Antidepressant compound Chinese medicine and preparation method thereof

Abstract

The present invention provides an antidepressant compound Chinese medicine and its preparation method, belonging to the technical field of traditional Chinese medicine. The present invention provides an antidepressant compound Chinese medicine, the components of which mainly include notopterygium root polysaccharide (NI) and Lycium ruthenicum acid polysaccharide (LA), which have excellent neuroprotective effects in mice. The prepared compound medicine has shown behavioral effects on mice with chronic unpredictable depression and LPS-induced depression, including reduced immobility time in tail suspension and swimming, and increased sucrose preference in the model mice. Furthermore, it has a protective effect on PC12 neural cells cultured in vitro. The notopterygium root polysaccharide extracted by this method has a purity of approximately 67% to 72%, and the Lycium ruthenicum acid polysaccharide has a purity of approximately 55% to 58%.

Description

Technical Field

[0001] The present invention belongs to the technical field of traditional Chinese medicine, and particularly relates to an antidepressant compound traditional Chinese medicine and a preparation method thereof. Background Art

[0002] Depression, a global mental illness, has become a leading cause of poor health and disability worldwide. Symptoms typically include low mood, anhedonia, anxiety, irritability, distractibility, loss of appetite, cognitive impairment, and even suicidal tendencies. Numerous theories exist regarding depression, but it is generally believed to be caused by multisystem interactions involving abnormal neurotransmitter expression, impaired neuroplasticity, hyperactivity of the hypothalamic-pituitary-adrenal (HPA) axis, and inflammation and oxidative stress.

[0003] Research on depression is still developing, and current drug treatments are associated with numerous side effects. Therefore, the development of safe and effective natural medicines has become a research hotspot. Natural medicines typically contain multiple active ingredients that can act on the pathogenesis of depression and improve depressive symptoms through various mechanisms, such as regulating neurotransmitter levels, improving antioxidant capacity, and reducing inflammatory responses. Therefore, the discovery of novel and highly effective antidepressant drugs is a pressing technical challenge in this field. Summary of the Invention

[0004] The present invention provides an antidepressant compound Chinese medicine and a preparation method thereof. Notopterygium incisum polysaccharide (NI) and Lycium ruthenicum acidic polysaccharide (LA) have high purity and better efficacy. The antidepressant compound Chinese medicine significantly enhances its neuroprotective effect through combined use of the drugs.

[0005] In order to achieve the above-mentioned purpose of the present invention, the following technical solutions are adopted:

[0006] An antidepressant compound traditional Chinese medicine, the components of which mainly include notopterygium wilfordii polysaccharide and lycium ruthenicum acid polysaccharide, and the weight ratio of the two is: notopterygium wilfordii polysaccharide: lycium ruthenicum acid polysaccharide = (1-2): (2-1).

[0007] Notopterygium wilfordii Notopterygium incisum Notopterygium wilfordii Notopterygium franchetii The main chemical components isolated from the dried rhizomes and roots include volatile oils and terpenes, coumarins, sugars and glycosides, phenolic acids, polyenes, alkaloids, etc. According to the Compendium of Materia Medica, Notopterygium incisum has anti-inflammatory, antibacterial, antioxidant, antipyretic and analgesic activities, and has significant effects on the cardiovascular system, digestive system, respiratory system and central nervous system. Notopterygium incisum polysaccharide is Notopterygium incisum ( Notopterygium incisumA polysaccharide compound extracted from the Chinese herb Rhizoma Notopterygii (Rhizoma Notopterygii). In traditional Chinese medicine, Notopterygii polysaccharide has multiple uses: dispelling wind and dampness, promoting blood circulation and removing blood stasis, and providing anti-inflammatory benefits.

[0008] Black wolfberry ( Lycium ruthenicum Lycium barbarum (Lycium ruthenicum) is a shrub belonging to the genus Lycium in the Solanaceae family. Black wolfberry (Lycium ruthenicum) has a long history of use in traditional Chinese medicine. According to the Uyghur Pharmacopoeia and the Jingzhu Materia Medica, its fruit has been shown to treat heart disease, strengthen the kidneys, moisten the liver, improve eyesight, invigorate the stomach, nourish the brain, combat aging, and regulate menstruation. In recent years, scientific research has further validated the pharmacological effects of black wolfberry, including its antioxidant and anti-inflammatory properties, leading to its widespread use in health foods. Lycium barbarum polysaccharides (Lycium barbarum polysaccharides) are the most studied group of water-soluble sugar complexes among the phytochemicals found in wolfberry. They are composed of branched polysaccharides, proteoglycans, galacturonic acid, and amino acids. The crude polysaccharides in black wolfberry are primarily composed of monosaccharides such as rhamnose, glucose, glucuronic acid, galactose, galacturonic acid, mannose, xylose, and arabinose. They exhibit a variety of biological activities, including antioxidant, immunomodulatory, neuroprotective, and prebiotic activities.

[0009] This application combines Notopterygium wilfordii polysaccharide with Lycium barbarum acid polysaccharide, and the antidepressant activity is significantly better than that of the single herb, showing obvious compatibility advantages.

[0010] The antidepressant compound Chinese medicine as described above mainly comprises Notopterygium wilfordii polysaccharide and Lycium ruthenicum acid polysaccharide, and the weight ratio of the two is: Notopterygium wilfordii polysaccharide: Lycium ruthenicum acid polysaccharide = 2:1.

[0011] The preparation method of the above-mentioned antidepressant compound Chinese medicine comprises the following steps:

[0012] 1) Preparation of Notopterygium wilfordii polysaccharide: Rhizome of Notopterygium wilfordii was provided by the Traditional Chinese Medicine Planting Cooperative of Makohe Township, Banma County, Golog Tibetan Autonomous Prefecture, Qinghai Province. Black wolfberry fruit was collected from Dongshagen, Dulan County, Haixi Mongol and Tibetan Autonomous Prefecture, Qinghai Province, China; identified by Professor Xue Fenglu of the Northwest Institute of Plateau Biology, Chinese Academy of Sciences (specimen number: LR20110719). Notopterygium wilfordii rhizome and black wolfberry were air-dried, ground, and sieved to obtain Notopterygium wilfordii rhizome powder and black wolfberry powder.

[0013] 2) soaking the Notopterygium wilfordii rhizome powder in pure water at room temperature, extracting with hot water, and subjecting it to ethanol graded precipitation, followed by deproteinization, and freeze-drying to obtain Notopterygium wilfordii crude polysaccharide NIWP.

[0014] 3) The ruthenium wolfberry powder was subjected to microwave treatment, hot water extraction, ethanol graded precipitation and vacuum drying to obtain ruthenium wolfberry total polysaccharide (LRWP); the ruthenium wolfberry total polysaccharide was further subjected to chromatography to obtain ruthenium wolfberry acidic polysaccharide LRWP-A.

[0015] 4) Determine the purity of the Notopterygium wilfordii polysaccharide and Lycium ruthenicum acidic polysaccharide obtained in steps 2) and 3). The purity of the Notopterygium wilfordii polysaccharide extracted by this method is 67-72%, and the purity of the Lycium ruthenicum acidic polysaccharide is 55-58%.

[0016] In the above-mentioned method for preparing the antidepressant compound Chinese medicine, the specific operations of step 1) include:

[0017] The Notopterygium wilfordii rhizome and Lycium ruthenicum var. ruthenicum rhizome are ground after being naturally dried, and then passed through a 70-100 mesh sieve to obtain Notopterygium wilfordii rhizome powder and Lycium ruthenicum var. ruthenicum powder.

[0018] Drying can remove excess moisture, making it easier to grind and increasing the solubility of polysaccharides. Grinding can destroy the cell walls of Notopterygium wilfordii rhizomes and Lycium ruthenicum, increasing the solubility of Notopterygium wilfordii polysaccharides and Lycium ruthenicum polysaccharides, making them easier to extract in subsequent processing.

[0019] In the above-mentioned method for preparing the antidepressant compound Chinese medicine, in step 2), the hot water extraction and ethanol graded precipitation treatment are specifically as follows:

[0020] 500 g of the Notopterygium wilfordii rhizome powder was weighed and added to 5 L of pure water and soaked at room temperature for 4 h. The powder was then boiled in boiling water for 2 h each time for a total of 3 extractions. After filtering through four layers of gauze, the filtrate was collected and concentrated to 4-5 L at 80° C. 95% ethanol was added until the ethanol concentration of the mixed solution reached 80%. The mixture was allowed to stand at room temperature overnight, and the precipitate was collected by centrifugation. The precipitate was freeze-dried to obtain Notopterygium wilfordii crude polysaccharide NIWP.

[0021] 95% ethanol solution is calculated based on volume percentage.

[0022] In the above-mentioned method for preparing the antidepressant compound Chinese medicine, in step 3), the crude extraction step is specifically as follows:

[0023] The ruthenium wolfberry powder was extracted by microwave at 370W for 30 minutes, and the solution and extract were soaked for 2.5 hours at a ratio of 25:1 to obtain ruthenium wolfberry total polysaccharide LRWP.

[0024] Microwave treatment can quickly heat the medicinal materials to ensure the activity of polysaccharides, and can also enhance the penetration of the solvent, thereby improving the extraction efficiency and yield of polysaccharides; at the same time, microwave treatment can make the heat inside the black fruit wolfberry evenly distributed, improving the consistency and repeatability of the extraction.

[0025] In the above-mentioned method for preparing the antidepressant compound Chinese medicine, in step 3), the chromatography step is specifically as follows:

[0026] The extracted total polysaccharides from Lycium ruthenicum were prepared into a 20-25 mg / mL aqueous solution. After centrifugation to remove impurities, the supernatant was collected. A DEAE-Cellulose column was equilibrated with three column volumes of 0.05 mol / L NaHCO₃ solution. The elution process was as follows: distilled water, then 0.05, 0.1, 0.25, and 0.5 mol / L NaHCO₃. The supernatant was applied to a DEAE-Cellulose column and eluted sequentially with distilled water and NaHCO₃ solution. The eluate was collected to obtain the neutral sugar fraction LRWP-N and the acidic polysaccharide LRWP-A.

[0027] Take 0.1 g of LRWP-A, dissolve it in ultrapure water to prepare a solution with a concentration of 10 mg / ml, pass it through a 0.45 μm filter membrane, and apply it to a DEAE-Sepharose Fast Flow column at a flow rate of 1 mL / min. Collect the eluate with a fraction collector, collecting one tube for every 5 mL, and elute until no polysaccharide is detected in the eluate. Dialyze and lyophilize to obtain the purified polysaccharide component LRWP-A.

[0028] Compared with the prior art, the present invention has the following beneficial effects:

[0029] 1. The antidepressant activity of Notopterygium wilfordii polysaccharide combined with Lycium barbarum acid polysaccharide on mice was significantly better than that of the single drug, showing obvious advantages of the combination. After the combination, the immobility time of mice in the tail suspension test and forced swimming test was significantly shortened, and the degree of sugar water preference of mice in the sugar water preference test was significantly improved.

[0030] 2. The extraction method of Notopterygium wilfordii polysaccharide and Lycium ruthenicum polysaccharide is easy to operate and has high extraction efficiency. The purity of Lycium ruthenicum acidic polysaccharide extracted by this method is about 55-58%, and the purity of Notopterygium wilfordii polysaccharide is about 67-72%.

[0031] Compared with the prior art, the antidepressant compound Chinese medicine of the present invention has the following beneficial effects:

[0032] (1) Significant neuroprotective effect: The combination of Notopterygium wilfordii polysaccharide and Lycium ruthenicum acid polysaccharide is significantly superior to single drugs, especially in the chronic unpredictable depression model and LPS-induced depression model mice, showing obvious compatibility advantages. Through behavioral experiments, it can effectively reduce the immobility time of tail suspension and swimming in model mice.

[0033] (2) Protective effect on nerve cells: The compound showed a good protective effect on PC12 nerve cells cultured in vitro, indicating its potential application value in the treatment of depression.

[0034] (3) The extraction process is simple and efficient: The extraction method of Notopterygium wilfordii polysaccharide and Lycium barbarum acid polysaccharide is simple to operate and has high extraction efficiency, which ensures the purity and activity of the finished product and provides the possibility for large-scale production. BRIEF DESCRIPTION OF THE DRAWINGS

[0035] Figure 1 Effects of NI, LA, and NILA on the behavior of LPS-induced depressed mice and CUMS-induced depressed mice. * P < 0.05 and ** P < 0.01 vs. the Control group. # P < 0.05 and ## P < 0.01 vs. the Model group.

[0036] Figure 2 Effects of different polysaccharides on the antioxidant activity of H2O2-induced PC12 cells, * P < 0.05 and ** P < 0.01 vs. the Control group. # P < 0.05 and ## P < 0.01 vs. the Model group.

[0037] Figure 3 The effects of different polysaccharides on the morphology of PC12 cells induced by H2O2;

[0038] Figure 4 Effects of different polysaccharides on the expression of Caspase 3 protein in PC12 cells induced by H2O2. DETAILED DESCRIPTION

[0039] The embodiments of the present invention will be described in detail below with reference to the examples, but those skilled in the art will appreciate that the following examples are intended only to illustrate the present invention and should not be construed as limiting the scope of the invention. Where specific conditions are not specified in the examples, the methods were performed according to conventional conditions or the conditions recommended by the manufacturer. Where the manufacturers of the reagents or instruments are not specified, they are all conventional products that can be purchased commercially. Example 1

[0040] Effects of Notopterygium wilfordii polysaccharide and Lycium ruthenicum polysaccharide on the behavior of LPS-induced depressed mice

[0041] Experimental animals: C57BL / 6J male mice (20 ± 2 g) were purchased from Changchun Yisi Laboratory Animal Technology Co., Ltd. (Experimental Animal License No.: SYXK (Ji) 2019-0015).

[0042] a. Experimental animals were given the drug by gavage for 10 consecutive days. The dosage is as follows:

[0043] Table 1 Animal gavage dosage table

[0044] Group dose volume Normal saline blank control group -- 0.2ml Fluoxetine positive control group 3.6 mg / kg 0.2ml Notopterygium wilfordii polysaccharide group 90 mg / kg 0.2ml Lycium barbarum acidic polysaccharide group 90 mg / kg 0.2ml Notopterygium wilfordii polysaccharide + Lycium ruthenicum acidic polysaccharide group 60+30 mg / kg 0.2ml

[0045] b. On days 9-11, the blank control group received an intraperitoneal injection of 10 ml / kg normal saline, and the other five groups received an intraperitoneal injection of the same volume of LPS (0.1 mg / ml).

[0046] c. Behavioral testing was performed on day 12. Tail suspension test (TST): Animals were suspended 50 cm above a table using tape, approximately 1 cm from the tip of the tail. The test lasted 6 minutes, and the time the mice remained immobile was recorded during the last 4 minutes of the test. Mice were considered immobile when they were passively suspended and completely motionless.

[0047] d. Forced Swim Test (FST): Mice were individually placed in a cylindrical container 25 cm high and 15 cm in diameter containing 10 cm of water at 25°C. The cumulative immobility time was recorded during the last 4 minutes of a 6-minute test. Mice were considered immobile when they stopped struggling or appeared to be floating.

[0048] e. Sugar Preference Test (SPT): Prior to testing, mice were given free access to a 1% sucrose solution for 24 hours and then to pure water for 24 hours. Testing was then conducted after a 12-hour food and water deprivation period. All mice were individually housed and had free access to 1% sucrose solution and pure water for 12 hours. The volumes of sucrose solution and pure water consumed were calculated. Sucrose preference was calculated as follows: Sucrose preference = (sucrose solution consumption / total consumption) × 100.

[0049] The experimental results are as follows Figure 1 As shown. In the tail suspension test, mice are suspended at an appropriate height and observed to see if they exhibit behaviors of weakness and despair. The results showed that compared with the control group, mice in the Notopterygium wilfordii polysaccharide + Lycium ruthenicum acid polysaccharide group exhibited significantly reduced tail suspension time, indicating that the combination of Notopterygium wilfordii polysaccharide and Lycium ruthenicum acid polysaccharide is more effective in improving depressive behavior. The Notopterygium wilfordii polysaccharide + Lycium ruthenicum acid polysaccharide group also showed a greater effect than mice given Notopterygium wilfordii polysaccharide or Lycium ruthenicum acid polysaccharide alone.

[0050] In the forced swim test, mice were placed in water and forced to swim to observe whether they displayed helplessness and negative behaviors. The results showed that the mice in the group receiving Notopterygium wilfordii polysaccharide plus Lycium ruthenicum acid polysaccharide significantly reduced their swimming time and number of dives, and displayed less depression and frustrated behavior compared to the control group. The effect of Notopterygium wilfordii polysaccharide plus Lycium ruthenicum acid polysaccharide was also more significant than that of mice receiving Notopterygium wilfordii polysaccharide or Lycium ruthenicum acid polysaccharide alone.

[0051] In a sugar water preference test, mice were offered two choices: sugar water or plain water. Results showed that mice in the Notopterygium wilfordii polysaccharide plus Lycium ruthenicum acid polysaccharide group showed a stronger preference for the sugar water and exhibited a stronger sense of reward and positive emotions compared to the control group. The Notopterygium wilfordii polysaccharide plus Lycium ruthenicum acid polysaccharide group also showed a greater effect than mice given either Notopterygium wilfordii polysaccharide or Lycium ruthenicum acid polysaccharide alone.

[0052] Experimental Example 2

[0053] Effects of Notopterygium wilfordii polysaccharide and Lycium ruthenicum polysaccharide on the behavior of CUMS depression mice

[0054] a. Experimental animals were given the drug by gavage for 4 consecutive weeks. The dosage is as follows:

[0055] Table 2 Oral dosing of experimental animals

[0056] Group dose volume Normal saline blank control group -- 0.2ml Fluoxetine positive control group 3.6 mg / kg 0.2ml Notopterygium wilfordii polysaccharide group 90 mg / kg 0.2ml Lycium barbarum acidic polysaccharide group 90 mg / kg 0.2ml Notopterygium wilfordii polysaccharide + Lycium ruthenicum acidic polysaccharide group 60+30 mg / kg 0.2ml

[0057] b. Except for the blank control group, the other groups were subjected to two different stressors daily. The stressors included food and water deprivation, night light, moist bedding, 45°C cage tilt, light / dark cycle, and tail clip stimulation. The order of stressors was changed every three days for 4 weeks.

[0058] c. After 4 weeks, conduct behavioral experiments similar to those in Experimental Example 1, including the tail suspension test (TST), forced swim test (FST), and sucrose preference test (SPT).

[0059] The experimental results are as follows Figure 1 In the tail suspension test, mice in the Notopterygium wilfordii polysaccharide + Lycium ruthenicum acid polysaccharide group showed significantly reduced tail suspension time compared to the control group, indicating that the combination of Notopterygium wilfordii polysaccharide and Lycium ruthenicum acid polysaccharide is more effective in improving depressive behavior. The Notopterygium wilfordii polysaccharide + Lycium ruthenicum acid polysaccharide group also showed a greater effect than mice given Notopterygium wilfordii polysaccharide or Lycium ruthenicum acid polysaccharide alone.

[0060] In the forced swim test, mice in the Notopterygium wilfordii polysaccharide + Lycium ruthenicum acid polysaccharide group showed significantly reduced swimming time and number of dives, and displayed less depression and frustrated behavior compared to the control group. The effect of the Notopterygium wilfordii polysaccharide + Lycium ruthenicum acid polysaccharide group was also more significant than mice given Notopterygium wilfordii polysaccharide or Lycium ruthenicum acid polysaccharide alone.

[0061] In a test of sugar water preference, mice in the group receiving Notopterygium wilfordii polysaccharide plus Lycium ruthenicum acid polysaccharide showed a greater preference for sugar water and exhibited a stronger sense of reward and positive emotions compared to the control group. The Notopterygium wilfordii polysaccharide plus Lycium ruthenicum acid polysaccharide group also had a better effect than mice given Notopterygium wilfordii polysaccharide or Lycium ruthenicum acid polysaccharide alone.

[0062] Experimental Example 3

[0063] Protective effect of Notopterygium wilfordii polysaccharide and Lycium ruthenicum polysaccharide on neuronal cells in vitro

[0064] a. PC12 cells were divided into the following 11 groups:

[0065] Table 3 PC12 cell grouping

[0066] Group <![CDATA[H2O2 dosage]]> Drug dosage Negative control group -- -- Model Group 100 μM -- Notopterygium wilfordii polysaccharide group 100 μM 75 μg / ml Lycium barbarum acidic polysaccharide group 100 μM 75 μg / ml Notopterygium wilfordii polysaccharide + Lycium ruthenicum acidic polysaccharide group (1:1) 100 μM 50+50 μg / ml Notopterygium wilfordii polysaccharide group 100 μM 100 μg / ml Lycium barbarum acidic polysaccharide group 100 μM 50 μg / ml Notopterygium wilfordii polysaccharide + Lycium ruthenicum acidic polysaccharide group (2:1) 100 μM 100+50μg / ml Notopterygium wilfordii polysaccharide group 100 μM 50 μg / ml Lycium barbarum acidic polysaccharide group 100 μM 100 μg / ml Notopterygium wilfordii polysaccharide + Lycium ruthenicum acidic polysaccharide group (1:2) 100 μM 50+100μg / ml

[0067] b. Seed PC12 cells in a 6-well plate and culture until the cell density reaches 70-80%. Wash the cells once with PBS to remove any residual culture medium. Add H2O2 (100 μM) and incubate for 24 hours to induce oxidative stress.

[0068] c. After injury induction, remove the culture medium and wash the cells with PBS to remove residual H₂O₂. Add culture medium containing different concentrations of the compound Chinese medicine and continue culturing the cells for 24 hours to observe the effects of the drug treatment on the cells.

[0069] d. The SOD activity kit and CAT activity kit were used to measure the activities of SOD and CAT in each group of cells.

[0070] e. The expression levels of Caspase 3 in cells of each group were analyzed by Western blot, using β-actin as an internal reference protein.

[0071] The experimental results are as follows Figure 2 As shown. SOD and CAT are two important antioxidant enzymes that can regulate the level of intracellular oxidative stress. In the model group, the activities of SOD and CAT were significantly reduced. However, in the group of Notopterygium wilfordii polysaccharide + Lycium ruthenicum acid polysaccharide, the activities of SOD and CAT were significantly increased, which was significantly better than the group treated with Notopterygium wilfordii polysaccharide or Lycium ruthenicum acid polysaccharide alone. The difference was statistically significant, indicating that the compound Chinese medicine has a significant antioxidant effect. The effect is most significant when the ratio of Notopterygium wilfordii polysaccharide and Lycium ruthenicum acid polysaccharide is 2:1. Therefore, the present invention selects Notopterygium wilfordii polysaccharide: Lycium ruthenicum acid polysaccharide in a ratio of 2:1 for subsequent research.

[0072] An inverted microscope was used to observe changes in cell morphology, and the morphology of cells in each group was recorded. Negative control group: cells showed normal morphology, with obvious protrusions, intact morphology and no damage. Cells in the model group showed obvious shrinkage and loss of normal morphology, and a large number of cells died. The cell morphology of the Notopterygium wilfordii polysaccharide-treated group and the Lycium ruthenicum acid polysaccharide-treated group improved, but some cells still shrank. The Notopterygium wilfordii polysaccharide + Lycium ruthenicum acid polysaccharide group (Notopterygium wilfordii polysaccharide: Lycium ruthenicum acid polysaccharide = 2:1) significantly improved cell morphology, and the cell morphology was significantly better than that of the group treated with Notopterygium wilfordii polysaccharide or Lycium ruthenicum acid polysaccharide alone. Cell shrinkage was reduced, and most cells returned to normal morphology.

[0073] Caspase 3 is an important enzyme that promotes cell apoptosis. In the model group, the expression level of Caspase 3 increased significantly, indicating that cell apoptosis was activated. However, in the group treated with Notopterygium wilfordii polysaccharide or Lycium ruthenicum acid polysaccharide alone, the expression level of Caspase 3 decreased significantly. In the Notopterygium wilfordii polysaccharide + Lycium ruthenicum acid polysaccharide group (Notopterygium wilfordii polysaccharide: Lycium ruthenicum acid polysaccharide = 2:1), the expression level of Caspase 3 decreased more significantly, which was lower than that in the group treated with Notopterygium wilfordii polysaccharide or Lycium ruthenicum acid polysaccharide alone. The difference was statistically significant, indicating that the compound Chinese medicine can effectively reduce cell apoptosis ( Figure 4 ).

[0074] In summary, the Chinese medicinal compound Notopterygium wilfordii polysaccharide + Lycium ruthenicum acid polysaccharide exhibited significant neuroprotective effects in a hydrogen peroxide-induced injury model for PC12 cells. Compared with the groups treated with either Notopterygium wilfordii polysaccharide or Lycium ruthenicum acid polysaccharide alone, the Notopterygium wilfordii polysaccharide + Lycium ruthenicum acid polysaccharide group demonstrated superior effects in terms of cell morphology protection, improvement of oxidative stress molecules, and reduction of cell apoptosis. These results suggest that the Notopterygium wilfordii polysaccharide + Lycium ruthenicum acid polysaccharide compound has a synergistic effect and can provide a stronger neuroprotective effect.

[0075] In summary, the research results of the present invention clearly demonstrate that the traditional Chinese medicine compound Notopterygium wilfordii polysaccharide and Lycium ruthenicum polysaccharide have excellent neuroprotective effects, with their antidepressant effects significantly superior to those of Notopterygium wilfordii polysaccharide or Lycium ruthenicum polysaccharide administered alone. This discovery provides strong scientific evidence for the potential application of the traditional Chinese medicine compound Notopterygium wilfordii polysaccharide and Lycium ruthenicum polysaccharide in neuroprotective treatments, and has important guiding significance for related clinical treatments and drug development.

[0076] It was concluded that the preparation method of the antidepressant compound Chinese medicine provided by the present invention not only improves the clinical application effect of Chinese medicine, but also provides a new idea and solution for the treatment of depression, and has good market prospects and social value.

[0077] Although the above embodiment provides a detailed description of the present invention, it is only a part of the embodiments of the present invention, not all of the embodiments. People can also obtain other embodiments based on this embodiment without creativity, and these embodiments all fall within the scope of protection of the present invention.

Claims

1. An antidepressant compound Chinese medicine, characterized in that: It is composed of Notopterygium wilfordii polysaccharide and Lycium ruthenicum acid polysaccharide, wherein the mass ratio of Notopterygium wilfordii polysaccharide to Lycium ruthenicum acid polysaccharide is 2:1; The preparation method of the Notopterygium wilfordii polysaccharide comprises the following steps: grinding dried Notopterygium wilfordii rhizomes into powder to obtain Notopterygium wilfordii rhizome powder; soaking the Notopterygium wilfordii rhizome powder in pure water at room temperature, extracting with hot water, and subjecting the powder to ethanol graded precipitation, followed by deproteinization, and freeze-drying to obtain Notopterygium wilfordii crude polysaccharide; wherein the soaking time at room temperature is 4 hours; The hot water extraction includes boiling water for 2 hours each time, for a total of 3 extractions; The ethanol graded precipitation comprises the steps of: filtering the filtrate obtained after hot water extraction, concentrating the filtrate, adding 95% ethanol until the ethanol concentration in the mixture reaches 80%, standing at room temperature overnight, collecting the precipitate by centrifugation, and freeze-drying the precipitate; The preparation method of the black wolfberry acid polysaccharide comprises the following steps: The black wolfberry powder was extracted by microwave at 370W for 30 minutes, and the solution and extract were soaked in a ratio of 25:1 for 2.5 hours to obtain the total polysaccharide of black wolfberry; The extracted total polysaccharide of Lycium ruthenicum was prepared into a 20-25 mg / mL aqueous solution, and the supernatant was collected after centrifugation to remove impurities; a DEAE-Cellulose column was equilibrated with 3 column volumes of 0.05 mol / L NaHCO3 solution; The supernatant was eluted with distilled water, 0.05, 0.1, 0.25 and 0.5 mol / L NaHCO3 solution in sequence; the supernatant was applied to a DEAE-Cellulose column and eluted with distilled water and NaHCO3 solution in sequence, and the eluate was collected to obtain the neutral sugar fraction and acidic polysaccharide; 0.1 g of acidic polysaccharide was dissolved in ultrapure water to prepare a solution with a concentration of 10 mg / ml. The solution was filtered through a 0.45 μm filter membrane and then loaded onto a DEAE-Sepharose Fast Flow column at a flow rate of 1 mL / min. The eluate was collected using a fraction collector, with 5 mL collected in one tube. The solution was eluted until no polysaccharide was detected in the eluate. The purified Lycium ruthenicum acidic polysaccharide was obtained by dialysis and freeze-drying. The purity of the Notopterygium wilfordii polysaccharide is 67-72%, and the purity of the Lycium ruthenicum acid polysaccharide is 55-58%.

2. The antidepressant compound Chinese medicine according to claim 1, characterized in that: After the powder is ground into powder, it is further passed through a 70-100 mesh sieve.

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