CN116920014A - Application of polyphenol extract of Chinese toon leaves - Google Patents
Application of polyphenol extract of Chinese toon leaves Download PDFInfo
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- CN116920014A CN116920014A CN202310959500.7A CN202310959500A CN116920014A CN 116920014 A CN116920014 A CN 116920014A CN 202310959500 A CN202310959500 A CN 202310959500A CN 116920014 A CN116920014 A CN 116920014A
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- 241000425037 Toona sinensis Species 0.000 title claims abstract description 73
- 239000000284 extract Substances 0.000 title claims abstract description 68
- 150000008442 polyphenolic compounds Chemical class 0.000 title claims abstract description 65
- 235000013824 polyphenols Nutrition 0.000 title claims abstract description 65
- 235000011783 Cedrela sinensis Nutrition 0.000 claims abstract description 56
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims abstract description 32
- 208000024827 Alzheimer disease Diseases 0.000 claims abstract description 30
- 239000003814 drug Substances 0.000 claims abstract description 13
- 238000000605 extraction Methods 0.000 claims abstract description 6
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- 241000244206 Nematoda Species 0.000 abstract description 57
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- 239000000243 solution Substances 0.000 description 12
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- MGJZITXUQXWAKY-UHFFFAOYSA-N diphenyl-(2,4,6-trinitrophenyl)iminoazanium Chemical compound [O-][N+](=O)C1=CC([N+](=O)[O-])=CC([N+]([O-])=O)=C1N=[N+](C=1C=CC=CC=1)C1=CC=CC=C1 MGJZITXUQXWAKY-UHFFFAOYSA-N 0.000 description 2
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- HQFLTUZKIRYQSP-UHFFFAOYSA-N 3-ethyl-2h-1,3-benzothiazole-6-sulfonic acid Chemical compound OS(=O)(=O)C1=CC=C2N(CC)CSC2=C1 HQFLTUZKIRYQSP-UHFFFAOYSA-N 0.000 description 1
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- ZTOJFFHGPLIVKC-CLFAGFIQSA-N abts Chemical compound S/1C2=CC(S(O)(=O)=O)=CC=C2N(CC)C\1=N\N=C1/SC2=CC(S(O)(=O)=O)=CC=C2N1CC ZTOJFFHGPLIVKC-CLFAGFIQSA-N 0.000 description 1
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- QKUSRAKPUWQSJS-UHFFFAOYSA-N diazanium 3-ethyl-2H-1,3-benzothiazole-6-sulfonate Chemical compound [NH4+].[NH4+].[O-]S(=O)(=O)C1=CC=C2N(CC)CSC2=C1.[O-]S(=O)(=O)C1=CC=C2N(CC)CSC2=C1 QKUSRAKPUWQSJS-UHFFFAOYSA-N 0.000 description 1
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- BASFCYQUMIYNBI-UHFFFAOYSA-N platinum Chemical compound [Pt] BASFCYQUMIYNBI-UHFFFAOYSA-N 0.000 description 1
- USHAGKDGDHPEEY-UHFFFAOYSA-L potassium persulfate Chemical compound [K+].[K+].[O-]S(=O)(=O)OOS([O-])(=O)=O USHAGKDGDHPEEY-UHFFFAOYSA-L 0.000 description 1
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Classifications
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/18—Magnoliophyta (angiosperms)
- A61K36/185—Magnoliopsida (dicotyledons)
- A61K36/58—Meliaceae (Chinaberry or Mahogany family), e.g. Azadirachta (neem)
-
- 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
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
- A61K2236/30—Extraction of the material
- A61K2236/33—Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones
- A61K2236/333—Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones using mixed solvents, e.g. 70% EtOH
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
- A61K2236/50—Methods involving additional extraction steps
- A61K2236/51—Concentration or drying of the extract, e.g. Lyophilisation, freeze-drying or spray-drying
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
- A61K2236/50—Methods involving additional extraction steps
- A61K2236/53—Liquid-solid separation, e.g. centrifugation, sedimentation or crystallization
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- Health & Medical Sciences (AREA)
- Natural Medicines & Medicinal Plants (AREA)
- Engineering & Computer Science (AREA)
- Life Sciences & Earth Sciences (AREA)
- Neurology (AREA)
- General Health & Medical Sciences (AREA)
- Neurosurgery (AREA)
- Chemical & Material Sciences (AREA)
- Biomedical Technology (AREA)
- Medicinal Chemistry (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Veterinary Medicine (AREA)
- Pharmacology & Pharmacy (AREA)
- Public Health (AREA)
- Animal Behavior & Ethology (AREA)
- Mycology (AREA)
- Biotechnology (AREA)
- Alternative & Traditional Medicine (AREA)
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- Medical Informatics (AREA)
- Hospice & Palliative Care (AREA)
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Abstract
The invention provides an application of a toona sinensis leaf polyphenol extract in preparing a medicine for treating Alzheimer's disease. The folium Toonnae sinensis is derived from Cedrela sinensis. The polyphenol extract of the Chinese toon leaves is ethanol extract of the Chinese toon leaves, and the extraction steps are as follows: weighing crushed Chinese toon leaves according to a feed liquid ratio of 1:30 to 1: adding 70% ethanol into 20, and ultrasonic extracting for 45-60min; filtering to obtain filtrate, centrifuging, collecting supernatant, purifying with column, collecting eluate, volatilizing ethanol, and lyophilizing to obtain folium Toonnae sinensis polyphenol extract. The cedrela sinensis polyphenol extract has remarkable treatment effect on AD nematodes, is shown to have remarkable inhibition effect on paralysis phenotype caused by AD caenorhabditis elegans intramuscular Abeta over-expression, and can remarkably delay the paralysis phenotype of the Alzheimer disease caenorhabditis elegans.
Description
Technical Field
The invention relates to the field of traditional Chinese medicine, in particular to application of a polyphenol extract of Chinese toon leaves.
Background
Alzheimer's Disease (AD) is a progressive degenerative disease of the nervous system with hidden disease progression. There is currently no effective method for predicting, preventing or reversing AD, and therefore the development of new, effective therapeutic agents has become a current worldwide research hotspot. Insoluble aβ peptide and aggregated aβ plaques are the major factors in the formation of AD, and thus the expression of aβ peptide is considered to be an important parameter in studying pathology of the caenorhabditis elegans AD model. AD has become a serious social problem as the most interesting aging-related disease, and if the aim of delaying AD can be achieved by adjusting daily diet, it is of great importance to the health of the aging population.
The Chinese toon is a plant of the genus Cedrela (Toona) of the family Meliaceae (Meliaceae) of the phylum Celastraceae (Angiospermae), belongs to perennial deciduous tree, has higher medicinal value, is a famous medicinal and edible homologous biological plant, and is used as a medicinal material for Chinese medicine. The separation and extraction of active ingredients in natural medicines has great potential in the treatment of AD.
Disclosure of Invention
After researching the induced toxicity of the caenorhabditis elegans AD pathological model by adopting the cedrela sinensis polyphenol extract, the invention discovers that the substance has in-vitro and in-vivo antioxidant activity and has obvious AD resisting effect. Therefore, the invention discloses application of the toona sinensis leaf polyphenol extract in preparing a medicament for treating Alzheimer's disease, and provides a new idea for treating Alzheimer's disease by natural products.
The invention provides an application of a toona sinensis leaf polyphenol extract in preparing a medicine for treating Alzheimer's disease.
In some embodiments, the toona sinensis leaves are from toona sinensis.
In some embodiments, the cedrela sinensis leaf polyphenol extract is an ethanol extract of cedrela sinensis leaves, the extraction steps are as follows: weighing crushed Chinese toon leaves according to a feed liquid ratio of 1:30 to 1: adding 70% ethanol into 20, and ultrasonic extracting for 45-60min; filtering to obtain filtrate, centrifuging, collecting supernatant, purifying with column, collecting eluate, volatilizing ethanol, and lyophilizing to obtain folium Toonnae sinensis polyphenol extract.
Experiments are carried out by taking caenorhabditis elegans as an Alzheimer disease pathological model, and the results show that: the toona sinensis leaf polyphenol extract has remarkable therapeutic effect on AD nematodes. The composition has remarkable inhibition effect on the paralytic phenotype caused by the over-expression of the Abeta in the muscle of the AD caenorhabditis elegans, and can remarkably delay the paralytic phenotype of the Alzheimer disease caenorhabditis elegans. In addition, the toona sinensis leaf polyphenol extract has in vitro and in vivo antioxidant activity, can improve the anti-stress capability of nematodes, and can relieve the toxicity induced by CL4176 nematode Abeta.
The invention discovers that the toona sinensis leaf polyphenol extract has obvious effect on the aspect of treating Alzheimer's disease and can be used for preparing medicines for treating Alzheimer's disease.
Drawings
FIG. 1A shows the scavenging effect of Vitamin C (VC) on 1, 1-diphenyl-2-trinitrophenylhydrazine (DPPH), wherein the abscissa shows the VC concentration and the ordinate shows the scavenging ability on DPPH.
FIG. 1B shows the DPPH radical scavenging effect of Toonae sinensis polyphenol extract.
FIG. 2A is a graph of VC scavenging on 2,2' -diazabis (3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt (ABTS) free radical, wherein the abscissa indicates VC concentration and the ordinate indicates ability to scavenge ABTS.
FIG. 2B shows the effect of Toona sinensis leaf polyphenol extract on ABTS free radical scavenging.
FIG. 3 shows the effect of Toona sinensis leaf polyphenol extract on the body bending frequency of caenorhabditis elegans, wherein the abscissa is the number of days, the ordinate is the body bending frequency of caenorhabditis elegans, the units are times/30 s, the concentration of the extract is shown in FIG. 3 for the columns at 3d, and the order of the concentrations is consistent at 3d for the columns at 6d and 9 d.
FIG. 4 is an effect of Toona sinensis leaf polyphenol extract on the paralysis time of CL4176 nematodes at 25℃with the abscissa being in hours and the ordinate being in percent unparalysis.
FIG. 5 shows the effect of Toona sinensis leaf polyphenol extract on exogenous 5-HT hypersensitivity to CL2355 nematodes, wherein the ordinate is the proportion of paralytic worms.
FIG. 6A is a graph showing the effect of Toona sinensis leaf polyphenol extract (TLPE) on survival time of CL4176 nematodes under heat stress, wherein the ordinate indicates nematode survival.
Fig. 6B is the effect of cedrela sinensis leaf polyphenol extract on survival time under oxidative stress of CL4176 nematodes.
FIGS. 7A and 7B are graphs showing the effect of Toona sinensis leaf polyphenol extract on Abeta deposition in CL4176 nematodes (FIG. 7A: representative images of ThS staining, arrow indicates Abeta fiber aggregation fluorescence; FIG. 7B: quantitative analysis of nematode fluorescence intensity, ordinate is fluorescence intensity).
FIGS. 8A and 8B are effects of Toona sinensis leaf polyphenol extract on the intracellular ROS accumulation in CL4176 nematodes (FIG. 8A: DCF fluorescence image of Cl4176 nematodes; FIG. 8B: quantitative analysis of nematode fluorescence intensity, ordinate: ROS level).
Detailed Description
The following examples will enable those skilled in the art to more fully understand the present invention and are not intended to limit the same in any way.
Drying and crushing Chinese toon leaves, and extracting by an alcohol extraction method to obtain an extract, wherein the specific extraction method is as follows: weighing crushed and sieved toona sinensis samples according to a feed liquid ratio of 1:30 to 1: adding 70% ethanol into 20, and ultrasonic extracting for 45-60min; mixing the filtrates to obtain crude extract, centrifuging, collecting supernatant, and purifying with column. And passing the centrifuged solution through an AB-8 ion exchange resin column. After the adsorption is completed, the column is washed by ultra-pure water until the effluent liquid is colorless, and then 70% ethanol is used for eluting until the effluent liquid is colorless. Collecting the eluent, volatilizing ethanol, and freeze-drying to obtain the total polyphenol extract of the Chinese toon leaves.
Example 1: the polyphenol extract of the Chinese toon leaves has stronger in-vitro antioxidation capability
The method comprises the following steps: the clearance of the polyphenol extract from the leaves of Toona sinensis on DPPH and ABTS free radicals was determined and calculated according to the following method, VC was used as a positive control, and the same treatment was performed three times per well.
(1) Scavenging of DPPH free radical
2mg of 2, 2-biphenyl-1-picrylhydrazyl (DPPH) is weighed, dissolved in absolute ethyl alcohol and the volume is fixed to 100mL, so as to obtain 0.02mg/mL of DPPH solution. And diluting the sample to be measured to a sample solution of 0-10 mug/mL. Different samples were taken and added to the test tubes, each with the following amounts:
A sample : sample 1mL+DPPH alcohol solution 4mL;
A blank : sample 1 ml+4 mL of absolute ethanol solution;
A control : 4mL of 50% absolute ethanol 1mL+DPPH alcohol solution;
VC is a positive control, the same treatment is carried out, after the mixture is uniformly mixed and placed in a dark place for 30min, absorbance at 517nm is measured, average value is taken, and the clearance rate of DPPH free radical of the sample is calculated according to the following formula:
(2) Scavenging of ABTS free radical
Mixing 7 mmol/L2, 2' -hydrazine-bis (3-ethylbenzothiazoline-6 sulfonic acid) diamine salt (ABTS) and 140mmol/L potassium persulfate in equal volume, reacting for 12 hours at room temperature in a dark place, diluting with absolute ethyl alcohol, and measuring the absorbance of the mixture to be 0.700+/-0.005 at 734nm wavelength to be used as an ABTS working solution. Diluting a sample to be measured to a sample solution of 0-10 mug/mL, taking test tubes, and adding different samples, wherein the addition amount of each tube is as follows:
A sample : sample 1mL+ABTS solution 4mL;
A blank : sample 1 ml+4 mL of absolute ethanol solution;
A control : absolute ethanol 1ml+abts solution 4mL;
VC is a positive control, the same treatment is carried out, after the mixture is uniformly mixed and placed in a dark place for 6min, absorbance at 517nm is measured, average value is taken, and the clearance rate of ABTS free radical of the sample is calculated according to the following formula:
results: FIG. 1B shows the DPPH radical scavenging ability of Toonae sinensis leaf polyphenol extract and its IC 50 6.716 + -0.012 μg/mL, FIG. 2B shows the ability of Toonae sinensis leaf polyphenol extract to scavenge ABTS free radicalsIC of extract to ABTS 50 44.777 + -0.206 μg/mL, shows that the Cedrela sinensis leaf extract can achieve stronger DPPH and ABTS free radical scavenging effect at low concentration, and has good in vitro antioxidant capacity.
Example 2: cedrela sinensis leaf polyphenol extract delays the decrease of CL4176 nematode body bending frequency
The method comprises the following steps: synchronizing CL4176 caenorhabditis elegans by sieving with common sieve, sterilizing at 121deg.C for 20min, adding 1mL CaCl after sterilization 2 、1mL MgSO 4 Shaking 1mL of cholesterol solution, pouring the solution into a plate, culturing at 37 ℃ until a layer of lawn grows on the surface), transferring the solution to a new culture medium with corresponding concentration every 2d, adding a proper amount (about 2 mL) of M9 buffer solution when the insect grows to 3d, 6d and 9d, measuring the number of times of sinusoidal movement of the insect in 30s under a microscope, and determining the change of the movement capacity of the nematodes. The experiment was independently repeated 3 times, and the number of samples in each group was not less than 30.
Results: FIG. 3 is a bar graph of body flexural frequencies measured for caenorhabditis elegans on days 3, 6 and 9, respectively. Over time, the body bending of caenorhabditis elegans gradually decreases, and the decrease of the body bending frequency of caenorhabditis elegans can be delayed after the treatment of the polyphenol extract of the cedrela sinensis leaves. 0.2-1.0 mg/mL Toona sinensis leaf polyphenol extract enhances the body bending frequency of nematodes and has significant difference (p < 0.05) when treated for 6d and 9 d. Overall, the Toona sinensis leaf polyphenol extract can delay the decrease of the body bending frequency of CL4176 nematodes, and the more obvious the effect is with the increase of days.
Example 3: experiments on the delay of the paralysis time of CL4176 nematodes by the Toona sinensis polyphenol extract
The method comprises the following steps: by utilizing the characteristic that nematodes have different body types in different periods, the CL4176 caenorhabditis elegans are synchronized by sieving through a sieve, and cultured on NGM plates containing a control (0.2% DMSO) and a cedrela sinensis leaf polyphenol extract (0.1, 0.2, 0.4, 0.6, 0.8 and 1.0 mg/mL) for 48 hours, then transferred to a constant temperature incubator at 25 ℃ for 36 hours, and the nematodes are induced to express A beta protein in a large amount, and the A beta aggregation can lead to CL4176 paralysis. Paralytic nematodes were observed, one time every 1 h. When lightly touched with a platinum wire, the nematodes were not able to move or were unable to make a complete sinusoidal movement and were then considered paralyzed. The experiment was independently repeated 3 times, and the number of samples in each group was not less than 30.
Results: the paralysis time of the concentration of the polyphenol extract from leaves of Toona sinensis screened against AD using caenorhabditis elegans is shown in FIG. 4. Compared with a control group, the Chinese toon leaf polyphenol extract can prolong the paralysis time of caenorhabditis elegans within the range of 0.1-1 mg/mL, and the Chinese toon leaf polyphenol extract with the concentration of 0.1mg/mL and 0.2mg/mL can obviously delay the paralysis of CL4176 nematodes, and compared with the control group, the average paralysis time is respectively delayed by 23.38% and 18.63%. Therefore, the toona sinensis leaf polyphenol extract has the activity of resisting AD, prolongs the paralysis time of CL4176 nematodes to a certain extent, and has no positive correlation with the concentration.
Example 4: experiments on the alleviation of the sensitivity of CL2355 nematodes to 5-HT by Toona sinensis polyphenol extracts
The method comprises the following steps: by utilizing the characteristic that nematodes have different body types in different periods, CL2355 caenorhabditis elegans are synchronized by sieving with a sieve, cultured for 48h on NGM plates containing control (0.2% DMSO) and cedrela sinensis leaf polyphenol extract (0.1, 0.2, 0.4, 0.6, 0.8 and 1.0 mg/mL), and then transferred to a constant temperature incubator at 25 ℃ for 36h. Insects were picked up and treated for 3min in 96-well plates containing 5-hydroxytryptamine (5-HT) solution and the percentage of paralytic nematodes per well was counted. The experiment was independently repeated 3 times, and the number of samples in each group was not less than 30.
Results: as shown in FIG. 5, 0.2mg/mL and 0.8mg/mL also reduced the paralysis of CL2355 to some extent by 36.7% and 31.71% (p < 0.01, p < 0.05), respectively, in the Toona sinensis leaf polyphenol extract treated group. The result shows that the toona sinensis leaf polyphenol extract can inhibit toxicity generated by the over-expression of Abeta in the nerve cells of the AD nematodes, lighten the hypersensitivity of CL2355 nematodes to exogenous 5-HT and has certain AD resisting activity.
Example 5: cedrela sinensis leaf polyphenol extract improves CL4176 nematode survival time under stress
The method comprises the following steps: heat stress: CL4176 nematodes were synchronized and cultured on NGM plates containing control (0.2% dmso) and cedrela sinensis leaf polyphenol extract (0.2 mg/mL), incubated at 16 ℃ for 36h and then transferred to 25 ℃ for 2h, and the number of nematode survivors was observed and counted for each group until all nematodes died. The experiment was independently repeated 3 times, and the number of samples in each group was not less than 30.
Oxidative stress: synchronizing CL4176 nematodes, culturing on NGM plate containing control (0.2% DMSO) and Toona sinensis leaf polyphenol extract (0.2 mg/mL), culturing at 16deg.C for 36H, washing with M9 buffer, sucking nematodes into 96-well plate, and adding M9 and H 2 O 2 Make H 2 O 2 The final concentration was 40mmol/L. The number of nematode survival per group was observed and counted every 2 hours until all nematodes died. The experiment was independently repeated 3 times, and the number of samples in each group was not less than 30.
Results: as shown in fig. 6A and 6B, the toona sinensis leaf polyphenol extract treatment group improved the survival rate of nematodes under heat stress, and CL4176 nematode mean life was prolonged by 17.65% (p < 0.01) compared with the control group, with a significant difference; meanwhile, the survival rate of nematodes under oxidative stress is increased, the average life is prolonged by 22.55% (p is less than 0.01) compared with a control group, and the nematode survival rate is remarkably different.
Example 6: toona sinensis leaf polyphenol extract for reducing Abeta deposition in CL4176 nematodes
The method comprises the following steps: CL4176 nematodes were synchronized and incubated on NGM plates containing control (0.2% DMSO) and Toona sinensis leaf polyphenol extract (0.2 mg/mL), incubated at 16℃for 36h and then transferred to 25℃for A.beta.induction for 48h. Fixation was performed using paraformaldehyde and the pellet was stained with thioflavin S (ThS) after incubation to reflect aβ aggregation fibrosis in nematodes. The number of samples in each group is not less than 30.
Results: as shown in FIGS. 7A and 7B, the decrease of Abeta fiber aggregation in nematodes after treatment with 0.2mg/mL Toona sinensis leaf polyphenol extract was observed after fluorescence aggregation by fluorescent spots. Compared with the control group, the fluorescence intensity of the treated cedrela sinensis leaf polyphenol extract with the concentration of 0.2mg/mL is reduced by 10.73 percent (p is less than 0.0001) by the quantitative analysis of the fluorescence intensity, and the difference is extremely remarkable. Therefore, the toona sinensis leaf polyphenol extract can reduce the deposition of Abeta in the nematode body so as to delay the paralysis effect of the nematode.
Example 7: cedrela sinensis leaf polyphenol extract reduces ROS accumulation in CL4176 nematodes
The method comprises the following steps: synchronizing CL4176 nematodes, culturing on NGM plate containing control (0.2% DMSO) and Toona sinensis leaf polyphenol extract (0.2 mg/mL), culturing at 16deg.C for 36H, transferring to 25deg.C, washing with M9 buffer, sucking nematodes into 96-well plate, and adding M9 and 2',7' -dichlorofluorescein diacetate (H) 2 DCF-DA), H 2 The nematodes were transferred to agar slides after incubation at 37℃at a final DCF-DA concentration of 50. Mu. Mol/L, observed with a fluorescence microscope and photographed. The number of samples in each group is not less than 30.
Results: fig. 8A and 8B are fluorescent plots of Reactive Oxygen Species (ROS) in the nematodes of each group after treatment with CL4176 with different drugs, the in vivo fluorescence intensity of the nematodes in the DMSO group is significantly stronger than that of the toona sinensis leaf polyphenol extract treatment group. Therefore, the toona sinensis leaf polyphenol extract treatment group reduced ROS levels in nematodes. Fluorescent quantification of ROS level shows that the fluorescence intensity of the treatment of the polyphenol extract of the Chinese toon leaves is reduced by 22.89% (p < 0.0001), and the fluorescence intensity is extremely remarkable; the toona sinensis leaf polyphenol extract can effectively relieve ROS accumulated in cells after oxidative stress.
The embodiment proves that the cedrela sinensis polyphenol extract provided by the invention plays a remarkable therapeutic role on the AD caenorhabditis elegans, not only improves the anti-stress capability of the nematodes and enhances the anti-oxidation defense system in the nematodes, but also relates to the inherent anti-oxidation capability of the cedrela sinensis polyphenol extract. The toona sinensis leaf polyphenol extract provided by the invention has the potential of treating AD, and can be applied to preparing medicines for preventing and treating Alzheimer's disease. In addition, the toona sinensis leaf polyphenol is nontoxic and has good tolerance as a medicine and food homologous food, and is expected to be a potential medicine for treating diseases related to Abeta abnormal aggregation as main pathological characteristics, such as AD, in daily diet.
It should be understood by those skilled in the art that the above embodiments are exemplary embodiments only and that various changes, substitutions and alterations can be made hereto without departing from the spirit and scope of the invention.
Claims (3)
1. An application of folium Toonnae sinensis polyphenol extract in preparing medicine for treating Alzheimer disease is provided.
2. The use according to claim 1, wherein the cedrela sinensis leaves are from cedrela sinensis.
3. The use according to claim 1, wherein the polyphenol extract of chinese toon leaves is an ethanol extract of chinese toon leaves, the extraction steps being as follows:
weighing crushed Chinese toon leaves according to a feed liquid ratio of 1:30 to 1: adding 70% ethanol into 20, and ultrasonic extracting for 45-60min; filtering to obtain filtrate, centrifuging, collecting supernatant, purifying with column, collecting eluate, volatilizing ethanol, and lyophilizing to obtain folium Toonnae sinensis polyphenol extract.
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