New use of rape pollen alkali A in neuroprotection and prevention and treatment of brain diseases

Abstract

The application discloses a new use of rape pollen alkali A in nerve protection and brain disease prevention and treatment, discloses beneficial effects and molecular mechanisms of the rape pollen alkali A after the rape pollen alkali A acts on Caenorhabditis elegans, and provides significant differences of three groups (young group, natural aging group and rape pollen alkali A group) of the Caenorhabditis elegans on a transcriptome and a proteome, and reveals potential effects of the rape pollen alkali A on neurodegenerative changes and brain diseases caused by inflammatory substances such as leukotriene B4. The application finds that the rape pollen alkali A improves the pharyngeal pump and body swing of the Caenorhabditis elegans which gradually deteriorate with growth, and significantly improves the oxidative stress capacity of the Caenorhabditis elegans, and preliminarily verifies the effects of the rape pollen alkali A on neurodegenerative changes and brain diseases caused by inflammatory substances such as leukotriene B4 on a model organism. The effects of the rape pollen alkali A in the application are clear, and the rape pollen alkali A has application prospects in nerve protection and brain disease prevention and treatment caused by leukotriene B4.

Description

TECHNICAL FIELD

[0001] The application belongs to the field of medicine and biotechnology, and relates to a new use of pollenopyrroside A, in particular to a molecular mechanism and remarkable effect of pollenopyrroside A in nerve protection and prevention and treatment of brain diseases. BACKGROUND

[0002] Pyrrolospiroketals alkaloids are a class of natural compounds, which are first isolated from Astragalus membranaceus, and then extracted from pollen of Brassica napus, Acorus gramineus and Leptopus chinensis. Existing studies show that the alkaloids have the effects of delaying cell aging, prolonging cell passage number and inhibiting ROS production in high-glucose-induced rat glomerular mesangial cells. Pollenopyrroside A (PA) is a pyrrolospiroketals hexose ring alkaloid isolated from pollen of Brassica napus. Its unique structure may have activity and mechanism of action worthy of exploration, compared with the remarkable activity of other such alkaloids.

[0003] Caenorhabditis elegans is a commonly used model organism, which is initially developed for understanding developmental biology and neurobiology problems. Due to its short life cycle, clear genetic background, easy breeding and preservation, and easy observation, research on C. elegans has been extended to different fields of modern biology, including research on basic functions and interactions, and research on some human diseases. SUMMARY

[0004] The purpose of the present application is to provide a use of pollenopyrroside A. The present application researches and finds beneficial effects and molecular mechanisms of pollenopyrroside A after acting on C. elegans, thereby providing a basis for further development and utilization of pollenopyrroside A.

[0005] The technical scheme adopted by the present application is as follows:

[0006] A new use of pollenopyrroside A in nerve protection and prevention and treatment of brain diseases, and application of the pollenopyrroside A in preparation of a medicine or a pharmaceutical composition for nerve protection, prevention and / or treatment of brain diseases; the medicine or the pharmaceutical composition comprises a pharmaceutically acceptable carrier.

[0007] When the pollenopyrroside A is used for nerve protection, the concentration of the pollenopyrroside A in the medicine or the pharmaceutical composition is 0.4-1.6 mM, and preferably the concentration is 0.8 mM;

[0008] When the pollenopyrroside A is used for prevention and / or treatment of brain diseases, the concentration of the pollenopyrroside A in the medicine or the pharmaceutical composition is 0.4-1.6 mM, and preferably the concentration is 0.8 mM.

[0009] Further, the "pharmaceutical composition" means a mixture containing one or more of the said sinapine A or its physiologically / pharmaceutically acceptable salts or prodrugs and other chemical components.

[0010] Further, the said brain disease is a brain disease caused by neurodegeneration or inflammatory substances.

[0011] Further, the said brain disease is a brain disease caused by inflammatory substances of the leukotriene B4 type.

[0012] The present application has been studied as follows:

[0013] Firstly, the present application provides the effect of sinapine A on the prolongation of the healthy lifespan of nematodes, i.e. the effect of sinapine A on the improvement of the body function is judged by the Caenorhabditis elegans lifespan experiment, wherein the effective concentration of the compound for prolonging the healthy lifespan of the nematodes is 0.4-1.6 mM, and preferably 0.8 mM. In the oxidative stress experiment, sinapine A prolongs the average lifespan of the nematodes cultured under the H2O2 condition by 30%, significantly improving the stress resistance of the nematodes, which indicates that sinapine A can protect the nematodes from oxidative damage, which is related to the up-regulation of the enzyme CYP35A involved in the oxidation reaction. The present application has found that the said sinapine A has a new use in the neuroprotection, prevention and / or treatment of brain diseases, which is as follows:

[0014] In the prevention and / or treatment of brain diseases:

[0015] The present application provides the significant differences in the transcriptome and proteome of the three groups of nematodes (young group, natural aging group and sinapine A group), which reveals the potential effect of sinapine A on the neurodegeneration and brain diseases caused by inflammatory substances such as leukotriene B4. The transcriptomic and proteomic analysis shows that sinapine A can reverse the expression changes of CYP35A2, CYP35A3 and CYP35A4 in the nematodes with age, which correspond to the human homolog CYP2U1, and has a potential preventive and therapeutic effect on brain diseases caused by substances of the leukotriene B4 type. The oxidative stress experiment results of the nematodes show that sinapine A significantly improves the stress resistance of the nematodes, which can protect the nematodes from oxidative damage, which is related to the up-regulation of the enzyme CYP35A involved in the oxidation reaction.

[0016] In the neuroprotection:

[0017] The proteomic analysis shows that sinapine A up-regulates the expression of the neuroprotective proteins ilys-3, pme-4, B0410.3 and ttr, which indicates that it can improve the motor ability decline by improving the neurodegeneration.

[0018] The results of the body swing experiment and the swallowing pump experiment of the nematode in the present application show that the rape pollen base A can improve the movement ability and the swallowing pump frequency of the nematode, and it is preliminarily proved that the rape pollen base has the neuroprotective effect and can improve the movement ability decline caused by the neurodegenerative changes.

[0019] The present application has the following beneficial effects:

[0020] (1) The present application selects Caenorhabditis elegans as a model organism, simulates natural aging, studies the significant changes in gene transcription and protein expression during the aging process, and thus reveals the diseases that can be improved and regulated by the rape pollen base A and the molecular mechanisms thereof.

[0021] (2) The present application screens the optimal concentration of the rape pollen base A for efficacy research based on the nematode lifespan experiment, analyzes the molecular mechanisms of the rape pollen base A and the relevance to human diseases through transcriptomics and proteomics, finds the protective effect of the rape pollen base A on the nervous system and the potential effect of the rape pollen base A on the brain diseases caused by the inflammatory substance leukotriene B4, and further verifies the effect of the rape pollen base A on the physiological functions of the nematode through the body swing experiment and the swallowing pump experiment, verifies the effect of the rape pollen base A on the stress resistance of the nematode through oxidative stress, and provides a strong theoretical basis for the further research and development of the rape pollen base A in neurodegenerative diseases or brain diseases caused by leukotriene B4. BRIEF DESCRIPTION OF DRAWINGS

[0022] Figure 1 is a survival curve result graph of Caenorhabditis elegans, ##, P<0.011 compared with the DMSO group; **, P<0.01 compared with the blank group, ****P<0.0001;

[0023] Figure 2 is a volcano plot of differential genes obtained after transcriptome analysis of Caenorhabditis elegans in the rape pollen base A group and the aging group;

[0024] Figure 3 is a Venn analysis of transcriptome gene data;

[0025] Figure 4 is a statistical analysis of transcriptome expression differential genes;

[0026] Figure 5 is a volcano plot of differential proteins obtained after proteome analysis of Caenorhabditis elegans in the rape pollen base A group and the aging group;

[0027] Figure 6 is a Venn analysis of proteome data;

[0028] Figure 7 is a statistical analysis of proteome expression differential genes;

[0029] Figure 8 Graph showing the results of C. elegans pharyngeal pumping frequency, ****P<0.0001, *P<0.005;

[0030] Figure 9 Graph showing the results of C. elegans motility, ****P<0.0001, ***P<0.05;

[0031] Figure 10 Graph showing the results of C. elegans survival curve under 10 mM H2O2, ****P<0.0001. DETAILED DESCRIPTION

[0032] The present application is further described in the following Examples and in association with the accompanying drawings, which are in no way intended to limit the present application. Specific materials used in the following Examples of the application are provided below. However, it is to be understood that these are only exemplary and that materials of the same or similar type, make, quality, property or function as the following reagents and instruments can be used to practice or carry out the present application. Unless otherwise indicated, the experimental methods used in the following Examples are routine methods. Unless otherwise indicated, the materials, reagents and the like used in the following Examples are commercially available.

[0033] In this specification, the terms (definitions) are as follows:

[0034] To facilitate the understanding of this disclosure, certain technical and scientific terms are specifically defined below. Unless otherwise defined herein, all other technical and scientific terms used herein have the meanings commonly understood by one of ordinary skill in the art to which this disclosure belongs.

[0035] The term "small molecule compound" refers to a compound having a relative molecular mass of no more than 1000, and is not a small peptide, oligopeptide, oligosaccharide and oligonucleotide.

[0036] The terms "inhibition" or "blockade" are used interchangeably and encompass both partial and complete inhibition / blockade. Inhibition / blockade of a ligand preferably reduces or alters the normal level or type of activity that occurs upon ligand binding in the absence of inhibition or blockade.

[0037] The term "treatment" means the administration of an internal or external therapeutic agent, such as a composition comprising any one of the compounds of the present application, to a patient who has one or more symptoms of a disease, where the therapeutic agent is known to have a therapeutic effect on those symptoms. Typically, the therapeutic agent is administered in an amount effective to alleviate one or more symptoms of the disease in the treated patient or population, whether by inducing regression of such symptoms or inhibiting the progression of such symptoms to any clinically measurable extent. The amount of therapeutic agent effective for the alleviation of any particular symptom of a disease (also referred to as the "therapeutically effective amount") can vary according to factors such as the disease state, age, and weight of the patient, and the ability of the drug to elicit the desired therapeutic effect in the patient. Whether a disease symptom has been alleviated can be assessed by any clinical measurement method typically used by a physician or other health care professional to assess the severity or progression of the symptom. It is possible that an embodiment of the present application (e.g., a method of treatment or article of manufacture) can not be effective in alleviating the target disease symptom in every patient suffering from the target disease symptom, but that a statistically significant number of patients determined according to any statistical test known in the art, such as the Student t-test, the Chi-square test, the U-test according to Mann and Whitney, the Kruskal-Wallis test (H-test), the Jonckheere-Terpstra test, and the Wilcoxon test, should have the target disease symptom alleviated.

[0038] The term "pharmaceutical composition" means a mixture of one or more compounds described in the present disclosure or a physiologically / pharmaceutically acceptable salt or prodrug thereof with other chemical components, such as physiologically / pharmaceutically acceptable carriers and excipients. The purpose of a pharmaceutical composition is to facilitate administration of the active ingredient to a subject and to facilitate absorption of the active ingredient to elicit a biological activity.

[0039] The term "effective amount" or "effective dose" refers to the amount of a drug, compound, or pharmaceutical composition necessary to achieve a desired therapeutic result. For prophylactic use, the desired therapeutic result includes eliminating or reducing risk, lessening the severity, or delaying the onset of a disease, including biochemical, histological, and / or behavioral symptoms of the disease, its complications, and intermediate pathological phenotypes presenting during the course of the disease. For therapeutic applications, the desired therapeutic result includes a clinical outcome.

[0040] The term "pharmaceutically acceptable carrier" refers to any inactive substance suitable for use in a formulation for delivering antibody or antigen-binding fragments. Carriers can be anti-adhesion agents, adhesives, coatings, disintegrants, fillers or diluents, preservatives (such as antioxidants, antibacterial agents, or antifungal agents), sweeteners, absorption delay agents, wetting agents, emulsifiers, buffers, etc. Examples of suitable pharmaceutically acceptable carriers include water, ethanol, polyols (such as glycerol, propylene glycol, polyethylene glycol, etc.), dextrose, vegetable oils (such as olive oil), saline, buffer solutions, buffered saline, and isotonic agents such as sugars, polyols, sorbitol, and sodium chloride.

[0041] The terms “administer” and “treatment” when applied to animals, humans, experimental subjects, cells, tissues, organs, or biological fluids refer to the contact of an exogenous drug, therapeutic agent, diagnostic agent, or composition with the animal, human, subject, cell, tissue, organ, or biological fluid. “Administer” and “treatment” can refer to, for example, therapeutic, pharmacokinetic, diagnostic, research, and experimental methods. Cellular treatment includes contact between a reagent and cells, as well as contact between a reagent and a fluid, wherein the fluid is in contact with the cells. “Administer” and “treatment” also mean, for example, the treatment of cells in vitro and in vitro with a reagent, diagnostic agent, conjugate composition, or another cell. When applied to humans, veterinary, or research subjects, “treatment” refers to therapeutic treatment, preventative or prophylactic measures, research, and diagnostic applications.

[0042] Example 1: Routine culture of nematodes

[0043] E. coli OP50 was evenly spread on NGM medium. After the bacterial culture dried, the nematodes were transferred to the medium and cultured in a 20°C constant temperature incubator. Subculture was carried out according to the food on the medium and the experimental needs.

[0044] Example 2: Synchronization of Nematodes

[0045] To ensure experimental accuracy, nematodes need to be synchronized before each experiment to guarantee consistent growth stages. Observe nematode growth; once the nematodes enter the oviposition stage, wash them into centrifuge tubes using M9 buffer, centrifuge at 5000 rpm for 1 min, discard the supernatant, and prepare a lysis buffer at a ratio of 5M sodium hydroxide: 5% sodium hypochlorite: M9 buffer of 1:1:1. Mix well and add to the centrifuge tubes. Pipette approximately 10 times to ensure sufficient contact between the nematodes and the lysis buffer. Carefully observe to avoid over-lysis that could destroy the eggs. Immediately centrifuge at 5000 rpm for 1 min, discard the supernatant, and wash twice with M9 buffer. If incomplete lysis is observed, pipette agitation can be used during buffer washing to ensure complete lysis. The precipitate is then aspirated and placed on NGM culture medium coated with OP50. The synchronized nematodes are observed the following day.

[0046] Example 3 C. elegans lifespan experiment

[0047] 1. Experimental method: The experiment was divided into blank group, rape pollen alkaline A group (0.4 mM, 0.8 mM, 1.6 mM, 3.2 mM), solvent control group (1% DMSO), positive control group (1 mM resveratrol), 150 nematodes in each group. Add the compound to the inactivated OP50 bacterial solution, and the final concentration is as described above (OP50 inactivation is to prevent the possible bacteriostatic effect of the compound from affecting the accuracy of the experiment). The synchronized nematodes were cultured to the L4 stage, and the nematodes were picked to NGM medium containing 100 μM FUDR (FUDR is used to inhibit nematode egg laying to prevent the influence of offspring), and placed in a 20°C incubator for culture. The nematodes were transferred to new medium every day, and the number of surviving, dead and lost nematodes was counted until all the nematodes died. During the experiment, the nematodes that died due to operational errors and the nematodes that crawled onto the wall of the dish were counted as lost. The nematodes were lightly touched with a picking needle, and the nematodes that did not respond to external stimuli were determined to be dead.

[0048] 2. Results: As shown in Table 1, compared with the blank group, the average lifespan of nematodes cultured at a rape pollen alkaline A concentration of 0.4 mM, 0.8 mM and 1.6 mM was extended by 33% (P<0.0001), 43% (P<0.0001) and 19% (P=0.0016), respectively. However, the average lifespan of nematodes cultured at a higher concentration (3.2 mM) was not significantly affected. In addition, compared with the DMSO group, the average lifespan of nematodes in the positive control group was extended by 12% (P=0.0061). The above data fully demonstrate that, under normal experimental conditions, within a certain concentration range, rape pollen alkaline A has a significant positive effect on extending the overall function of C. elegans. Figure 1

[0049] Example 4 Transcriptome and proteome analysis of C. elegans

[0050] 1. Experimental method: Synchronized nematodes were divided into three groups: young group: 3d nematodes fed with OP50; old group: 13d nematodes fed with OP50; rape pollen alkaline A group: 13d nematodes fed with OP50 containing 0.8 mM PA, and the nematodes were cultured under the same conditions. Among them, 3-day-old nematodes represent young nematodes; 13-day-old nematodes are in the transition period from middle age to old age, representing aging nematodes. Three parallel samples were set up in each group. The nematode samples were collected, rinsed twice with M9 buffer, centrifuged to discard the supernatant, the precipitate was dried as much as possible, then quickly frozen with liquid nitrogen, and stored at -80°C. Further detection was assisted by Shanghai Euilight Biotechnology Co., Ltd.

[0051] ​2. Transcriptome analysis results: statistically analyze the differences in the expression of Caenorhabditis elegans genes, and screen according to the threshold (|log2Fold Change|>1, p-value<0.05). Compared with the old group, 320 genes are up-regulated and 48 genes are down-regulated in the Brassica pollen alkali A group ( Figure 2 ). Venn diagram analysis is performed on the Brassica pollen alkali A group vs. the old group, the Brassica pollen alkali A group vs. the young group, and the young group vs. the old group ( Figure 3 ). It can be seen that there are 36 transcriptome genes that are significantly changed by Brassica pollen alkali A, in addition to the differentially expressed genes shared by the young group. Brassica pollen alkali A can reverse the transcription of some changes with age ( Figure 4 ). Among them, the genes CELE_C49G7.8 and CELE_K09D9.2 are of concern. The expression of these two genes decreases to about 20% of the young group with aging, and can be adjusted to the level of the young group under the intervention of Brassica pollen alkali A. It is found through reactome retrieval embedded on Uniprot that the proteins expressed by these two genes are CYP35A3 and CYP35A4, which are homologues of human CYP4502U1. It is found through retrieval that human CYP4502U1 is highly expressed in the brain and can metabolize inflammatory substances leukotriene B4. The accumulation of leukotriene B4 in the brain is closely related to brain diseases such as ischemic stroke [Nouri K, Pietrancosta N, Le Corre L, et al. Human Orphan Cytochrome P450 2U1 Catalyzes the ω-Hydroxylation of Leukotriene B4 [J]. International Journal of Molecular Sciences, 2022, 23(23): 14615.]. In subsequent proteome data analysis, the protein expression shows difference (up-regulation) is CYP35A2, which is also a homologue of human CYP4502U1. Both transcriptome and proteome data show that Brassica pollen alkali A can reverse the down-regulation of CYP35A class genes in nematodes with age, and has potential prevention and treatment effects on human brain diseases induced by inflammatory mediators such as leukotriene B4.

[0052] 3. Proteome analysis results: There are significant differences in protein expression between the Brassica pollen alkali A group and the aging group. According to the difference screening condition Fold change≥1.2 or Fold change≤1 / 1.2 and p-value<0.05, 174 differential proteins are screened out in the Brassica pollen alkali A group and the old group, including 105 up-regulated proteins and 59 down-regulated proteins ( Figure 5). Venn diagram analysis of Brassica pollen allergen A group vs. old group, Brassica pollen allergen A group vs. young group, and young group vs. old group Figure 6 It can be seen that, in addition to the differentially expressed proteins shared with the young group, 52 proteins were significantly changed by Brassica pollen allergen A. Brassica pollen allergen A can effectively reverse the protein expression changes with aging Figure 7

[0053] ​By analyzing the top 20 proteins up-regulated, we found that the proteins related to nerves were significantly up-regulated (see Table 1): ilys-3 protein is essential for grinding in the pharynx during the growth of nematodes [Gravato-Nobre M J, Vaz F, Filipe S, et al. The invertebrate lysozyme effector ILYS-3 is systemically activated in response to danger signals and confers antimicrobial protection in C. elegans [J]. PLoS pathogens, 2016, 12(8): e1005826.], the pumping of the pharynx in nematodes is a behavior controlled by multiple independent nervous system cells that contract muscles to pump food into the intestine, which is a typical index of C. elegans motor nerve behavior [Avery L, Shtonda B B. Food transport in the C. elegans pharynx [J]. Journal of Experimental Biology, 2003, 206(14): 2441-2457.]. pme-4 encodes a poly(ADP-ribose) glycohydrolase (PARG) homologous to human PARG, mainly expressed in nerve cells and embryos, and poly(ADP-ribose) metabolism is essential for maintaining the normal function of neurons [St-Laurent J F, Gagnon S N, Dequen F, et al. Altered DNA damage response in Caenorhabditis elegans with impaired poly(ADP-ribose) glycohydrolases genes expression [J]. DNA repair, 2007, 6(3): 329-343.]; B0410.3 is a neuropeptide-like protein, and neuropeptides are distributed throughout the nervous system, can be used as neurotransmitters, neuromodulators or neurohormones, play an important physiological role in the brain and have neuroprotective effects [Dam D V, Dijck AV, Janssen L, et al. Neuropeptides in Alzheimer's disease: from pathophysiological mechanisms to therapeutic opportunities [J]. Current Alzheimer Research, 2013, 10(5): 449-468.].Therefore, the effect of canola pollenine A on the pharyngeal pumping frequency of nematodes was further evaluated on the nematode model, and the verification of the neuroprotective effect of canola pollenine A was carried out. Figure 8 )

[0054] Table 1. Expression of nerve-related differential proteins

[0055]

[0056] In addition, ttr-32 and ttr-30 are transthyretin-like genes, which are closely related to the abnormal motor behavior of nematodes. Transthyretin TTR plays an important protective role in typical neurodegenerative diseases such as Alzheimer's disease, mainly by interacting with beta-amyloid protein (Aβ) to slow the occurrence of the disease [Alemi M, Silva S C, Santana I, et al. Transthyretin stability is critical in assisting beta amyloid clearance-Relevance of transthyretin stabilization in Alzheimer's disease [J]. CNS neuroscience & therapeutics, 2017, 23(7): 605-619.]. Further screening of all differentially expressed genes found that a series of transthyretin-like proteins were up-regulated in expression (Table 2). It is shown that canola pollenine A up-regulates the expression of transthyretin-like differential proteins, which may improve the movement ability of nematodes, and the subsequent body swing experiment of nematodes is used for verification.

[0057] Table 2. Expression of transthyretin-like differential proteins

[0058]

[0059] Example 5: Pharyngeal pump experiment of Caenorhabditis elegans

[0060] 1. Experimental method: synchronized L4 stage nematodes were transferred to NGM medium coated with inactivated OP50, and inactivated OP50 containing 0, 0.8 mM canola pollenine A was used as blank control group and canola pollenine A group, respectively. On the 5th, 7th and 9th day, not less than 10 nematodes were picked to new medium without OP50, and the number of pharyngeal pump beats in 15s was counted.

[0061] 2. Results: The pharyngeal pump of C. elegans is similar to the human heart. The pharyngeal pumping represents the feeding behavior, and the frequency is related to the degree of nerve injury. The frequency of the pharyngeal pumping of the nematodes decreases with age. The frequency of the pharyngeal pumping of the rapeseed alkaloid A group of nematodes at 7 and 9 days is significantly different from that of the control group (P < 0.0001), as shown in Figure 8 . It is proved that rapeseed alkaloid A can delay the decrease of the frequency of the pharyngeal pumping with age and has a protective effect on the nervous system.

[0062] Example 6 Experiment of the movement ability of C. elegans

[0063] 1. Experimental method: The synchronized L4 stage nematodes were transferred to NGM medium coated with inactivated OP50 containing 0, 0.8 mM rapeseed alkaloid A. At 5, 7, and 9 days, respectively, no less than 10 nematodes were picked to new medium without OP50 and allowed to move freely for 2 min, and then the number of body swings of the nematodes within 20 s was counted.

[0064] 2. Results: As shown in Figure 9 , the number of body swings of the nematodes decreases with the increase of the life span of the nematodes. The administration of rapeseed alkaloid A can significantly increase the number of body swings of the nematodes (P < 0.05). It is proved that rapeseed alkaloid A can significantly improve the movement ability of the nematodes, improve the neurodegenerative changes, and improve the survival state of the nematodes.

[0065] Example 7 Oxidative stress experiment of C. elegans

[0066] 1. Experimental method: The experiment was divided into a blank group, a rapeseed alkaloid A group (0.8 mM), a solvent control group (1% DMSO), and a positive control group (1 mM resveratrol), with 90 nematodes in each group. The synchronized L4 stage nematodes were picked to NGM medium coated with inactivated OP50 containing the above-mentioned concentrations of compounds. After being cultured at 20°C for 3 days, the nematodes were transferred to NGM medium containing 10 mM H2O2 and cultured at 20°C, and the number of dead and surviving nematodes was counted every 1 h until all the nematodes died.

[0067] 2. Results: Oxidative stress is mainly due to the imbalance of redox balance in vivo, resulting in excessive reactive oxygen species, making the body face damage. Oxidative stress can cause direct damage to the central nervous system. Neurons have a high oxygen demand, but the level of antioxidant is relatively low, so neurons are at high risk of oxidative stress [Di Pietro V, Lazzarino G, Amorini AM, et al. Neuroglobin expression and oxidant / antioxidant balance after graded traumatic brain injury in the rat [J]. Free Radical Biology and Medicine, 2014, 69: 258-264.]. When the active oxygen increases significantly, it will damage dopamine-neurons [Shukla V, Mishra S K, Pant H C. Oxidative stress in neurodegeneration [J]. Advances in pharmacological sciences, 2011, 2011.], which is an important pathogenesis of Parkinson's disease, and also one of the molecular mechanisms of Alzheimer's disease [Kemppainen S, Lindholm P, Galli E, et al. Cerebral dopamine neurotrophic factor improves long-term memory in APP / PS1 transgenic mice modeling Alzheimer's disease as well as in wild-type mice [J]. Behavioural brain research, 2015, 291: 1-11.]. As shown in Table 2, compared with the blank control group, the average lifespan was prolonged by 30% at a concentration of 0.8 mM PA. The effect of prolonging lifespan was significant (P < 0.0001). This indicates that rape pollen base A significantly improves the stress resistance of nematodes and can protect nematodes from oxidative damage, which is related to the up-regulation of the enzyme CYP35A involved in oxidative reactions. Figure 10

[0068] In summary, combined with the effects of rape pollen base A on the healthy lifespan of C. elegans, gene transcription and protein expression, it is found that rape pollen base A has a potential role in neuroprotection and prevention of intracerebral diseases.

[0069] ​The above embodiments only express the implementation ways of the present application, and cannot be understood as the limitation to the scope of the present application patent. It should be pointed out that, for those skilled in the art, several modifications and improvements can be made without departing from the concept of the present application, and these all belong to the protection scope of the present application.

Claims

1. The use of rapeseed pollen alkaloid A in the preparation of a medicament or pharmaceutical composition for the prevention and / or treatment of brain diseases; characterized in that, The brain disease mentioned is Alzheimer's disease, which is caused by neurodegenerative changes.

2. The application according to claim 1, characterized in that, The drug or drug composition includes a pharmaceutically acceptable carrier.

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