deviate the essence of the corresponding technical solutions from the spirit and scope of the technical solutions of the embodiments of the present invention.

Claims

1. A Chinese medicine essential oil for improving or treating insomnia, characterized in that: The traditional Chinese medicine formula essential oil is composed of the following components in parts by weight: 1-10 parts of bupleurum essential oil, 1-10 parts of longan pulp essential oil, 1-10 parts of tuberose essential oil, 1-10 parts of Allium macrostemon essential oil, 1-10 parts of angelica essential oil and 1-6 parts of borneol.

2. A Chinese medicine essential oil for improving or treating insomnia according to claim 1, characterized in that: The traditional Chinese medicine formula essential oil is composed of the following components in parts by weight: 4-8 parts of bupleurum essential oil, 4-8 parts of longan pulp essential oil, 4-8 parts of tuberose essential oil, 3-7 parts of Allium macrostemon essential oil, 4-8 parts of angelica essential oil and 2-5 parts of borneol.

3. A Chinese medicine essential oil for improving or treating insomnia according to claim 1, characterized in that: The traditional Chinese medicine formula essential oil is composed of the following ingredients in parts by weight: 6 parts of bupleurum essential oil, 6 parts of longan pulp essential oil, 6 parts of tuberose essential oil, 5 parts of Allium macrostemon essential oil, 6 parts of angelica essential oil and 3 parts of borneol.

4. A Chinese medicine essential oil for improving or treating insomnia according to claim 1, characterized in that: Bupleurum essential oil, longan meat essential oil, flos cholerae essential oil, Allium macrostemon essential oil and angelica essential oil are extracted from original plants using conventional essential oil extraction methods, including pressing extraction, supercritical carbon dioxide extraction, steam distillation extraction, immersion method, chemical solvent extraction method and oil separation method.

5. A Chinese medicine essential oil for improving or treating insomnia according to claim 3, characterized in that: The distillation concentration of the above-mentioned single essential oil is controlled to be above 99%.

6. The use of the Chinese medicine essential oil according to any one of claims 1 to 3 in the preparation of a medicine for improving or treating insomnia, characterized in that: The traditional Chinese medicine essential oil is added with medically acceptable auxiliary materials to prepare an inhalation dosage form for administration by inhalation and sniffing.

7. The use of the Chinese medicine essential oil in the preparation of a medicine for improving or treating insomnia according to claim 5, characterized in that: The effective content of the essential oil of the traditional Chinese medicine formula in the medicine is 5-20%.

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