A traditional Chinese medicine composition for treating depression

Abstract

The present invention discloses a traditional Chinese medicine composition for treating depression, belonging to the technical field of traditional Chinese medicine. Calculated by mass parts, the traditional Chinese medicine composition is prepared from the following raw materials: 8-15 parts of ginseng, 7-12 parts of morinda officinalis, 8-13 parts of ligusticum wallichii, 8-12 parts of gardenia jasminoides, 8-16 parts of medicated leaven, 7-12 parts of cyperus rotundus, and 6-13 parts of atractylodes lancea. The combination of components of the traditional Chinese medicine composition of the present invention is scientific and reasonable, has the effects of cultivating primordial qi and supplementing qi, promoting qi movement and dredging stagnation, is safe in taking medicine, has good curative effects, can be effectively used for the treatment of depression, is a major innovation in the drugs for treating depression, and has remarkable economic and social benefits.

Description

Technical Field

[0001] The present invention relates to the technical field of traditional Chinese medicine, and particularly to a traditional Chinese medicine composition for treating depression. Background Art

[0002] Depression is a common mental disorder with an increasing incidence rate year by year. It is characterized by high incidence, high recurrence rate, high disability rate, and high suicide rate, and the main clinical manifestations include low mood, anhedonia, reduced energy, etc. At present, the pathogenesis of depression has not been fully clarified.

[0003] Traditional Chinese medicine is good at analyzing things systematically, holistically, macroscopically, and dynamically, grasping the spatio-temporal development law of things, and has a unique understanding of emotional diseases. It believes that the body and mind are integrated, nourishing the body with the mind and nourishing the mind with the body. When the mind is ill, the body will be ill; when the body is ill, the mind will be ill. Depression belongs to the category of "depressive disorder" and "lily disease" in traditional Chinese medicine in China. According to different causes, it is divided into various types of depression, and its basic pathogenesis is mainly liver qi stagnation. With the continuous in-depth research on depression and clinical practice, it has been shown that the onset of depression is not only related to liver depression, but also closely related to the deficiency of primordial qi.

[0004] With the vigorous development of traditional Chinese medicine, traditional Chinese medicine has shown excellent advantages in the treatment of depression, such as less side effects, long-lasting efficacy, high patient compliance, and the ability to act on multiple ion channels and multiple targets, significantly improving the clinical symptoms and prognosis of patients. At present, the research on the treatment of depression with traditional Chinese medicine is relatively lacking. Therefore, providing a traditional Chinese medicine composition that can effectively treat depression will be of great significance for the clinical treatment of depression.

[0005] Chinese patent document CN115737745A discloses a traditional Chinese medicine composition for treating depression, which is characterized by including 5 - 10 parts of gardenia, 6 - 9 parts of lily, 5 - 7 parts of ginseng, 3 - 7 parts of tangerine peel, 5 - 10 parts of wild jujube seed, 10 - 15 parts of poria cocos, 8 - 12 parts of dragon bone, 10 - 12 parts of mother-of-pearl, and 9 - 15 parts of rehmannia glutinosa, solving the problems of relatively large toxic and side effects and being unable to take into account both the psychological and physical aspects.

[0006] Chinese patent document CN115919982A discloses a pharmaceutical composition for treating post-stroke depression, which is characterized by including 10 - 12 parts of bupleurum, 10 - 12 parts of turmeric, 12 - 15 parts of poria cocos, 10 - 12 parts of angelica dahurica, 10 - 15 parts of angelica sinensis, 10 - 12 parts of ligusticum wallichii, 10 - 12 parts of gastrodia elata, 10 - 12 parts of pinellia ternata, 10 - 12 parts of safflower, 15 - 20 parts of earthworm, 12 - 15 parts of acorus tatarinowii, 15 - 30 parts of clerodendrum bungei, 12 - 15 parts of albizia julibrissin, 10 - 12 parts of rose, 12 - 15 parts of cyathula officinalis, 15 - 30 parts of millettia reticulata, 10 - 12 parts of polygala tenuifolia honey-fried, and 3 - 6 parts of roasted licorice. It solves the problems of drug resistance and relatively high treatment cost of existing western medicines.

[0007] Chinese patent document CN115487285A discloses a traditional Chinese medicine composition for treating depression caused by insufficient heart yin. The traditional Chinese medicine composition, by weight, comprises: 8-10 parts of liquorice, 28-32 parts of fructus tritici, 5-10 parts of fructus ziziphi jujubae, 25-35 parts of astragalus membranaceus, 1-5 parts of fructus amomi villosi, 1-5 parts of bupleuri radix, 13-18 parts of puerariae lobatae radix, 15-20 parts of arillus longan, 15-20 parts of fructus mume and 28-32 parts of fresh ginger. It solves the problem of high toxic and side effects of existing antidepressants.

[0008] However, there has been no report on a traditional Chinese medicine composition with good curative effect for treating depression. Summary of the Invention

[0009] The purpose of the present invention is to provide a traditional Chinese medicine composition for treating depression to solve the problems existing in the above-mentioned prior art. The composition and compatibility of the traditional Chinese medicine composition are scientific and reasonable, and it has the effects of nourishing primordial qi and regulating qi and relieving stagnation. It is safe to take, has good curative effect and small side effects, can be effectively used for the treatment of depression, is a major innovation in the field of depression drugs, and has significant economic and social benefits.

[0010] To achieve the above purpose, the present invention provides the following solutions:

[0011] The present invention provides a traditional Chinese medicine composition for treating depression. By mass, the traditional Chinese medicine composition comprises the following components: 8-15 parts of ginseng, 7-12 parts of morinda officinalis, 8-13 parts of chuanxiong rhizome, 8-12 parts of gardenia jasminoides, 8-16 parts of medicated leaven, 7-12 parts of cyperus rotundus and 6-13 parts of atractylodes lancea.

[0012] Preferably, by mass, the traditional Chinese medicine composition is made from the following raw materials: 9-14 parts of ginseng, 8-10 parts of morinda officinalis, 9-12 parts of chuanxiong rhizome, 9-12 parts of gardenia jasminoides, 10-14 parts of medicated leaven, 8-10 parts of cyperus rotundus and 8-12 parts of atractylodes lancea.

[0013] Preferably, by mass, the traditional Chinese medicine composition is made from the following raw materials: 10 parts of ginseng, 10 parts of morinda officinalis, 10 parts of chuanxiong rhizome, 10 parts of gardenia jasminoides, 10 parts of medicated leaven, 10 parts of cyperus rotundus and 10 parts of atractylodes lancea.

[0014] The present invention also provides a preparation method of the traditional Chinese medicine composition for treating depression, including any one of the following methods (1)-(3):

[0015] (1) After crushing each raw material in the formula, mix them to make the traditional Chinese medicine composition for treating depression;

[0016] (2) The extract obtained by mixing or separately extracting each raw material in the formula is the traditional Chinese medicine composition for treating depression;

[0017] (3) The extract obtained by mixing or separately extracting each raw medicinal material in the formula, and then adding pharmaceutically acceptable excipients to prepare a conventional dosage form is the traditional Chinese medicine composition for treating depression.

[0018] Preferably, the extraction method includes decocting or refluxing.

[0019] Preferably, the dosage form includes granules, capsules or oral liquids.

[0020] The present invention also provides the application of the traditional Chinese medicine composition in the preparation of a drug for treating depression.

[0021] The present invention discloses the following technical effects:

[0022] The traditional Chinese medicine composition of the present invention is scientifically and rationally formulated according to the compatibility principle of monarch, minister, assistant and envoy in traditional Chinese medicine, and supports each other. Among them, ginseng is used as the monarch drug to invigorate the spleen and replenish qi; morinda officinalis and cyperus rotundus are used as the minister drugs to tonify the kidney and assist yang and regulate qi and relieve depression respectively; ligusticum wallichii is used to regulate qi and activate blood circulation, gardenia jasminoides is used to clear heat and cool blood and remove vexation, and atractylodes lancea is used to dry dampness and strengthen the spleen as the assistant drugs, and medicated leaven is used to strengthen the spleen and harmonize the stomach as the envoy drug. The whole formula regulates both the mind and body, treats both the symptoms and the root causes, and can be modified according to the syndrome, and is applicable to different pathological stages of depression. It has a good curative effect on a series of symptoms such as low mood, listlessness, reluctance to talk and move, restlessness and panic, insomnia and dreaminess, pale complexion, and fatigue. More specifically, through animal experiments, it is found that the traditional Chinese medicine composition has a good therapeutic effect on high-salt + depression model rats, and can significantly improve the general condition, body weight, blood pressure and behavioral evaluation of rats. In terms of the curative mechanism, the traditional Chinese medicine composition of the present invention can play an antidepressant role by improving neuronal cell damage, reducing neuronal apoptosis, restoring the vascular neuron structure of the hippocampal tissue, reducing the permeability of the blood-brain barrier, reducing the expression levels of serum-related inflammatory factors (LPS, TLR4, TNF-α, NF-κB, IL-6, IL-1β) in the hippocampal tissue and serum, reducing the contents of VEGF, MMP-9 and ET-1 in the hippocampal tissue, increasing the contents of Occludin, ZO-1, Claudin-5 and eNOS in the hippocampal tissue, and improving the genus level of the intestinal flora of rats.

[0023] Using the traditional Chinese medicine composition of the present invention to treat depression may reduce the economic burden of patients and the national medical cost compared with the current treatment means for depression. In terms of the action mechanism, the traditional Chinese medicine composition of the present invention can be improved simultaneously from multiple mechanisms, and finally achieve the purpose of improving depression, broaden the current possibilities for treating depression, and further promote the vigorous development of traditional Chinese medicine in the treatment of depression, with great social value. Description of the Drawings

[0024] To more clearly illustrate the technical solutions in the embodiments of the present invention or the prior art, the following will briefly introduce the accompanying drawings required in the embodiments. Obviously, the accompanying drawings in the following description are only some embodiments of the present invention. For those of ordinary skill in the art, without creative efforts, other accompanying drawings can be obtained based on these drawings.

[0025] Figure 1 Effect of Peiyuan Tongzhi Formula on the body weight of rats with hypertension accompanied by depression;

[0026] Figure 2 Effect of Peiyuan Tongzhi Formula on the systolic blood pressure of rats with hypertension accompanied by depression;

[0027] Figure 3 Effect of Peiyuan Tongzhi Formula on the diastolic blood pressure of rats with hypertension accompanied by depression;

[0028] Figure 4 Comparison results of the sucrose preference rate of rats in each group;

[0029] Figure 5 Comparison results of the stationary time of rats in the open field test in each group;

[0030] Figure 6 Comparison results of the stationary time of rats in the forced swimming test in each group;

[0031] Figure 7 Comparison results of Nissl staining in the CA1 region of the hippocampus in the brain tissue of rats in each group under light microscopy;

[0032] Figure 8 Comparison results of immunofluorescence staining of neurons in the CA1 region of the hippocampus in the brain tissue of rats in each group;

[0033] Figure 9 Comparison results of the ratio of positive cells in the CA1 region of the hippocampus in the brain tissue of rats in each group;

[0034] Figure 10 Comparison results of electron micrographs of vascular nerve units in the hippocampal tissue of rats in each group; Scale bar: 2 μm;

[0035] Figure 11 Comparison results of LFB staining in the CA1 region of the hippocampus of rats in each group;

[0036] Figure 12 Comparison results of the blood-brain barrier permeability of rats in each group;

[0037] Figure 13 Comparison results of the expression levels of serum-related inflammatory factors of rats in each group;

[0038] Figure 14 Comparison results of the expression levels of hippocampal tissue-related inflammatory factors of rats in each group;

[0039] Figure 15 Results of comparison of VEGF and MMP-9 contents in the hippocampal tissues of rats in each group;

[0040] Figure 16 Results of comparison of Occludin, ZO-1, and Claudin-5 contents in the hippocampal tissues of rats in each group;

[0041] Figure 17 Results of comparison of eNOS and ET-1 contents in the hippocampal tissues of rats in each group;

[0042] Figure 18 Results of comparison of α-diversity of intestinal flora in rats in each group;

[0043] Figure 19 PCA comparative analysis chart;

[0044] In the above figures, LS represents the low-salt group; HS represents the high-salt group; M represents the high-salt + CUMS group; PYKY represents the high-salt + CUMS + Peiyuan Tongzhi Formula group; EO represents the high-salt + CUMS + citalopram group. Detailed implementation manners

[0045] Now, various exemplary implementation manners of the present invention will be described in detail. This detailed description should not be considered as a limitation to the present invention, but should be understood as a more detailed description of certain aspects, characteristics, and implementation schemes of the present invention.

[0046] It should be understood that the terms described in the present invention are only for describing specific implementation manners and are not used to limit the present invention. Additionally, for the numerical ranges in the present invention, it should be understood that each intermediate value between the upper and lower limits of the range is also specifically disclosed. Any intermediate value within any stated value or stated range, as well as each smaller range between any other stated value or intermediate value within the stated range, is also included in the present invention. The upper and lower limits of these smaller ranges can be independently included or excluded from the range.

[0047] Unless otherwise specified, all technical and scientific terms used herein have the same meaning as commonly understood by those of ordinary skill in the art to which the present invention pertains. Although the present invention only describes preferred methods and materials, any methods and materials similar or equivalent to those described herein can also be used in the implementation or testing of the present invention. All documents mentioned in this specification are incorporated by reference to disclose and describe the methods and / or materials related to the documents. In case of conflict with any incorporated document, the content of this specification shall prevail.

[0048] Without departing from the scope or spirit of the present invention, various improvements and changes can be made to the specific embodiments of the description of the present invention, which will be obvious to those skilled in the art. Other embodiments obtained from the description of the present invention will be obvious to those skilled in the art. The description and examples of the present invention are merely exemplary.

[0049] Regarding the use of "comprising", "including", "having", "containing", etc. herein, they are all open-ended terms, meaning including but not limited to.

[0050] The technical concept of the present invention:

[0051] Traditional Chinese medicine believes that depression (depressive disorder) is mostly caused by congenital deficiency, improper diet, excessive or insufficient work and rest, and emotional injury, resulting in deficiency of vital qi in the body, generating pathological products such as dampness, phlegm, stasis, heat, and toxin in the body. Then, due to deficiency, stagnation occurs, and the combination of deficiency and stagnation further aggravates the vicious cycle of deficiency and stagnation in the pathological process. The pathogenesis of depression has the characteristics of the pathogenesis of "stagnation-deficiency-stagnation", emphasizing the pathological characteristics of taking qi deficiency as the root and retention and stagnation as the target, forming the basic law of pathogenesis changes of retention and stagnation due to deficiency, transforming from qi to blood, phlegm turbidity gradually increasing, stasis blood being prominent, and toxin heat causing damage.

[0052] The theoretical basis of the traditional Chinese medicine composition of the present invention is: under the guidance of the pathogenesis theory of "deficiency of qi with retention and stagnation", taking the deficiency of primordial qi as the root, and combined with excessive seven emotions, depression finally occurs. "Deficiency of qi" refers to the deficiency of primordial qi in the elderly, congenital deficiency, or the decline of qi in obese people with a strong build. Since the heart is the residence of the spirit and the brain is the residence of the original spirit, damage to the heart and brain directly injures the spirit, leading to "stagnation". In addition, after the onset of the disease, the psychological burden of the patient is relatively heavy, and social support is often insufficient, thus aggravating "stagnation", and finally forming a situation where "deficiency-stagnation-stagnation" coexist. Therefore, the treatment method is to replenish the yuan and promote qi stagnation, and the medication is mainly to supplement qi, and at the same time use qi-promoting and stagnation-passing drugs. Taking supplementing qi as the key point, aiming to strengthen the root of "deficiency of qi". When the healthy qi is restored, the qi movement and transportation of the body can function, and the distribution of qi, blood, and body fluids can be smooth; at the same time, taking into account the treatment of the target of "retention and stagnation", treating both the root and the branch simultaneously to achieve the adjustment of both the body and the mind.

[0053] Based on the above theoretical research results and symptom manifestations, the traditional Chinese medicine composition for treating depression of the present invention is developed. The traditional Chinese medicine composition of the present invention is formulated according to the principle of monarch, minister, assistant, and envoy, and is composed of 8-15 parts of ginseng, 7-12 parts of morinda officinalis, 8-13 parts of chuanxiong rhizome, 8-12 parts of gardenia, 8-16 parts of medicated leaven, 7-12 parts of cyperus rotundus, and 6-13 parts of atractylodes lancea. According to the compatibility principle of monarch, minister, assistant, and envoy of traditional Chinese medicine, the present invention makes a scientific and reasonable compatibility, supporting each other. Ginseng replenishes the yuan, strengthens the spleen, and supplements qi as the monarch drug; morinda officinalis tonifies the kidney and aids yang, and cyperus rotundus regulates qi and relieves depression together as the minister drugs; chuanxiong rhizome regulates qi and activates blood circulation, gardenia clears heat and cools blood to remove vexation, and atractylodes lancea dries dampness and strengthens the spleen as the assistant drugs, and medicated leaven strengthens the spleen and harmonizes the stomach as the envoy drug. The whole formula adjusts both the body and the mind, treats both the root and the branch simultaneously, and can be added or subtracted according to the syndrome, and is applicable to different pathological stages of depression.

[0054] Ginseng: sweet, slightly bitter, slightly warm, attributing to the spleen and lung meridians. It greatly tonifies primordial qi, invigorates the spleen and benefits the lung, promotes fluid production to quench thirst, and soothes the mind and improves intelligence.

[0055] Morinda officinalis: pungent, sweet, slightly warm, attributing to the kidney meridian. It tonifies the kidney and boosts yang, dispels wind and dampness.

[0056] Chuanxiong: pungent, warm. Attributing to the liver, gallbladder, and pericardium meridians. It promotes blood circulation and qi movement, dispels wind and alleviates pain.

[0057] Gardenia jasminoides: bitter, cold. Attributing to the heart, lung, stomach, and triple energizer meridians. It purges fire and relieves vexation, clears heat and promotes diuresis, cools blood and detoxifies.

[0058] Medicated leaven: sweet, pungent, warm. Attributing to the spleen and stomach meridians. It promotes digestion and harmonizes the stomach.

[0059] Cyperus rotundus: pungent, slightly bitter, slightly sweet, flat. Attributing to the liver and triple energizer meridians. It soothes the liver and regulates qi, regulates menstruation and alleviates pain.

[0060] Atractylodes lancea: pungent, bitter, warm, attributing to the spleen and stomach meridians. It dries dampness and strengthens the spleen, dispels wind and dampness.

[0061] Example 1

[0062] The traditional Chinese medicine composition comprises the following components in the given dosages: 8 g of ginseng, 7 g of Morinda officinalis, 8 g of Chuanxiong, 8 g of Gardenia jasminoides, 8 g of medicated leaven, 7 g of Cyperus rotundus, and 6 g of Atractylodes lancea.

[0063] Preparation method: Weigh each raw material medicine according to the dosages in the formula, soak in cold water for 30 minutes, add water exceeding the drug surface by 3 cm, decoct twice. For the first decoction, first bring to a boil over high heat, then keep boiling over low heat for 25 minutes, filter to obtain the first medicinal liquid; add water to the drug residue again to submerge the drug surface, bring to a boil over high heat, then keep boiling over low heat for 25 minutes, filter to obtain the second medicinal liquid; combine the two medicinal liquids, concentrate into a clear paste, add the auxiliary material dextrin (added according to the mass ratio of the clear paste to dextrin of 1:2), mix evenly, dry, and make into granules, thus obtaining the granule preparation of the traditional Chinese medicine composition for treating depression. 1 g of the granule is equivalent to 5 g of the raw material medicine. Take 4 g each time, take it with warm water, 2 - 3 times a day.

[0064] Example 2

[0065] The traditional Chinese medicine composition comprises the following components in the given dosages: 8 g of ginseng, 10 g of Morinda officinalis, 9 g of Chuanxiong, 10 g of Gardenia jasminoides, 12 g of medicated leaven, 8 g of Cyperus rotundus, and 7 g of Atractylodes lancea.

[0066] Preparation method: Weigh the ginseng slices according to the formula dosage, soak them in cold water for 30 minutes, add water 3 cm above the drug surface, decoct twice. For the first decoction, first bring to a boil over high heat, then keep boiling over low heat for 30 minutes, filter to obtain the first medicinal liquid; add water to the drug residue again to submerge the drug surface, bring to a boil over high heat, then keep boiling over low heat for 30 minutes, filter to obtain the second medicinal liquid; combine the two medicinal liquids and concentrate them into a clear paste, add the auxiliary material dextrin (the mass ratio of the clear paste to dextrin is 1:2), mix evenly, dry, granulate, and package to obtain the ginseng granules;

[0067] According to the same method, prepare Morinda officinalis How granules, Ligusticum wallichii Franch granules, Gardenia jasminoides Ellis granules, Medicated leaven granules, Cyperus rotundus L. granules and Atractylodes lancea (Thunb.) DC. granules.

[0068] Take the granule combination prepared from different slices according to the dosage of each component in the formula (8 g of ginseng, 10 g of Morinda officinalis How, 9 g of Ligusticum wallichii Franch, 10 g of Gardenia jasminoides Ellis, 12 g of Medicated leaven, 8 g of Cyperus rotundus L. and 7 g of Atractylodes lancea (Thunb.) DC.), and obtain the traditional Chinese medicine composition granule preparation for treating depression.

[0069] Example 3

[0070] The traditional Chinese medicine composition includes the following components in the following dosages: 9 g of ginseng, 8 g of Morinda officinalis How, 9 g of Ligusticum wallichii Franch, 9 g of Gardenia jasminoides Ellis, 10 g of Medicated leaven, 8 g of Cyperus rotundus L. and 8 g of Atractylodes lancea (Thunb.) DC.

[0071] Preparation method: Weigh the ginseng slices according to the formula dosage, soak them in cold water for 30 minutes, add water 3 cm above the drug surface, for the first decoction, first bring to a boil over high heat, then keep boiling over low heat for 30 minutes, filter to obtain the first medicinal liquid; add water to the drug residue again to submerge the drug surface, bring to a boil over high heat, then keep boiling over low heat for 30 minutes, filter to obtain the second medicinal liquid; combine the two medicinal liquids and concentrate them into a clear paste, add the auxiliary material dextrin (the mass ratio of the clear paste to dextrin is 1:2), mix evenly, dry, granulate, and package to obtain the ginseng granules;

[0072] According to the same method, prepare Morinda officinalis How granules, Ligusticum wallichii Franch granules, Gardenia jasminoides Ellis granules, Medicated leaven granules, Cyperus rotundus L. granules and Atractylodes lancea (Thunb.) DC. granules.

[0073] Take the granule combination prepared from different slices according to the dosage of each component in the formula (9 g of ginseng, 8 g of Morinda officinalis How, 9 g of Ligusticum wallichii Franch, 9 g of Gardenia jasminoides Ellis, 10 g of Medicated leaven, 8 g of Cyperus rotundus L. and 8 g of Atractylodes lancea (Thunb.) DC.), and obtain the traditional Chinese medicine composition granule preparation for treating depression.

[0074] Example 4

[0075] The traditional Chinese medicine composition includes the following components in the following dosages: 10 g of ginseng, 10 g of Morinda officinalis How, 10 g of Ligusticum wallichii Franch, 10 g of Gardenia jasminoides Ellis, 10 g of Medicated leaven, 10 g of Cyperus rotundus L. and 10 g of Atractylodes lancea (Thunb.) DC.

[0076] Preparation method: Weigh the ginseng slices according to the formula dosage, soak them in cold water for 30 minutes, add water with a depth 3 cm above the drug surface, decoct twice. For the first decoction, first bring to a boil over high heat, then keep boiling over low heat for 30 minutes, filter to obtain the first medicinal liquid; add water to the drug residue again to submerge the drug surface, bring to a boil over high heat, then keep boiling over low heat for 30 minutes, filter to obtain the second medicinal liquid; combine the two medicinal liquids and concentrate them into a clear paste, add the auxiliary material dextrin (the mass ratio of the clear paste to dextrin is 1:2), mix evenly, dry, granulate, and package to obtain the ginseng granules;

[0077] According to the same method, prepare Morinda officinalis How granules, Ligusticum wallichii Franch granules, Gardenia jasminoides Ellis granules, Medicago sativa L. granules, Cyperus rotundus L. granules and Atractylodes lancea (Thunb.) DC. granules.

[0078] Take the granule combination prepared from different slices according to the dosage of each component in the formula (10 g of ginseng, 10 g of Morinda officinalis How, 10 g of Ligusticum wallichii Franch, 10 g of Gardenia jasminoides Ellis, 10 g of Medicago sativa L., 10 g of Cyperus rotundus L. and 10 g of Atractylodes lancea (Thunb.) DC.), and obtain the traditional Chinese medicine composition granule preparation for treating depression.

[0079] Example 5

[0080] The traditional Chinese medicine composition includes the following components in the following dosages: 12 g of ginseng, 9 g of Morinda officinalis How, 9 g of Ligusticum wallichii Franch, 11 g of Gardenia jasminoides Ellis, 11 g of Medicago sativa L., 9 g of Cyperus rotundus L. and 10 g of Atractylodes lancea (Thunb.) DC.

[0081] Preparation method: Weigh the ginseng slices according to the formula dosage, soak them in cold water for 30 minutes, add water with a depth 3 cm above the drug surface, decoct twice. For the first decoction, first bring to a boil over high heat, then keep boiling over low heat for 35 minutes, filter to obtain the first medicinal liquid; add water to the drug residue again to submerge the drug surface, bring to a boil over high heat, then keep boiling over low heat for 35 minutes, filter to obtain the second medicinal liquid; combine the two medicinal liquids and concentrate them into a clear paste, add the auxiliary material dextrin (the mass ratio of the clear paste to dextrin is 1:2), mix evenly, dry, granulate, and package to obtain the ginseng granules;

[0082] According to the same method, prepare Morinda officinalis How granules, Ligusticum wallichii Franch granules, Gardenia jasminoides Ellis granules, Medicago sativa L. granules, Cyperus rotundus L. granules and Atractylodes lancea (Thunb.) DC. granules.

[0083] Take the granule combination prepared from different slices according to the dosage of each component in the formula (12 g of ginseng, 9 g of Morinda officinalis How, 9 g of Ligusticum wallichii Franch, 11 g of Gardenia jasminoides Ellis, 11 g of Medicago sativa L., 9 g of Cyperus rotundus L. and 10 g of Atractylodes lancea (Thunb.) DC.), and obtain the traditional Chinese medicine composition granule preparation for treating depression.

[0084] Example 6

[0085] The traditional Chinese medicine composition includes the following components in the following dosages: 14 g of ginseng, 10 g of Morinda officinalis How, 12 g of Ligusticum wallichii Franch, 12 g of Gardenia jasminoides Ellis, 14 g of Medicago sativa L., 10 g of Cyperus rotundus L. and 12 g of Atractylodes lancea (Thunb.) DC.

[0086] Preparation method: Weigh the ginseng slices according to the formula dosage, decoct them twice with water. For the first decoction, bring to a boil over high heat first, then keep boiling over low heat for 25 minutes, filter to obtain the first medicinal liquid; add water to the medicinal residues again to submerge the surface of the medicine, bring to a boil over high heat, then keep boiling over low heat for 25 minutes, filter to obtain the second medicinal liquid; combine the two medicinal liquids and concentrate them into a clear paste, add the auxiliary material dextrin (the mass ratio of the clear paste to dextrin is 1:2), mix evenly, dry, granulate, and package to obtain ginseng granules;

[0087] According to the same method, prepare morinda officinalis granules, chuanxiong granules, gardenia granules, medicated leaven granules, cyperus rotundus granules and atractylodes lancea granules.

[0088] Take the granule combination prepared from different slices according to the dosage of each component in the formula (14 g of ginseng, 10 g of morinda officinalis, 12 g of chuanxiong, 12 g of gardenia, 14 g of medicated leaven, 10 g of cyperus rotundus and 12 g of atractylodes lancea), and obtain the traditional Chinese medicine composition granule preparation for treating depression.

[0089] Example 7

[0090] The traditional Chinese medicine composition includes the following components in the following dosages: 10 g of ginseng, 7 g of morinda officinalis, 11 g of chuanxiong, 10 g of gardenia, 10 g of medicated leaven, 7 g of cyperus rotundus and 11 g of atractylodes lancea.

[0091] Preparation method: Weigh each raw material medicine according to the formula dosage, soak it in cold water for 30 minutes, add water 3 cm above the surface of the medicine, decoct it twice. For the first decoction, bring to a boil over high heat first, then keep boiling over low heat for 25 minutes, filter to obtain the first medicinal liquid; add water to the medicinal residues again to submerge the surface of the medicine, bring to a boil over high heat, then keep boiling over low heat for 25 minutes, filter to obtain the second medicinal liquid; combine the two medicinal liquids, concentrate, add the auxiliary materials for capsules, and make into capsules.

[0092] Example 8

[0093] The traditional Chinese medicine composition includes the following components in the following dosages: 12 g of ginseng, 9 g of morinda officinalis, 10 g of chuanxiong, 11 g of gardenia, 12 g of medicated leaven, 10 g of cyperus rotundus and 9 g of atractylodes lancea.

[0094] Preparation method: Weigh each raw material medicine according to the formula dosage, soak it in cold water for 30 minutes, add water 3 cm above the surface of the medicine, decoct it twice separately. For the first decoction, bring to a boil over high heat first, then keep boiling over low heat for 25 minutes, filter to obtain the first medicinal liquid; add water to the medicinal residues again to submerge the surface of the medicine, bring to a boil over high heat, then keep boiling over low heat for 25 minutes, filter to obtain the second medicinal liquid; combine the two filtrates, divide them into two equal parts on average, each part is 200 - 300 mL. Take it twice a day.

[0095] Example 9

[0096] The traditional Chinese medicine composition comprises the following components in the following dosages: 13 g of ginseng, 9 g of morinda root, 8 g of chuanxiong rhizome, 9 g of gardenia fruit, 8 g of medicated leaven, 10 g of cyperus rhizome and 6 g of atractylodes rhizome.

[0097] Preparation method: Weigh each raw material medicine according to the formula ratio, extract by refluxing with water twice, for 50 min for the first time and 30 min for the second time, filter, combine the filtrates, and concentrate to about 400 mL to obtain. It is taken twice a day.

[0098] Example 10

[0099] The traditional Chinese medicine composition comprises the following components in the following dosages: 14 g of ginseng, 11 g of morinda root, 10 g of chuanxiong rhizome, 10 g of gardenia fruit, 13 g of medicated leaven, 11 g of cyperus rhizome and 12 g of atractylodes rhizome.

[0100] Preparation method: Weigh each raw material medicine according to the formula ratio, decoct with water twice respectively, for 30 min for the first time and 20 min for the second time, filter, combine the filtrates, and concentrate to about 400 mL to obtain. It is taken twice a day.

[0101] Example 11

[0102] The traditional Chinese medicine composition comprises the following components in the following dosages: 15 g of ginseng, 12 g of morinda root, 13 g of chuanxiong rhizome, 12 g of gardenia fruit, 16 g of medicated leaven, 12 g of cyperus rhizome and 13 g of atractylodes rhizome.

[0103] Preparation method: Weigh each raw material medicine according to the formula ratio, decoct with water twice respectively, for 30 min for the first time and 20 min for the second time, filter, combine the filtrates, and concentrate to about 400 mL to obtain. It is taken twice a day.

[0104] Animal experiment:

[0105] 1. Experimental drugs

[0106] Blank control group: Intragastric administration with equal-dose normal saline, lasting for 6 weeks in total.

[0107] Model control group: Intragastric administration with equal-dose normal saline, lasting for 6 weeks in total.

[0108] Experimental group: The traditional Chinese medicine composition prepared in Example 4 (i.e., 10 g of ginseng, 10 g of morinda root, 10 g of cyperus rhizome, 10 g of chuanxiong rhizome, 10 g of atractylodes rhizome, 10 g of gardenia fruit and 10 g of medicated leaven, using formula granules, called "Peiyuan Tongzhi Formula"), provided by Beijing Kangrentang Pharmaceutical Co., Ltd., Peiyuan Tongzhi Formula was given at 8.40 g / kg / d, lasting for 6 weeks in total. Escitalopram oxalate tablets: Specification 10 mg × 10 tablets / box, produced by Sichuan Kelun Pharmaceutical Co., Ltd., escitalopram was intragastrically administered at 1.05 mg / kg / d, lasting for 6 weeks in total.

[0109] 2. Experimental animals

[0110] One hundred and twenty male SHR rats, 12 weeks old, and thirty male WKY rats, 8 weeks old, were provided by Beijing Vital River Laboratory Animal Technology Co., Ltd.

[0111] 3. Main reagents and instruments

[0112] 3.1 Reagents: 10% chloral hydrate, glutaraldehyde, 4% paraformaldehyde, 2% Evans blue solution, formamide solution.

[0113] 3.2 Instruments:

[0114] Object recognition test: The experimental chamber was purchased from Shanghai Xinruan Information Technology Co., Ltd., with the product model XR-XX117, a plastic square box of 75 cm × 75 cm × 40 cm, with a black bottom and transparent walls on all sides.

[0115] Open field test: It was modified from the experimental chamber of the object recognition test, a plastic square box of 75 cm × 75 cm × 40 cm with a black bottom and black walls on all sides.

[0116] Animal behavior analysis software: SuperMaze animal behavior video analysis system developed by Shanghai Xinruan Information Technology Co., Ltd.; BP-2000 non-invasive blood pressure analysis system for animals (USA, Visitech, model: BP-2000); Color cards: Color scale card and gray scale card of Tiffen Q13 of Tianfen in the USA, with a size of 6 cm × 12 cm, used for color correction during digital photography. Nikon D90 single-lens reflex camera and AF-S VR105 mm f / 2.8G IF ED lens [Nikon Imaging (China) Co., Ltd.].

[0117] American Beckman automatic biochemical analyzer AU480, reagents are from Intech (Xiamen) Technology Co., Ltd. Qilin Bell vortex mixer (model: GL-88B), Hengnuo instrument centrifuge (model: AXTGL16 M), American Corning EP tube (model: 1.5mL, 5mL, 10mL); fully automatic multifunctional microplate reader: MULTISKANMK3, Thermo, USA; electric constant temperature incubator: DH4000A, Tianjin Test MINI shaker: MH-1, Kylin-Bell Lab Instruments QILINBEIER. ACS-2EAS electronic scale (Beijing Feimus Technology Development Company); AE160 electronic analytical balance (Swiss Metele Company), in addition, infrared camera, digital illuminance meter, etc. are all domestically produced. The 7.0T small animal MRI system was purchased from Varian, Palo Alto, CA, USA; the vortex oscillator (MIX-28) was purchased from Ningbo Yinzhou Qunan Experimental Instrument Co., Ltd.; the low-temperature desktop centrifuge (TGL-16) was purchased from Hunan Xiangyi Laboratory Instrument Development Co., Ltd.; the enzyme label analyzer (DNM-9602) was purchased from Beijing Pulang New Technology Co., Ltd.; the electrophoresis instrument (Mini Ge Tank) was purchased from Thermo Scientific; and the membrane transfer instrument (eBlot TM L1) GenScriPt; shaker (NYC-80) Taizhou Nomi Medical Technology Co., Ltd.; exposure instrument (eBlot) Shanghai Yibote; automatic dehydration machine (ASP200S) Leica; paraffin slicer (RM2235) Leica; baking table (HI1220) Leica; water bath (HI1220) Leica; heated paraffin embedding system (G1150H) Leica; upright fluorescence microscope (DM3000) Leica; PCR amplifier purchased from ABI, USA, NovaSeq PE 250 sequencing platform was purchased from Illumina, ultrasonic cleaner (SB-5200DT, Ningbo Xinzhi Biotechnology Co., Ltd.), sample grinder (JXFSTPRP-24 / 32, Shanghai Jingxin Industrial Development Co., Ltd.), vortex oscillator (TYXH-I, Shanghai Hannuo Instrument Co., Ltd.), desktop high-speed refrigerated centrifuge (TGL-16MS, Shanghai Luxiangyi Centrifuge Instrument Co., Ltd.), gas chromatography-mass spectrometry (7890B-5977B, Agilent), and chromatographic column (DB-WAX, 30m×0.25mm×0.25μm, Agilent).

[0118] 4. Experimental methods

[0119] 4.1 Animal model preparation and drug administration method

[0120] Referring to the internationally recognized chronic unpredictable mild stress (CUMS), a total of 12 kinds of stimuli were arranged into the stress time by the random number table method. One or two kinds of stress were given daily, and the same kind of stimulus did not appear continuously. The stress lasted for 12 weeks. WKY rats were used as the healthy control group (LS, low-salt group), and SHR rats were randomly divided into the blank group (HS, high-salt group), the model group (M, high-salt + CUMS group), the Peiyuan Tongzhi Fang group (PYKY, high-salt + CUMS + Peiyuan Tongzhi Fang group), and the citalopram group (EO, high-salt + CUMS + citalopram group), with 30 rats in each group. The healthy control group, the blank group, and the model group were intragastrically administered with normal saline. Administration concentration and dose: The equivalent dose ratio calculated according to the body surface area between humans and animals. Calculated by the dose per unit body weight, the equivalent dose of rats is 6.3 times that of humans. Citalopram was intragastrically administered at 1.05 mg / kg / d, and Peiyuan Tongzhi Fang was administered at 8.40 g / kg / d.

[0121] 4.2 General observation of rats

[0122] Including mental state, activity, hair color, etc. The stress duration and degree of spontaneous neurological deficit symptoms in rats were detected. The rats were scored for neurological function using the Bederson method (0-7 point method). The more severe the neurological function damage, the higher the score.

[0123] 4.3 Evaluation of depressive-like behaviors in rats

[0124] 4.3.1 Sucrose preference test

[0125] Before the experiment, the rats were subjected to adaptation training. In the first 24 h, two bottles of 1% sucrose solution were placed in each cage. In the following 24 h, one of the bottles was replaced with pure water, that is, one bottle of 1% sucrose solution and one bottle of pure water. After the adaptation was completed, the rats were fasted and water-deprived for 24 h for the experiment. During the experiment, one bottle of 1% sucrose solution and one bottle of pure water were placed in each cage at the same time. The positions of the bottles were exchanged at 1 h of the test, and the bottles were removed after 2 h. The water bottles were weighed before and after the experiment respectively, and the consumption of sucrose solution and pure water by the rats within 2 h was calculated.

[0126] Sucrose preference rate (%) = sucrose consumption / (sucrose consumption + pure water consumption) × 100%.

[0127] 4.3.2 Open field test

[0128] All rats were placed in a quiet, ventilated, light-proof barrier environment. The experiment was carried out using a special rat open field platform (100 cm × 100 cm × 45 cm). The first day of the experiment was for adaptation training. All rats were placed in the central grid of the experimental box for 10 minutes. After each experiment, the platform was wiped with alcohol and dried with a paper towel. Each rat was experimented for 5 minutes on the second day. The immobile time, rearing frequency, and crossing frequency of the rats were analyzed using the Labmaze animal behavior analysis software.

[0129] 4.3.3 Forced swimming experiment

[0130] Place the rats in a swimming bucket (a transparent round bucket with an inner diameter of 35 cm and a height of 60 cm, the water depth in the bucket is about 35 cm, and the water temperature is about 23 °C), observe for 5 min, and record the immobile state time in the last 4 min with a camera system. The immobile state refers to the state where the rats give up struggling, their bodies float without twisting, and the center of gravity is stable.

[0131] 4.4 Observation of rat brain microcirculation

[0132] Use a dynamic visualization and continuous microcirculation research technology platform to record the dynamic changes in blood flow velocity and blood vessel diameter of the brain microcirculation system, the dynamics of leukocyte and platelet adhesion to vascular endothelium, and the dynamics of vascular endothelial injury and repair, etc.

[0133] 4.5 Observation of rat brain tissue pathology

[0134] Use Nissl staining to observe the pathological morphology of brain tissue, TTC staining to observe the ischemic infarction of brain tissue, LFB staining to observe the degree of white matter damage, Evans blue penetration method to quantitatively analyze the change in blood-brain barrier permeability, and transmission electron microscopy to observe the structural changes of neurovascular units in the hippocampal region.

[0135] 4.6 Detection of rat intestinal flora

[0136] Use 16S rRNA high-throughput sequencing to detect fecal intestinal flora.

[0137] 4.7 Detection of rat serum and brain tissue

[0138] Use ELISA and Western blot to detect the protein expression levels of the LPS / TLR4 / NF-κB inflammatory signaling pathway and the VEGF / MMP-9 endothelial function signaling pathway in serum and hippocampus, detect the expression levels of tight junction proteins occludin, ZO-1, and claudin-5 in the hippocampal region, and use ELISA to detect the expression levels of hippocampal endothelial function markers eNOS and ET-1.

[0139] 5. Results and analysis

[0140] 5.1 Effects of Peiyuan Tongzhi Formula on the general condition of rats

[0141] In the low-salt group, the rats in the cage were quiet, huddled together, docile when grabbed, and had white and moist hair. In the high-salt group and the high-salt + CUMS group, increased urine volume occurred at 6 weeks, the urine color was basically normal, the bedding was wet, the claws were dry and desquamated, and the hair was dry, standing upright, withered and yellow, accompanied by varying degrees of irritability; in the high-salt + CUMS group, slow weight gain, significantly reduced activity, and easier urination and defecation occurred. At 12 weeks, compared with the high-salt group, the high-salt + CUMS group gradually showed significantly slower weight gain, mental fatigue, significantly reduced resistance, decreased activity, curled up and lethargic, reduced water intake, increased urine volume, light urine color, dry and withered hair, and irregular stool consistency. At 12 weeks, compared with the high-salt + CUMS group, the high-salt + CUMS + Buyuan Tongzhi Formula group and the high-salt + CUMS + citalopram group had relatively more activity, increased water intake, still resisted restraint stimulation, dry and withered hair, and basically normal stools. The limb movements of the rats in each group were normal and coordinated, and their steps were steady, and no neurological deficits occurred.

[0142] 5.2 Effect of Buyuan Tongzhi Formula on the body weight of rats

[0143] As Figure 1 shown, at the 6th week of stress, compared with the high-salt group, the body weights of the high-salt + CUMS group, the high-salt + CUMS + Buyuan Tongzhi Formula group, and the high-salt + CUMS + citalopram group decreased significantly (P < 0.05), and there was no statistical difference among the three groups (P > 0.05). At the 12th week of stress, after 6 weeks of administration, compared with the high-salt + CUMS group, the body weights of the high-salt + CUMS + Buyuan Tongzhi Formula group and the high-salt + CUMS + citalopram group increased (P < 0.05), and there was no statistical difference between the two groups (P > 0.05).

[0144] 5.3 Effect of Buyuan Tongzhi Formula on the blood pressure of rats

[0145] As Figure 2 and Figure 3 shown, at the 12th week of stress, after 6 weeks of administration, compared with the high-salt + CUMS group, the systolic blood pressure and diastolic blood pressure of the high-salt + CUMS + Buyuan Tongzhi Formula group and the high-salt + CUMS + citalopram group decreased significantly (P < 0.05).

[0146] 5.4 Behavioral evaluation

[0147] 5.4.1 Sucrose preference test

[0148] As Figure 4 shown, at the 12th week of stress, after 6 weeks of administration, compared with the low-salt group and the high-salt group, the sucrose preference rate of the high-salt + CUMS group decreased (P < 0.01). Compared with the high-salt + CUMS group, the sucrose preference rates of the high-salt + CUMS + Buyuan Tongzhi Formula group and the high-salt + CUMS + citalopram group increased (P < 0.05).

[0149] 5.4.2 Open field test

[0150] As Figure 5 shown, after 12 weeks of stress and 6 weeks of drug administration, compared with the low-salt group and the high-salt group, the immobility time of the high-salt + CUMS group increased significantly (P < 0.01). Compared with the high-salt + CUMS group, the immobility time of the high-salt + CUMS + Peiyuan Tongzhi Formula group and the high-salt + CUMS + citalopram group decreased significantly (P < 0.05).

[0151] 5.4.3 Forced swimming test

[0152] As Figure 6 shown, after 12 weeks of stress and 6 weeks of drug administration, compared with the low-salt group and the high-salt group, the immobility time of the high-salt + CUMS group increased significantly (P < 0.01). Compared with the high-salt + CUMS group, the immobility time of the high-salt + CUMS + Peiyuan Tongzhi Formula group and the high-salt + CUMS + citalopram group decreased significantly (P < 0.05).

[0153] 5.5 Nissl staining of hippocampal CA1 region

[0154] The results of Nissl staining of the hippocampal CA1 region of rats in each group observed under light microscopy are as Figure 7 shown. After 12 weeks of stress and 6 weeks of drug administration, in the low-salt group, the number of neuronal cells in the hippocampal CA1 region of rats was relatively large, arranged closely, of uniform size, regular in shape, with intact cell membranes, deeply stained cytoplasm, abundant Nissl bodies visible in the cytoplasm, clear and full cell nuclei, round in shape, with intact nuclear membrane boundaries, and clearly visible nucleoli (LS). In the high-salt group, the number of neuronal cells in the hippocampal CA1 region of rats decreased slightly, arranged more loosely, but the cytoplasm was still deeply stained, and the nuclear membrane and nucleoli were relatively clear (HS). In the high-salt + CUMS group, neuronal cells were damaged, with unequal cell sizes, irregular shapes, scattered arrangements, deeply stained cytoplasm, and reduced Nissl bodies in the cytoplasm (M). In the high-salt + CUMS + Peiyuan Tongzhi Formula group and the high-salt + CUMS + citalopram group, the damage of neuronal cells recovered more than that in the high-salt + CUMS group, the number of neuronal cells increased significantly, arranged closely, with mostly normal-shaped cells, intact cell membranes, clear and full cell nuclei, and clear nucleoli (EO, PYKY).

[0155] 5.6 Immunofluorescence

[0156] As Figure 8 and Figure 9 shown, compared with the high-salt group, the apoptosis of neuronal cells in the high-salt + CUMS group increased (P < 0.05). Compared with the high-salt + CUMS group, the apoptosis of neuronal cells in the high-salt + CUMS + Peiyuan Tongzhi Formula group and the high-salt + CUMS + citalopram group decreased significantly (P < 0.05).

[0157] 5.7 Electron microscopy of vascular nerve units in hippocampal tissue

[0158] As Figure 10As shown in the figure, a transmission electron microscope was used to observe the pathological changes of the vascular nerve units in the hippocampal tissue. It was found that in the low-salt group, the capillary morphology was regular, the basement membrane (BM) of the capillary was clear and intact; the astrocytes (ASC) outside the capillary basement membrane were normal in morphology; the endothelial cells (NC) inside the basement membrane were closely attached to the capillary wall, with regular morphology, the endothelial cell nucleus (N) had a regular morphology, and the mitochondria (M) in the cytoplasm were oval or short rod-shaped, with clear inner cristae and intact structure in the mitochondria, and the tight junctions (LJ) between endothelial cells were clear; the axons of the neurons (NEU) around the capillaries had regular morphology, and the mitochondria in the cytoplasm had clear structure.

[0159] In the high-salt group, the capillary morphology was irregular, and some segments of the capillary basement membrane were thin and the structure was blurred; the astrocytes outside the capillary basement membrane were swollen; the endothelial cells inside the basement membrane were closely attached to the capillary wall, the endothelial cells were swollen, the mitochondria in the cytoplasm were oval, the mitochondria were slightly swollen and the inner cristae were blurred, and the tight junction structure between endothelial cells was blurred; the axons of the neurons around the capillaries were swollen, and there was swelling in the cytoplasm.

[0160] In the high-salt + CUMS group, the capillary morphology was extremely irregular, and most segments of the capillary basement membrane were thin and the structure was blurred; the astrocytes outside the capillary basement membrane were swollen; the endothelial cells inside the basement membrane were closely attached to the capillary wall, the endothelial cells were swollen, the mitochondria in the cytoplasm were oval, the mitochondria were slightly swollen and the inner cristae were blurred, and the tight junction structure between endothelial cells was blurred; the axons of the neurons around the capillaries were swollen, and there were swelling and vacuolization in the cytoplasm.

[0161] In the high-salt + CUMS + citalopram group, the capillary morphology was slightly irregular, and most segments of the capillary basement membrane had clear structure and a few segments were slightly blurred; the astrocytes outside the capillary basement membrane were slightly swollen; the endothelial cells inside the basement membrane were closely attached to the capillary wall, with regular morphology, the endothelial cell nucleus had a regular morphology, the mitochondria in the cytoplasm were oval or short rod-shaped, the inner cristae of the mitochondria were slightly blurred, and the tight junction structure between endothelial cells was slightly blurred; the axons of the neurons around the capillaries had basically regular morphology, and most of the mitochondria in the cytoplasm had clear structure.

[0162] In the high-salt + CUMS + Peiyuan Tongzhi formula group, the capillary morphology was slightly irregular, and most segments of the capillary basement membrane had clear structure and a few segments were slightly blurred; the astrocytes outside the capillary basement membrane were slightly swollen; the endothelial cells inside the basement membrane were closely attached to the capillary wall, with regular morphology, the endothelial cell nucleus had a regular morphology, the mitochondria in the cytoplasm were oval or short rod-shaped, the inner cristae of the mitochondria were slightly blurred, and the tight junction structure between endothelial cells was slightly blurred; the axons of the neurons around the capillaries had basically regular morphology, and most of the mitochondria in the cytoplasm had clear structure.

[0163] 5.8 LFB staining of hippocampal CA1 region

[0164] AsFigure 11 As shown in the figure, white matter lesions in the hippocampal tissue area were observed. It was found that in the low-salt group, myelin fibers were strongly stained by LFB, the myelin fibers were dense and uniform, the morphology of the myelin fibers was intact, arranged neatly and compactly, and there was no vacuole structure. In the high-salt group and the high-salt + CUMS group, the LFB staining was lighter, the myelin sheath gap became larger, a large number of vacuoles were visible, and the myelin arrangement was irregular and more chaotic, which was a typical demyelinating lesion, and the degree in the high-salt + CUMS group was more severe. Compared with the high-salt + CUMS group, in the high-salt + CUMS + Peiyuan Tongzhi formula group, the myelin was denser, the disordered arrangement was improved, and the staining degree of LFB increased significantly. In the high-salt + CUMS + citalopram group, the improvement was not obvious.

[0165] 5.9 Blood-brain barrier permeability

[0166] As Figure 12 shown, after 12 weeks of stress and 6 weeks of drug administration, compared with the high-salt group, the blood-brain barrier permeability in the high-salt + CUMS group increased (P < 0.05). Compared with the high-salt + CUMS group, the blood-brain barrier permeability in the high-salt + CUMS + Peiyuan Tongzhi formula group and the high-salt + CUMS + citalopram group decreased (P < 0.05).

[0167] 5.10 Expression levels of serum-related inflammatory factors

[0168] As Figure 13 shown, after 12 weeks of stress and 6 weeks of drug administration, compared with the low-salt group, the expression levels of LPS, TLR4, TNF-α, NF-κB, IL-6, and IL-1β in the high-salt group increased (P < 0.01). Compared with the high-salt group, the expression levels of LPS, TLR4, TNF-α, NF-κB, IL-6, and IL-1β in the high-salt + CUMS group increased (P < 0.05). Compared with the high-salt + CUMS group, the expression levels of LPS, TLR4, TNF-α, NF-κB, IL-6, and IL-1β in the high-salt + CUMS + Peiyuan Tongzhi formula group and the high-salt + CUMS + citalopram group decreased (P < 0.01), and there was no statistical difference between the two groups.

[0169] 5.11 Expression levels of hippocampal tissue-related inflammatory factors

[0170] As Figure 14As shown, after 12 weeks of stress and 6 weeks of drug administration, compared with the low-salt group, the expression levels of LPS, TLR4, TNF-α, NF-κB, IL-6, and IL-1β in the high-salt group increased (P<0.01). Compared with the high-salt group, the expression levels of LPS, TLR4, TNF-α, NF-κB, IL-6, and IL-1β in the high-salt + CUMS group increased (P<0.05). Compared with the high-salt + CUMS group, the expression levels of LPS, TLR4, TNF-α, NF-κB, IL-6, and IL-1β in the high-salt + CUMS + Peiyuan Tongzhi Formula group and the high-salt + CUMS + citalopram group decreased (P<0.01), and there was no statistical difference between the two groups.

[0171] 5.12 Contents of VEGF and MMP-9 in hippocampal tissue

[0172] As Figure 15 shown, after 12 weeks of stress and at the 6th week of drug administration, compared with the low-salt group, the contents of VEGF and MMP-9 in the hippocampal tissue of the high-salt group increased significantly (P<0.01). Compared with the high-salt group, the contents of VEGF and MMP-9 in the hippocampal tissue of the high-salt + CUMS group increased significantly (P<0.01). Compared with the high-salt + CUMS group, the contents of VEGF and MMP-9 in the hippocampal tissue of the high-salt + CUMS + Peiyuan Tongzhi Formula group and the high-salt + CUMS + citalopram group decreased significantly (P<0.01), and there was no statistical difference between the two groups.

[0173] 5.13 Contents of Occludin, ZO-1, and Claudin-5 in hippocampal tissue

[0174] As Figure 16 shown, after 12 weeks of stress and at the 6th week of drug administration, compared with the low-salt group, the contents of Occludin, ZO-1, and Claudin-5 in the hippocampal tissue of the high-salt group decreased significantly (P<0.05). Compared with the high-salt group, the contents of Occludin, ZO-1, and Claudin-5 in the hippocampal tissue of the high-salt + CUMS group decreased significantly (P<0.01). Compared with the high-salt + CUMS group, the contents of Occludin, ZO-1, and Claudin-5 in the hippocampal tissue of the high-salt + CUMS + Peiyuan Tongzhi Formula group and the high-salt + CUMS + citalopram group increased significantly (P<0.01), and there was no statistical difference between the two groups.

[0175] 5.14 Contents of eNOS and ET-1 in hippocampal tissue

[0176] As Figure 17As shown, compared with the low-salt group, the content of eNOS in the hippocampal tissue of the high-salt group decreased (P<0.01), and the content of ET-1 increased (P<0.01). Compared with the high-salt group, the content of eNOS in the hippocampal tissue of the high-salt + CUMS group decreased (P<0.05), and the content of ET-1 increased (P<0.05). Compared with the high-salt + CUMS group, the content of eNOS in the high-salt + CUMS + Peiyuan Tongzhi Formula group and the high-salt + CUMS + citalopram group increased (P<0.05), and the content of ET-1 decreased (P<0.05).

[0177] 5.15 α-diversity analysis

[0178] α-diversity analysis can reflect the diversity of microorganisms in the detected samples. Common indicators include the Chao1 index, Shannon index, and Simpson index. As Figure 18 shown, the present invention found that compared with the low-salt group, the Chao1 index, Shannon index, and Simpson index in the high-salt group decreased significantly (P<0.05), indicating a reduction in the diversity of the microbial community. Compared with the high-salt group, the Chao1 index, Shannon index, and Simpson index in the high-salt + CUMS group decreased (P<0.05), indicating a reduction in the diversity of the microbial community. Compared with the high-salt + CUMS group, there was no statistical significance in the Chao1 index, Shannon index, and Simpson index in the high-salt + CUMS + Peiyuan Tongzhi Formula group, indicating that Peiyuan Tongzhi Formula may not increase the diversity of the microbial community in depressive rats.

[0179] 5.16 β-diversity analysis

[0180] β-diversity analysis mainly reflects the similarity degree of microbial communities between groups. By comparing the OTU sequence similarity, evolutionary relationship, and community structure of each group, the differences between samples of different groups are compared. The present invention uses principal component analysis (PCA) for analysis. As Figure 19 shown, when the species composition of the samples is more similar, the closer their distances are in the graph, and vice versa, the greater the differences. The results found that the difference between the low-salt group and the high-salt group was small, and there were differences between the high-salt group and the high-salt + CUMS group, and the high-salt + CUMS + Peiyuan Tongzhi Formula group.

[0181] The embodiments described above are only descriptions of the preferred embodiments of the present invention, and do not limit the scope of the present invention. Without departing from the design spirit of the present invention, various deformations and improvements made by those of ordinary skill in the art to the technical solutions of the present invention should all fall within the protection scope determined by the claims of the present invention.

Claims

1. A Chinese medicine composition for treating depression, characterized in that: The Chinese medicine composition is composed of the following components by weight: 10 parts of ginseng, 10 parts of Morinda officinalis, 10 parts of Chuanxiong, 10 parts of Gardenia jasminoides, 10 parts of Shenqu, 10 parts of Cyperus rotundus and 10 parts of Atractylodes lancea; The depression has the pathological characteristics of "depression-deficiency-stagnation"; The preparation method of the Chinese medicine composition is selected from any one of the following methods (1) to (3): (1) crushing the raw materials and mixing them to prepare the traditional Chinese medicine composition for treating depression; (2) The raw materials are mixed and extracted or extracted separately to obtain an extract, which is the Chinese medicine composition for treating depression; (3) The raw materials are mixed and extracted or extracted separately, and the obtained extract is added with pharmaceutically acceptable excipients to prepare a conventional dosage form, which is the Chinese medicine composition for treating depression.

2. A method for preparing a Chinese medicine composition for treating depression as claimed in claim 1, characterized in that: Select any one of the following methods (1)-(3): (1) crushing the raw materials and mixing them to prepare the traditional Chinese medicine composition for treating depression; (2) The raw materials are mixed and extracted or extracted separately to obtain an extract, which is the Chinese medicine composition for treating depression; (3) The raw materials are mixed and extracted or extracted separately, and the obtained extract is added with pharmaceutically acceptable excipients to prepare a conventional dosage form, which is the Chinese medicine composition for treating depression.

3. The preparation method according to claim 2, characterized in that: The extraction method is decoction or reflux.

4. The preparation method according to claim 2, characterized in that: The dosage form is granules, capsules or oral liquid.

5. Use of the Chinese medicine composition as claimed in claim 1 in preparing a medicine for treating depression.

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