CN105079098A - Application of dracocephalum moldavica total flavonoids to preparation of drug for preventing and treating AD (Alzheimer's disease) - Google Patents
Application of dracocephalum moldavica total flavonoids to preparation of drug for preventing and treating AD (Alzheimer's disease) Download PDFInfo
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Abstract
The invention discloses an application of dracocephalum moldavica total flavonoids to preparation of a drug for preventing and treating the AD (Alzheimer's disease). An AD mouse model is established through hypodermic injection of sodium nitrite/D-galactose, the learning and memory capacity and biochemical indexes are scientifically assessed through mouse positioned navigation tests and space exploration tests, it proves that the dracocephalum moldavica total flavonoids can improve the space learning and memory capacity of AD model mice, dose dependence is formed, escape incubation of the AD mice is shortened, and platform crossing times in the space exploration tests are significantly increased; meanwhile, the content of MDA in cortical tissue in the AD mice can be significantly decreased, the content of SOD can be significantly increased, and the oxidative stress level in brain tissue can be reduced; also, through reduction of expression of pro-inflammatory factors IL-1beta and TNF-alpha, inflammatory injuries to the brain tissue can be relieved, the neuron protection role can be played, and the learning and memory capacity of the AD mice can be improved finally. Thus, a reference basis can be provided for clinic application of the dracocephalum moldavica total flavonoids in the future.
Description
Technical field
The present invention relates to a kind of application of active constituents of medicine Herba Dracocephali total flavones, be specifically related to the application of a kind of Herba Dracocephali total flavones in preparation control Alzheimer disease drugs.
Background technology
Alzheimer (alzheimer'sdisease, AD), also known as alzheimer disease, it is a kind of common neurodegenerative diseases, being the more multiple a kind of commonly encountered diseases of old people after heart disease, tumor and cerebrovascular disease, is also cause dull-witted most common cause in old people.Clinically with hypomnesis, cognitive dysfunction for feature, the state of an illness is that Progressive symmetric erythrokeratodermia increases the weight of, and loses the ability of living on one's own life in several years, Chang Yin accompanying infection in about 10 years and dead.Epidemiological study shows, and the sickness rate of over-65s AD is 5-10%, doubles every 5 annual morbidities later, and within more than 80 years old, AD sickness rate can be increased to more than 50%.Enter 21 century, along with the raising of medical level, the average life span greatly extends, and the problem of an aging population that the mankind face is increasingly sharpened.Current China aging population have exceeded 10% of total population, and indicate that China has become aged country, to the year two thousand twenty whole world, more than 60 years old old man will reach more than 10 hundred million, and huge aging population radix, makes old dementia patients obviously increase.According to " report of world's Alzheimer " display that 2013 announce, about there are 4,400 ten thousand AD patients in the current whole world, and China is about 800-1000 ten thousand.The research such as Delphi shows, to the year two thousand forty, global AD patient will reach 8,110 ten thousand.In western countries, dull-witted first of having occupied neuropathy, and 2/3 is diagnosed as AD, has a strong impact on the Health and Living quality of old people, and bring heavy psychology and financial burden to country, society and family.Meanwhile, as a great public health problem, the control of alzheimer disease has become the focal issue of current medical domain research.
Current, several hypothesis is existed to the pathogenesis understanding of AD, comprises Tau abnormal protein phosphorylation hypothesis, amyloid-beta cascade hypothesis, oxidative stress hypothesis, inflammation hypothesis, cholinergic neuron damage hypothesis etc.Research shows, oxidative stress is one of change the earliest in onset of Alzheimer disease mechanism, the oxidative damage of active oxygen can cause apoptosis, the interphase interaction of excessive active oxygen, lipid peroxidation and calcium overload, finally cause irreversible neuronal damage, cause old and feeble and ofneurodegenerative change.More and more study discovery in recent years, inflammatory reaction plays an important role in onset of Alzheimer disease mechanism.The display of research report, the activation of astrocyte and microglia is had near senile plaque caused by A β deposits, discharge a large amount of inflammatory cytokine, as nitric oxide (nitricoxide, NO), interleukin-1 (interleukin-1, IL-1), tumor necrosis factor-alpha (tumornecrosisfactor-α, TNF-α) etc., the medium-term and long-term chronic inflammatory reaction of cerebral tissue finally causes neuronic a large amount of loss, causes the generation of AD.
The dry aerial parts of Herba Dracocephali system labiate Herba Dracocephali (DrococephalummoldivacaL.), for annual herb plant, it is China's Chinese medicine, mainly be distributed in the provinces and regions such as North China, northeast, northwest, Xinjiang cultivation is more, Uygur medicine is referred to as the right Ji Buya of Badische, and the useful heart protects brain, open the function such as inaccessible in brain, is usually used in the treatment of heart disease, hypertension, tracheitis etc.Herba Dracocephali total flavones is active substance in Herba Dracocephali, and research shows, Herba Dracocephali total flavones has antioxidation, resists myocardial ischemia, blood fat reducing, protect the liver, antiinflammatory, antiviral, the pharmacological action such as antibacterial.So far not yet find that Herba Dracocephali total flavones is applied to the report in the ability of learning and memory disorder remedies of control Alzheimer initiation.
Summary of the invention
Technical problem to be solved by this invention is, provides the application of a kind of Herba Dracocephali total flavones in preparation control Alzheimer disease drugs.
The technical scheme that the present invention solves the employing of its technical problem is, Herba Dracocephali total flavones is preparing as active constituents of medicine the application prevented and/or treated in Alzheimer disease drugs.
Further, every day, effective dose was 25 ~ 100mg/kg with oral or non-oral routes mode administration, and above-mentioned dosage can a dosage unit or be divided into several dosage unit administration.Dosage of the present invention depends on many factors, as the sex of patient or animal, the age, body weight, suffer from the disease the order of severity, self Liver and kidney function state and route of administration etc.Therefore, dosage of the present invention can have the change of certain limit.
Further, described Herba Dracocephali total flavones exists with the solid-state form of tablet, capsule, pill, injection, slow release, controlled-release pharmaceutical formulation as sole active agent.
Further, described Herba Dracocephali total flavones exists with the solid-state form of tablet, capsule, pill, injection, slow release, controlled release compound medicinal formulation as wherein a kind of active component.
The present invention sets up dementia mice model by internationally recognized subcutaneous injection sodium nitrite/D-galactose, through the test of mice orientation navigation and space exploration test, ability of learning and memory, biochemical indicator carry out the assessment of science, demonstrate Herba Dracocephali total flavones and can improve Alzheimer disease model mice Spatial memory ability, and one-tenth dose dependent, shorten dementia mice escape latency, significantly increase the number of times of spanning platform in space exploration experiment; Significantly reduce MDA content in dementia mice cortical tissue simultaneously, and can significantly increase SOD content, reduce oxidative stress level in cerebral tissue; Also by lowering the expression of proinflammatory inflammation factor IL-1 β, TNF-α, alleviating the inflammation damnification of cerebral tissue, playing neuroprotective, finally improving the learning and remembering ability of dementia mice.Visible, the present invention is applied to clinically provides reference frame for treating Alzheimer from now on.
Detailed description of the invention
Below in conjunction with embodiment, the present invention is illustrated further.
Embodiment 1: Herba Dracocephali total flavones is on the impact of dementia mice learning and memory
1, experiment material
(1) trial drug: Herba Dracocephali total flavones (pharmaceutical college of Shihezi Univ makes by oneself, and purity is 82.3%).
(2) animal: Kunming mouse 50, male, body weight (23 ± 2) g, is provided by Shihezi Univ's Experimental Animal Center.
(3) reagent: sodium nitrite, D-galactose are commercially available analytical pure;
(4) instrument: Morris water maze (institute of Materia Medica,Chinese Academy of Medical Sciences development);
2, experimental technique
(1) dementia mice model is set up
Kunming mice 50 is divided into 5 groups at random, often organizing 10, had both been dosage group (50mg/kg), Herba Dracocephali total flavones high dose group (100mg/kg) in blank group, model group, Herba Dracocephali total flavones low dose group (25mg/kg), Herba Dracocephali total flavones.Except Normal group intraperitoneal injection of saline (equal-volume), all the other 4 groups of mouse subcutaneous injection sodium nitrite (90mg/kg) and D-galactose (120mg/kg), successive administration 60 days, sets up dementia mice model.The 30th day of modeling, except Normal group, model group gavage isodose normal saline every day, other respectively organizes the testing drug that equal gavage gives corresponding dosage.
(2) test of ability of learning and memory
Morris water maze laboratory: give testing drug and carry out Morris water maze laboratory after 30 days, continuous 5 days Morris water maze tests, are divided into orientation navigation to test and space exploration test.Within first 4 days, position sea trial, water maze is divided into four quadrants, and Southwest Quadrant center is located at by platform, and mice enters water from Northeast Quadrant and Northwest Quadrant every day, and record finds plateau time.Then 120 seconds are recorded as incubation period as do not found platform.Within 5th day, carry out space exploration test, remove platform, mice enters water from selected quadrant, record mice spanning platform position number of times.
3, statistical procedures
Experimental data with
represent, comparison in difference use two-way analysis of variance incubation period (TWO-wayANOVA) between each group in orientation navigation experiment.Compare between explorative experiment group and adopt one factor analysis of variance (one-wayANOVA), check in conjunction with Dunnett ' sposthoctest, more each group difference, p < 0.05 thinks to there is significant difference.
4, experimental result
Morris water maze laboratory result shows, compared with blank group, model group mice escape latency significant prolongation (p < 0.01), spanning platform number of times obviously reduces (p < 0.05), confirms that mice Model of Dementia copies successfully.Compared with model group, Herba Dracocephali total flavones various dose group all can shorten dementia mice in various degree and find the platform preclinical time, and wherein high dose group has significant difference (p < 0.05) compared with model group.In space exploration experiment, compared with model group mice, Herba Dracocephali total flavones high dose group obviously can increase dementia mice spanning platform number of times (p < 0.05) (see table 1-2).
Table 1 Herba Dracocephali total flavones on the impact of dementia mice escape latency (
n=10)
Compare with blank group:
##p<0.01; Compare with model group:
*p<0.05
Table 2 Herba Dracocephali total flavones on the impact of dementia mice spanning platform number of times (
n=10)
| Group | Dosage (mg/kg) | Spanning platform number of times (t) |
| Blank group | 4.88±0.59 | |
| Model group | 2.3±0.47 # | |
| Herba Dracocephali total flavones | 25 | 2.78±0.67 |
| Herba Dracocephali total flavones | 50 | 2.60±0.53 |
| Herba Dracocephali total flavones | 100 | 3.94±0.38 * |
Compare with blank group:
#p<0.05; Compare with model group:
*p<0.05
Embodiment 2: Herba Dracocephali total flavones is on the impact of dementia mice brain tissue oxidizing stress level
1, experiment material
(1) reagent: malonaldehyde (MDA), superoxide dismutase (SOD) and glutathion peroxidase (GSH-PX) test kit all build up Bioengineering Research Institute purchased from Nanjing;
(2) instrument: microplate reader (Thermo3001, Thermo company of the U.S.); TGL-16G freezing centrifuge (Anting Scientific Instrument Factory, Shanghai);
2, experimental technique
Water maze laboratory terminates rear execution animal, pluck full brain and clean bloodstain with ice-cold normal saline, isolate cerebral cortex at low ambient temperatures, the normal saline adding 9 times of volumes after weighing makes 10% homogenate, the centrifugal 10min of 3500r/min, gets supernatant and measures malonaldehyde, superoxide dismutase, glutathion peroxidase content in cerebral tissue according to test kit description.
3, statistical procedures
Experimental data with
represent, compare between group and adopt one factor analysis of variance (one-wayANOVA), check in conjunction with Dunnett ' sposthoctest, more each group difference, p < 0.05 thinks to there is significant difference.
4, experimental result
Result shows, and compare with blank group, in model group mouse cortex tissue, MDA content obviously raises (p<0.05), SOD and GSH-PX content significantly reduces (p<0.05).Compare with model group, Herba Dracocephali total flavones various dose group can reduce MDA content in mouse cortex tissue in various degree, and middle and high dosage group has significant difference (p<0.05) compared with model group.Can raise the content of SOD in dementia mice cortical tissue after the administration of Herba Dracocephali total flavones, wherein high dose group has significant difference (p<0.05) compared with model group; Compare with model group, Herba Dracocephali total flavones has no significant effect (see table 3-5) GSH-PXH content in dementia mice cortex.
Table 3 Herba Dracocephali total flavones on the impact of MDA content in mouse cortex tissue (
n=10)
| Group | Dosage (mg/kg) | MDA(nmol/mg) |
| Blank group | 5.36±0.73 | |
| Model group | 9.02±0.87 # | |
| Herba Dracocephali total flavones | 25 | 6.75±0.58 |
| Herba Dracocephali total flavones | 50 | 5.83±0.98 * |
| Herba Dracocephali total flavones | 100 | 5.48±0.73 * |
Note: compare with blank group
#p<0.05; Compare with model group
*p<0.05.
Table 4 Herba Dracocephali total flavones on the impact of SOD content in mouse cortex tissue (
n=10)
| Group | Dosage (mg/kg) | SOD(U/mgp) |
| Blank group | 18.82±3.18 | |
| Model group | 11.09±2.61 # | |
| Herba Dracocephali total flavones | 25 | 14.36±4.01 |
| Herba Dracocephali total flavones | 50 | 13.20±4.63 |
| Herba Dracocephali total flavones | 100 | 19.32±5.23 * |
Note: compare with blank group
#p<0.05; Compare with model group
*p<0.05.
Table 5 Herba Dracocephali total flavones on the impact of GSH-PX content in mouse cortex tissue (
n=10)
| Group | Dosage (mg/kg) | GSH-PX(U) |
| Blank group | 324.16±36.90 | |
| Model group | 193.74±32.86 # | |
| Herba Dracocephali total flavones | 25 | 200.35±29.89 |
| Herba Dracocephali total flavones | 50 | 215.83±36.01 |
| Herba Dracocephali total flavones | 100 | 207.30±30.12 |
Note: compare with blank group
#p<0.05; Compare with model group
*p<0.05.
Embodiment 3: Herba Dracocephali total flavones is on the impact of dementia mice brain tissue inflammation level
1, experiment material
(1) reagent: interleukin-11 β (1L-1 β), tumor necrosis factor α (TNF-α) ELISA kit (Wuhan Boster Biological Technology Co., Ltd.).Inducible nitric oxide (NOS2) detection kit (Wuhan excellent Er Sheng commerce and trade company limited);
(2) instrument: microplate reader (Thermo3001, Thermo company of the U.S.); TGL-16G freezing centrifuge (Anting Scientific Instrument Factory, Shanghai);
2, experimental technique
Water maze laboratory terminates rear execution animal, pluck full brain and clean bloodstain with ice-cold normal saline, isolate cerebral cortex at low ambient temperatures, the normal saline adding 9 times of volumes after weighing makes 10% homogenate, the centrifugal 10min of 3500r/min, gets supernatant and measures interleukin-11 β, tumor necrosis factor α and inducible nitric oxide content in cerebral tissue according to test kit description.
3, statistical procedures
Experimental data with
represent, compare between group and adopt one factor analysis of variance (one-wayANOVA), check in conjunction with Dunnett ' sposthoctest, more each group difference, p < 0.05 thinks to there is significant difference.
4, experimental result
Compared with blank group, in model group mouse cortex tissue, the expression of IL-1 β, TNF-α, iNOS all raises, and difference has significance (p<0.01), demonstrates inflammatory reaction in model group mouse cortex tissue comparatively obvious.Can reduce the expression of IL-1 β in cortical tissue after Herba Dracocephali total flavones administration process, wherein middle and high dosage group has significant difference (p<0.05, p<0.01) compared with model group.Herba Dracocephali total flavones high dose group also significantly can reduce the expression of TNF-α in dementia mice cortex (p<0.05), it can lower the expression of iNOS to a certain extent, but does not reach significant difference (see table 6-8).
Table 6 Herba Dracocephali total flavones on the impact of IL-1 β content in mouse cortex tissue (
n=10)
| Group | Dosage (mg/kg) | IL-1β(pg·mg -1) |
| Blank | 114.68±25.73 | |
| Model | 268.36±30.42 ## | |
| Herba Dracocephali total flavones | 25 | 230.36±32.06 |
| Herba Dracocephali total flavones | 50 | 189.74±46.31 * |
| Herba Dracocephali total flavones | 100 | 164.46±30.03 ** |
Note: compare with blank group
#p<0.05,
##p<0.01; Compare with model group
*p<0.05,
*p<0.01.
Table 7 Herba Dracocephali total flavones on the impact of TNF-alpha content in mouse cortex tissue (
n=10)
| Group | Dosage (mg/kg) | TNF-α(pg·mg -1) |
| Blank | 7.32±1.10 | |
| Model | 15.02±0.92 ## | |
| Herba Dracocephali total flavones | 25 | 15.30±1.36 |
| Herba Dracocephali total flavones | 50 | 13.33±1.10 |
| Herba Dracocephali total flavones | 100 | 10.32±1.89 * |
Note: compare with blank group
#p<0.05,
##p<0.01; Compare with model group
*p<0.05.
Table 8 Herba Dracocephali total flavones on the impact of iNOS content in mouse cortex tissue (
n=10)
| Group | Dosage (mg/kg) | iNOS(pg·mg -1) |
| Blank | 23.31±4.16 | |
| Model | 40.31±5.21 ## | |
| Herba Dracocephali total flavones | 25 | 41.72±5.20 |
| Herba Dracocephali total flavones | 50 | 38.71±6.17 |
| Herba Dracocephali total flavones | 100 | 34.16±4.01 |
Note: compare with blank group
#p<0.05,
##p<0.01.
Claims (4)
1. Herba Dracocephali total flavones is as the application of active constituents of medicine in preparation control Alzheimer disease drugs.
2. according to asking application according to claim 1, it is characterized in that, every day, effective dose was 25 ~ 100mg/kg with oral or non-oral routes mode administration.
3. application according to claim 1 and 2, is characterized in that, described Herba Dracocephali total flavones exists with the solid-state form of tablet, capsule, pill, injection, slow release, controlled-release pharmaceutical formulation as sole active agent.
4. application according to claim 1 and 2, is characterized in that, described Herba Dracocephali total flavones exists with the solid-state form of tablet, capsule, pill, injection, slow release, controlled release compound medicinal formulation as wherein a kind of active component.
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| KR20210148533A (en) * | 2020-05-29 | 2021-12-08 | 성이바이오(주) | Composition for preventing or treating cognitive disorder or psychiatric disorder comprising extracts of Dracocephalum moldavica |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102138967A (en) * | 2010-01-29 | 2011-08-03 | 孙雅煊 | Application of dracocephalum moldavica L. general flavone in preparing medicament for preventing and treating cerebrovascular disease |
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| CN102138967A (en) * | 2010-01-29 | 2011-08-03 | 孙雅煊 | Application of dracocephalum moldavica L. general flavone in preparing medicament for preventing and treating cerebrovascular disease |
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| KR20210148533A (en) * | 2020-05-29 | 2021-12-08 | 성이바이오(주) | Composition for preventing or treating cognitive disorder or psychiatric disorder comprising extracts of Dracocephalum moldavica |
| KR102451200B1 (en) * | 2020-05-29 | 2022-10-07 | 성이바이오(주) | Composition for preventing or treating cognitive disorder or psychiatric disorder comprising extracts of Dracocephalum moldavica |
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