Application of Kushui rose essential oil in preparation of medicine for treating depression

Abstract

The invention relates to the technical field of biology, in particular to application of Kushui rose essential oil in preparation of a medicine for treating depression, and main effective components of the Kushui rose essential oil are citronellol and geraniol; according to the Kushui rose essential oil, in two biochemical indexes of BDNF and Cort, along with the increase of the concentration of the Kushui rose essential oil, the anti-depression drug effect is more obvious, and a certain dose-effect relationship is shown. It is indicated that the Kushui rose essential oil possibly increases the BDNF and 5-HT level in the hippocampus by inhibiting HPA axial hyperfunction, and therefore hippocampus neurons are protected against stress damage; the Kushui rose essential oil can increase the ERK1 / 2 phosphorylation level by activating a BDNF receptor Tr kappa B and a downstream signal channel, so that the anti-depression effect is achieved; in a certain concentration range, the Kushui rose essential oil has no toxic effect on PC12 cells and has a wide application prospect.

Description

Technical Field

[0001] The invention relates to the field of biotechnology, and in particular to application of rosa tataricus essential oil in preparing a drug for treating depression. Background Art

[0002] Depression is a common mood disorder characterized by persistent sadness and a lack of pleasure. As the condition worsens, patients may experience feelings of guilt, worthlessness, and suicidal tendencies. Severe depression can even lead to cognitive impairment, including difficulty thinking, making decisions, and concentrating. Changes in appetite, weight, and sleep can also cause psychomotor retardation or agitation, severely impacting daily life and work. Existing treatments for depression primarily include three main types: psychotherapy, medication, and physical therapy. Mild depression can be significantly alleviated through psychological counseling. Commonly used psychotherapies include cognitive behavioral therapy, interpersonal therapy, acceptance and commitment therapy, and mindfulness-based cognitive therapy. Each therapy is designed based on different therapeutic concepts and has slightly different therapeutic goals. For patients with severe depression, clinicians generally use a combination of psychotherapy and medication to fully alleviate depressive symptoms. Commonly used antidepressants are second-generation antidepressants, which increase the levels of monoamine neurotransmitters in synapses and enhance signaling in monoaminergic neurons. However, these drugs have significant side effects: slow onset of action, limited effectiveness in some patients, numerous adverse reactions, and a high risk of relapse after discontinuation. Researchers have recently focused their attention on ketamine, a general anesthetic. Numerous studies have shown that ketamine can produce rapid antidepressant effects by antagonizing N-methyl-D-aspartate receptors, providing new insights into the development of fast-acting and effective antidepressants. Additionally, the anti-inflammatory drugs celecoxib and minocycline act on neuroimmune targets and are being used for patients with primary immune diseases and depression. Mifepristone antagonizes glucocorticoid receptors, inhibiting HPA axis hyperactivity and exhibiting some antidepressant effects. For patients with refractory depression, particularly those with comorbid psychiatric conditions or severe suicidal tendencies, electroconvulsive therapy (ECT) may be considered in certain circumstances. Transcranial magnetic stimulation (TMS) and transcranial electrical stimulation (TES) are recent developments in physical science. TES treats depression by stimulating specific brain regions to alter the excitability of corresponding neurons, while TMS uses magnetic fields of specific strength to induce currents in the central nervous system, thereby modulating neuronal electrophysiology to treat depression. In addition, there are some complementary treatments, such as some compounds with antidepressant activity, omerga-3 fatty acids, S-adenosyl-L-methionine, St. John's wort extract, 5-hydroxytryptophan, etc., regular physical exercise, acupuncture, etc.; but because the cause of depression has not been clearly explored, the current treatment methods have limited effects.

[0003] Essential oils are natural products extracted from aromatic plants. They contain numerous active ingredients that can be used to prevent and treat diseases, playing a significant role in modern alternative medicine. Rose dahliae essential oil, extracted from the rose, is a mixture. Rose dahliae essential oil contains a variety of aroma components, such as citronellol, nerol, geraniol, and linalool. Compared with commonly used damask rose essential oil, rose dahliae essential oil contains fewer aroma components like rose oxide and linalool, while containing higher levels of citronellol and geraniol, suggesting differences in the pharmacological activities of rose dahliae and damask rose essential oils. Previous research by our group has demonstrated the anti-Alzheimer's disease activity of rose dahliae essential oil using a nematode model. KERO inhibits abnormal Aβ deposition and downregulates the levels of highly toxic Aβ oligomers in nematodes, demonstrating superior pharmacological activity compared to the individual effects of citronellol and geraniol. Rose dahliae essential oil may exert its anti-AD effects by affecting the SKN-1 signaling pathway, demonstrating promising pharmacological activity in the treatment of psychiatric disorders. However, there are currently no studies showing that rose hip essential oil has antidepressant effects.

[0004] In response to the above technical problems, the present invention provides a novel use of Rosa rugosa essential oil in the preparation of antidepressant drugs. The antidepressant efficacy of the Rosa rugosa essential oil in two biochemical indicators, BDNF and Cort, increases with increasing concentration of the Rosa rugosa essential oil, demonstrating a dose-response relationship. This suggests that Rosa rugosa essential oil may protect hippocampal neurons from stress damage by inhibiting HPA axis hyperfunction and increasing BDNF and 5-HT levels in the hippocampus. Rosa rugosa essential oil may exert its antidepressant effect by activating the BDNF receptor TrκB and downstream signaling pathways, increasing ERK1 / 2 phosphorylation levels. Within a certain concentration range, Rosa rugosa essential oil has no toxic effect on PC12 cells. Summary of the Invention

[0005] The primary purpose of the present invention is to provide an application of bitter water rose essential oil in the preparation of antidepressant drugs, wherein the active ingredients of the bitter water rose essential oil include one or more of citronellol, geraniol, linalool, farnesol and pinoresinol.

[0006] Preferably, the active ingredients of the bitter water rose essential oil include citronellol, geraniol, linalool, farnesol and pinesinol.

[0007] Preferably, the effective ingredients of the bitter water rose essential oil account for more than 65% of the total amount of the bitter water rose essential oil.

[0008] Preferably, the effective ingredients of the bitter water rose essential oil account for more than 69% of the total amount of the bitter water rose essential oil.

[0009] The second object of the present invention is to provide a pharmaceutical composition comprising essential oil of Rosa rugosa and other active ingredients for treating depression.

[0010] Preferably, the pharmaceutical composition is added with pharmaceutically acceptable excipients to be prepared into an oral dosage form or an injection.

[0011] Preferably, the oral dosage form is tablets, capsules, granules, films, or sustained-release preparations.

[0012] The beneficial effects of the present invention are as follows: the present invention provides the use of bitter rose essential oil in the preparation of antidepressant drugs, and the bitter rose essential oil can improve the levels of 5-HT and Cort, and a high dose of bitter rose essential oil (0.05 mL / kg) can effectively reverse the decrease in BDNF. In the two biochemical indicators of BDNF and Cort, as the concentration of bitter rose essential oil increases, the antidepressant effect becomes more obvious, showing a certain dose-effect relationship. At the same time, the expression of BDNF signaling pathway proteins related to neuroprotection in the hippocampus tissue of mice was detected: compared with the blank group, the levels of TrκB and ERK1 / 2 in the hippocampus tissue of mice in the model group were significantly decreased; compared with the model group, the TrκB level of the bitter rose essential oil high-dose group was significantly increased, and the phosphorylation level of ERK1 / 2 was increased, but it could not reverse the decrease in ERK; in addition, there was no difference in CREB and its phosphorylation levels among the experimental groups. Moreover, within a certain concentration range, rose essential oil has no toxic effect on PC12 cells; cell experiment results show that the concentration of rose essential oil during the experiment should be less than or equal to 1 μL / mL, and rose essential oil has a protective effect on damage caused by Cort. The bitter water rose essential oil described in the present invention has broad application prospects in the field of antidepressant. BRIEF DESCRIPTION OF THE DRAWINGS

[0013] In order to more clearly illustrate the technical solution of the present invention, the following briefly introduces the drawings required for use in the description of the present invention. Obviously, the drawings described below are only some embodiments of the present invention. For ordinary technicians in this field, other drawings can be obtained based on these drawings without paying any creative work.

[0014] Figure 1 Effects of Rose hippocampus essential oil on 5-HT levels in the hippocampus of CUMS mice

[0015] Note: n=5, **p<0.01, compared with the blank group; #p<0.05, compared with the model group

[0016] Figure 2 Effects of Rosehip Oil on BDNF Levels in Hippocampus of CUMS Mice

[0017] Note: n=5, ** p<0.01, compared with the blank group; # p<0.05, ##p<0.01 compared with the model group

[0018] Figure 3 Effect of Rosehip Oil on Serum Cort Level in CUMS Mice

[0019] Note: n=8, * p<0.05, compared with the blank group; ### p<0.001 compared with the model group

[0020] Figure 4 Effect of Rose hippocampus essential oil on TrκB protein expression in hippocampus of CUMS mice Note: (A) Representative bands; (B) TrκB expression levels; n = 3, ** p<0.01, compared with the blank control group; # p<0.05, compared with the model group

[0021] Figure 5 Effects of Rose hippocampus essential oil on ERK1 / 2 and pERK1 / 2 protein expression in the hippocampus of CUMS mice Note: (A) Representative bands; (B) ERK1 / 2 expression levels; (C) pERK1 / 2 expression levels; n = 3, ** p<0.01, compared with the blank control group; # p<0.05, compared with the model group

[0022] Figure 6 Effects of Rose hippocampus essential oil on CREB and pCREB protein expression in the hippocampus of CUMS mice Note: (A) Representative bands; (B) CREB expression level; (C) pCREB expression level; n = 3

[0023] Figure 7 MTT assay was used to detect the cell viability of PC12 cells treated with 25-1600 μmol / L Cort in 10% HS + 5% FBS and 5% FBS; n = 3, compared with the control group: * p<0.05, ** p<0.01 Figure 8 The MTT assay was used to determine the effects of different doses of rose essential oil on the cell viability of PC12 cells cultured in 10% HS + 5% FBS and 5% FBS; n = 3, compared with the control group: * p<0.05, ** p<0.01 Figure 9 Effects of Rose hippocastanum essential oil on the viability of PC12 cells damaged by Cort; n=3, compared with the control group: * p<0.05, ** p<0.01, compared with the model group, #p<0.05, ## p<0.01. DETAILED DESCRIPTION

[0024] To make the objectives, features, and advantages of the present invention more readily apparent, the present invention is further described below in detail with reference to the accompanying drawings and embodiments. It should be understood that the embodiments described are only a portion of the embodiments of the present invention, not all of them. All other embodiments derived by persons of ordinary skill in the art based on the embodiments of the present invention without inventive effort are intended to fall within the scope of protection of the present invention.

[0025] The bitter water rose essential oil of the present invention is produced by Gansu Oriental Tianrun Rose Technology Co., Ltd.

[0026] Example 1: Determination of the components of Rose hippocastanum essential oil

[0027] The main components and their contents of the bitter water rose essential oil were detected by gas chromatography-mass spectrometry (GC-MS), providing a basis for the next step of exploring the main antidepressant active ingredients in the bitter water rose essential oil.

[0028] Table 1 Main components and contents of bitter water rose essential oil

[0029]

[0030]

[0031]

[0032]

[0033] The above tests show that the main components of bitter water rose essential oil are citronellol and geraniol; in addition, higher content of other essential oils include citronellol, cubebene, farnesol, cadinol, pinoresinol, etc.

[0034] Example 2: Antidepressant Effect of Rosehip Oil

[0035] 1. Experimental Materials and Instruments

[0036] 1.1 Animals

[0037] Male, SPF-grade Kunming mice, 6-8 weeks old, weighing 18-20 g, were purchased from the Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, with certificate number SCXK(GS)2020-0002. All mice were housed in an SPF-grade cleanroom at the College of Life Sciences, Lanzhou University, at room temperature of 24 ± 2°C, relative humidity of 50%-60%, with a 12-h light-dark cycle (8:00 AM–8:00 PM). They had free access to food and water and were caged six to a cage for one week before the experiment began.

[0038] 1.2 Reagents and Materials

[0039] Cort ELISA kit: Jiangsu ELISA Industrial Co., Ltd., MM-0061M1;

[0040] TrκB antibody: ABclonal, A2099;

[0041] ERK1 / 2 antibody: Proteintech, 11257-1-AP;

[0042] p-ERK1 / 2 antibody: Immunoway, YP0101;

[0043] CREB antibody: Proteintech, 12208-1-AP;

[0044] p-CREB antibody: Proteintech, 28792-1-AP;

[0045] GAPDH antibody: Proteintech, 10494-1-AP;

[0046] Rabbit-IgG antibody: Proteintech, SA00001-2;

[0047] High-efficiency RIPA tissue / cell lysis buffer: Beijing Solebow Technology Co., Ltd., R0010;

[0048] Protease inhibitor cocktail: AbMole Bioscience, M5283;

[0049] Phosphatase Inhibitor Cocktail (A, B): M7528;

[0050] BCA protein detection kit: Beijing Solebow Technology Co., Ltd., PC0020;

[0051] Universal antibody diluent: NCM Biotechnology Co., Ltd., WB100D;

[0052] ECL chemiluminescent solution: Millpore, Germany, WBKLS0100;

[0053] Membrane regeneration fluid: Kangwei Century Biotechnology Co., Ltd., CW0056.

[0054] 1.3 Experimental instruments

[0055] Fully automatic chemiluminescence imaging analysis system: Beijing Yuanpinghao Biotechnology Co., Ltd., model Tanon-5200.

[0056] 2. Experimental Methods

[0057] 2.1 Grouping and treatment

[0058] After one week of cohabitation, the mice were randomly divided into six groups: a normal group, a model group, a positive control group, a low-dose Rose d'Argentia essential oil group, a medium-dose Rose d'Argentia essential oil group, and a high-dose Rose d'Argentia essential oil group. Except for the normal group, the other five groups of mice were housed in solitary cages and subjected to a CUMS-induced model for seven weeks, with an unpredictable mild stimulus administered daily. Drug administration and stimulation were administered concurrently starting in the third week of modeling, with experiments conducted one hour after drug administration.

[0059] The groups and treatment methods are shown in Table 2.

[0060] Table 2 Grouping and treatment methods

[0061]

[0062] The model group was gavaged daily with normal saline; the positive control group was gavaged daily with 10 mg / kg of fluoxetine (fluoxetine dissolved in normal saline to form a suspension); and each group receiving R. truncatum essential oil was gavaged daily with a specific dose of essential oil (0.05 mL / kg for the low-dose group, 0.1 mL / kg for the medium-dose group, and 0.2 mL / kg for the high-dose group. The R. truncatum essential oil was dissolved in normal saline containing 1% Tween-80 and diluted to the corresponding concentration for each dose. The solution was shaken and pipetted before gavage to form a milky white emulsion). The dose was administered once daily, with each gavage volume being 0.1 mL / 10 g, for a total of 35 days.

[0063] 2.2 CUMS model establishment

[0064] Mice in the model and fluoxetine groups were housed individually in cages and randomly assigned to a mild stimulus daily according to an Excel-generated random number table. These included 24-hour food deprivation (A), 24-hour water deprivation (B), 24-hour day / night reversal (C), 24-hour crowding (D), 24-hour exposure to foreign objects (E), 5-minute swimming in 45°C hot water (F), 2-minute tail clamping (G), 2-hour restraint (H), 5-minute swimming in 4°C ice water (I), and 24-hour humidity (J). The same stimulus was not presented on two consecutive days. All experiments were conducted between 9:00 and 11:00 AM daily. Detailed experimental procedures are shown in Table 3.

[0065] Table 3 CUMS experimental operation

[0066]

[0067] 2.3 Sample collection and preservation

[0068] On the 53rd day after the start of the experiment, the mice were weighed, blood was collected from the eyeballs, placed in labeled 2mL sterile centrifuge tubes, and placed in a 4°C refrigerator overnight. During the process, avoid shaking and oscillation to prevent hemolysis. The next day, the separated serum was aspirated with a small-scale pipette, aliquoted, and stored in a -80°C refrigerator. The mice were killed by cervical dislocation, and the hippocampal tissue of the mice was quickly dissected and removed. The removed hippocampal tissue was placed in a labeled sterile centrifuge tube and placed on ice. After all the hippocampi of the mice were collected, the hippocampal tissue was placed in a -80°C refrigerator overnight.

[0069] 2.4ELISA test

[0070] (1) Preparation before testing: prepare enough crushing beads in advance, clean them with methanol and distilled water, dry them, and pre-cool them in a -80℃ refrigerator. Open the freezer grinder and pre-cool it to 4℃ before crushing. Add 0.5mL of 0.9% saline to each centrifuge tube containing hippocampal tissue samples, place a crushing bead, set the frequency to 60Hz, grind for 45s, and start grinding. Centrifuge the ground tissue homogenate at 4℃, 12000rpm for 5min, collect the supernatant and divide it into packages, and store it in a -80℃ refrigerator. Before starting the ELISA test, take out the detection reagents, samples and standards in advance and equilibrate them at room temperature for 15-30min.

[0071] (2) ELISA test steps: Dilute the sample and standard according to the instructions and add them to the wells of the enzyme-labeled plate. After sealing the plate with a film, place it in a 37°C oven for 30 minutes; after pouring out the liquid in the wells, wash the plate 5 times with the diluted washing solution (add 400 μl of washing solution to each well, let it stand for 30 seconds, pour it out, and tap the liquid in the wells clean). Then, add 100 μl of enzyme-labeled reagent to each well and place it in a 37°C oven for 30 minutes; pour out the liquid in the wells and wash the plate 5 times with the diluted washing solution. Then, add 50 μl of color development solution A and 50 μl of color development solution B, place it in a 37°C oven for color development for 10 minutes, add 50 μl of stop solution, and use a microplate reader to read the optical density (OD value) of each well at a wavelength of 450 nm within 15 minutes. A standard curve is established by the OD value and concentration of the standard to determine the concentration of the analyte in each well sample. Multiply it by the sample dilution factor to obtain the final concentration of the analyte in the sample.

[0072] 2.5 Western Blot detection of changes in the TrκB-ERK-CREB signaling pathway

[0073] (1) Total protein extraction and preparation: Prepare RIPA lysis buffer containing a final concentration of 1 mM protease inhibitor and phosphatase inhibitor (Phosphatase Inhibitor Cocktail A, B) in advance, place the mouse hippocampus tissue in a 2 mL EP tube, add an appropriate amount of lysis buffer, set the freeze grinder to 60 Hz and grind for 45 seconds, then place the tissue homogenate on ice for 30 minutes, vortex once every 10 minutes to ensure that the tissue is fully lysed. Subsequently, place the tissue homogenate in a low-temperature ultracentrifuge, centrifuge at 4°C, 12000 rpm for 30 minutes, collect the supernatant, use the BCA method to detect protein concentration, and mix the supernatant with the loading buffer at a ratio of 4:1. Heat in a metal bath at 100°C for 10 minutes to denature the protein. After the deformation is completed and the temperature is lowered, vortex the protein sample again, divide it into 1.5 mL EP tubes, and store it in a -80°C refrigerator.

[0074] (2) Western Blot Step: Prepare 12% separation gel and 4% stacking gel, load 20-40 μg of protein, and perform SDS-PAGE electrophoresis. Set the starting voltage to 80 V. After all samples are flattened to the same line, adjust the voltage to 120 V to separate proteins of different molecular weights. After the electrophoresis is completed, refer to the molecular weight indicated by the marker, cut off the gel containing the test protein, and cut a PVDF membrane of the same size. Soak the membrane in methanol for 30 seconds to activate it. Immerse the lab equipment required for transfer in transfer solution. Assemble the transfer cassette using the sandwich method (positive plate, two layers of sponge pad, two layers of filter paper, PVDF membrane, SDS-PAGE gel, two layers of filter paper, two layers of sponge pad, negative plate), removing any air bubbles during stacking. Place the assembled transfer cassette into the transfer tank, aligned with the positive and negative electrodes. Pour the transfer solution in. Pre-cool the entire transfer apparatus in ice water. Transfer the membrane at low temperatures to offset the exothermic heat generated during transfer. Transfer conditions are 100V or 300mA for 1 hour 45 minutes. After transfer, soak the PVDF membrane in 5% skim milk powder in TBST and block it for 1 hour with gentle agitation on a shaker. After blocking, wash the membrane twice with TBST for 5 minutes each, then soak the PVDF membrane in the corresponding primary antibody and incubate overnight at 4°C. After incubation, remove the primary antibody, wash the membrane three times with TBST for 10 minutes each, and then incubate with the secondary antibody at room temperature for 1 hour. After incubation, the membrane was washed again with TBST three times for 10 minutes each time. ECL luminescent solution was then added dropwise to evenly cover the membrane surface. The bands were exposed using an automated chemiluminescence analyzer to observe the depth of the protein bands and take photos. The grayscale values ​​of the bands were measured using Image J software.

[0075] 3. Statistical methods

[0076] Experimental data were recorded as mean ± SEM and processed using SPSS 19.0 statistical software. One-way ANOVA was used to compare data between groups, with group status as the intergroup factor. Statistical differences were considered significant when p < 0.05. GraphPad Prism 8.0 was used for plotting.

[0077] 4. Experimental Results

[0078] 4.1 Effect of Rose hippocampus essential oil on 5-HT content in the hippocampus of depressed mice

[0079] The hippocampus tissue of mice was taken and the 5-HT level was detected. The results were as follows: Figure 1 As shown. Compared with the blank group, the model group showed a significant decrease in 5-HT levels (p < 0.01), and the model group mice exhibited monoamine neurotransmitter disorders. Compared with the model group, 5-HT levels in all treatment groups were significantly increased. 5-HT levels in the low-, medium-, and high-dose fluoxetine and rosa tataricus essential oil groups increased by 51.16%, 51.68%, 68.56%, and 50.86%, respectively (p < 0.05). 5-HT levels in all rosa tataricus essential oil dose groups were similar to those in the fluoxetine group, and 5-HT levels were similar across dose groups, with no dose-response relationship observed. This suggests that rosa tataricus essential oil can improve the decrease in 5-HT levels caused by long-term stress, with an effect similar to that of fluoxetine.

[0080] 4.2 Effect of Rosehip Oil on BDNF Content in Hippocampus of Depressed Mice

[0081] The BDNF level in the mouse hippocampus was detected. Figure 2 As shown. Compared with the blank group, the model group showed a significant decrease in BDNF levels (p < 0.01). Compared with the model group, the fluoxetine group and the high-dose Rose dahliae essential oil group showed significant increases in BDNF levels, by 112.94% (p < 0.05) and 140.79% (p < 0.01), respectively. The medium- and low-dose Rose dahliae essential oil groups showed no significant differences, and the high-dose Rose dahliae essential oil group showed higher BDNF levels than the fluoxetine group. This suggests that 0.05 mL / kg of Rose dahliae essential oil can significantly reverse the decrease in BDNF levels caused by long-term stress, and its efficacy is superior to fluoxetine.

[0082] 4.3 Effect of Rosehip Oil on Serum Cort Content in Depressed Mice

[0083] We took the serum of mice after drug treatment and tested the Cort concentration. The results are as follows Figure 3 shown.

[0084] Compared with the blank group, the model group showed a significant increase in Cort levels (p < 0.05), indicating that the model group mice exhibited HPA axis hyperfunction. Compared with the model group, Cort levels decreased by 23.42%, 24.51%, 25.32%, and 35.50% in the low-, medium-, and high-dose groups of fluoxetine and R. truncatum essential oil, respectively (p < 0.001). The high-dose R. truncatum essential oil group (0.05 mL / kg) showed the most significant improvement. Cort levels gradually decreased with increasing R. truncatum essential oil concentrations. Compared with the fluoxetine group, the 0.05 mL / kg R. truncatum essential oil group showed lower Cort levels. This suggests that R. truncatum essential oil can improve elevated Cort levels caused by long-term stress, exhibiting a dose-response relationship and superior efficacy to fluoxetine.

[0085] 4.4 Effects of Rosehip Oil on the BDNF Signaling Pathway in the Hippocampus of Depressed Mice

[0086] The expression of BDNF receptor TrκB protein in the hippocampus of mice in each group was detected. Figure 4 As shown in the results, compared with the blank group, TrκB levels in the hippocampus of mice in the model group were significantly decreased (p < 0.01); compared with the model group, TrκB levels in the high-dose group treated with R. oleracea essential oil were significantly increased (p < 0.05). This suggests that the BNDF receptor TrκB levels in the hippocampus of depressed mice are significantly decreased, and administration of 0.05 mL / kg of R. oleracea essential oil can reverse the decrease in TrκB and restore it to normal levels.

[0087] The expression and phosphorylation of ERK1 / 2 proteins in the BDNF downstream pathway in the hippocampus of mice in each group were detected. Figure 5 As shown in the figure, compared with the blank group, ERK1 / 2 levels in the hippocampus of mice in the model group were significantly decreased (p<0.01). Compared with the model group, ERK1 / 2 phosphorylation levels were significantly increased in the medium- and high-dose groups of R. oleracea essential oil. This suggests that ERK1 / 2 and its phosphorylation levels are decreased in depressed mice. Administration of 0.005 kg / mL and 0.05 kg / mL of R. oleracea essential oil increased ERK phosphorylation levels and restored them to normal levels, but did not reverse the decrease in ERK1 / 2.

[0088] The expression and phosphorylation of CREB protein in the BDNF downstream pathway in the hippocampus of mice in each group were detected. Figure 6 As shown in the figure, the changes in CREB and its phosphorylation levels in each experimental group were not statistically significant, indicating that there were no differences in CREB and its phosphorylation levels among the experimental groups.

[0089] Example 3: Cell experiment on the antidepressant effect of rose hip essential oil

[0090] 1. Construction of an in vitro cell model of depression

[0091] In order to select the optimal concentration of corticosterone to induce PC12 cells, the in vitro cell model was constructed and PC12 cells were treated with different concentrations of corticosterone for 24 hours. Figure 7 As shown, the inhibitory effect of 25-1600 μmol / L Cort on PC12 cell viability in a dose-dependent manner was observed in PC12 cells cultured in 10% HS + 5% FBS. Cell viability decreased to 32.05% ± 20.53% (p < 0.01) after exposure to 800 μmol / L. Furthermore, after exposure to different concentrations of corticosterone for 24 hours in PC12 cells cultured in 5% FBS, cell viability showed a dose-dependent relationship with corticosterone concentration, with 800 μmol / L decreasing to 43.93% ± 4.73% (p < 0.01). Based on experimental data and references, 600 μmol / L Cort was selected as the modeling concentration.

[0092] 2. Safe concentration of bitter water rose essential oil

[0093] MTT assay of PC12 cells treated with different concentrations of rose essential oil (0.01-1 μL / mL) Figure 8 As shown in the results, after 24 hours, there was no significant difference in the survival rate of PC12 cells compared to the blank control group (p>0.05), indicating that rose essential oil had no significant toxic effect on PC12 cells. Statistical analysis showed that when the concentration of rose essential oil was greater than 2 μL / mL, the presence of rose essential oil had an effect on cell viability, and the difference was statistically significant (p<0.01), indicating that within a certain concentration range, rose essential oil has no toxic effect on PC12 cells. Therefore, the concentration of rose essential oil in the experiment should be less than or equal to 1 μL / mL.

[0094] 3. Effects of Rosehip Oil on an In Vitro Cell Model of Depression

[0095] The MTT results of rose essential oil on the damage caused by Cort are as follows Figure 9 As shown in the results, compared with the control group, the survival rate of PC12 cells in the 600 μmol / L Cort model group decreased significantly to 40.37%±20.46% (p<0.01), indicating that corticosterone can cause damage to PC12 cells and the model was successfully established. Compared with the Cort model group, the survival rates of PC12 cells in the 0.01, 0.05, 0.1, 0.5, and 1 μL / mL rose essential oil treatment groups increased from the model group to (65.50±14.65)%, (83.41±7.91)%, (85.71±4.72)%, (83.97±13.97)%, and (68.02±15.75)%, respectively. The results showed that rose essential oil had a protective effect on the damage caused by Cort (p>0.05).

[0096] In summary, the present invention provides the use of bitter water rose essential oil in the preparation of antidepressant drugs. The bitter water rose essential oil can improve 5-HT and Cort levels, and a high dose of bitter water rose essential oil (0.05mL / kg) can effectively reverse the decrease in BDNF. In the two biochemical indicators of BDNF and Cort, as the concentration of bitter water rose essential oil increases, the antidepressant effect becomes more obvious, showing a certain dose-effect relationship. At the same time, the expression of BDNF signaling pathway proteins related to neuroprotection in mouse hippocampal tissue was detected: compared with the blank group, the levels of TrκB and ERK1 / 2 in the hippocampal tissue of mice in the model group were significantly decreased; compared with the model group, the TrκB level in the bitter water rose essential oil high-dose group was significantly increased, and the phosphorylation level of ERK1 / 2 was increased, but it could not reverse the decrease in ERK; in addition, CREB and its phosphorylation levels did not show differences in the experimental groups. Moreover, within a certain concentration range, rose essential oil has no toxic effect on PC12 cells; cell experiment results show that the concentration of rose essential oil during the experiment should be less than or equal to 1 μL / mL, and rose essential oil has a protective effect on damage caused by Cort. The bitter water rose essential oil described in the present invention has broad application prospects in the field of antidepressant.

Claims

1. The application of rosa ovata essential oil in the preparation of antidepressant drugs, characterized in that: The effective ingredients of the bitter water rose essential oil include one or more of citronellol, geraniol, linalool, farnesol and pinoresinol.

2. The use according to claim 1, characterized in that The effective ingredients of the bitter water rose essential oil include citronellol, geraniol, linalool, farnesol and pinoresinol.

3. The use according to claim 2, characterized in that The effective components of the bitter water rose essential oil account for more than 65% of the total amount of the bitter water rose essential oil.

4. The use according to claim 3, characterized in that The effective components of the bitter water rose essential oil account for more than 69% of the total amount of the bitter water rose essential oil.

5. A pharmaceutical composition, characterized in that The pharmaceutical composition comprises rosa tataricus essential oil and other active ingredients for treating depression.

6. The pharmaceutical composition according to claim 5, characterized in that The pharmaceutical composition is added with pharmaceutically acceptable excipients to be prepared into an oral dosage form or an injection.

7. The pharmaceutical composition according to claim 6, characterized in that The oral dosage forms are tablets, capsules, granules, films and sustained-release preparations.

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