Anti-depression compound traditional Chinese medicine neuroprotective agent and preparation method thereof

Abstract

The invention provides an anti-depression compound traditional Chinese medicine neuroprotective agent and a preparation method thereof, and belongs to the technical field of traditional Chinese medicines. The invention provides an anti-depression compound traditional Chinese medicine neuroprotective agent which mainly comprises notopterygium root polysaccharide (NI) and lycium ruthenicum (LA) polysaccharide and has a good neuroprotective effect on mice. The prepared compound medicine shows that the behavioristics of mice in a chronic unpredictable depression model and an LPS-induced depression model comprise the reduction of tail suspension and swimming immobility time of the model mice and the increase of sucrose preference of the model mice, and in addition, the compound medicine has a protection effect on PC12 nerve cells cultured in vitro. The purity of the notopterygium root polysaccharide extracted by the method is about 67%-72%, and the purity of the lycium ruthenicum polysaccharide is about 55%-58%.

Description

Technical Field

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

[0002] Currently, there are many theories about depression, but it is generally believed that depression is caused by the interaction of multiple systems, involving many aspects such as abnormal expression of neurotransmitter levels, impaired neural plasticity, hyperfunction of the hypothalamus-pituitary-adrenal (HPA) axis, as well as inflammatory response and oxidative stress.

[0003] Research on depression is in the development stage, and current drug treatments are accompanied by a variety of side effects. Therefore, the development of safe and effective natural medicines has become a research hotspot. Natural medicines usually contain a variety of active ingredients, which can act on the pathological mechanism of depression and improve depressive symptoms through multiple mechanisms such as regulating neurotransmitter levels, improving antioxidant capacity and reducing inflammatory responses. Therefore, exploring new and effective antidepressant drugs is a technical problem that needs to be solved urgently in this field. Summary of the invention

[0004] The invention provides an antidepressant compound Chinese medicine neuroprotectant and a preparation method thereof. Notopterygium wilfordii polysaccharide (NI) and Lycium ruthenicum polysaccharide (LA) have high purity and better efficacy. The antidepressant compound Chinese medicine neuroprotectant can significantly enhance its neuroprotective effect through drug combination application.

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

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

[0007] Notopterygium incisum is the dried rhizome and root of Notopterygium incisum or Notopterygium franchetii of the Umbelliferae family. The main chemical components isolated from it 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 and cerebrovascular systems, digestive systems, respiratory systems and central nervous systems. Notopterygium incisum polysaccharide is a polysaccharide compound extracted from Notopterygium incisum, a Chinese herbal medicine. In traditional Chinese medicine, Notopterygium incisum polysaccharide has a variety of applications: dispelling wind and dampness, promoting blood circulation and removing blood stasis, and anti-inflammatory effects.

[0008] Lycium ruthenicum Murr. is a shrub of the Lycium genus of the Solanaceae family. Lycium ruthenicum has a long history of application in traditional Chinese medicine. According to the Uyghur Pharmacopoeia and Jingzhu Materia Medica, its fruit has the effects of treating heart disease, strengthening the kidneys, moistening the liver, improving eyesight, strengthening the stomach, nourishing the brain, resisting aging, and regulating menstruation. In recent years, the pharmacological effects of Lycium ruthenicum have been further verified through scientific research, including its antioxidant and anti-inflammatory effects, and it has been more widely used in health foods. Lycium barbarum polysaccharides are the most studied group of water-soluble sugar complexes among the phytochemical components contained in wolfberry. They are composed of branched polysaccharides, proteoglycans, galacturonic acid and amino acids. The crude polysaccharides in Lycium ruthenicum are mainly composed of monosaccharides such as rhamnose, glucose, glucuronic acid, galactose, galacturonic acid, mannose, xylose, arabinose, etc., and have multiple biological activities, such as antioxidant, immunomodulatory, neuroprotective and prebiotic activities.

[0009] In this application, the antidepressant activity of Notopterygium wilfordii combined with Lycium ruthenicum is significantly better than that of single herbs, showing obvious advantages of the combination.

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

[0011] The method for preparing the antidepressant compound Chinese medicine neuroprotectant as described above comprises the following steps:

[0012] 1) Preparation of Notopterygium wilfordii polysaccharide rhizome was provided by the Chinese medicinal materials planting cooperative in Makohe Township, Banma County, Golog Tibetan Autonomous Prefecture, Qinghai Province. Lycium ruthenicum fruit was collected from Dongshagen, Dulan County, Haixi Mongolian and Tibetan Autonomous Prefecture, Qinghai Province, China; it was identified by Professor Xue Fenglu of the Northwest Plateau Institute of Biology, Chinese Academy of Sciences (specimen number: LR20110719). Notopterygium wilfordii rhizome and Lycium ruthenicum were naturally dried, ground, and sieved to obtain Notopterygium wilfordii rhizome powder and Lycium ruthenicum powder.

[0013] 2) The Notopterygium wilfordii rhizome powder is soaked in pure water at room temperature, extracted with hot water, and precipitated with ethanol for deproteinization, and freeze-dried to obtain Notopterygium wilfordii crude polysaccharide NIWP.

[0014] 3) The lycium ruthenicum powder is subjected to microwave treatment, hot water extraction, ethanol graded precipitation and vacuum drying to obtain lycium ruthenicum total polysaccharide (LRWP); the lycium ruthenicum total polysaccharide is further subjected to chromatography to obtain lycium ruthenicum acidic sugar fraction LRWP-Ap.

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

[0016] The preparation method of the above-mentioned antidepressant compound Chinese medicine neuroprotectant, the specific operation of step 1) comprises:

[0017] The notopterygium root rhizome and ruthenic wolfberry are ground after being naturally dried, and then sieved through a 70-100 mesh sieve to obtain notopterygium root rhizome powder and ruthenic wolfberry powder.

[0018] Drying can remove excess moisture, making it easier to grind, and at the same time increase the solubility of polysaccharides. The grinding process can destroy the cell walls of the Notopterygium wilfordii rhizomes and Lycium ruthenicum, increase the solubility of Notopterygium wilfordii polysaccharides and Lycium ruthenicum polysaccharides, and make them easier to extract in subsequent processing.

[0019] In the above-mentioned method for preparing the antidepressant compound Chinese medicine neuroprotectant, 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, added into 5 L of pure water and soaked at room temperature for 4 h, then boiled in boiling water for 2 h / time for a total of 3 times, filtered through four layers of gauze, the filtrate was collected, concentrated to 4-5 L at 80°C, 95% ethanol was added until the ethanol concentration of the mixed solution reached 80%, and allowed to stand at room temperature overnight. The precipitate was collected by centrifugation, and the precipitate was freeze-dried to obtain the Notopterygium wilfordii crude polysaccharide NIWP.

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

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

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

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

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

[0026] The extracted total polysaccharide of Lycium barbarum was prepared into an aqueous solution of 20-25 mg / mL, and the supernatant was collected after centrifugation to remove impurities. The DEAE-Cellulose column was balanced with 3 column volumes of 0.05 mol / L NaHCO3 solution. The elution process was as follows: eluted with distilled water, 0.05, 0.1, 0.25 and 0.5 mol / L NaHCO3 in sequence. The supernatant was loaded onto 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 LRWP-N and the acidic polysaccharide LRWP-A.

[0027] Take 0.1 g of LRWP-A, dissolve it in ultrapure water to make a solution with a concentration of 10 mg / ml, pass it through a 0.45 μm filter membrane, and then apply it to a DEAE-Sepharose FastFlow column at a flow rate of 1 mL / min. Use a partial collector to collect the eluate, collect 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 Qianghuo combined with Ruoguo Lycium barbarum on mice is significantly better than that of a single herb, 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 increased.

[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 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 neuroprotectant of the present invention has the following beneficial effects:

[0032] (1) Significant neuroprotective effect: The combination of Notopterygium wilfordii and Lycium barbarum is significantly better than single drugs, especially in the chronic unpredictable depression model and LPS-induced depression model mice, showing obvious advantages of combination. 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 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 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. *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 in conjunction with the examples, but it will be appreciated by those skilled in the art that the following examples are only used to illustrate the present invention and should not be construed as limiting the scope of the present invention. If specific conditions are not specified in the examples, they are carried out according to conventional conditions or conditions recommended by the manufacturer. If the manufacturer is not specified for the reagents or instruments used, they are all conventional products that can be purchased commercially.

[0040] Example 1

[0041] Effect of Notopterygium wilfordii + Lycium ruthenicum on the behavior of LPS-induced depressed mice

[0042] 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).

[0043] a. Experimental animals were given the drug by gavage for 10 consecutive days, and the dosage was as follows:

[0044] Table 1 Animal gavage dosage table

[0045]

[0046]

[0047] b. On days 9-11, the blank control group was intraperitoneally injected with 10 ml / kg normal saline, and the other five groups were intraperitoneally injected with the same volume of LPS (0.1 mg / ml).

[0048] c. Behavioral experiments were performed on day 12. Tail suspension test (TST): The animals were suspended 50 cm above the table with tape, and the tape was about 1 cm away from the tip of the tail. The test lasted for 6 min, and the time the mice remained motionless was recorded in the last 4 min of the test. When the mice were passively suspended and completely motionless, they were considered to be immobile.

[0049] d. Forced swimming test (FST): The mice were placed individually in a cylindrical container with a height of 25 cm and a diameter of 15 cm, containing 10 cm of water, and tested at 25°C. The cumulative immobility time of the mice in the last 4 minutes of the 6-minute test was recorded. When the mice stopped struggling or floated, they were considered immobile.

[0050] e. Sugar preference test (SPT): Before the test, mice were allowed to drink 1% sucrose solution freely for 24 hours, and then drink pure water freely for 24 hours. Then the test was conducted after deprivation of food and water for 12 hours. All mice were housed individually and had free access to 1% sucrose solution and pure water for 12 hours. The volume of sucrose solution and pure water consumed was calculated. Sucrose preference was calculated as follows: Sucrose preference = (sucrose solution consumption / total consumption) × 100.

[0051] The experimental results are as follows Figure 1 As shown. In the tail suspension test, mice were suspended at an appropriate height to observe whether they showed weakness and despair behavior. The results showed that compared with the control group, the mice in the Notopterygium wilfordii + black wolfberry group showed a significantly reduced tail suspension time, indicating that the combination of Notopterygium wilfordii and black wolfberry had a more significant effect on improving depressive behavior. The Notopterygium wilfordii + black wolfberry group also had a better effect than mice given Notopterygium wilfordii or black wolfberry alone.

[0052] In the forced swimming test, mice were placed in water and forced to swim to see if they showed helplessness and negative behavior. The results showed that the mice in the Notopterygium wilfordii + Lycium ruthenicum group had significantly reduced swimming time and diving times, and showed less depression and frustrated behavior compared to the control group. The effect of the Notopterygium wilfordii + Lycium ruthenicum group was also more significant than that of mice given Notopterygium wilfordii or Lycium ruthenicum alone.

[0053] In the sugar water preference test, mice were given two choices: sugar water and ordinary water. The results showed that the mice in the group of Notopterygium wilfordii + black wolfberry had a higher preference for sugar water and showed a stronger sense of reward and positive emotions compared with the control group. The effect of the group of Notopterygium wilfordii + black wolfberry was also better than that of mice given Notopterygium wilfordii or black wolfberry alone.

[0054] Experimental Example 2

[0055] Effect of Notopterygium wilfordii + Lycium ruthenicum on the behavior of CUMS depression mice

[0056] a. Experimental animals were given the drug by gavage for 4 weeks continuously. The dosage is as follows:

[0057] Table 2 Oral dosing table for experimental animals

[0058] Group dose volume Normal saline blank control group -- 0.2ml Fluoxetine positive control group 3.6mg / kg 0.2ml Notopterygium group 90mg / kg 0.2ml Black wolfberry group 90mg / kg 0.2ml Notopterygium wilfordii + Lycium ruthenicum group 60+30mg / kg 0.2ml

[0059] b. Except for the blank control group, the other groups were applied with two different stressors every day. The stressors included food and water deprivation, night light, moist bedding, 45°C cage tilt, light / dark continuity and tail clip stimulation. The order of stressors was changed every three days for 4 weeks.

[0060] c. After 4 weeks, behavioral experiments were carried out in the same manner as in Experimental Example 1, including the tail suspension test (TST), the forced swim test (FST), and the sucrose preference test (SPT).

[0061] The experimental results are as follows Figure 2 As shown. In the tail suspension test, compared with the control group, the mice in the Notopterygium wilfordii + Lycium ruthenicum group showed a significantly reduced tail suspension time, indicating that the combination of Notopterygium wilfordii and Lycium ruthenicum has a more significant effect on improving depressive behavior. Compared with mice given Notopterygium wilfordii or Lycium ruthenicum alone, the Notopterygium wilfordii + Lycium ruthenicum group also had a better effect.

[0062] In the forced swimming test, mice in the Notopterygium wilfordii + Lycium ruthenicum group had significantly reduced swimming time and diving times, and showed less depression and frustrated behavior compared with the control group. The effect of Notopterygium wilfordii + Lycium ruthenicum group was also more significant than that of mice given Notopterygium wilfordii or Lycium ruthenicum alone.

[0063] In the test of the preference for sugar water, mice in the group of Notopterygium wilfordii + black wolfberry showed a higher preference for sugar water and a stronger sense of reward and positive emotions compared with the control group. The effect of the group of Notopterygium wilfordii + black wolfberry was also better than that of mice given Notopterygium wilfordii or black wolfberry alone.

[0064] Experimental Example 3

[0065] Protective effect of Notopterygium wilfordii + Lycium ruthenicum on neuronal cells in vitro

[0066] a. PC12 cells were divided into the following 11 groups:

[0067] Table 3 PC12 cell grouping

[0068] Group <![CDATA[H2O2 dosage]]> Drug dosage Negative control group -- -- Model Group 100μM -- Notopterygium group 100μM 75μg / ml Black wolfberry group 100μM 75μg / ml Notopterygium wilfordii + Lycium ruthenicum group (1:1) 100μM 50+50μg / ml Notopterygium group 100μM 100μg / ml Black wolfberry group 100μM 50μg / ml Notopterygium wilfordii + Lycium ruthenicum group (2:1) 100μM 100+50μg / ml Notopterygium group 100μM 50μg / ml Black wolfberry group 100μM 100μg / ml Notopterygium root + black wolfberry group (1:2) 100μM 50+100μg / ml

[0069] b. PC12 cells were seeded in 6-well plates and cultured until the cell density reached 70-80%. The cells were washed once with PBS to remove the residues in the culture medium. H2O2 (100 μM) was added and incubated for 24 hours to induce oxidative stress damage.

[0070] c. After inducing injury, the culture medium was removed and the cells were washed with PBS to remove residual H2O2. The culture medium containing different concentrations of compound Chinese medicine was added, and the cells were cultured for another 24 hours to observe the effects of drug treatment on the cells.

[0071] d. The activities of SOD and CAT in each group of cells were determined using SOD activity kit and CAT activity kit.

[0072] e. The expression level of Caspase 3 in each group of cells was analyzed by Western blot, using β-actin as the internal reference protein.

[0073] The experimental results are as follows Figure 3 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 + Lycium ruthenicum, the activities of SOD and CAT were significantly increased, which was significantly better than the group treated with Notopterygium wilfordii or Lycium ruthenicum alone, and 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 and Lycium ruthenicum is 2:1, so the present invention selects Notopterygium wilfordii: Lycium ruthenicum as a ratio of 2:1 for subsequent research.

[0074] 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 and Lycium barbarum treatment groups improved, but some cells were still wrinkled. The Notopterygium and Lycium barbarum + Lycium barbarum group (Notopterygium and Lycium barbarum: Lycium barbarum = 2:1) significantly improved cell morphology, and the cell morphology was significantly better than that of the treatment groups using Notopterygium or Lycium barbarum alone, with less cell shrinkage and most cells returning to normal morphology.

[0075] 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 or Lycium barbarum alone, the expression level of Caspase 3 decreased significantly. In the Notopterygium + Lycium barbarum group (Notopterygium: Lycium barbarum = 2:1), the expression level of Caspase 3 decreased more significantly, which was lower than that of the group treated with Notopterygium or Lycium barbarum alone. The difference was statistically significant, indicating that the compound Chinese medicine can effectively reduce cell apoptosis ( Figure 4 ).

[0076] In summary, the compound Chinese medicine Notopterygium wilfordii + Lycium ruthenicum showed significant neuroprotective effects in the hydrogen peroxide-induced injury model of PC12 cells. Compared with the treatment groups using Notopterygium wilfordii or Lycium ruthenicum alone, the Notopterygium wilfordii + Lycium ruthenicum group showed superior effects in cell morphology protection, improvement of oxidative stress molecules, and reduction of cell apoptosis. These results indicate that the compound Chinese medicine of Notopterygium wilfordii + Lycium ruthenicum has a synergistic effect and can provide a stronger neuroprotective effect.

[0077] In summary, the research results of the present invention clearly show that the Chinese herbal compound drugs Notopterygium and Lycium ruthenicum have excellent neuroprotective effects, and their antidepressant effects are significantly better than those of Notopterygium or Lycium ruthenicum alone. This discovery provides a strong scientific basis for the potential application of the Chinese herbal compound drugs Notopterygium and Lycium ruthenicum in neuroprotective treatment, and has important guiding significance for related clinical treatments and drug development.

[0078] It is concluded that the preparation method of the antidepressant compound Chinese medicine neuroprotectant 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.

[0079] Although the above embodiment describes the present invention in detail, 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 protection scope of the present invention.

Claims

1. An antidepressant compound Chinese medicine neuroprotectant, characterized in that: The invention comprises notopterygium wilfordii polysaccharide and lycium ruthenicum polysaccharide, wherein the mass ratio of notopterygium wilfordii polysaccharide to lycium ruthenicum polysaccharide is (1-2): (1-2).

2. The antidepressant compound Chinese medicine neuroprotectant according to claim 1, characterized in that: The preparation method of the Notopterygium wilfordii polysaccharide comprises the following steps: grinding dried Notopterygium wilfordii rhizomes into powder to obtain Notopterygium wilfordii rhizomes powder; soaking the Notopterygium wilfordii rhizomes powder in pure water at room temperature, extracting with hot water, and subjecting the Notopterygium wilfordii rhizomes powder to ethanol graded precipitation, followed by deproteinization, and freeze-drying to obtain Notopterygium wilfordii crude polysaccharide NIWP.

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

4. The antidepressant compound Chinese medicine neuroprotectant according to claim 2, characterized in that: The room temperature soaking time is 4h; The hot water extraction includes boiling water for 2 hours each time, and extracting 3 times in total; The ethanol graded precipitation comprises filtering the filtrate after hot water extraction, concentrating it, adding 95% ethanol until the ethanol concentration in the mixed solution reaches 80%, standing at room temperature overnight, centrifuging to collect the precipitate, and freeze-drying the precipitate.

5. The antidepressant compound Chinese medicine neuroprotectant according to claim 1, characterized in that: The preparation method of black wolfberry polysaccharide comprises the following steps: drying black wolfberry and grinding it into powder to obtain black wolfberry powder; The black wolfberry powder is treated with microwave, extracted with hot water, precipitated with ethanol and dried in vacuum to obtain the black wolfberry total polysaccharide; the black wolfberry total polysaccharide is further chromatographed to obtain the black wolfberry acidic sugar fraction LRWP-Ap.

6. The antidepressant compound Chinese medicine neuroprotectant according to claim 5, characterized in that: The preparation method of the black wolfberry polysaccharide comprises the following steps: The black wolfberry powder was extracted by microwave at 370W for 30 minutes, and the solution and the extract were soaked for 2.5 hours at a ratio of 25:1 to obtain the black wolfberry total polysaccharide LRWP; The chromatography steps are as follows: preparing the extracted total polysaccharide of Lycium ruthenicum into a 20-25 mg / mL aqueous solution, centrifuging to remove impurities and taking the supernatant; balancing the DEAE-Cellulose column with 3 column volumes of 0.05 mol / L NaHCO3 solution; The elution process is as follows: eluting with distilled water, 0.05, 0.1, 0.25 and 0.5 mol / L NaHCO3 in sequence; applying the supernatant to a DEAE-Cellulose column, eluting with distilled water and NaHCO3 solution in sequence, collecting the eluate to obtain the neutral sugar fraction LRWP-N and the acidic polysaccharide LRWP-A; Take 0.1 g of LRWP-A, dissolve it in ultrapure water to make a solution with a concentration of 10 mg / ml, pass it through a 0.45 μm filter membrane, and then apply it to a DEAE-Sepharose FastFlow column at a flow rate of 1 mL / min. Use a partial collector to collect the eluate, collect 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.

7. The antidepressant compound Chinese medicine neuroprotectant according to claim 1, characterized in that: The purity of the Notopterygium wilfordii polysaccharide is 67-72%, and the purity of the Lycium ruthenicum polysaccharide is 55-58%.

8. Use of the antidepressant compound Chinese medicinal neuroprotectant according to any one of claims 1 to 7 in the preparation of neuroprotective drugs.

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