CN116509870A - New use of rape pollen alkaloid A in neuroprotection and brain disease prevention and treatment - Google Patents

New use of rape pollen alkaloid A in neuroprotection and brain disease prevention and treatment Download PDF

Info

Publication number
CN116509870A
CN116509870A CN202310318285.2A CN202310318285A CN116509870A CN 116509870 A CN116509870 A CN 116509870A CN 202310318285 A CN202310318285 A CN 202310318285A CN 116509870 A CN116509870 A CN 116509870A
Authority
CN
China
Prior art keywords
rape pollen
nematodes
alkaloid
neuroprotection
treatment
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Pending
Application number
CN202310318285.2A
Other languages
Chinese (zh)
Inventor
李悦青
左雨薇
孟庆伟
杨澳
高志刚
王世盛
郭修晗
赵伟杰
王帅
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Dalian University of Technology
Ningbo Research Institute of Dalian University of Technology
Original Assignee
Dalian University of Technology
Ningbo Research Institute of Dalian University of Technology
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Dalian University of Technology, Ningbo Research Institute of Dalian University of Technology filed Critical Dalian University of Technology
Priority to CN202310318285.2A priority Critical patent/CN116509870A/en
Publication of CN116509870A publication Critical patent/CN116509870A/en
Pending legal-status Critical Current

Links

Classifications

    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/535—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with at least one nitrogen and one oxygen as the ring hetero atoms, e.g. 1,2-oxazines
    • A61K31/5375—1,4-Oxazines, e.g. morpholine
    • A61K31/5386—1,4-Oxazines, e.g. morpholine spiro-condensed or forming part of bridged ring systems
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P25/00—Drugs for disorders of the nervous system
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P25/00—Drugs for disorders of the nervous system
    • A61P25/14—Drugs for disorders of the nervous system for treating abnormal movements, e.g. chorea, dyskinesia
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P25/00—Drugs for disorders of the nervous system
    • A61P25/28—Drugs for disorders of the nervous system for treating neurodegenerative disorders of the central nervous system, e.g. nootropic agents, cognition enhancers, drugs for treating Alzheimer's disease or other forms of dementia
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P39/00—General protective or antinoxious agents
    • A61P39/06—Free radical scavengers or antioxidants
    • Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
    • Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
    • Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
    • Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change

Abstract

A new application of rape pollen alkaloid A in neuroprotection and brain disease prevention and treatment is disclosed, which discloses the beneficial effect and molecular mechanism of rape pollen alkaloid A after acting on caenorhabditis elegans. The caenorhabditis elegans is adopted as model organism, and the obvious difference of three groups of nematodes (young group, natural aging group and rape pollen base A group) in transcriptome and proteome is provided, so that the potential effect of the rape pollen base A on neurodegenerative change and brain diseases caused by inflammatory substances such as leukotriene B4 is revealed. The invention discovers that the improvement of the rape pollen alkali A on the swallowing pump and the body swing of nematodes which gradually worsen along with the growth, obviously improves the oxidative stress capability of the nematodes, and preliminarily verifies the effect of the rape pollen alkali A on neurodegenerative changes and brain diseases caused by inflammatory substances such as leukotriene B4 in model organisms. The rape pollen alkaloid A has definite effect and has application prospect in the aspects of neuroprotection and brain disease prevention and treatment caused by leukotriene B4 substances.

Description

New use of rape pollen alkaloid A in neuroprotection and brain disease prevention and treatment
Technical Field
The invention belongs to the field of medicine and biotechnology, relates to a new application of rape pollen alkaloid A, and in particular relates to a molecular mechanism and remarkable effect of rape pollen alkaloid A in the aspect of neuroprotection and brain disease prevention and treatment.
Background
The pyrrole spiroketal alkaloid is a natural compound, is initially separated from astragalus, and is sequentially extracted from rape pollen, grassleaf sweelflag rhizome and caper. The existing researches show that the alkaloid has the effects of delaying cell aging, prolonging cell passage times and inhibiting the generation of ROS in rat glomerular mesangial cells induced by high sugar. Rape pollen alkaloid A (Pollenopyrroside A, PA for short) is pyrrole spiroketal six-membered sugar ring alkaloid separated from rape pollen, and the unique structure of the rape pollen alkaloid A possibly has activity and action mechanism worthy of exploration compared with other obvious activities of the alkaloid.
Caenorhabditis elegans (c.elegans) is a common model organism originally developed for understanding developmental biological and neurobiological problems. Because of its characteristics of short life cycle, clear genetic background, easy propagation and preservation, easy observation, etc., the research on caenorhabditis elegans has been expanded to explore different fields of modern biology, including the research on basic functions and interactions, and the research on certain diseases in humans.
Disclosure of Invention
The invention aims to provide the application of the rape pollen alkali A, and researches show that the rape pollen alkali A has beneficial effects and molecular mechanisms after acting on caenorhabditis elegans, thereby providing basis for further development and utilization of the rape pollen alkali A.
The invention adopts the technical scheme that:
a new application of rape pollen alkaloid A in the aspects of neuroprotection and brain disease prevention and treatment, and the application of rape pollen alkaloid A in the preparation of medicaments or pharmaceutical compositions for neuroprotection, prevention and/or treatment of brain diseases; the medicament or the pharmaceutical composition comprises a pharmaceutically acceptable carrier.
When canola pollen alkaloid A is used in neuroprotection, its concentration in the medicament or pharmaceutical composition is 0.4-1.6mM, preferably 0.8mM;
when canola pollen alkaloid A is used in the prevention and/or treatment of brain diseases, its concentration in the medicament or pharmaceutical composition is 0.4-1.6mM, preferably 0.8mM.
Further, the "pharmaceutical composition" means a mixture containing one or more of the canola pollen alkaloid a or a physiologically/pharmaceutically acceptable salt or prodrug thereof and other chemical components.
Further, the brain disease is a brain disease caused by neurodegenerative changes or inflammatory substances.
Further, the brain diseases are brain diseases caused by inflammatory substances of leukotriene B4.
The present invention was studied as follows:
firstly, the invention provides the effect of the rape pollen base A on prolonging the healthy life of the insects, namely the effect of the rape pollen base A on improving the function of the organism is judged by the caenorhabditis elegans life experiment, wherein the effective concentration of the compound for prolonging the healthy life of the caenorhabditis elegans is 0.4-1.6mM, preferably 0.8mM. In the oxidative stress experiment, brassinosteroids A and H 2 O 2 The average life span of the nematodes cultured under the condition is prolonged by 30%, and the anti-stress capability of the nematodes is obviously improved, which indicates that the rape pollen base A can protect the nematodes from oxidative damage and is related to the enzyme CYP35A which up-regulates the nematodes to participate in the oxidation reaction. The research of the invention discovers that the rape pollen alkaloid A has new application in the aspects of neuroprotection, prevention and/or treatment of brain diseases, and the application is as follows:
in the prevention and/or treatment of brain diseases:
the present invention provides significant differences in transcriptome and proteome among three groups of nematodes (young, naturally senescent and canola pollen base a), revealing the potential effect of canola pollen base a on neurodegenerative changes and inflammatory substances such as leukotriene B4. Transcriptomics and proteomics analysis show that the rape pollen alkaloid A can reverse the expression change of CYP35A2, CYP35A3 and CYP35A4 in nematodes with age, and the rape pollen alkaloid A corresponds to human homolog CYP2U1 and has potential prevention and treatment effects on brain diseases caused by leukotriene B4 substances. The experimental result of the oxidative stress of the nematodes shows that the rape pollen alkali A remarkably improves the anti-stress capability of the nematodes, can protect the nematodes from oxidative damage, and is related to up-regulating enzyme CYP35A participating in the oxidative reaction.
In neuroprotection:
proteomic analysis showed that oilseed rape pollen base A up-regulates the expression of neuroprotective proteins ilys-3, pme-4, B0410.3 and ttr, indicating that it can improve motor capacity by improving neurodegenerative changes.
The results of the body swing experiment and the swallowing pump experiment show that the rape pollen alkali A can improve the exercise capacity and the swallowing pump frequency of nematodes, and the rape pollen alkali has the neuroprotection effect and can improve the exercise capacity decline caused by neurodegenerative change.
The invention has the beneficial effects that:
(1) The invention selects caenorhabditis elegans as model organism, simulates natural aging, and researches the obvious change of rape pollen alkali A on gene transcription and protein expression accompanying the aging process, thereby revealing the diseases and molecular mechanism of the rape pollen alkali A possibly improved and regulated.
(2) The invention screens out the optimal concentration of the rape pollen alkali A for efficacy research based on nematode life experiments, analyzes the molecular mechanism of the rape pollen alkali A and the relevance of the rape pollen alkali A and human diseases through transcriptomics and proteomics, and discovers the protection effect on a nervous system and the potential effect on inflammatory substances leukotriene B4 to cause brain diseases. Further verifying the effect of the rape pollen alkali A on the physiological function of the insects through a body swing experiment and a swallowing pump experiment; verifying the effect of the rape pollen alkali A on the nematode anti-stress capability through oxidative stress; provides a powerful theoretical basis for further research and development of brain diseases caused by rape pollen alkali A in neurodegenerative diseases or leukotriene B4.
Drawings
FIG. 1 is a graph showing the results of the survival curves of caenorhabditis elegans, #, with P <0.011 compared to the DMSO group; * P <0.01, < P <0.0001;
FIG. 2 is a diagram of differential gene volcanic obtained after transcriptome analysis of the rape pollen base A group and the caenorhabditis elegans of the senescence group;
FIG. 3is Venn analysis of transcriptome gene data;
FIG. 4 is a statistical analysis of transcriptome expression differential genes;
FIG. 5 shows a protein profile of C.elegans from the pollinosis A and B.elegans from the aging group after proteomic analysis;
FIG. 6 is Venn analysis of proteomic data;
FIG. 7 is a statistical analysis of proteomic expression differential genes;
fig. 8 is a graph of caenorhabditis elegans swallowing pump frequency results with P <0.0001 and P <0.005;
fig. 9 is a graph of caenorhabditis elegans exercise capacity results with P <0.0001 and P <0.05;
FIG. 10 is 10mM H 2 O 2 Graph of the survival curve of caenorhabditis elegans under conditions<0.0001。
Detailed Description
The invention is further illustrated by the following examples, which are not to be construed as limiting the invention, in conjunction with the accompanying drawings. Specific materials and sources thereof used in embodiments of the present invention are provided below. However, it should be understood that these are merely exemplary and are not intended to limit the present invention, as materials that are the same as or similar to the type, model, quality, nature, or function of the reagents and instruments described below may be used in the practice of the present invention. The experimental methods used in the following examples are conventional methods unless otherwise specified. Materials, reagents and the like used in the examples described below are commercially available unless otherwise specified.
In this specification, the terms (definitions) are as follows:
for easier understanding of the present disclosure, certain technical and scientific terms are specifically defined below. Unless defined otherwise herein, all other technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this disclosure belongs.
The term "small molecule compound" refers to: the relative molecular mass is not more than 1000, and is not small peptide, oligopeptide, oligosaccharide and oligonucleotide.
The terms "inhibit" or "block" are used interchangeably and encompass both partial and complete inhibition/blocking. Inhibition/blocking of the ligand preferably reduces or alters the normal level or type of activity that occurs when ligand binding occurs without inhibition or blocking.
The term "treatment" means administration of an internally or externally applied therapeutic agent, such as a composition comprising any of the compounds of the invention, to a patient having one or more symptoms of a disease for which the therapeutic agent is known to have a therapeutic effect. Typically, the therapeutic agent is administered to the subject patient or population in an amount effective to alleviate one or more symptoms of the disease, whether by inducing regression of such symptoms or inhibiting the development of such symptoms to any clinically measurable extent. The amount of therapeutic agent (also referred to as a "therapeutically effective amount") effective to alleviate any particular disease symptom can vary depending on a variety of factors, such as the disease state, age, and weight of the patient, and the ability of the drug to produce a desired therapeutic effect in the patient. Whether a disease symptom has been reduced can be assessed by any clinical test method that a physician or other healthcare professional typically uses to assess the severity or progression of the symptom. While embodiments of the present invention (e.g., therapeutic methods or articles of manufacture) may be ineffective in alleviating symptoms of each of the disease of interest, they should alleviate symptoms of the disease of interest in a statistically significant number of patients, as determined by any statistical test methods known in the art, such as Student t test, chi-square test, U test according to Mann and Whitney, kruskal-Wallis test (H test), jonckheere-Terpstra test, and Wilcoxon test.
The term "pharmaceutical composition" means a mixture comprising one or more compounds of the present disclosure or a physiologically/pharmaceutically acceptable salt or prodrug thereof, and other chemical components, such as physiologically/pharmaceutically acceptable carriers and excipients. The purpose of the pharmaceutical composition is to promote the administration to organisms, facilitate the absorption of active ingredients and thus exert biological activity.
The terms "effective amount", "effective dose" and "effective amount" refer to the amount of a drug, compound or pharmaceutical composition necessary to achieve any one or more beneficial or desired therapeutic results. For prophylactic use, beneficial or desired results include elimination or reduction of risk, lessening the severity, or delaying the onset of a disorder, including biochemical, histological and/or behavioral symptoms of the disorder, its complications, and intermediate pathological phenotypes that are exhibited during the development of the disorder. For therapeutic applications, beneficial or desired results include clinical results.
The term "pharmaceutically acceptable carrier" refers to any inactive substance suitable for use in a formulation for delivery of an antibody or antigen-binding fragment. The carrier may be an anti-adherent, binder, coating, disintegrant, filler or diluent, preservative (e.g., antioxidant, antimicrobial or antifungal), sweetener, absorption delaying agent, wetting agent, emulsifier, buffer, etc. Examples of suitable pharmaceutically acceptable carriers include water, ethanol, polyols (e.g., glycerol, propylene glycol, polyethylene glycol, and the like) dextrose, vegetable oils (e.g., olive oil), saline, buffers, buffered saline, and isotonic agents such as sugars, polyols, sorbitol, and sodium chloride.
The terms "administering" and "treating" when applied to an animal, human, experimental subject, cell, tissue, organ, or biological fluid, refer to contacting an exogenous pharmaceutical, therapeutic, diagnostic, or composition with the animal, human, subject, cell, tissue, organ, or biological fluid. "administration" and "treatment" may refer to, for example, therapeutic, pharmacokinetic, diagnostic, research, and experimental methods. Treatment of a cell includes contacting a reagent with the cell, and contacting the reagent with a fluid, wherein the fluid is in contact with the cell. "administration" and "treatment" also mean in vitro and ex vivo treatment of, for example, a cell by an agent, diagnosis, binding composition, or by another cell. "treatment" when applied to a human, veterinary or research subject refers to therapeutic treatment, prophylactic or preventative measures, research and diagnostic applications.
EXAMPLE 1 routine cultivation of nematodes
E.coli OP50 is uniformly coated on the NGM culture medium, after bacterial liquid is dried, nematodes are transferred to the culture medium and cultured in a constant temperature incubator at 20 ℃, and passage is carried out according to food on the culture medium and experimental requirements.
EXAMPLE 2 synchronization of nematodes
To ensure the accuracy of the experiment, the nematodes need to be subjected to a synchronization treatment before each experiment is performed, so as to ensure the consistency of the growth stages. Observing the growth condition of the nematodes, flushing the nematodes into a centrifuge tube by using M9 buffer solution when the nematodes enter the spawning period, centrifuging for 1min at 5000r/min, discarding the supernatant, and then adding 5M sodium hydroxide: 5% sodium hypochlorite: preparing lysate by using M9 buffer solution in a ratio of 1:1:1, mixing uniformly, and adding into a centrifuge tube. And (5) blowing the mixture by a pipetting gun for about 10 times to fully contact the insect body with the lysate. Note that failure to lyse excessively leads to destruction of the eggs. Centrifugation was then performed at 5000r/min for 1min, the supernatant was discarded, and M9 buffer was added for 2 washes. If the lysis is found to be incomplete, the insect body can be completely lysed by blowing with a liquid-transferring gun during the washing of the buffer solution. And then sucking out the sediment, placing the sediment on NGM culture coated with OP50, and observing the sediment on the next day to obtain the synchronized nematode.
EXAMPLE 3 caenorhabditis elegans longevity test
1. The experimental method comprises the following steps: the experiments were divided into a blank group, a rape pollen base A group (0.4 mM, 0.8mM, 1.6mM, 3.2 mM), a solvent control group (1% DMSO), a positive control group (1 mM resveratrol), 150 nematodes per group. The compounds were added to the inactivated OP50 bacterial liquid at the final concentrations as described above (OP 50 inactivation is to prevent the compounds from affecting experimental accuracy by the bacteriostatic effect that may be present). The synchronized nematodes were cultured to stage L4, the nematodes were picked onto NGM medium containing 100. Mu.M FUDR (FUDR was used to suppress nematode oviposition, prevent the effects of progeny) and placed in a 20℃incubator. Nematodes were transferred daily to new medium and the number of surviving, dying and lost nematodes was counted until the nematodes all died. During the experiment, the nematodes that die due to misoperation were all counted as missing nematodes that had climbed onto the dish wall. The nematodes were lightly touched with a pick to determine that the nematodes did not respond to the external stimulus.
2. Results: as shown in FIG. 1, the average life span of nematodes was prolonged by 33% (P < 0.0001), 43% (P < 0.0001) and 19% (P=0.0016) respectively at concentrations of brassinosteroids A of 0.4mM, 0.8mM and 1.6mM, compared with the blank group. While the mean life of the nematodes cultivated at higher concentrations (3.2 mM) had no significant effect. In addition, the mean nematode life was 12% longer (p=0.0061) compared to the DMSO group for the positive control group. The data fully demonstrate that, under normal experimental conditions, the rape pollen alkaloid A has obvious positive influence on prolonging the overall function of caenorhabditis elegans within a certain concentration range.
EXAMPLE 4 transcriptome and proteome analysis of C.elegans
1. The experimental method comprises the following steps: synchronous nematodes are divided into 3 groups: young group: 3d nematodes feeding OP 50; aged group: nematodes fed OP50 d; oilseed rape pollen alkaloid A group: nematodes fed OP50 d containing 0.8mM PA were all cultured under the same conditions. Wherein the 3-day nematodes represent young nematodes; the 13-day nematodes are the transition period from middle-aged to elderly and represent senescent nematodes. Three parallel samples were set up for each group. Nematode samples were collected separately, washed 2 times with M9 buffer, centrifuged to discard supernatant, the pellet was blotted as dry as possible, and then flash frozen with liquid nitrogen and stored at-80 ℃. Further testing was aided by Shanghai European biotechnology company.
2. Transcriptome analysis results: statistical analysis of differences in caenorhabditis elegans expression genes was performed according to a threshold (|log) 2 Fold Change|>1，p-value<0.05 Screening, up-regulation of 320 gene expression and down-regulation of 48 gene expression in rape pollen alkali A group compared with the aged group (figure 2); analysis of Venn plots for the aged, young and young groups of brassinosteroids a vs (fig. 3) showed that the removal of differentially expressed genes shared by the young groups resulted in 36 transcriptome genes significantly altered by brassinosteroids a, which effectively reversed some of the transcripts altered by senescence (fig. 4). Of interest are the nematode genes CELE_ C49G7.8 and CELE_ K09D9.2 whose expression decreases to about 20% of that in young, which can be regulated back to young levels with the intervention of brassinosteroids A, and the proteins expressed by these two genes, CYP35A3 and CYP35A4, are found to be homologs to human CYP4502U1, which is found to be highly expressed in the brain to metabolize the inflammatory substance leukotriene B4, which is accumulated in brain diseases such as ischemic stroke [ Nouri K, pietrancosta N, le Corre L, et al Human Orphan Cytochrome P4502U1 Catalyzes the omega-Hydroxylation of Leukotriene B4[ J].International Journal of Molecular Sciences，2022，23(23):14615.]. In subsequent proteomic data analysis, protein expression exhibited variability (upregulation) of CYP35A2, also a homolog with human CYP4502U 1. Both transcriptome and proteome data indicate that brassinosteroids A can reverse the down-regulation of the nematode CYP 35A-like gene with age, for inflammatory diseases such as leukotriene B4The medium-induced human brain diseases have potential prevention and treatment effects.
3. Proteomic analysis results: the expression of the rape pollen alkaloid A group and the expression of the aging histone are obviously different, and 174 differential proteins comprising 105 up-regulating proteins and 59 down-regulating proteins are screened out according to the differential screening condition that the Fold change is more than or equal to 1.2 or the Fold change is less than or equal to 1/1.2 and the p-value is less than 0.05 compared with the aging group. Analysis of Venn plots for the aged, young and young groups of brassinosteroids a vs (fig. 6) showed that the removal of the differentially expressed proteins shared with the young groups resulted in 52 protein expressions evident from brassinosteroids a changes, and brassinosteroids a effectively reversed the protein expression that changed with age (fig. 7).
By analyzing the first 20 proteins up-regulated proteins, it was found that the proteins associated with nerves were significantly up-regulated (see table 1): the ilys-3 protein is essential for pharyngeal abrasion during nematode growth [ Gravat-Nobre M J, vaz F, filipe S, et al invertebrate lysozyme effector ILYS-3is systemically activated in response to danger signals and confers antimicrobial protection in C.elegans[J ], PLoS pathens, 2016, 12 (8): e1005826 ], and nematode pump pharyngeal pumping is a behavior in which food is pumped into the gut by multiple nerve cells independent of the systemic nervous system, a typical motor nerve behavior-related index for C.elegans [ Avery L, shtonda 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, expressed primarily in neurons and embryos, [ St-Laurent J F, gagnon S N, deque F, et al, altered DNAdamage response in Caenorhabditis elegans with impaired poly (ADP-ribose) glycohydrolases genes expression [ J ]. DNArepair,2007,6 (3): 329-343 ] necessary to maintain the normal function of neuronal cells; b0410.3 is a neuropeptide-like protein, which extends throughout the nervous system and acts as a neurotransmitter, neuromodulator or neurohormone, playing an important physiological role in the brain and neuroprotection [ 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 influence of the rape pollen alkali A on the swallowing frequency of nematodes is further evaluated on a nematode model, and the verification of the neuroprotection effect of the rape pollen alkali A is carried out. (FIG. 8)
TABLE 1 expression of neural-related differential proteins
In addition, ttr-32 and ttr-30 are transthyretin genes, and are closely related to abnormal motor nerve behaviors of nematodes. Transthyretin TTR plays an important protective role in the typical neurodegenerative disease Alzheimer 'S disease, and slows down the occurrence of the disease mainly through interaction with beta-amyloid (Abeta) [ Alemi M, silva S C, santana I, et al, transthretin 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 revealed that there was also a series of upregulation of transthyretin expression levels (Table 2). The expression of transthyretin-like differential protein is regulated by rape pollen alkali A, so that the nematode movement capacity can be improved, and the follow-up verification is carried out by adopting a nematode body swing experiment.
TABLE 2 transthyretin-like differential protein expression
EXAMPLE 5 caenorhabditis elegans swallowing Pump test
1. The experimental method comprises the following steps: the L4-phase nematodes which are synchronized are transferred to NGM medium coated with inactivated OP50, and the inactivated OP50 contains 0mM rape pollen base A and 0.8mM rape pollen base A as blank control group and rape pollen base A group respectively. And (3) picking up at least 10 nematodes on the 5 th, 7 th and 9 th days to a new culture medium without OP50, allowing the nematodes to freely move for 2min, and counting the number of beats of the nematode swallowing pump within 15 s.
2. Results: the pharyngeal pumping tissue of caenorhabditis elegans resembles the human heart. Pharyngeal pumping represents a feeding behavior whose frequency is related to the extent of nerve damage. The rate of pump beat by nematodes decreases with age. The frequency of swallowing pump beating by the colza pollen base group a nematodes was significantly different from that of the control group on days 7, 9 (P < 0.0001), as shown in fig. 8. Proved by the experiment, the rape pollen alkaloid A can delay the decline of the frequency of a swallowing pump along with the age, and has the protection effect on the nervous system.
EXAMPLE 6 caenorhabditis elegans exercise ability test
1. The experimental method comprises the following steps: the contemporaneous L4 nematodes were transferred to NGM medium coated with inactivated OP50, the inactivated OP50 containing 0, 0.8mM canola pollen base a. And (3) picking up at least 10 nematodes on a new culture medium without OP50 on days 5, 7 and 9 respectively, allowing the nematodes to freely move for 2min, and counting the swinging times of the body of the nematode within 20 s.
2. Results: as shown in fig. 9, the number of body oscillations of the nematodes tended to decrease as the life of the nematodes increased. The administration of rape pollen alkaloid A can significantly increase the number of body swings of nematodes (P < 0.05). The rape pollen alkali A can obviously improve the movement ability of nematodes, improve neurodegenerative changes and improve the survival state of the nematodes.
EXAMPLE 7 caenorhabditis elegans oxidative stress experiment
1. The experimental method comprises the following steps: the experiments were divided into a blank group, a rape pollen base A group (0.8 mM), a solvent control group (1% DMSO), a positive control group (1 mM resveratrol), and 90 nematodes per group. The contemporaneous L4 nematodes were picked up onto NGM medium coated with inactivated OP50, the OP50 containing the compounds at the concentrations indicated above. After 3 days of incubation at 20℃the nematodes were transferred to a culture containing 10mM H 2 O 2 On NGM medium and at 20deg.C, counting the number of nematodes dead and surviving every 1h until all nematodes die.
2. Results: oxidative stress is mainly caused by the imbalance of redox balance in the body, and excessive active oxygen is generated, so that the body is damaged. Oxidative stress can cause direct damage to the central nervous system. Neurons have a large oxygen demand but relatively low levels of antioxidant, and therefore 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 active oxygen is significantly increased, it destroys 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 is also one of the molecular mechanisms of Alzheimer ' S disease [ Kemppanen 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 fig. 10, the average lifetime was extended by 30% at 0.8 mpa concentration compared to the blank group. The life prolonging effect is remarkable (P < 0.0001). This indicates that cole pollen base A significantly improves the anti-stress ability of nematodes, can protect nematodes from oxidative damage, and is related to up-regulating the enzyme CYP35A involved in oxidation reaction.
In conclusion, the potential effects of the rape pollen alkali A on neuroprotection and brain disease prevention and treatment are found by combining the effects of the rape pollen alkali A on the health life of caenorhabditis elegans and the effects of gene transcription and protein expression.
The examples described above represent only embodiments of the invention and are not to be understood as limiting the scope of the patent of the invention, it being pointed out that several variants and modifications may be made by those skilled in the art without departing from the concept of the invention, which fall within the scope of protection of the invention.

Claims (4)

1. The novel application of the rape pollen alkali A in the aspects of neuroprotection and brain disease prevention and treatment is characterized in that the application of the rape pollen alkali A in the preparation of medicaments or pharmaceutical compositions for neuroprotection, prevention and/or treatment of brain diseases.
2. The novel use of brassinosteroids a according to claim 1 for neuroprotection and brain disease control, wherein said medicament or pharmaceutical composition comprises a pharmaceutically acceptable carrier.
3. The novel use of brassinosteroids a according to claim 1 for neuroprotection and brain disease control, wherein said brain disease is a brain disease caused by neurodegenerative changes or inflammatory substances.
4. The novel use of brassinosteroids a according to claim 3 for neuroprotection and brain disease control, wherein said brain disease is a brain disease caused by leukotriene B4-like inflammatory substances.
CN202310318285.2A 2023-03-29 2023-03-29 New use of rape pollen alkaloid A in neuroprotection and brain disease prevention and treatment Pending CN116509870A (en)

Priority Applications (1)

Application Number Priority Date Filing Date Title
CN202310318285.2A CN116509870A (en) 2023-03-29 2023-03-29 New use of rape pollen alkaloid A in neuroprotection and brain disease prevention and treatment

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
CN202310318285.2A CN116509870A (en) 2023-03-29 2023-03-29 New use of rape pollen alkaloid A in neuroprotection and brain disease prevention and treatment

Publications (1)

Publication Number Publication Date
CN116509870A true CN116509870A (en) 2023-08-01

Family

ID=87396646

Family Applications (1)

Application Number Title Priority Date Filing Date
CN202310318285.2A Pending CN116509870A (en) 2023-03-29 2023-03-29 New use of rape pollen alkaloid A in neuroprotection and brain disease prevention and treatment

Country Status (1)

Country Link
CN (1) CN116509870A (en)

Citations (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5856331A (en) * 1993-11-12 1999-01-05 Cell Therapeutics, Inc. Method for preventing tissue injury from hypoxia
US20030177507A1 (en) * 2000-03-22 2003-09-18 Marius Hoener Nematodes as model organisms for the investigation of neurodegenerative diseases, in particular parkinsons disease, uses and methods for the discovery of substances and genes which can used in the treatment of the above disease states and identification of anematode gene
CN102241719A (en) * 2011-04-22 2011-11-16 大连理工大学 Glycopyrrolate compounds and synthesis method thereof
CN104628803A (en) * 2015-02-06 2015-05-20 大连理工大学 Total synthesis method for rape pollen alkali A and caper alkali B and analogues thereof
US20170239235A1 (en) * 2014-05-09 2017-08-24 The Regents Of The University Of California Small-molecule ut-a-selective urea transport inihibitors
KR101887631B1 (en) * 2017-06-01 2018-08-13 우석대학교 산학협력단 Composition for delaying senescence, lifespan extension, or preventing, treating or improving of diseases caused by lipofuscin accumulation comprising sauchinone as an active ingredient of Saururus chinensis

Patent Citations (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5856331A (en) * 1993-11-12 1999-01-05 Cell Therapeutics, Inc. Method for preventing tissue injury from hypoxia
US20030177507A1 (en) * 2000-03-22 2003-09-18 Marius Hoener Nematodes as model organisms for the investigation of neurodegenerative diseases, in particular parkinsons disease, uses and methods for the discovery of substances and genes which can used in the treatment of the above disease states and identification of anematode gene
CN102241719A (en) * 2011-04-22 2011-11-16 大连理工大学 Glycopyrrolate compounds and synthesis method thereof
US20170239235A1 (en) * 2014-05-09 2017-08-24 The Regents Of The University Of California Small-molecule ut-a-selective urea transport inihibitors
CN104628803A (en) * 2015-02-06 2015-05-20 大连理工大学 Total synthesis method for rape pollen alkali A and caper alkali B and analogues thereof
KR101887631B1 (en) * 2017-06-01 2018-08-13 우석대학교 산학협력단 Composition for delaying senescence, lifespan extension, or preventing, treating or improving of diseases caused by lipofuscin accumulation comprising sauchinone as an active ingredient of Saururus chinensis

Non-Patent Citations (3)

* Cited by examiner, † Cited by third party
Title
曹志: "吡咯并噁嗪类生物碱的合成研究", 中国博士学位论文全文数据库 工程科技Ⅰ辑, no. 06, 15 June 2017 (2017-06-15), pages 014 - 46 *
蔡志友: "阿尔茨海默病血管基础", vol. 1, 31 January 2021, 科学技术文献出版社, pages: 271 *
陈盛禄: "中国蜜蜂学", vol. 1, 31 July 2001, 中国农业出版社, pages: 743 *

Similar Documents

Publication Publication Date Title
KR101740893B1 (en) COMPOSITION COMPRISING EXTRACELLULAR VESICLES DERIVED FROM Akkermansia muciniphila AS AN ACTIVE INGREDIENT FOR TREATING OR PREVENTING METABOLIC DISEASE
US20180055896A1 (en) Compositions and methods for increasing lifespan and health span
Harun-Or-Rashid et al. Structural and functional rescue of chronic metabolically stressed optic nerves through respiration
CN109843279A (en) Pharmaceutical composition comprising SSAO/VAP-1 inhibitor and SGLT2 inhibitor, its purposes
KR102257130B1 (en) Composition for Prevention or Treatment of Neurological or Mental Disorders Comprising Extracellular Vesicles Derived from Lactobacillus paracasei
Rao et al. Brugia malayi: effects of antibacterial agents on larval viability and development in vitro
Rushita et al. Effect of Pleurotus citrinopileatus on blood glucose, insulin and catalase of streptozotocin-induced type 2 diabetes mellitus rats.
CN105517546A (en) Methods of treating fragile X syndrome and related disorders
Liu et al. Apelin-12 exerts neuroprotective effect against ischemia-reperfusion injury by inhibiting JNK and P38MAPK signaling pathway in mouse.
CN112088837A (en) Construction method of non-alcoholic fatty liver mouse model
Chen et al. LncRNA SNHG1 inhibits neuronal apoptosis in cerebral infarction rats through PI3K/Akt signaling pathway.
Cao et al. Susceptibility and immune responses of hybrid snakehead (Channa maculata♀× Channa argus♂) following infection with snakehead fish vesiculovirus
CN111135177A (en) Application of iron death inhibitor Liproxstatin-1 in preparation of medicine for treating brain trauma
Zhang et al. Autophagy plays a protective role in sodium hydrosulfide-induced acute lung injury by attenuating oxidative stress and inflammation in rats
Xu et al. Secretory autophagosomes from alveolar macrophages exacerbate acute respiratory distress syndrome by releasing IL-1β
CN108379302B (en) Application of double-split sea sedge ethyl acetate extract in preparation of anti-inflammatory and analgesic drugs
Zheng et al. Neutrophil elastase inhibitor suppresses oxidative stress in obese asthmatic rats by activating Keap1/Nrf2 signaling pathway.
Zhang et al. Prenatal exposure to di (2-ethylhexyl) phthalate causes autism-like behavior through inducing Nischarin expression in the mouse offspring
Catalani et al. Neuroprotective role of plumbagin on eye damage induced by high-sucrose diet in adult fruit fly Drosophila melanogaster
CN116509870A (en) New use of rape pollen alkaloid A in neuroprotection and brain disease prevention and treatment
He et al. Cordyceps proteins alleviate lupus nephritis through modulation of the STAT3/mTOR/NF-кB signaling pathway
CN114159447A (en) Application of 18 beta-glycyrrhetinic acid in preparation of medicine for treating depression-related neuron protection
CN104582730B (en) A kind of method improving Metabolism of Mitochondria function and application
Pereira et al. White bean flour (Phaseolus vulgaris): Therapeutic and toxicological research in Wistar rats
CN102258527A (en) Application of stimulating agent CRX-675 of Toll-like receiver in resisting pulmonary fibrosis

Legal Events

Date Code Title Description
PB01 Publication
PB01 Publication
SE01 Entry into force of request for substantive examination
SE01 Entry into force of request for substantive examination