Agilawood compound traditional Chinese medicine composition for relieving depression and preparation method thereof

Abstract

The invention relates to the technical field of plant extraction, and particularly provides an agilawood compound traditional Chinese medicine composition for relieving depression and a preparation method of the agilawood compound traditional Chinese medicine composition. The agilawood compound traditional Chinese medicine composition comprises the following components in parts by mass: 20-30 parts of an agilawood extract, 5-8 parts of a mountain radish flower extract, 5-10 parts of a radix bupleuri extract and 5-10 parts of a radix scutellariae extract. Compared with the prior art, the Chinese eaglewood compound traditional Chinese medicine composition disclosed by the invention has a remarkable inhibition effect on pressure index cortisol. The embodiment of the invention proves that compared with a contrast, the agilawood compound traditional Chinese medicine composition has a better effect of inhibiting cortisol, and the inhibition rate reaches 69.7-71.6%. Therefore, the agilawood compound traditional Chinese medicine composition can be used as an active component to be widely applied to various foods and drinks subsequently, and the pressure problem caused by working pressure, staying up late, computer radiation, negative emotion and the like is relieved.

Description

Technical Field

[0001] The present invention relates to the technical field of plant extraction, and more specifically to an agarwood compound Chinese medicine composition for relieving depression and a preparation method thereof. Background Art

[0002] Agarwood is one of the commonly used medicines in traditional Chinese medicine. It comes from the resinous wood of Aquilaria agallocha Roxb. and Aquilaria sinensis (Lour.) Gilg. Agarwood has a pungent and bitter flavor and is slightly warm in nature. It has the effects of promoting qi and relieving pain, warming the middle and stopping vomiting, and relieving asthma.

[0003] Qi stagnation is a physical condition that is formed due to long-term emotional depression and qi stagnation, with introverted and unstable personality, melancholy and fragility, sensitivity and suspicion as the main manifestations. The tendency of Qi stagnation is prone to depression, visceral agitation, lily disease, etc., and is easily stimulated by the mind. Such patients are in a state of Qi stagnation for a long time, which is also a normal state. However, when in a disease state, the tendency of the physical constitution will make the patient pathologically more prone to liver Qi stagnation, phlegm stagnation turning into heat, Qi stagnation and blood stasis, Qi stagnation and phlegm accumulation, liver depression and spleen deficiency, etc.

[0004] Cortisol is a hormone secreted by the adrenal glands and is often referred to as the "stress hormone." It plays an important role in coping with stress and regulating a variety of physiological functions. High levels of cortisol may interfere with the balance of neurotransmitters (such as serotonin and norepinephrine) that are closely related to mood regulation, leading to depression. Cortisol's regulatory effects on the immune system may also affect the development of depression, as chronic inflammation is often associated with depressive symptoms.

[0005] Long-term excessive secretion of cortisol can have adverse effects on health. Currently, there is no Chinese herbal composition that can inhibit the production of cortisol. Summary of the invention

[0006] In view of this, the object of the present invention is to provide an agarwood compound Chinese medicine composition for relieving depression and a preparation method thereof. The agarwood compound Chinese medicine composition provided by the present invention can effectively inhibit cells from secreting cortisol, thereby relieving depression.

[0007] In order to achieve the above object, the present invention adopts the following technical solutions:

[0008] A composite agarwood Chinese medicinal composition for relieving depression, comprising, by weight, 20 to 30 parts of agarwood extract, 5 to 8 parts of chervil flower extract, 5 to 10 parts of bupleurum extract and 5 to 10 parts of scutellaria baicalensis extract.

[0009] Mountain Radish, a Chinese herbal medicine. This product is Scabiosa comosa Fisch.ex Roem.et Schult. of the Sichuan Dipsacus family, and its flowers are used as medicine. Functions and indications: clearing away heat and purging fire. It is mainly used to treat liver-fire headache, fever, lung-heat cough, and jaundice.

[0010] Preferably, the preparation method of the agarwood extract comprises:

[0011] (1) extracting agarwood with an extraction solvent, drying, and obtaining agarwood powder;

[0012] (2) Using supercritical fluid to extract the agarwood powder, drying it, and obtaining the agarwood extract.

[0013] Preferably, the solid-liquid ratio of the agarwood and the extraction solvent is 1: (25-30) g / ml.

[0014] Preferably, powdered agarwood is extracted with an extraction solvent; the agarwood powder is preferably passed through a 60-80 mesh sieve, preferably a 80 mesh sieve; the solid-liquid ratio of the agarwood and the extraction solvent is 1: (25-30) g / ml; the extraction temperature is 80-100°C, and the time is 60-80 min; the extraction solvent includes an alcohol extraction solvent, preferably ethanol.

[0015] In one embodiment of the present invention, agarwood is mixed with an extraction solvent, and the solid and liquid are separated to obtain a filter residue and a first filtrate, the filter residue is mixed with an extraction solvent, and the solid and liquid are separated to obtain a second filtrate, and the first filtrate and the second filtrate are mixed to obtain an agarwood extract.

[0016] Preferably, before mixing the agarwood with the extraction solvent, the agarwood is dried; the drying temperature is preferably 50-60°C, more preferably 60°C, and the drying time is preferably 15-20 min, more preferably 15 min.

[0017] Preferably, after extracting agarwood with an extraction solvent to obtain an agarwood extract, the agarwood extract is dried at a temperature of 40 to 50° C. for 5 to 6 hours.

[0018] Preferably, a supercritical fluid is used to extract the agarwood powder, and the supercritical fluid includes carbon dioxide; the extraction temperature is 30-50°C, the pressure is 30-40MPa, and the time is 2-5h.

[0019] Preferably, the preparation method of the chervil flower extract comprises:

[0020] The serrulata flower is crushed, mixed with water, distilled under reduced pressure, and dried to obtain a serrulata flower extract.

[0021] Preferably, the preparation method of the chervil flower extract comprises:

[0022] Use dried scabiosa flowers (Scabiosa comosa Fisch.ex Roem.et Schult.), rinse the flowers gently with clean water to remove impurities and dust, and then coarsely grind them. Put the grinded flowers into an extractor, add water 10 times the mass of the scabiosa flowers, heat to boiling, keep it for 1 hour, then filter out the liquid, distill the extract under reduced pressure, and dry it at 80°C to obtain a scabiosa flower extract.

[0023] Preferably, the preparation method of the Bupleurum extract comprises:

[0024] Mix bupleurum root with water, soak for 50-70 minutes to obtain the medicine to be decocted; decoct the medicine to be decocted for 50-70 minutes, filter and obtain the filtrate; centrifuge the filtrate, take the supernatant, and dry to obtain the bupleurum root extract.

[0025] Preferably, the preparation method of the Bupleurum extract comprises:

[0026] Take 5g of Bupleurum chinensie DC., wash impurities with ultrapure water, add 200g of ultrapure water, and soak for 60 minutes. Boil in an electric stove, and then decoct for 60 minutes. Filter through 8 layers of gauze, transfer the filtrate to a 50mL centrifuge tube, centrifuge the filtrate at 3000rpm for 35 minutes, take the supernatant, filter through a 0.22μm microporous filter into a sterile centrifuge tube, and dry at 80°C to obtain Bupleurum extract.

[0027] Preferably, the preparation method of the Scutellaria baicalensis extract comprises:

[0028] Mix scutellaria baicalensis with water, soak for 50-70 minutes to obtain the medicine to be decocted; decoct the medicine to be decocted for 50-70 minutes, filter and obtain the filtrate; centrifuge the filtrate, take the supernatant, and dry to obtain the scutellaria baicalensis extract.

[0029] In one embodiment of the present invention, the method for preparing the scutellaria baicalensis extract comprises:

[0030] Take 2g of Scutellaria baicalensis Georgi, wash impurities with ultrapure water, add 100g of ultrapure water, and soak for 60min. Boil in an electric stove, and then decoct for 60min. Filter through 8 layers of gauze, transfer the filtrate to a 50mL centrifuge tube, centrifuge the filtrate at 3000rpm for 35min, take the supernatant, filter through a 0.22μm microporous filter into a sterile centrifuge tube, and dry at 80℃ to obtain the Scutellaria baicalensis extract.

[0031] The present invention also provides the use of the agarwood compound Chinese medicinal composition for relieving depression in the preparation of a product for preventing and / or treating the inhibition of cortisol secretion.

[0032] Preferably, the concentration of the agarwood compound traditional Chinese medicine composition is 10-20wt%, preferably 10wt%.

[0033] The present invention also provides the use of the agarwood compound Chinese medicine composition for relieving depression in the preparation of products for preventing and / or treating anti-inflammatory diseases.

[0034] Preferably, the anti-inflammatory effect includes inhibiting inflammatory factors; the inflammatory factors include IL-6.

[0035] The agarwood compound Chinese medicine composition of the present invention has a significant inhibitory effect on the stress index cortisol. It is confirmed by the examples of the present invention that compared with the comparative example, the agarwood compound Chinese medicine composition has a better effect of inhibiting cortisol, and the inhibition rate reaches 69.7% to 71.6%. Therefore, the agarwood compound Chinese medicine composition of the present invention can be widely used as an active ingredient in various foods and beverages to relieve stress problems caused by work pressure, staying up late, computer radiation, and negative emotions. DETAILED DESCRIPTION

[0036] The technical solution of the present invention will be clearly and completely described below in conjunction with the embodiments of the present invention. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0037] In order to further illustrate the present invention, the following examples are used to explain it in detail. The raw materials used in the following examples and comparative examples of the present invention are all commercially available products.

[0038] The preparation method of the agarwood extract of the embodiment of the present invention and the comparative example comprises the following steps:

[0039] (1) Extraction: After drying agarwood (Aquilaria spp.) in a 50°C oven for 20 min, the agarwood was crushed using a traditional Chinese medicine grinder and passed through an 80-mesh sieve. Ethanol was added to a certain mass of the powder with a solid-liquid ratio of 1:25. The powder was then extracted in an 80°C oil bath for 80 min, and then filtered using a circular filter paper. The filtrate was set aside. Ethanol was added to the residue again, and the extraction was repeated once under the above conditions. The residue was then filtered using a circular filter paper. The two extraction filtrates were combined and placed in a constant temperature drying oven and dried at 50°C for 6 h to obtain agarwood powder.

[0040] (2) In a supercritical extraction device, agarwood powder was extracted using carbon dioxide supercritical fluid at an extraction temperature of 40°C and a pressure of 30 MPa. The flow rate of the supercritical carbon dioxide fluid was set to 300 L / h. After 3 hours of extraction, the agarwood extract was obtained by drying at 80°C.

[0041] The preparation method of the chervil flower extract of the embodiment and the comparative example of the present invention comprises the following steps:

[0042] Use dried scabiosa flowers (Scabiosa comosa Fisch.ex Roem.et Schult.), rinse the flowers gently with clean water to remove impurities and dust, and then coarsely grind them. Put the grinded flowers into an extractor, add water 10 times the mass of the scabiosa flowers, heat to boiling, keep it for 1 hour, then filter out the liquid, distill the extract under reduced pressure, and dry it at 80°C to obtain a scabiosa flower extract.

[0043] The preparation method of the Bupleurum extract of the embodiment and the comparative example of the present invention comprises the following steps:

[0044] Take 5g of Bupleurum chinensie DC., wash impurities with ultrapure water, add 200g of ultrapure water, and soak for 60 minutes. Boil in an electric stove, and then decoct for 60 minutes. Filter through 8 layers of gauze, transfer the filtrate to a 50mL centrifuge tube, centrifuge the filtrate at 3000rpm for 35 minutes, take the supernatant, filter through a 0.22μm microporous filter into a sterile centrifuge tube, and dry at 80°C to obtain Bupleurum extract.

[0045] The preparation method of the scutellaria baicalensis extract of the embodiment and the comparative example of the present invention comprises the following steps:

[0046] Take 2g of Scutellaria baicalensis Georgi, wash impurities with ultrapure water, add 100g of ultrapure water, and soak for 60min. Boil in an electric stove, and then decoct for 60min. Filter through 8 layers of gauze, transfer the filtrate to a 50mL centrifuge tube, centrifuge the filtrate at 3000rpm for 35min, take the supernatant, filter through a 0.22μm microporous filter into a sterile centrifuge tube, and dry at 80℃ to obtain the Scutellaria baicalensis extract.

[0047] Example 1

[0048] The agarwood compound Chinese medicine composition of this embodiment is composed of the following components: 20g of agarwood extract, 8g of chervil flower extract, 10g of bupleurum extract and 10g of scutellaria baicalensis extract.

[0049] The preparation method of the above-mentioned agarwood compound Chinese medicine composition comprises: mixing the various raw materials according to proportions, stirring evenly, and obtaining the agarwood compound Chinese medicine composition.

[0050] Comparative Example 1

[0051] The agarwood compound Chinese medicine composition of this comparative example is different from that of Example 1 in that this comparative example only includes agarwood extract, mountain radish flower extract and bupleurum extract, and the missing amount of scutellaria baicalensis extract is distributed to the corresponding components according to the proportion of other substances.

[0052] Comparative Example 2

[0053] The agarwood compound Chinese medicine composition of this comparative example is different from that of Example 1 in that this comparative example only includes agarwood extract, mountain radish flower extract and scutellaria baicalensis extract, and the missing amount of bupleurum extract is distributed to the corresponding components according to the proportion of other substances.

[0054] Comparative Example 3

[0055] The agarwood compound Chinese medicine composition of this comparative example is different from that of Example 1 in that this comparative example only includes agarwood extract, scutellaria baicalensis extract and bupleurum extract, and the missing amount of chervil flower extract is distributed to the corresponding components according to the proportion of other substances.

[0056] Comparative Example 4

[0057] The agarwood compound Chinese medicine composition of this comparative example is different from that of Example 1 in that this comparative example only includes Scutellaria baicalensis extract, Serissa serrata flower extract and Bupleurum chinense extract, and the missing amount of agarwood extract is distributed to the corresponding components according to the proportion of other substances.

[0058] Example 2

[0059] The agarwood compound Chinese medicine composition of this embodiment is composed of the following components: 30g of agarwood extract, 5g of chervil flower extract, 5g of bupleurum extract and 5g of scutellaria baicalensis extract.

[0060] The preparation method of the above-mentioned agarwood compound Chinese medicine composition comprises: mixing the various raw materials according to proportions, stirring evenly, and obtaining the agarwood compound Chinese medicine composition.

[0061] Example 3

[0062] The agarwood compound Chinese medicine composition of this embodiment is composed of the following components: 25g of agarwood extract, 6g of chervil flower extract, 7g of bupleurum extract and 10g of scutellaria baicalensis extract.

[0063] The preparation method of the above-mentioned agarwood compound Chinese medicine composition comprises: mixing the various raw materials according to proportions, stirring evenly, and obtaining the agarwood compound Chinese medicine composition.

[0064] Test Example 1

[0065] 1. Culture medium and solution preparation

[0066] (1) DMEM complete medium: Add FBS 10% / double antibody 1% to DMEM medium

[0067] (2) LPS: Use 1× PBS buffer as diluent to prepare a 1 mg / ml LPS solution.

[0068] 2. Effects of test samples on cortisol secretion by HaCat cells

[0069] (1) Cell seeding: 10,000 HaCat cells were seeded into a 96-well plate and cultured in an incubator.

[0070] (2) Experimental grouping: a negative control group, an agonist positive control group, an inhibitor positive control group and a sample group were set up. The sample group included the agarwood compound Chinese medicine composition obtained in the examples and comparative examples, and each group was set up with 2-6 replicate wells.

[0071] (3) Cortisol conversion reaction: dilute 50mM cortisone solution to 100μM and add 50μl / well to the well plate. Dilution and addition of samples to be tested: use complete culture medium as diluent, prepare sample working solutions of different concentrations according to the sample test concentration table, add 50μl / well, a total of 100μl / well reaction system, the experimental concentration of the agarwood compound Chinese medicine composition is 5.0wt% (the mass percentage of the agarwood compound Chinese medicine composition in the system).

[0072] Negative control group: add an equal amount of complete culture medium;

[0073] Agonist positive control group: add 100 μM Cortisone 100 μl / well;

[0074] Inhibitor positive control group: add 100 μl / well of 200 μM Metyrapone.

[0075] The above groups were cultured in an incubator for 24 h, and the cell morphology was observed under a microscope.

[0076] (4) Take the supernatant from the well plate and detect the cortisol content by ELISA method. The operation steps are as follows:

[0077] A. Serial dilution of standard.

[0078] B. Collect the cell culture supernatant into a sterile centrifuge tube and centrifuge to obtain the supernatant.

[0079] C. Add 50 μl of standard working solution and test sample to each reaction well. Each group has 2 duplicate wells. Add 50 μl of biotin-labeled antibody working solution to each reaction well and incubate.

[0080] D. Discard the liquid, spin dry, add washing solution to each reaction well, soak and spin dry.

[0081] E. Add 100 μl of HRP-labeled streptavidin working solution to each reaction well and incubate.

[0082] F. Add 300 μL of washing solution to each reaction well, and spin dry the washing solution after 30 seconds.

[0083] G. Add 90 μL of color developer to each reaction well and develop the color at 37°C in the dark for about 15 minutes.

[0084] H. Add 50 μL of stop solution to each reaction well and immediately measure the OD value at 450 nm using a microplate reader. The OD value is inversely proportional to the cortisol content.

[0085] I. Calculate the standard curve regression equation (R) based on the known concentration of the standard and the measured OD value. 2 >0.99), substitute the OD value of the sample well into the calculated concentration of the sample, and then multiply by the dilution factor to obtain the actual cortisol concentration of the original sample. The experimental results are shown in Table 1.

[0086] Table 1

[0087] concentration Cortisol content (ng / ml) Cortisol suppression rate (%) Negative / -67.64 / Positive / 248.31 / Metyrapone 200μM 126.51 49.1 Example 1 5.0wt% 75.29 69.7 Example 2 5.0wt% 70.63 71.6 Example 3 5.0wt% 72.85 70.7 Comparative Example 1 5.0wt% 147.23 40.7 Comparative Example 2 5.0wt% 150.36 39.4 Comparative Example 3 5.0wt% 149.62 39.7

[0088] It can be seen from Table 1 that, compared with the comparative example, the agarwood compound Chinese medicine composition provided by the embodiment of the present invention can significantly reduce the production of cortisol.

[0089] Test Example 2

[0090] This test example explores the inhibitory effect of the agarwood compound Chinese medicine composition obtained in the embodiment and the comparative example on the inflammatory factor IL-6, specifically comprising the following steps:

[0091] (1) Cell seeding: RAW264.7 cells were seeded at 4500 cells / well in a 96-well plate and cultured in a 37° C., 5% CO 2 incubator for 24 h.

[0092] (2) Experimental grouping: a blank group and a sample group were set up, and the test substances in the sample group were the compositions prepared in the examples and comparative examples.

[0093] (3) LPS-induced inflammatory response: 1 mg / mL LPS solution (solvent: PBS buffer) was diluted to 2 μg / mL, added to the well plate at 50 μL / well, and placed in an incubator for culture.

[0094] (4) Sample addition: Prepare sample working solution using complete culture medium as diluent, 100 μL / well. Add equal amount of complete culture medium to negative control group and positive control group and culture in an incubator for 24 h.

[0095] (5) Take the supernatant from the well plate and detect the IL-6 content by ELISA method. The operation steps are as follows:

[0096] A. Calculate the required enzyme label strips in advance, take out the kit and restore it to room temperature before the experiment;

[0097] B. Serial dilution of standard: dilute the standard with standard & sample diluent to 500, 250, 125, 62.5, 0 ng / mL;

[0098] C. Collect the cell culture supernatant into a sterile centrifuge tube and centrifuge to obtain the supernatant;

[0099] D. Add standard working solution and test samples to each reaction well, set up 2 replicate wells for each group, and incubate;

[0100] E. Discard the liquid, spin dry, add biotin-labeled antibody working solution to each reaction well, and incubate;

[0101] F. Discard the liquid, spin dry, add washing solution to each reaction well, soak and spin dry;

[0102] G. Add HRP-labeled streptavidin working solution to each reaction well and incubate;

[0103] H. Add washing solution to each reaction well, and spin dry the washing solution after 30 seconds.

[0104] I. Add the color developing agent to each reaction well and develop the color at 37°C in the dark for 15 minutes;

[0105] J. Add stop solution to each reaction well and measure the OD value at 450nm using a microplate reader;

[0106] K. Calculate the standard curve regression equation (R) based on the known concentration of the standard and the measured OD value. 2 >0.99), the OD value of the sample well was substituted into the concentration of the sample to be measured, and then multiplied by the dilution factor to obtain the actual IL-6 concentration of the original sample. The test results are shown in Table 2.

[0107] Table 2

[0108] Group IL-6 secretion (ng / mL) Blank Group 1.5 Example 1 0.54 Example 2 0.51 Example 3 0.52 Comparative Example 1 1.06 Comparative Example 2 1.09 Comparative Example 3 1.14 Comparative Example 4 1.05

[0109] It can be seen from Table 2 that, compared with the comparative example, the agarwood compound Chinese medicine composition provided in the embodiment has a significant inhibitory effect on the inflammatory factor IL-6.

[0110] The above description of the disclosed embodiments enables those skilled in the art to implement or use the present invention. Various modifications to these embodiments will be apparent to those skilled in the art, and the general principles defined herein may be implemented in other embodiments without departing from the spirit or scope of the present invention. Therefore, the present invention will not be limited to the embodiments shown herein, but rather to the widest scope consistent with the principles and novel features disclosed herein.

Claims

1. A compound agarwood Chinese medicine composition for relieving depression, characterized in that: The invention comprises, by weight, 20 to 30 parts of agarwood extract, 5 to 8 parts of chervil flower extract, 5 to 10 parts of bupleurum extract and 5 to 10 parts of scutellaria baicalensis extract.

2. The agarwood compound Chinese medicine composition for relieving depression according to claim 1, characterized in that: The preparation method of the agarwood extract comprises: (1) extracting agarwood with an extraction solvent, drying, and obtaining agarwood powder; (2) Using supercritical fluid to extract the agarwood powder, drying it, and obtaining the agarwood extract.

3. The agarwood compound Chinese medicine composition for relieving depression according to claim 2, characterized in that: The solid-liquid ratio of the agarwood and the extraction solvent is 1: (25-30) g / ml.

4. The agarwood compound Chinese medicine composition for relieving depression according to claim 1, characterized in that: The preparation method of the chervil flower extract comprises: The serrulata flower is crushed, mixed with water, distilled under reduced pressure, and dried to obtain a serrulata flower extract.

5. The agarwood compound Chinese medicine composition for relieving depression according to claim 1, characterized in that: The preparation method of the Bupleurum extract comprises: Mix bupleurum root with water, soak for 50-70 minutes to obtain the medicine to be decocted; decoct the medicine to be decocted for 50-70 minutes, filter and obtain the filtrate; centrifuge the filtrate, take the supernatant, and dry to obtain the bupleurum root extract.

6. The agarwood compound Chinese medicine composition for relieving depression according to claim 1, characterized in that: The preparation method of the scutellaria baicalensis extract comprises: Mix scutellaria baicalensis with water, soak for 50-70 minutes to obtain the medicine to be decocted; decoct the medicine to be decocted for 50-70 minutes, filter and obtain the filtrate; centrifuge the filtrate, take the supernatant, and dry to obtain the scutellaria baicalensis extract.

7. Use of the agarwood compound Chinese medicine composition for relieving depression according to any one of claims 1 to 6 in the preparation of a product for preventing and / or treating the inhibition of cortisol secretion.

8. The use according to claim 7, characterized in that: The concentration of the agarwood compound traditional Chinese medicine composition is 10-20wt%.

9. Use of the agarwood compound Chinese medicine composition for relieving depression according to any one of claims 1 to 6 in the preparation of products for preventing and / or treating anti-inflammatory diseases.

10. The use according to claim 9, characterized in that: The anti-inflammatory effect includes inhibiting inflammatory factors; the inflammatory factors include IL-6.