Traditional Chinese medicine compound essential oil and application thereof in preparation of anti-depression product

Abstract

The invention provides traditional Chinese medicine compound essential oil and application thereof in preparation of anti-depression products, and relates to the technical field of traditional Chinese medicine essential oil pharmacy. The traditional Chinese medicine compound essential oil is prepared from 3-15 parts of benzoin essential oil, 2-10 parts of China rose essential oil, 2-10 parts of immature bitter orange essential oil, 1-8 parts of amomum cardamomum essential oil and 1-8 parts of fennel essential oil. The traditional Chinese medicine compound essential oil disclosed by the invention is administered in a sniffing manner, and improves cerebral depression symptoms by stimulating olfactory nerves and influencing cerebral cortex edges and neurotransmitters. After a CUMS depression animal model is treated by the traditional Chinese medicine compound essential oil disclosed by the invention, the symptom of pleasant sensation disappearance is well reversed and improved, and the state of animal movement reduction and the behavior state of despair and struggling abandoning of the depression animal model are obviously improved; meanwhile, the levels of DA, 5-HT and NE in brain tissues of the depression animal model are remarkably improved, and the pathological change of the brain-sea horse area of the depression animal model is remarkably improved. The traditional Chinese medicine compound essential oil disclosed by the invention can be used for preparing medicines for aromatherapy, has no adverse reaction and high acceptability, and provides a new way for treating depression.

Description

Technical Field

[0001] The invention relates to the technical field of Chinese medicine essential oil pharmaceutical manufacturing, and in particular to a Chinese medicine compound essential oil and application thereof in preparing an antidepressant product. Background Art

[0002] With the continuous development of society, people's life pressure is increasing, and the incidence of depression is gradually increasing. As a clinical type of mental illness with affective disorders, depression is specifically manifested as difficulty sleeping, loss of interest, decreased appetite, emotional instability, etc. In the treatment of depression, most of the antidepressants used in clinical practice are synthetic antidepressants. The first generation of such drugs has been gradually abandoned due to safety issues, and the transition has been to the second generation of synthetic antidepressants. However, the second generation of drugs still have shortcomings such as full onset and relatively single target. These factors have prompted scientists to develop new third-generation antidepressants with higher targeting, higher efficiency and higher safety. However, the current third-generation antidepressants still have problems such as insufficient efficacy and many adverse reactions in the long-term and short-term treatment of patients. Therefore, finding natural antidepressants with less toxicity and higher efficacy has become a new development hotspot.

[0003] In the theory of traditional Chinese medicine, Chinese medicine with aromatic smell is collectively called aromatic medicine. Aromatic medicines account for a large proportion of Chinese medicinal materials and play an indispensable role in clinical treatment. Using aromatic medicines as a medium can improve the traditional "aromatherapy". Aromatic substances can be made into inhalants, essential oils, etc., which enter the body through breathing or skin to relieve mental stress, treat diseases, and promote human health.

[0004] Volatile oil is a general term for oily substances with volatile aromatic smell, also known as essential oil. Most aromatic Chinese medicines contain volatile oil components. Studies have shown that aromatic Chinese medicines regulate human functions by affecting people's emotions and psychological states. After the volatile oil of medicinal plants enters the nasal cavity through inhalation, part of it is absorbed into the blood circulation, and the other part combines with the olfactory receptors in the olfactory cells of the nose and is transmitted to the cerebral cortex through the olfactory nerves, producing either a soothing and stress-relieving effect or an exciting and pleasant effect, thereby alleviating the symptoms of depression. At present, the intervention research on volatile oil of medicinal plants for depression mainly focuses on the application of volatile oil extracted from a single plant, and there are few reports on the anti-depressant Chinese medicine compound essential oil. Summary of the invention

[0005] 1. Technical issues to be resolved

[0006] In view of the deficiencies in the prior art, the present invention provides a traditional Chinese medicine compound essential oil and application thereof in the preparation of an antidepressant product.

[0007] (II) Technical solution

[0008] To achieve the above objectives, the present invention is implemented through the following technical solutions:

[0009] Firstly, the present invention provides a traditional Chinese medicine compound essential oil, which is composed of the following traditional Chinese medicine plant essential oils in parts by volume: 3-15 parts of benzoin essential oil, 2-10 parts of rose essential oil, 2-10 parts of citrus aurantium essential oil, 1-8 parts of white cardamom essential oil, and 1-8 parts of fennel essential oil.

[0010] Preferably, the Chinese herbal compound essential oil is composed of the following Chinese herbal plant essential oils in the following proportions by volume: 5-10 parts of benzoin essential oil, 4-8 parts of rose essential oil, 4-8 parts of Citrus aurantium essential oil, 1-5 parts of white cardamom essential oil, and 1-5 parts of fennel essential oil.

[0011] Preferably, the Chinese herbal compound essential oil is composed of the following Chinese herbal plant essential oils in the following parts by volume: 6 parts of benzoin essential oil, 5 parts of rose essential oil, 5 parts of Citrus aurantium essential oil, 3 parts of white cardamom essential oil, and 3 parts of fennel essential oil.

[0012] The above-mentioned single Chinese medicinal plant essential oil is extracted from the corresponding Chinese medicinal plant as raw material by distillation or supercritical extraction, and the purity of the essential oil is above 99%.

[0013] The above-mentioned single Chinese medicinal plant essential oils can also be purchased commercially.

[0014] Secondly, the present invention also provides an application of the above-mentioned Chinese herbal compound essential oil in the preparation of an antidepressant product, wherein the antidepressant product is a product used for aromatherapy, specifically, each single Chinese herbal plant essential oil is weighed according to the above-mentioned volume fractions, mixed evenly, and medically acceptable excipients are added to prepare a dosage form for administration by inhalation.

[0015] In the antidepressant product, the effective content of the Chinese medicine compound essential oil is 5-15%.

[0016] Benzoin is the dried resin of Styrax tonkinensis (Pierre) Craib ex Hart., a plant of the Styracaceae family. It is pungent, fragrant, flat, non-toxic, and has the effects of refreshing the mind, promoting qi and blood circulation, and relieving pain. It is used for stroke, phlegm syncope, sudden syncope due to qi stagnation, coma due to poisoning, abdominal pain, postpartum blood loss, and infantile convulsions. Benzoin essential oil mainly contains ingredients such as benzyl benzoate, benzyl cinnamate, vanillin, and styrax. Modern pharmacological studies have shown that benzoin essential oil has the effects of antibacterial and anti-inflammatory, expectorant, skin repair, and mood regulation.

[0017] Rose is the dried flower of the rose plant Rosa chinensis Jacq. of the Rosaceae family. It is sweet in taste, warm in nature, and enters the liver meridian; it has the effects of promoting blood circulation and regulating menstruation, soothing the liver and relieving depression. It is often used for qi stagnation and blood stasis, irregular menstruation, dysmenorrhea, amenorrhea, and chest and flank pain. Rose essential oil mainly contains terpenoid compounds, aromatic compounds, aliphatic compounds, etc., which have pharmacological effects such as antibacterial and anti-inflammatory, antioxidant, immunomodulatory, cardiovascular and nerve protection, anti-tumor, and mood regulation.

[0018] Citrus aurantium L. is a dried young fruit of the Rutaceae plant Citrus aurantium L. It is bitter, pungent, sour, slightly cold, and enters the spleen and stomach meridians. It has the effects of breaking up qi and eliminating stagnation, resolving phlegm and dispersing lumps. It is used for stagnation of internal stagnation, fullness and pain, heaviness after diarrhea, constipation, phlegm stagnation and qi obstruction, chest pain, chest congestion, and organ ptosis. Citrus aurantium essential oil mainly contains monoterpenes, sesquiterpenes, aldehydes and ketones, esters and other compounds. Modern pharmacological studies have shown that it has pharmacological effects such as regulating the digestive system, antibacterial and anti-inflammatory, antioxidant, anti-aging, protecting cardiovascular, and regulating immune function.

[0019] White cardamom is the dried mature fruit of Amomum kravanh Picrre cx Gagnep., a plant of the ginger family. It is pungent in taste and warm in nature. It enters the lung, spleen and stomach meridians. It has the effects of removing dampness and promoting qi, warming the middle and stopping vomiting, and stimulating appetite and digestion. It is mainly used to treat dampness blocking qi stagnation, spleen and stomach disharmony, abdominal distension, loss of appetite, initial damp-heat, chest tightness without hunger, stomach cold vomiting, and indigestion. White cardamom essential oil mainly contains compounds such as monoterpenes, sesquiterpenes, esters and ethers. Modern pharmacological studies have shown that it has pharmacological effects such as regulating the digestive system, antibacterial and anti-inflammatory, antioxidant, anti-aging, regulating immune function, and improving the respiratory system.

[0020] Fennel is the dried mature fruit of Foeniculum vdgare Mill., a plant of the Apiaceae family. It is pungent in taste and warm in nature. It enters the liver, kidney, spleen, and stomach meridians. It has the effects of dispelling cold and relieving pain, regulating qi and harmonizing the stomach. It is used for abdominal pain caused by cold hernia, testicular prolapse, dysmenorrhea, cold pain in the lower abdomen, abdominal distension and pain, and vomiting and diarrhea due to poor appetite. Fennel essential oil mainly contains compounds such as monoterpenes, sesquiterpenes, aldehydes and ketones. Modern pharmacological studies have shown that it has pharmacological effects such as regulating the digestive system, antibacterial and anti-inflammatory, antioxidant, anti-aging, improving the respiratory system, regulating immune function, and regulating endocrine.

[0021] (III) Beneficial effects

[0022] The present invention is based on the principle of TCM syndrome differentiation and treatment, and is aimed at the pathological mechanism of depression. It follows the principle of monarch, minister, assistant and envoy compatibility, and uses benzoin as the monarch drug. Its pungent, dispersing, bitter and descending nature can soothe the liver and relieve depression, open the orifices and refresh the mind, and directly targets the core pathological mechanism of depression syndrome, qi stagnation, and phlegm and blood stasis blocking the orifices. Rose is sweet and warm in nature, and has the effects of promoting blood circulation and regulating menstruation, soothing the liver and regulating qi, helping the monarch drug to enhance the power of qi circulation and relieving depression, and promoting the smooth flow of qi and blood to relieve liver depression; Citrus aurantium can break up qi and eliminate accumulation, resolve phlegm and disperse knots, and not only enhance the power of qi circulation and stagnation, but also descend qi and resolve phlegm to eliminate phlegm turbidity and obstruction, and the two are minister drugs. Fennel is used to warm the middle and promote qi circulation, lead the medicine into the liver meridian, and enhance the effect of soothing the liver and relieving depression. White cardamom is used to eliminate dampness and promote qi circulation, and to harmonize the spleen and stomach, which not only prevents the pungent and fragrant qi regulating medicine from damaging the spleen and stomach, but also harmonizes the liver and spleen by "cultivating soil and nourishing wood". The whole prescription takes "soothing the liver - promoting qi - resolving phlegm - strengthening the spleen" as the treatment principle, benzoin relieves depression and awakens the mind to regulate the mind, rose softens the liver, immature bitter orange breaks stagnation, fennel warms the middle, and white cardamom harmonizes the stomach, which is consistent with the treatment idea of ​​"qi depression first, taking into account phlegm and dampness" in the theory of six depressions in "Danxi's Heart Method", and at the same time reflects the treatment characteristics of depression syndrome of "regulating qi" and "protecting the middle jiao" in "Clinical Guide Medical Cases". The compatibility of various medicines plays the role of soothing the liver and relieving depression, strengthening the spleen and calming the mind, and is suitable for depression of qi stagnation and phlegm obstruction and mental malnutrition type. Based on the above-mentioned prescription, benzoin, rose, immature bitter orange, white cardamom, and fennel plants are used as raw materials, and their volatile oil components are extracted to make a Chinese medicine compound essential oil, which has the effects of soothing the liver and relieving depression, strengthening the spleen and calming the mind. The Chinese medicine compound essential oil of the present invention is administered by inhalation and sniffing, and stimulates the olfactory nerves to affect the edge of the cerebral cortex and neurotransmitters, thereby improving the brain depression-like symptoms. The traditional Chinese medicine compound essential oil of the present invention can be used for preparing medicines used for aromatherapy, has no adverse reactions, is highly acceptable, and provides a new approach for the treatment of depression.

[0023] The results of animal experiments show that after the treatment of the Chinese herbal compound essential oil of the present invention, the symptoms of anhedonia in the CUMS animal depression model are well reversed and improved, and the state of animal movement reduction and the behavior of despair and giving up struggle in the depressed animal model are significantly improved; at the same time, the levels of DA, 5-HT, and NE in the brain tissue of the depressed animal model are significantly increased, and the pathological changes in the hippocampus of the depressed animal model are significantly improved, and the therapeutic effect of the Chinese herbal relapse essential oil of the present invention on the depressed animal model is better than that of Western medicine. The Chinese herbal compound essential oil of the present invention is expected to become a new drug for aromatherapy to treat depression. BRIEF DESCRIPTION OF THE DRAWINGS

[0024] Figure 1 HE staining of hippocampal tissues of rats in each group, HE×400. 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 compound essential oil for anti-depression, which consists of the following traditional Chinese medicine plant essential oils in parts by volume: 3 parts of benzoin essential oil, 2 parts of rose essential oil, 2 parts of immature bitter orange essential oil, 1 part of white cardamom essential oil and 1 part of fennel essential oil.

[0028] The above-mentioned single Chinese medicinal plant essential oil is extracted from the corresponding Chinese medicinal plant as raw material by distillation or supercritical extraction, and the purity of the essential oil is above 99%.

[0029] Each single Chinese medicinal plant essential oil is weighed according to the above volume fractions, mixed evenly, and medically acceptable auxiliary materials are added to prepare a dosage form for administration by inhalation, wherein the effective content of the Chinese medicinal compound essential oil is 5-15%.

[0030] Example 2

[0031] The difference between this embodiment and embodiment 1 is that the Chinese herbal compound essential oil is composed of the following Chinese herbal plant essential oils in parts by volume: 13 parts of benzoin essential oil, 9 parts of rose essential oil, 9 parts of Citrus aurantium essential oil, 6 parts of white cardamom essential oil, and 6 parts of fennel essential oil.

[0032] Example 3

[0033] The difference between this embodiment and embodiment 1 is that the Chinese herbal compound essential oil is composed of the following Chinese herbal plant essential oils in parts by volume: 4 parts of benzoin essential oil, 3 parts of rose essential oil, 3 parts of Citrus aurantium essential oil, 2 parts of white cardamom essential oil, and 1 part of fennel essential oil.

[0034] Example 4

[0035] The difference between this embodiment and embodiment 1 is that the Chinese herbal compound essential oil is composed of the following Chinese herbal plant essential oils in parts by volume: 10 parts of benzoin essential oil, 6 parts of rose essential oil, 6 parts of Citrus aurantium essential oil, 4 parts of white cardamom essential oil, and 3 parts of fennel essential oil.

[0036] Example 5

[0037] The difference between this embodiment and embodiment 1 is that the Chinese herbal compound essential oil is composed of the following Chinese herbal plant essential oils in parts by volume: 5 parts of benzoin essential oil, 4 parts of rose essential oil, 4 parts of Citrus aurantium essential oil, 1 part of white cardamom essential oil, and 1 part of fennel essential oil.

[0038] Example 6

[0039] The difference between this embodiment and embodiment 1 is that the Chinese herbal compound essential oil is composed of the following Chinese herbal plant essential oils in parts by volume: 10 parts of benzoin essential oil, 8 parts of rose essential oil, 8 parts of Citrus aurantium essential oil, 5 parts of white cardamom essential oil, and 5 parts of fennel essential oil.

[0040] Example 7

[0041] The difference between this embodiment and embodiment 1 is that the Chinese herbal compound essential oil is composed of the following Chinese herbal plant essential oils in parts by volume: 7 parts of benzoin essential oil, 5 parts of rose essential oil, 5 parts of Citrus aurantium essential oil, 2 parts of white cardamom essential oil, and 3 parts of fennel essential oil.

[0042] Example 8

[0043] The difference between this embodiment and embodiment 1 is that the Chinese medicine compound essential oil is composed of the following Chinese medicinal plant essential oils in parts by volume: 8 parts of benzoin essential oil, 6 parts of rose essential oil, 4 parts of Citrus aurantium essential oil, 3 parts of white cardamom essential oil, and 2 parts of fennel essential oil.

[0044] Example 9

[0045] The difference between this embodiment and embodiment 1 is that the Chinese herbal compound essential oil is composed of the following Chinese herbal plant essential oils in parts by volume: 6 parts of benzoin essential oil, 5 parts of rose essential oil, 5 parts of Citrus aurantium essential oil, 3 parts of white cardamom essential oil, and 3 parts of fennel essential oil.

[0046] Test Example 1

[0047] Effects of different components of traditional Chinese medicine essential oils on depressive-like behavior in mice

[0048] 1 Experimental Materials

[0049] 1.1 Experimental animals

[0050] 60 KM mice, all male, weighing 19-22g, SPF grade. Animal qualification certificate number: No.370726211100923115; provided by Shandong Jinan Pengyue Experimental Animal Breeding Co., Ltd., license number SCXK(Lu)20190003; laboratory use license number: SYXK(Yu)2020-0004. Raised in the Animal Experiment Center of Henan University of Traditional Chinese Medicine, unit use license number: SYXK(Yu)2020-0004.

[0051] 1.2 Main drugs

[0052] The Chinese medicine compound essential oil of the present invention: benzoin essential oil 6mL, rose essential oil 5mL, citrus aurantium essential oil 5mL, white cardamom essential oil 3mL, fennel essential oil 3mL

[0053] Control essential oil 1: benzoin essential oil 6mL, white cardamom essential oil 3mL, fennel essential oil 3mL

[0054] Control compound essential oil 2: 5 mL of rose essential oil, 5 mL of Citrus aurantium essential oil, 3 mL of white cardamom essential oil, 3 mL of fennel essential oil

[0055] Control essential oil 3: benzoin essential oil 6mL, rose essential oil 5mL, citrus aurantium essential oil 5mL

[0056] The essential oils are prepared from benzoin, rose, immature bitter orange, white cardamom and fennel as raw materials respectively by supercritical carbon dioxide extraction method. The purity of the essential oils is above 99%. The essential oils are accurately weighed according to the above volume fractions and mixed evenly to obtain the essential oils.

[0057] 2 Experimental methods

[0058] 2.1 Model preparation

[0059] Ten of 70 SPF ICR male mice were randomly selected as the blank group (Control), and the remaining 60 mice were used to establish the CUMS depression model using solitary housing combined with chronic unpredictable mild stress (CUMS). The specific method is: 10 different stimuli were selected: cage tilt (24h), moist bedding (24h), day and night reversal, ice water swimming (4℃, 5min), fasting (24h), water deprivation (24h), heat stimulation (50℃, 5min), tail clamping (1min), electric shock to the sole (voltage 50mV, 1 stimulation every 30s, each lasting 10s, a total of 15 times), noise stimulation (100Hz, 2h), one stimulus was given every day, the order was random, and each stimulus was not used for 2 consecutive days, so that the animals could not anticipate the occurrence of the stimulus, and the stress lasted for 6 weeks. The blank control group was not given any stimulus.

[0060] 2.2 Experimental groups and drug administration

[0061] Ten mice were randomly selected for the blank group (Control) before CUMS stimulation. After 6 weeks of CUMS treatment, mice with abnormal sugar water preference rate were eliminated according to the results of the sugar water preference rate experiment, and 50 mice with successful model preparation and good condition were selected and randomly divided into 5 groups, with 10 mice in each group:

[0062] Model group: no drug treatment, only pure water;

[0063] Test group 1: the traditional Chinese medicine compound essential oil of the present invention, the dosage is 5 μL / 20g;

[0064] Experimental group 2: control compound essential oil 1, dose 5 μL / 20 g;

[0065] Experimental group 3: control compound essential oil 2, dose 5 μL / 20 g;

[0066] Experimental group 4: control compound essential oil 3, dose 5 μL / 20 g;

[0067] The above compound essential oils were all administered by sniffing. When sniffing, the mice were first placed in a self-made closed sniffing box (45×30×25cm), and then the Chinese medicine compound essential oil of the present invention was mixed with 5mL of distilled water according to the above dosage and placed in the atomization cup. The air compression pump was started to atomize the essential oil solution into fine mist particles. After it diffused throughout the sniffing box, the timing was started, and the sniffing administration lasted for 30min. Each administration group began to be administered after the end of CUMS, once a day, for 2 weeks, that is, the 43rd-57th day of the experiment.

[0068] 2.3 Neurobehavioral index detection

[0069] The neurobehavioral tests of mice in each group were performed before and after the experiment. The specific test items are as follows:

[0070] (1) Sucrose preference test (SPT)

[0071] The mice were subjected to a continuous sugar water preference rate experiment. The mice were trained with sugar water two days before the experiment. After fasting and depriving of water for 24 hours, the test was carried out. The experimental operation started at 9 am every day. The operation steps are detailed in the table:

[0072] Table 2 Experimental design of sugar water preference rate

[0073]

[0074]

[0075] (2) Open field test (OFT)

[0076] The OFT open field activity experimental system for mice was used to observe the movement distance and movement time of mice in each group within 5 minutes (open field size: 50×50×40 cm).

[0077] (3) Tail suspension test (TST)

[0078] Using the mouse tail suspension video analysis system, the mice were suspended in the tail suspension device for 6 minutes, and the immobility time of the mice after 4 minutes was recorded.

[0079] 2.4 Statistical methods

[0080] SPSS 25.0 software was used for statistical analysis. Measurement data were expressed as mean ± standard deviation (x ± s), and one-way analysis of variance was used for comparison between groups. The Games-Howell method was used for unequal variances, and the LSD method was used for equal variances. Ridit test was used for rank data.

[0081] 3 Results Effects of the Chinese herbal essential oil on the neurobehavior of the CUMS mouse depression model

[0082] Table 1 shows the results of the sugar water preference rate of each group of mice before and after treatment. Before treatment, compared with the blank group, the sugar water preference rate of mice in each model group was significantly reduced (P<0.01), and there was no significant difference in the sugar water preference rate among the model groups (P>0.05). After treatment, compared with the blank group, the sugar water preference rate of mice in the model group was significantly reduced (P<0.01); compared with the model group, the sugar water preference rate of test groups 1-4 was significantly increased (P<0.01), among which the increase in the sugar water preference rate of test group 1 (the Chinese medicine compound essential oil of the present invention) was significantly higher than that of test groups 2-3.

[0083] Table 1 Sugar water preference rate of mice in each group before and after treatment

[0084] Group Before treatment (%) After treatment (%) Blank Group 81.2±3.6 81.4±2.7 Model Group <![CDATA[62.4±3.9 △△ ]]> <![CDATA[65.7±2.6 △△ ]]> Experimental Group 1 <![CDATA[62.3±3.1 △△ ]]> <![CDATA[75.1±2.8 ** ]]> Experimental Group 2 <![CDATA[62.4±2.8 △△ ]]> <![CDATA[69.8±2.6 ** ]]> Experimental Group 3 <![CDATA[62.6±2.8 △△ ]]> <![CDATA[69.1±2.4 ** ]]> Experimental Group 4 <![CDATA[62.5±2.9 △△ ]]> <![CDATA[70.2±3.0 ** ]]>

[0085] Note: Compared with the blank group, △△ P<0.01, △ P<0.05; compared with the model group, **P<0.01, *P<0.05

[0086] Table 2 shows the open field test results of each group of mice before and after treatment. Before treatment, compared with the blank group, the movement distance and movement time of each group of mice in the model were significantly reduced (P<0.01), and there was no significant difference in movement distance and movement time between the groups in the model (P>0.05). After treatment, compared with the model group, test group 1 (the Chinese medicine compound essential oil of the present invention) can significantly increase the total movement distance of mice (P<0.01), test group 4 (control compound essential oil 3) can significantly increase the total movement distance of mice (P<0.05), and test groups 2 and 3 only have a trend of increasing the movement distance of mice (P>0.05); test group 1 can significantly prolong the movement time of mice (P<0.01), and test groups 2 and 4 can significantly prolong the movement time of mice (P<0.05).

[0087] Table 2 Open field test results of mice in each group before and after treatment

[0088]

[0089] Note: Compared with the blank group, △△P<0.01, △ P<0.05; compared with the model group, **P<0.01, *P<0.05

[0090] Table 3 shows the results of the tail suspension test of mice in each group. Before treatment, compared with the blank group, the immobility time of each model group was significantly prolonged (P<0.01), and there was no significant difference among the model groups (P>0.05). Compared with the model group, test group 1 can significantly shorten the immobility time of mice (P<0.01), test group 4 can significantly shorten the immobility time of mice (P<0.05), and test groups 2 and 3 only have a tendency to shorten the immobility time of mice (P>0.05).

[0091] Table 3 shows the test results of the tail suspension test for each group of mice

[0092] Group Immobility time before treatment (s) Immobility time after treatment (s) Blank Group 115.5±22.3 124.8±18.3 Model Group <![CDATA[177.5±19.4 △△ ]]> <![CDATA[183.2±16.0 △△ ]]> Experimental Group 1 <![CDATA[177.1±19.5 △△ ]]> <![CDATA[142.9±21.7 ** ]]> Experimental Group 2 <![CDATA[177.2±18.8 △△ ]]> 161.2±23.1 Experimental Group 3 <![CDATA[177.5±19.1 △△ ]]> 165.9±22.5 Experimental Group 4 <![CDATA[177.3±19.4 △△ ]]> <![CDATA[154.3±20.9 * ]]>

[0093] Note: Compared with the blank group, △△ P<0.01, △ P<0.05; compared with the model group, **P<0.01, *P<0.05

[0094] 4 Conclusion

[0095] After the treatment of the Chinese herbal compound essential oil of the present invention, the symptoms of anhedonia in the CUMS mouse depression model were well reversed and improved, and the state of reduced movement of mice and the behavior of despair and giving up struggle of the depression model mice were significantly improved; the improvement effect of the Chinese herbal compound essential oil of the present invention on the depressive-like behavior of mice was significantly better than that of other compound essential oils, and the various components of the Chinese herbal compound essential oil of the present invention cooperated with each other to synergize. The Chinese herbal compound essential oil of the present invention is expected to become a new drug for aromatherapy to treat depression.

[0096] Test Example 2

[0097] Effect of the traditional Chinese medicine compound essential oil of the present invention on the depression model of rats induced by CUMS

[0098] 1 Experimental Materials

[0099] 1.1 Experimental animals

[0100] Male SD rats, SPF grade, weighing 180-200 g, were provided by Jinan Pengyue Experimental Animal Breeding Co., Ltd., license number: SCXK(Lu)20190003; animal qualification certificate number: 1107262011002341; the animals were raised in the Animal Experiment Center of Henan University of Traditional Chinese Medicine, unit use license number: SYXK(Yu)2020-0004.

[0101] 1.2 Main drugs

[0102] The Chinese medicinal compound essential oil comprises: 6 mL of benzoin essential oil, 5 mL of rose essential oil, 5 mL of immature citrus aurantium essential oil, 3 mL of white cardamom essential oil and 3 mL of fennel essential oil. The essential oils are respectively prepared from benzoin, rose, immature citrus aurantium, white cardamom and fennel as raw materials by adopting a supercritical carbon dioxide extraction method. The purity of the essential oil is above 99%. The essential oils are accurately weighed according to the above volume fractions and uniformly mixed to obtain the essential oil.

[0103] Fluoxetine hydrochloride (Manufacturer: Changzhou Siyao Pharmaceutical Co., Ltd., product batch number: 20201110).

[0104] 2 Experimental design and methods

[0105] 2.1 Experimental design

[0106] After all rats were purchased, they were adaptively raised for 3 days, and then housed in single cages for 2 weeks. Then, they were modeled (housed in single cages + CUMS) for 6 weeks, and drug intervention lasted for 4 weeks. After sampling, the indexes were tested. Every 2 weeks, the rats were tested for sugar water preference rate. Before sampling, the sugar water preference rate test, open field test, and forced swimming test were performed in sequence. The behavioral experimental tests all started at 9 am.

[0107] 2.2 Experimental methods

[0108] 2.2.1 Model preparation

[0109] Ten rats were randomly selected from 50 SD rats (180-200g, male, SPF grade) as the blank group, and the remaining 40 rats were used for modeling. Before CUMS, each rat was kept in a single cage for 2 weeks. From the third week, CUMS (continued to be kept in a single cage) was started. Different stimulations were randomly given every day. Each stimulation could not appear continuously. Ten stimulation methods were randomly applied within 6 weeks to prepare the CUMS depression model. The 10 stimulations included: ① fasting (24h), ② water deprivation (24h), ③ day and night reversal, ④ wet bedding, ⑤ heat stress (45℃, 5min), ⑥ ice water swimming (4℃, 5min), ⑦ tail clamp (1min), ⑧ noise stimulation (100Hz, 2h), ⑨ odor stimulation (appropriate amount of pepper, 12h), and ⑩ flicker stimulation (color light high-frequency flicker stimulation for 12h).

[0110] 2.2.2 Experimental Grouping and Treatment

[0111] The rats with successful modeling were divided into 4 groups, with 10 rats in each group, and the groups were as follows:

[0112] Model group: Oral administration of pure water;

[0113] Fluoxetine hydrochloride group: dose 3.33 mg / kg, concentration 0.333 mg / ml;

[0114] The high-dose group of the Chinese medicine compound essential oil of the present invention (hereinafter referred to as the high-dose group): the dose is 100 μL / kg;

[0115] The low-dose group of the Chinese medicine compound essential oil of the present invention (hereinafter referred to as the low-dose group): the dose is 50 μL / kg;

[0116] A blank group was set up: no CUMS modeling was performed, and pure water was administered;

[0117] The drug was administered from the 4th week onwards, for a total of 4 weeks, i.e., the 28th to 56th day of the experiment. In the rat experiment, fluoxetine hydrochloride was converted to 10 times the clinical dosage, and the dosage of the Chinese medicine compound essential oil of the present invention was determined according to the preliminary experimental results. Rats were weighed weekly, and the dosage was adjusted according to their body weight.

[0118] The Chinese medicinal compound essential oil of the present invention is administered by sniffing. When sniffing, the rats are first placed in a self-made closed sniffing box (45×30×25 cm), then the Chinese medicinal compound essential oil of the present invention is mixed with 5 mL of distilled water according to the above dosage and placed in an atomizing cup, and an air compression pump is started to atomize the essential oil solution into fine mist particles. After the mist particles are diffused throughout the sniffing box, timing is started, and the sniffing administration lasts for 30 minutes.

[0119] 2.3 Observation indicators and detection

[0120] 2.3.1 Behavioral index detection

[0121] (1) Sugar Preference Test (SPT)

[0122] The experimental operation refers to Test Example 1, and the test time is 1 hour.

[0123] (2) Open field test (OFT)

[0124] The experimental operation was carried out as in Experimental Example 1, and a rat open field test system (100×100×40 cm) was selected for the test and the test was performed for 5 minutes.

[0125] (3) Forced swim test (FET)

[0126] Rats were placed in a forced swimming bucket (21×21×50cm, water depth of about 33cm) with a water temperature of about 25°C. The test time was 6 minutes, and the immobility time of the rats was recorded in the last 4 minutes. Behavioral standards: swimming—the rats swam around in the water; climbing—the rats paws paddled the water surface; struggling—the rats struggled violently with their limbs / the forelimbs were motionless, and the hind limbs kept paddling; immobility—the rats floated on the water surface with their limbs motionless / occasionally slid their limbs, with only the head above the water surface to breathe.

[0127] 2.3.2 Biochemical index detection

[0128] The rats were fasted but allowed to drink water for 12 hours the night before sampling. They were weighed 1 hour after the last administration. The rats were killed by dislocating the cervical vertebrae and dissected. The brains were taken out and 10% brain tissue homogenate was prepared. The 5-HT, DA, and NE in the brain homogenate were detected by enzyme-linked immunosorbent assay (ELISA).

[0129] 2.3.3 Histopathological examination

[0130] HE staining was used to detect the pathological changes of hippocampal tissues in each group of rats.

[0131] 2.4 Statistical analysis

[0132] SPSS 25.0 software was used for statistical analysis. Measurement data were expressed as mean ± standard deviation (x ± s), and one-way analysis of variance was used for comparison between groups. The Games-Howell method was used for unequal variances, and the LSD method was used for equal variances. Ridit test was used for rank data.

[0133] 3 Results

[0134] 3.1 Effect of the Chinese herbal essential oil of the present invention on the behavior of CUMS-induced depression model in rats

[0135] 3.1.1 Effect on body weight in CUMS-induced depression rat model

[0136] The results are shown in Table 4. Compared with the blank group, the body weight of rats in the model group was significantly reduced (P<0.01); compared with the model group, the body weight of rats in the high-dose group was significantly increased (P<0.01), and there was a trend of increasing the body weight of rats in the low-dose group (P>0.05).

[0137] Table 4 Changes in body weight of rats in each group before and after the experiment

[0138] Group Before drug intervention (g) After drug intervention (g) Blank Group 299.1±5.4 381.4±13.0 Model Group <![CDATA[281.2±8.8 △△ ]]> <![CDATA[336.9±21.6 △△ ]]> Fluoxetine hydrochloride group <![CDATA[281.8±6.8 △△ ]]> 343.2±20.8 High dose group <![CDATA[281.0±7.2 △△ ]]> <![CDATA[360.9±19.9 ** ]]> Low dose group <![CDATA[282.2±7.3 △△ ]]> 352.3±20.3

[0139] Note: Compared with the blank group, △△ P<0.01, △ P<0.05; compared with the model group, **P<0.01, *P<0.05

[0140] 3.1.2 Effects on sugar water preference rate in CUMS-induced depression model of rats

[0141] The sugar water preference rate of each group of SD rats was tested every two weeks, and the test results are shown in Table 5. In the second week of intervention, the sugar water preference rate of rats in the model group and each drug-treated group was significantly reduced compared with the blank group (P<0.01). In the fourth week of intervention, the sugar water preference rate of rats in the model group was significantly reduced compared with the blank group (P<0.01); compared with the model group, the sugar water preference rate of rats in the fluoxetine hydrochloride group, high-dose group, and low-dose group was significantly increased (P<0.01).

[0142] Table 5 Test results of sugar water preference rate of rats in each group

[0143] Group Intervention Week 2 (%) Intervention Week 4 (%) Blank Group 84.79±6.97 85.47±5.43 Model Group <![CDATA[63.76±9.30 △△ ]]> <![CDATA[59.65±10.31 △△ ]]> Fluoxetine hydrochloride group <![CDATA[66.61±6.27 △△ ]]> <![CDATA[75.13±8.71 ** ]]> High dose group <![CDATA[69.33±5.61 △△ ]]> <![CDATA[75.39±6.20 ** ]]> Low dose group <![CDATA[67.84±5.78 △△ ]]> <![CDATA[73.56±6.29 ** ]]>

[0144] 3.1.3 Effects on the open field test of CUMS-induced depression model in rats

[0145] On the morning of the 52nd-53rd day of the experiment (24th-25th day after administration), the rats in each group were tested in an open field test box of 100×100×40cm. The movement of the rats was judged by counting the movement distance and time of the rats in 5 minutes. The results are shown in Table 6. Compared with the blank group, the movement distance and time of the rats in the model group were significantly reduced (P<0.01). Compared with the model group, the high-dose and low-dose groups could significantly prolong the movement distance of the rats (P<0.05), and the fluoxetine hydrochloride group could significantly prolong the movement distance of the rats (P<0.01); compared with the model group, the high-dose group and the fluoxetine hydrochloride group could significantly increase the movement time of the rats (P<0.01), and the low-dose group could significantly increase the movement time of the rats (P<0.01).

[0146] Table 6 Test results of open field test for rats in each group

[0147] Group Movement distance (cm) Exercise time(s) Blank Group 3190.3±236.6 264.6±14.3 Model Group <![CDATA[2501.4±405.9 △△ ]]> <![CDATA[237.6±19.7 △△ ]]> Fluoxetine hydrochloride group <![CDATA[3159.0±347.2 ** ]]> <![CDATA[257.0±13.7 * ]]> High dose group <![CDATA[2911.9±375.5 * ]]> <![CDATA[261.8±13.9 ** ]]> Low dose group <![CDATA[2897.4±408.8 * ]]> <![CDATA[258.6±13.7 * ]]>

[0148] 3.1.4 Effects on the forced swimming test in the CUMS-induced depression model in rats

[0149] On the 54th day of the experiment (26th day after administration), each group of rats was subjected to forced swimming training, each forced swimming training for 15 minutes, and the rats rested for 24 hours (27th day after administration) before the formal test. The results are shown in Table 7. Compared with the blank group, the immobility time of the model group rats in the forced swimming test was significantly prolonged (P<0.01); compared with the model group, the high-dose group and the fluoxetine hydrochloride group could significantly shorten the immobility time of the rats (P<0.01), and the low-dose group could significantly shorten the immobility time of the rats (P<0.05). It is suggested that the Chinese medicine compound essential oil of the present invention can improve the desperate struggle of rats caused by CUMS.

[0150] Table 7 Test results of forced swimming test for rats in each group

[0151] Group Immobility time(s) Blank Group 62.5±14.4 Model Group <![CDATA[139.3±22.2 △△ ]]> Fluoxetine hydrochloride group <![CDATA[107.5±17.4 ** ]]> High dose group <![CDATA[105.9±24.9 ** ]]> Low dose group <![CDATA[111.4±26.2 * ]]>

[0152] 3.2 Effects of the Chinese herbal compound essential oil of the present invention on brain tissue biochemical indices in the CUMS-induced depression model of rats

[0153] The results are shown in Table 8. Compared with the blank group, the levels of DA, 5-HT, and NE in the brain tissue homogenate of the model group rats were significantly reduced (P<0.01), indicating that the monoamine neurotransmitters in the brain tissue of the model group rats were reduced. Compared with the model group, the high-dose group, the low-dose group and the fluoxetine hydrochloride group could significantly increase the DA level in the brain tissue of the rats (P<0.01); the high-dose group and the fluoxetine hydrochloride group could significantly increase the 5-HT and NE levels in the brain tissue of the rats (P<0.01), and the low-dose group could significantly increase the 5-HT and NE levels in the brain tissue of the rats (P<0.05).

[0154] Table 8 DA, 5-HT and NE levels in brain tissue homogenates of rats in each group

[0155] Group <![CDATA[DA(pg·ml -1 )]]> <![CDATA[5-HT(ng·ml -1 )]]> <![CDATA[NE(ng·ml -1 )]]> Blank Group 1483.8±119.7 131.59±5.62 122.17±6.72 Model Group <![CDATA[1310.8±71.9 △△ ]]> <![CDATA[110.55±3.18 △△ ]]> <![CDATA[103.14±7.00 △△ ]]> Fluoxetine hydrochloride group <![CDATA[1486.2±97.0 ** ]]> <![CDATA[124.56±5.76 ** ]]> <![CDATA[118.68±3.90 ** ]]> High dose group <![CDATA[1480.8±82.4 ** ]]> <![CDATA[126.48±6.08 ** ]]> <![CDATA[118.54±7.99 ** ]]> Low dose group <![CDATA[1428.1±84.7 ** ]]> <![CDATA[117.37±5.81 * ]]> <![CDATA[111.31±7.64 * ]]>

[0156] 3.3 Effect of the Chinese herbal essential oil of the present invention on pathological changes in the hippocampus of the rat depression model induced by CUMS

[0157] The arrangement and distribution of nerve cells in the hippocampus of each group of rats, as well as the degree of cell changes, were observed, and the pathological tissues were graded according to semi-quantitative standards. The results are shown in Table 9. Figure 1 As shown in the figure, Ridit test showed that compared with the blank group, the neurons in the hippocampus of the model group were scattered, with fewer cells, sunken and deformed neurons, cell lysis, blurred edges, and cytoplasmic staining (P<0.01); compared with the model group, the high-dose group and fluoxetine hydrochloride group could significantly improve the morphological and pathological changes of neurons in the hippocampus of the rats induced by CUMS (P<0.01), and the low-dose group could significantly improve the morphological and pathological changes of neurons in the hippocampus of the rats induced by CUMS (P<0.05).

[0158] Table 9 Results of hippocampal pathological changes in rats in each group

[0159] Group - + ++ +++ P Blank Group 10 0 0 0 <0.01 Model Group 0 1 5 4 - Fluoxetine hydrochloride group 1 8 1 0 <0.01 High dose group 2 5 3 0 <0.01 Low dose group 1 4 5 0 <0.05

[0160] 4 Conclusion

[0161] The above animal experimental results show that after the CUMS depression model rats are treated with the Chinese herbal compound essential oil of the present invention, the weight of the depression model rats can be significantly increased, the symptoms of anhedonia are well reversed and improved, the state of reduced rat movement and the behavioral state of despair and giving up struggle in the depression model rats are significantly improved; at the same time, the levels of DA, 5-HT and NE in the brain tissue of the depression model rats are significantly increased, the pathological changes in the hippocampus of the depression model rats are significantly improved, and the therapeutic effect of the Chinese herbal relapse essential oil of the present invention on the depression model rats is better than that of Western medicine.

[0162] 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 compound essential oil, characterized in that: The traditional Chinese medicine compound essential oil is composed of the following traditional Chinese medicine plant essential oils in parts by volume: 3-15 parts of benzoin essential oil, 2-10 parts of rose essential oil, 2-10 parts of citrus aurantium essential oil, 1-8 parts of white cardamom essential oil and 1-8 parts of fennel essential oil.

2. A Chinese medicine compound essential oil as claimed in claim 1, characterized in that: The traditional Chinese medicine compound essential oil is composed of the following traditional Chinese medicine plant essential oils in parts by volume: 5-10 parts of benzoin essential oil, 4-8 parts of rose essential oil, 4-8 parts of citrus aurantium essential oil, 1-5 parts of white cardamom essential oil and 1-5 parts of fennel essential oil.

3. A Chinese medicine compound essential oil as claimed in claim 1, characterized in that: The traditional Chinese medicine compound essential oil is composed of the following traditional Chinese medicine plant essential oils in parts by volume: 6 parts of benzoin essential oil, 5 parts of rose essential oil, 5 parts of citrus aurantium essential oil, 3 parts of white cardamom essential oil and 3 parts of fennel essential oil.

4. The Chinese medicinal compound essential oil according to any one of claims 1 to 3, characterized in that: Benzoin essential oil, rose essential oil, Citrus aurantium essential oil, white cardamom essential oil and fennel essential oil are extracted from corresponding Chinese medicinal plants by distillation or supercritical extraction, and the purity of the essential oil is above 99%.

5. Use of the Chinese herbal compound essential oil according to any one of claims 1 to 3 in the preparation of antidepressant products.

6. The use of the Chinese medicine compound essential oil as claimed in claim 5 in the preparation of an antidepressant product, characterized in that: The antidepressant product is a product used for aromatherapy, specifically, each single Chinese medicinal plant essential oil is weighed according to the above-mentioned volume fractions, mixed evenly, and then medically acceptable excipients are added to prepare a dosage form for administration by inhalation.

7. The use of the Chinese medicine compound essential oil as claimed in claim 5 in the preparation of an antidepressant product, characterized in that: In the antidepressant product, the effective content of the Chinese medicine compound essential oil is 5-15%.

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