Application of mimosa pudica extract in preparation of medicine for treating cervical spondylotic radiculopathy

Abstract

The invention belongs to the field of biological medicine, and particularly relates to a composition for treating cervical spondylotic radiculopathy and application thereof. The composition disclosed by the invention is prepared from a mimosa pudica extract. The extract can promote M2 type polarization of microglial cells, and has protection and repair effects on damaged nerves.

Description

Technical Field

[0001] The present invention belongs to the field of biomedicine, and particularly relates to a composition drug for treating nerve root type cervical spondylosis and its application. Background Art

[0002] Nerve root type cervical spondylosis is a general term for a series of symptoms and signs caused by factors such as degenerative changes of the cervical intervertebral disc, bone hyperplasia, or protrusion of the cartilage plate, which compress the cervical nerve roots. Clinically, patients often present with neck pain, discomfort in the shoulder and back, as well as radiating pain, sensory abnormalities, and muscle strength reduction in the area innervated by the corresponding nerve roots. As a typical clinical manifestation of nerve root type cervical spondylosis, limb paralysis is usually related to a specific cervical level. For example, a herniated C5 / 6 intervertebral disc may cause muscle weakness and sensory abnormalities in the upper limb. Since there is no effective drug to relieve paralysis like pain symptoms that can be emergently relieved by painkillers, the long-term limb paralysis caused by nerve root type cervical spondylosis seriously affects the quality of life of patients. Even the symptoms may worsen as the disease progresses, severely affecting the daily life and working ability of patients.

[0003] Currently, the means for treating limb paralysis in nerve root type cervical spondylosis mainly include conservative treatment and surgical treatment. Conservative treatment usually includes drug treatment, physical therapy, and functional exercise, etc., aiming to relieve symptoms and improve function. Surgical treatment directly relieves the compression of the nerve roots through decompression surgery in the hope of restoring nerve function. However, these treatment means can only partially relieve symptoms. Conservative treatment requires a long treatment cycle to have partial effects, and the treatment principle is only to repair the nerves. However, none of the above treatments can fundamentally solve pathological changes such as demyelination. In addition, surgical treatment has certain risks and complications, and not all patients are suitable for surgery.

[0004] Therefore, developing new treatment methods related to the pathogenic mechanism of limb paralysis in nerve root type cervical spondylosis, such as drug treatment targeting molecular targets in the demyelination process, may provide more effective treatment options for patients. Summary of the Invention

[0005] The present invention mainly aims at the above technical problems and provides a drug for treating nerve root type cervical spondylosis and its preparation method.

[0006] The present invention provides the following technical solutions:

[0007] In the first aspect, the present invention provides a composition for treating nerve root type cervical spondylosis, characterized in that the drug contains Mimosa hostilis extract.

[0008] Further, the drug may contain a pharmaceutically acceptable carrier or excipient.

[0009] On the other hand, another object of the present invention is to provide the use of Mimosa hostilis extract in the preparation of a medicament for treating nerve root type cervical spondylosis.

[0010] Furthermore, the Mimosa hostilis extract is an aqueous extract of Mimosa hostilis, and the extraction method of the Mimosa hostilis extract comprises the following steps:

[0011] 1) Pre-freezing Mimosa hostilis under low temperature conditions;

[0012] 2) Freeze-drying and dehydrating the pre-frozen Mimosa hostilis;

[0013] 3) After pulverizing the freeze-dried Mimosa hostilis, adding water for reflux extraction, with the reflux temperature being 60 - 100°C and the reflux time being 1 - 3 h. After obtaining the extract, filter and concentrate it;

[0014] 4) Freeze-drying the concentrate.

[0015] Correspondingly, another object of the present invention is to provide a method for preparing a medicament for treating nerve root type cervical spondylosis, and the method comprises the above-mentioned extraction steps of the Mimosa hostilis extract.

[0016] On the other hand, another object of the present invention is to provide the use of Mimosa hostilis extract in the preparation of promoting the activation of microglial M2.

[0017] On the other hand, the present invention provides a medicament for promoting the activation of microglial M2, characterized in that the medicament contains Mimosa hostilis extract.

[0018] Furthermore, the medicament may contain a pharmaceutically acceptable carrier or excipient.

[0019] On the other hand, another object of the present invention is to provide a method for preparing a medicament for promoting the activation of microglial M2, and the method comprises the following steps:

[0020] 1) Pre-freezing Mimosa hostilis under low temperature conditions;

[0021] 2) Freeze-drying and dehydrating the pre-frozen Mimosa hostilis;

[0022] 3) After pulverizing the freeze-dried Mimosa hostilis, adding water for reflux extraction, with the reflux temperature being 60 - 100°C and the reflux time being 1 - 3 h. After obtaining the extract, filter and concentrate it;

[0023] 4) Freeze-drying the concentrate.

[0024] Preferably, in the extraction method of the aforementioned Mimosa tenuiflora extract or the drug preparation method, the mass-volume ratio of Mimosa tenuiflora to water in step 3) is 1 g:10 ml to 1 g:40 ml (w / v). The pre-freezing temperature in step 1) is 80 °C, and the pre-freezing time is 2 h.

[0025] Preferably, the extraction method of the aforementioned Mimosa tenuiflora extract or the specific steps of the aforementioned drug preparation method are as follows:

[0026] 1. Raw material pretreatment

[0027] Washing: The fresh whole Mimosa tenuiflora is gently rinsed with distilled water to remove soil and impurities, avoiding mechanical damage.

[0028] Initial drying: The fresh product is directly freeze-dried.

[0029] 2. Freeze-drying

[0030] Pre-freezing: Cut the material into small pieces and place it in a -80 °C ultra-low temperature refrigerator for pre-freezing for 12 hours.

[0031] Freeze-drying parameters: Vacuum freeze-dryer (cold trap temperature ≤ -50 °C, vacuum degree ≤ 0.1 mbar) for 24 - 48 hours until completely dehydrated.

[0032] 3. Crushing and extraction

[0033] Crushing: The freeze-dried material is crushed into 80 - 100 mesh powder by a ball mill.

[0034] Water extraction: The powder and distilled water are in a ratio of 1 g:20 ml (w / v), and reflux at 80 °C for 2 hours.

[0035] Filtration and concentration: Centrifuge (8000 rpm, 10 minutes) to take the supernatant, and rotary evaporate (50 °C) to obtain an extract.

[0036] 4. Secondary freeze-drying

[0037] The extract is redissolved in deionized water, pre-frozen and then vacuum freeze-dried to obtain a freeze-dried powder, which is stored in the dark at -20 °C.

[0038] 5. Quality control

[0039] Microbial limit: It needs to meet the cell experiment level standard of the Chinese Pharmacopoeia (bacteria < 100 CFU / g, mold / yeast < 10 CFU / g).

[0040] Optionally, the aforementioned drug contains a pharmaceutically acceptable carrier or excipient.

[0041] On the other hand, the present invention provides a method for promoting the M2 polarization of microglia for non-therapeutic purposes, which is characterized in that the method comprises the following steps:

[0042] 1) Culture microglia;

[0043] 2) Treat the cells cultured in step 1) with the aforementioned drug.

[0044] Preferably, the main active ingredient of the aforementioned drug is Mimosa hostilis extract.

[0045] Furthermore, Mimosa hostilis extract can be selected as the only active ingredient.

[0046] On the other hand, the present invention provides a method for screening drugs for treating nerve root type cervical spondylosis or promoting M2 polarization of microglia, and the method includes the following steps:

[0047] 1) Culture microglia;

[0048] 2) Treat the cells cultured in step 1) with the aforementioned drug and a candidate drug;

[0049] 3) Determine whether the candidate drug can treat nerve root type cervical spondylosis or promote M2 polarization of microglia by comparing the effects of the aforementioned drug and the candidate drug on cell treatment.

[0050] Preferably, the main active ingredient of the aforementioned drug is Mimosa hostilis extract.

[0051] Furthermore, Mimosa hostilis extract can be selected as the only active ingredient.

[0052] The present invention has the following beneficial effects compared with the prior art:

[0053] 1. It is found for the first time that Mimosa hostilis extract can treat nerve root type cervical spondylosis.

[0054] 2. The specific extraction method of Mimosa hostilis extract that can treat nerve root type cervical spondylosis is disclosed for the first time.

[0055] 3. It is found for the first time that Mimosa hostilis extract can promote M2 polarization of microglia. In addition to being used for treating nerve root type cervical spondylosis, it can also be used for culturing cells with M2 polarization of microglia or as a positive control drug for promoting M2 polarization of microglia. Brief Description of the Drawings

[0056] The products, methods and their beneficial effects of the present invention will be described in detail below in conjunction with the drawings and specific embodiments.

[0057] Figure 1 are the results of modeling and treatment detection of nerve root type cervical spondylosis mice; wherein Figure 1 A is the process of modeling nerve root type cervical spondylosis mice; Figure 1B is to detect the change process of mechanical pain threshold in rats using von Frey filaments; Figure 1 C is to detect somatosensory evoked potential (SEP) on the 14th day of treatment; Figure 1 D is HE staining of histopathological sections; Figure 1 E is the result of transmission electron microscopy examination.

[0058] Figure 2 are the results of transcriptome and flow cytometry detection under different treatment conditions; among them Figure 2 A is the result of transcriptome pathway enrichment analysis combined with immune infiltration analysis; Figure 2 B is the heat map of gene expression related to BTregs infiltration; Figure 2 C is to detect the change of microglial M2 by flow cytometry.

[0059] Figure 3 are the results of metabolome detection under different treatment conditions; among them Figure 3 A is the KEGG enrichment analysis of the metabolome; Figure 3 B is the heat map of genes interacting with differential metabolites, where H is the Mimosa pudica treatment group and C is the control group; Figure 3 C is the Venn diagram of targets in different databases; Figure 3 D is the Venn diagram of metabolic target genes and gene target genes; Figure 3 F is the result of multi-color immunofluorescence verification; Figure 3 E is the interaction network diagram of core target genes.

[0060] Figure 4 are the results of microglial M2 polarization detection under different treatment conditions; among them Figure 4 A is the result of behavioral detection; Figure 4 B is somatosensory evoked potential detection; Figure 4 C is HE staining; Figure 4 D is transmission electron microscopy; Figure 4 E is western detection of CD206 protein expression; Figure 4 F is gene expression level detection; Figure 4 G is biomarker expression level detection; Figure 4 H is the result of flow cytometry detection.

[0061] Figure 5 are the results of pathological tissue section detection of kidney, lung and liver under different treatment conditions. Specific implementation mode

[0062] The technical solutions in the embodiments of the present invention will be clearly and completely described below. Apparently, the described embodiments are only a part of the embodiments of the present invention, rather than all of them. All other embodiments obtained by those of ordinary skill in the art based on the embodiments of the present invention without creative efforts shall fall within the protection scope of the present invention.

[0063] Unless otherwise defined, all technical and scientific terms used in this application have the same meaning as commonly understood by those of ordinary skill in the technical field to which this application belongs.

[0064] Example 1. Preparation process of freeze-dried powder of Mimosa pudica

[0065] 1. Pretreatment of raw materials

[0066] Cleaning: The fresh whole plant of Mimosa pudica is gently rinsed with distilled water to remove soil and impurities, avoiding mechanical damage.

[0067] Initial drying: The fresh product is directly freeze-dried.

[0068] 2. Freeze-drying

[0069] Pre-freezing: Cut the material into small pieces and place it in an ultra-low temperature refrigerator at -80°C for pre-freezing for 12 hours.

[0070] Freeze-drying parameters: Vacuum freeze-dryer (cold trap temperature ≤ -50°C, vacuum degree ≤ 0.1 mbar) for 24 - 48 hours until completely dehydrated.

[0071] 3. Crushing and extraction

[0072] Crushing: The freeze-dried material is crushed into powder with 80 - 100 meshes by a ball mill.

[0073] Water extraction: The powder and distilled water are in a ratio of 1 g:20 mL (w / v), and refluxed at 80°C for 2 hours to obtain an extract.

[0074] Filtration and concentration: The extract is centrifuged (8000 rpm, 10 minutes) to take the supernatant, and rotary evaporated (50°C) to obtain an extract paste.

[0075] 4. Secondary freeze-drying

[0076] The extract paste is redissolved in deionized water, pre-frozen and then vacuum freeze-dried to obtain freeze-dried powder, which is stored in the dark at -20°C.

[0077] 5. Quality control

[0078] Microbial limit: It needs to meet the standards of the Chinese Pharmacopoeia for cell experiment level (bacteria < 100 CFU / g, molds / yeasts < 10 CFU / g).

[0079] Example 2 Therapeutic effect of the whole plant of Mimosa pudica on nerve root type cervical spondylosis

[0080] Model establishment of nerve root compression:

[0081] Anesthetize the rats: Intraperitoneally inject 10% chloral hydrate (0.4 ml / 100 g).

[0082] All Sprague-Dawley (SD) rats were subjected to right-sided somatosensory evoked potential (SLSEP) detection of the median nerve before surgery to obtain reference values, and then the nerve root compression method was used to establish the model (L4D): After general anesthesia and disinfection, the cervical vertebra was exposed through an incision. Use a mosquito forceps bone nibbler to remove the lateral vertebral arch of one side of C6 and C7. Place a nylon thread about 1 cm long and 0.5 mm in diameter along the longitudinal axis of the spinal canal down to the C6-T1 nerve root, and place the second one upward at the C7-C5 nerve root. Close the surgical wound and suture layer by layer. After surgery, apply gentamicin powder to the wound regularly for anti-infection for 3 consecutive days. Evaluate the surviving rats on the 5th day after surgery and screen them for inclusion in the group (the increase in E-C6Lat on the operative side before and after modeling is 0.2 - 0.4 ms); randomly divide the successfully modeled rat models into 7 groups: The control group was given intragastric administration of normal saline containing 1% DMSO, and the experimental group was given intragastric administration of drugs. Right-sided SLSEP (somatosensory evoked potential of the median nerve) detection was performed 1 week and 2 weeks after treatment. After 2 weeks, take the C6-T1 segment of the spinal cord to make tissue pathological sections for observation. (Dou Xiarui, Sun Jianning, Wang Wei, et al. Establishment of a model of nerve root type cervical spondylosis in the acute stage [J]. Journal of Beijing University of Traditional Chinese Medicine, 2006, 29(5): 332 - 334, 343, 361).

[0083] Schematic diagram of nerve root compression model establishment is as Figure 1 shown in

[0084] Intragastric administration was performed on the rats with nerve root type cervical spondylosis model at a dose of 100 mg / kg / day (HXC group). The Control group (wild type) was only given intragastric administration of normal saline. The Model group was given intragastric administration of normal saline after modeling. The Positive group was given intragastric administration of mecobalamin at 1 mg / kg after modeling.

[0085] Von Frey filaments were used to detect the changes in mechanical pain threshold of the rats ( Figure 1B). The results showed that, compared with the control group, the mechanical pain threshold of the rats in the model group was significantly decreased on the 5th day after surgery (HXC group: 9.85 ± 0.86 g, Model group: 8.55 ± 0.76, positive group: 9.68 ± 0.78, control group: 12.73 ± 1.02 g, P < 0.01), suggesting that nerve root compression led to decreased pain sensitivity. After intervention with the aqueous extract of HXC (215 μg / mL), the mechanical pain threshold increased by 8.98 ± 0.73 g and 10.34 ± 0.90 g on the 7th and 14th days of treatment respectively, which was significantly improved compared with the Model group and the positive group, indicating that the aqueous extract of HXC could effectively relieve the pain abnormality caused by nerve root compression.

[0086] Two weeks later, the somatosensory evoked potential (SEP) detection showed that ( Figure 1 C), compared with the model group, the amplitude of P1-N1 wave in the HXC treatment group was significantly restored, the latency was shortened, and the nerve conduction velocity was improved on the 14th day, suggesting that the aqueous extract of HXC could promote the regression of nerve edema and the repair of myelin sheath. Two weeks later, histopathological observation found that ( Figure 1 D), the number of spinal cord motor neurons in the model group was significantly reduced, the nuclear staining became lighter, and the morphology was irregular; while the number of neurons in the HXC treatment group increased significantly, the nuclear staining deepened, and the morphological characteristics were basically restored. The results of transmission electron microscopy showed that ( Figure 1 E), pathological changes such as axonal distortion, myelin sheath loss and neuronal vacuolization appeared in the model group, while the myelin sheath thickness was restored and the degree of neuronal damage was significantly reduced in the HXC treatment group, confirming that the aqueous extract of HXC had a protective and reparative effect on the damaged nerve.

[0087] Example 3 Transcriptome and metabolome analysis of the components of the whole herb entering the blood

[0088] To analyze the in vivo metabolic characteristics and action mechanisms of the active components of HXC, this study integrated serum metabolomics and peripheral blood mononuclear cell (PBMC) transcriptomics analysis. When sampling after 15 days of treatment, serum samples of the control group and the HXC intervention group were obtained by abdominal aortic blood collection. The enrichment analysis of all differential gene pathways in the transcriptome combined with immune infiltration analysis showed that HXC intervention significantly regulated the macrophage polarization state in the CSR rat model ( Figure 2 A). Peripheral immune analysis showed that the aqueous extract of HXC could induce macrophages to transform from the pro-inflammatory M1 phenotype to the reparative M2 phenotype. Further research found that regulatory T cells (Tregs) played a key role in the immune remodeling mediated by HXC. The infiltration level of Tregs in the HXC treatment group was significantly increased compared with the control group ( Figure 2 B), and Tregs might inhibit the M1 polarization of microglia and at the same time promote its transformation to the M2 phenotype.

[0089] Metabolomics detection found that genes related to differential metabolites in the serum of the HXC intervention group were intersected with the target genes screened by network pharmacology and then subjected to enrichment analysis. The results showed significant enrichment in the microglial activation pathway (KEGG analysis, Figure 3 A), in which CXCR3 was predicted to be a key regulatory gene (differential metabolite-gene interaction heatmap, Figure 3 B). Further integrating the disease targets of the GeneCards, DisGeNet, and OMIM databases ( Figure 3 C), four core targets, COMT, CXCR3, SCN9A, and TRPV1, were screened out (Venn intersection analysis, Figure 3 D; gene interaction network, Figure 3 E). Multicolor immunofluorescence verification showed that the expression of CXCR3 in the anterior horn of the spinal cord in the HXC treatment group was significantly upregulated ( Figure 3 F), and it showed a co-localization feature with the microglial M2 polarization marker CD206, suggesting that it plays a neuroprotective role by regulating CXCR3-mediated glial cell phenotype transformation.

[0090] Example 4 Detection of changes in microglial M2 by flow cytometry

[0091] Control group Con, model group (Model), low concentration group of Mimosa pudica (HXC-L, 143 μg / mL), high concentration group of Mimosa pudica (HXC-H, 322 μg / mL). After 7 days of treatment, abdominal aortic blood was taken to isolate PBMCs for flow cytometry detection. The flow cytometry results showed that indeed M2 was higher than M1 ( Figure 2 C).

[0092] Example 5 HXC promotes microglial M2 polarization

[0093] To verify that the aqueous extract of HXC exerts a therapeutic effect through M2-polarized microglia, this study used the M2 polarization-specific inhibitor Stattic (10 mg / kg) for a mechanism blocking experiment (STAT3 plays an important role in M2 polarization. By inhibiting the activity of STAT3, the transformation of microglia into the M2 type can be inhibited. Stattic is a classical STAT3 inhibitor that directly binds to STAT3, preventing its phosphorylation and activation, thereby inhibiting its transcriptional activity). Four groups were designed for the experiment: the model group (Model), the control group (Control), the positive drug group (Positive), and the HXC+Stattic group (HXC+Stattic). Behavioral tests showed that compared with the group treated with the positive drug alone (BBB score: 18.3±1.2), the Stattic combined intervention group failed to reverse the limb paralysis symptoms (BBB score of the HXC+Stattic group: 12.1±1.6 vs the positive drug group, P<0.001), and the degree of loss of its efficacy reached 56%( Figure 4 A). Somatosensory evoked potential, HE staining, and transmission electron microscopy results( Figure 4 B-D) found that it could not protect and repair the damaged nerves, indicating that HXC treats CSR limb paralysis through the activation of microglial M2. These evidences suggest that the M2 polarization of spinal cord anterior horn microglia is a possible mechanism for HXC to improve CSR limb paralysis.

[0094] Example 6 Promote the M2 activation of HMC3 cells (human microglial cell line) by Mimosa pudica L.

[0095] Grouping design:

[0096] Negative control group: Treated with complete medium.

[0097] Positive control group: Treated with IL-4 (20 ng / mL).

[0098] HXC drug treatment group: Select 3 non-toxic concentrations (such as 50, 100, 215 μM).

[0099] Inhibitor group: Pretreated with the STAT3 inhibitor Stattic (5.1 μM) for 1 hour and then added with the HXC drug.

[0100] Treatment time: 48 hours.

[0101] The results showed that HXC-H drug significantly upregulated the M2 markers: the expressions of Arg1 and Ym1 / 2 increased (compared with the negative control group), and the protein level of CD206 and the secretion amount of IL-10 increased( Figure 4 E~ Figure 4G). Flow cytometry detection in the cell line after drug treatment showed an increase in the amount of cells positive for the M2 polarization cell marker CD206. The M2 polarization effect of HXC drug could be partially reversed by using the STAT3 inhibitor Stattic. Figure 4 H).

[0102] Example 7 Toxic and Side Effects Study of HXC

[0103] The evaluation method referred to Greaves P. Histopathology of preclinical toxicity studies: interpretation and relevance in drug safety evaluation, Book·Fourth Edition·2012. Histopathological evaluations of liver, lung, and kidney tissues showed that Figure 5 ), no significant organizational structure abnormalities were observed in the control group, model group, and HXC treatment group (215 μg / mL, gavage for 15 days). In HE-stained sections, the hepatic cell cords in the three groups were arranged regularly, and no fatty degeneration or necrosis foci were seen; the glomerular structure was intact, and the morphology of renal tubular epithelial cells was normal; the alveolar lumen was clear, and there was no inflammatory infiltration or fibrosis in the interstitium, indicating that HXC did not cause toxic damage to parenchymal organs at this dose.

[0104] The above description of the disclosed embodiments enables those skilled in the art to implement or use the present invention. Various modifications to the above embodiments will be obvious to those skilled in the art, and the general principles defined herein can be implemented in other embodiments without departing from the spirit or scope of the present invention. Therefore, the present invention will not be limited to these embodiments shown herein, but rather to the widest scope consistent with the principles and novel features disclosed herein.

Claims

1. Application of Brazilian Mimosa pudica extract in the preparation of medicines for treating cervical spondylotic radiculopathy.

2. The use according to claim 1, characterized in that The Brazilian Mimosa extract is a water extract of Brazilian Mimosa, and the extraction method of the Brazilian Mimosa extract comprises the following steps: 1) pre-freezing Brazilian Mimosa pudica under low temperature conditions; 2) freeze-drying and dehydrating the pre-frozen Brazilian Mimosa pudica; 3) crushing the freeze-dried Brazilian mimosa, adding water for reflux extraction, the reflux temperature is 60-100° C., the reflux time is 1-3 h, and the extract is filtered and concentrated; 4) The concentrate is freeze-dried.

3. A drug for treating cervical spondylotic radiculopathy, characterized in that: The medicine comprises Brazilian mimosa pudica extract and a pharmaceutically acceptable carrier or excipient.

4. A method for preparing a drug for treating cervical spondylotic radiculopathy, characterized in that: The method comprises the following steps: 1) pre-freezing Brazilian mimosa under low temperature conditions; 2) freeze-drying and dehydrating the pre-frozen Brazilian Mimosa pudica; 3) crushing the freeze-dried Brazilian mimosa, adding water for reflux extraction, the reflux temperature is 60-100° C., the reflux time is 1-3 h, and the extract is filtered and concentrated; 4) The concentrate is freeze-dried.

5. Application of Brazilian Mimosa pudica extract in the preparation of drugs that promote M2 polarization of microglia.

6. A drug for promoting microglial M2 polarization, characterized in that: The medicine comprises Brazilian mimosa pudica extract and a pharmaceutically acceptable carrier or excipient.

7. A method for preparing a drug for promoting microglial M2 polarization, characterized in that: The method comprises the following steps: 1) pre-freezing Brazilian mimosa under low temperature conditions; 2) freeze-drying and dehydrating the pre-frozen Brazilian Mimosa pudica; 3) crushing the freeze-dried Brazilian mimosa, adding water for reflux extraction, the reflux temperature is 60-100° C., the reflux time is 1-3 h, and the extract is filtered and concentrated; 4) The concentrate is freeze-dried.

8. The use according to claim 2 or the method according to claim 4 or the method according to claim 7, characterized in that: In step 3), the mass volume ratio of Brazilian mimosa to water is 1g:10ml to 1g:40ml.

9. A method for promoting microglial M2 polarization for non-therapeutic purposes, characterized in that: The method comprises the following steps: 1) Cultivate microglia; 2) treating the microglia cultured in step 1) with a drug comprising the Brazilian Mimosa pudica extract according to claim 1.

10. A method for screening a drug for treating cervical spondylotic radiculopathy or promoting microglial M2 polarization, the method comprising the following steps: 1) Cultivate microglia; 2) treating the cells cultured in step 1) with the Mimosa pudica extract according to claim 1 and the candidate drug; 3) Determine whether the candidate drug can treat cervical spondylotic radiculopathy or promote microglial M2 type by comparing the effects of the Brazilian Mimosa pudica extract and the candidate drug on cell treatment.

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