CN110393799A - The application of Nrf2 agonist - Google Patents
The application of Nrf2 agonist Download PDFInfo
- Publication number
- CN110393799A CN110393799A CN201810375101.5A CN201810375101A CN110393799A CN 110393799 A CN110393799 A CN 110393799A CN 201810375101 A CN201810375101 A CN 201810375101A CN 110393799 A CN110393799 A CN 110393799A
- Authority
- CN
- China
- Prior art keywords
- nrf2
- cell
- stem cell
- application
- agonist
- 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
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
- A61K31/045—Hydroxy compounds, e.g. alcohols; Salts thereof, e.g. alcoholates
- A61K31/05—Phenols
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K45/00—Medicinal preparations containing active ingredients not provided for in groups A61K31/00 - A61K41/00
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P3/00—Drugs for disorders of the metabolism
- A61P3/08—Drugs for disorders of the metabolism for glucose homeostasis
- A61P3/10—Drugs for disorders of the metabolism for glucose homeostasis for hyperglycaemia, e.g. antidiabetics
-
- 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
Landscapes
- Health & Medical Sciences (AREA)
- Medicinal Chemistry (AREA)
- Veterinary Medicine (AREA)
- Public Health (AREA)
- General Health & Medical Sciences (AREA)
- Animal Behavior & Ethology (AREA)
- Life Sciences & Earth Sciences (AREA)
- Pharmacology & Pharmacy (AREA)
- Chemical & Material Sciences (AREA)
- Diabetes (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Epidemiology (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Organic Chemistry (AREA)
- General Chemical & Material Sciences (AREA)
- Obesity (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Engineering & Computer Science (AREA)
- Endocrinology (AREA)
- Emergency Medicine (AREA)
- Hematology (AREA)
- Toxicology (AREA)
- Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
- Acyclic And Carbocyclic Compounds In Medicinal Compositions (AREA)
Abstract
The present invention relates to field of biotechnology, the in particular to application of Nrf2 agonist.White reed alcohol is the agonist of Nrf2; hUMSC is handled with white reed alcohol; a series of cytoprotection genes with up-regulation Nrf2 and Nrf2 regulation increase cell viability, enhance the activity of the stemness of stem cell and the class pancreas islet corpusculum β cell of induction and the function of excreting insulin.Since white reed alcohol is natural products, living to have source wide, toxicity is low.The hUMSC treatment diabetes mice handled with hUMSC and white reed alcohol can prevent and repair Pancreas Islet Structure caused by STZ and destroy, and increase insulin secretion obviously, the hUMSC hypoglycemic effect of white reed alcohol processing is more preferable.The present invention is clear with the mechanism of action, and function and effect are significant, securely and reliably, and it is simple and easy.The present invention is of great significance to the clinical therapeutic efficacy for improving mescenchymal stem cell.
Description
Technical field
The present invention relates to field of biotechnology, the in particular to application of Nrf2 agonist.
Background technique
The morbidity of diabetes in recent years is in explosion type ascendant trend in the whole world, it is contemplated that the number meeting of the year two thousand forty whole world diabetes
Reach 6.4 hundred million, wherein there are about 1.5 hundred million for China.Therefore it is badly in need of effective method to go to control this disease.Diabetes are treated at present
Method is mainly chemical Antidiabetic Drug and insulin injection to control hyperglycemia caused by diabetes, however these methods can not be from
The fundamentally damage of body multiple organ caused by reverting diabetes.Pancreas islet organ transplant pancreatic islets transplantation is that treatment diabetes illustrate
New hope, but due to organ donor there is a serious shortage of and tissue matching ask body, be not at present able to satisfy and to be growing completely
Patient population needs.
The discovery of multipotential stem cell and stem cell biological progress of research answer the clinic of cell therapy and neomorph
With being possibly realized.Mescenchymal stem cell is a member of stem cell line, it is present in the almost all of organ of human body, is had certainly
I updates and the cell of powerful differentiation potential, can be divided into different tissues, organ under given conditions.Meanwhile it filling
Matter stem cell possesses the unexistent advantage of other stem cells, that is, it, which also has to injury tissue, migrates, and carries out tissue repair
And have the function of to carry out immunoregulation.In addition, mescenchymal stem cell itself has low immunogenicity, even if allosome or making across kind
With being difficult to cause immunological rejection.Therefore, mescenchymal stem cell is the important tool of organizational project and cell therapy, is
The treatment method of the core mechanism reverting diabetes natural history of diabetes can be uniquely directed in current all hypoglycemic schemes,
Huge potentiality are demonstrated by clinical application.But mescenchymal stem cell in vitro can not infinite multiplication differentiation, after a certain period of time
It just will appear aging death.
The method to improve stem cell function reported at present have 1) enhanced with 3D culture technique and lure from fat stem cell
The survival and function for the class pancreas islet corpusculum led.Ciprofloxacin (ciprofaxcin) can improve the skin progenitor cell of in vitro culture
Clone formation, the expression of stemness the marker Nanog and Oct4 of stem cell.Zhongjun Zhou research team finds white reed
Chenopodiaceae alcohol (resveratrol) can increase Zmpste24-/- senescence accelerated mouse adult stem cell quantity, significant extension senescence accelerated mouse
Service life.4) Oltipraz (Oltipraz) can delay the ageing process of human child's early ageing disease mescenchymal stem cell, and improve
Its activity in vivo.However there are the following problems: 1) 3D culture be simulate human body three-dimensional structure progress relative ideal situation it is thin
Born of the same parents' cultivating system, but the system of Neena Aloysious and Prabha Nair with a series of chemical methodes by dextrose
Glycosides is crosslinked with gelatin, although this 3D system is cultivated in vitro achieves better result, the chemical reagent introduced is difficult to dispel
It removes, from clinical application, there are also longer roads.2) Ciprofloxacin (ciprofaxcin) is a kind of broad-spectrum antibiotic, although external place
The activity that cell has enhancing stem cell stemness well is managed, but prolonged application can cause human flora's deficiency disorder or induction in vivo
Generate super antibody-resistant bacterium.3) Liu et al has found combination of the white reed alcohol by regulation LaminA/SIRT1 to increase
SIRT1's goes acyl ability to improve the function of senile cell.These experiments only carry out in the mouse of gene defect, do not say
Bright white reed alcohol is to regulate and control mice age process by Nrf2.4) Oltipraz (Oltipraz) is the small molecule of the synthesis seventies
Medicine most goes out to be mainly used for snail fever, but effect not as good as praziquantel and has serious pay to act on, and has been eliminated.
Transcription regulatory factor Nrf2 is the key factor for regulating and controlling internal cellular oxidation emergency reaction, belongs to CNC-bZiP transcription
The nuclear factor of factor family.It improves cell by regulating and controlling in vivo a series of anti-oxidant albumen and removing toxic substances Protecting gene
Survival ability.It is a large amount of research shows that oxidation emergency reaction be internal many disease developments originate or key factor.
Nrf2 passes through the reaction of regulation oxidation emergency and diabetes, tumour, liver diseases, kidney trouble, respiratory disease, nervous system
The occurrence and development of a variety of diseases of the human bodies such as disease, skin disease, metabolic disease and heavy metal poisoning have close relationship.Fixed point
Knock out the diabetes that the mouse of Nrf2 gene induces poisonous substance, hepatic injury, tumour etc. has very strong neurological susceptibility and more serious
Pathologic process.Therefore, Nrf2 is the potential target spot of disease treatment.Nrf2 inducer is likely to become potential gene target and controls
Treat drug.
Summary of the invention
In view of this, the present invention provides the applications of Nrf2 agonist.The function and effect of the Nrf2 agonist are significant, safety
Reliably, and it is simple and easy.The present invention is of great significance to the clinical therapeutic efficacy for improving mescenchymal stem cell.
In order to achieve the above-mentioned object of the invention, the present invention the following technical schemes are provided:
The present invention provides application of the Nrf2 agonist in the drug that preparation improves the bioactivity of mescenchymal stem cell.
The present invention provides Nrf2 agonists to improve the anti-drug at programmed cell death ability of mescenchymal stem cell in preparation
In application.
The present invention provides application of the Nrf2 agonist in the drug that preparation improves mescenchymal stem cell anti-aging ability.
The present invention provides application of the Nrf2 agonist in the drug that preparation extends the service life of mescenchymal stem cell.
The present invention provides Nrf2 agonists in the protective agent of p-Isletβ cell for preparing mescenchymal stem cell induction
Using.
The present invention provides Nrf2 agonist answering in the survival of preparation enhancing class pancreas islet corpusculum and/or the drug of function
With.
The present invention provides Nrf2 agonists in the drug that preparation improves the function of class pancreas islet corpusculum excreting insulin
Using.
The present invention provides application of the Nrf2 agonist in the drug of preparation treatment diabetes.
In some specific embodiments of the invention, the Nrf2 agonist includes tea polyphenols, curcumin, radish sulphur
Mixture more than one or both of alkane, resveratrol, tanshinone or feverfew.
White reed alcohol (resveratrol) also known as resvertrol, 3,4', 5- resveratrol, (E) -5-2- (4- hydroxyphenyl)-second
Alkenyl-Resorcinol, chemical formula (molecular formula): C14H12O3 is a kind of polyphenols with important physiological activity, is
Grape, main composition in the various plants body such as uranium skin, with antitumor, protection is cardiovascular, prevents the function of organism aging process
Effect.Pharmacological action: the generation of anti-bacteria and anti-virus, antitumor, anti-hepatitis, inhibition platelet aggregation and thromboxane B2.It can be hindered
The only oxidation of low-density lipoprotein has potential prevention and cure of cardiovascular disease, anti-cancer, antiviral and immunoregulation effect, makees
With being mainly shown as its anti-oxidation characteristics.Safety: the index for discharging lactic dehydrogenase as cellular damage using leucocyte is come
The safety for detecting white reed alcohol finds that its LD50 is per kilogram of body weight 1000mg.5×10-7To point 10-3The white reed alcohol of M is drawn
The release of the lactic dehydrogenase risen is lower than 7%.Illustrate that white reed alcohol has very high safety.Can be widely applied to food,
Health care product, medicine and other fields.The discovery of molecular drug Study on mechanism, the cytosis target spot of white reed alcohol be transcriptional control because
Sub- Nrf2.White reed alcohol processing cell can raise the expression of Nrf2 gene for 6 hours, increase a series of work of antioxidases of cell
Property and cell detoxification revision points expression, improve cell viability.
In some specific embodiments of the invention, the effective dose of the Nrf2 agonist is 1~20 μM/(1~20
μM/5x105-1x106Cell) or 20~50 μ g/kg the weight of animals (rat or mouse)/d, also there is close effect to the mankind.
The present invention provides a kind of bioactivity for improving mescenchymal stem cell and/or anti-at programmed cell death and/or anti-
The ability of aging and/or the service life of extension mescenchymal stem cell and/or the method for increasing cell therapy effect, with Nrf2 agonist
Act on mescenchymal stem cell, human or animal body.
In some specific embodiments of the invention, the effective dose of the Nrf2 agonist is 1~20 μM/(1~20
μM/5x105-1x106Cell) or 20~50 μ g/kg the weight of animals (rat or mouse)/d, also there is close effect to the mankind.
The present invention also provides a kind of methods of survival and/or function for enhancing class pancreas islet corpusculum, with Nrf2 agonist effect
In human or animal body.
In some specific embodiments of the invention, the effective dose of the Nrf2 agonist is 1~20 μM/(1~20
μM/5x105-1x106 cell) or 20~50 μ g/kg the weight of animals (rat or mouse)/d, also there is close effect to the mankind.
The present invention also provides a kind of methods of function of improving class pancreas islet corpusculum excreting insulin, with Nrf2 agonist work
For human or animal body.
In some specific embodiments of the invention, the effective dose of the Nrf2 agonist is 1~20 μM/(1~20
μM/5x105-1x106Cell) or 20~50 μ g/kg the weight of animals (rat or mouse)/d, also there is close effect to the mankind.
The present invention also provides the treatment methods of diabetes, with Nrf2 agonist effect in human or animal body.
In some specific embodiments of the invention, the effective dose of the Nrf2 agonist is 1~20 μM/(1~20
μM/5x105-1x106Cell) or 20~50 μ g/kg the weight of animals (rat or mouse)/d, also there is close effect to the mankind.
In specific embodiments of the present invention, Nrf2 agonist is white reed alcohol.
Transcription regulatory factor Nrf2 is anti-oxidant in vivo, anti-cell aging and the master switch for protecting cell viability.To experiment
Property animal diabetes occurrence and development have important protective effect.White reed alcohol is the agonist of Nrf2, is handled with white reed alcohol
HUMSC has a series of cytoprotection genes of up-regulation Nrf2 and Nrf2 regulation, increases cell viability, enhances the dry of stem cell
The function of the activity and excreting insulin of property and the class pancreas islet corpusculum β cell of induction.Since white reed alcohol is natural products, tool living
There is source wide, toxicity is low.The hUMSC treatment diabetes mice handled with hUMSC and white reed alcohol can prevent and repair STZ and cause
Pancreas Islet Structure destroy, increase insulin secretion obviously, white reed alcohol processing hUMSC hypoglycemic effect it is more preferable.
Therefore, the present invention is clear with the mechanism of action, and function and effect are significant, securely and reliably, and it is simple and easy.The present invention couple
The clinical therapeutic efficacy for improving mescenchymal stem cell is of great significance.
Detailed description of the invention
In order to more clearly explain the embodiment of the invention or the technical proposal in the existing technology, to embodiment or will show below
There is attached drawing needed in technical description to be briefly described.
Fig. 1 shows that 1-20 μM of white reed alcohol processing hUMSCI cell can raise the expression of Nrf2 gene for 6 hours, increases cell
A series of activity of antioxidases and the expression of cell detoxification revision points, improve oxidability caused by the high sugar of cell confrontation and
The anti-vigor at sequenceization death；Wherein, A: white reed alcohol induces 24 hours up-regulation Nrf2；B: and antioxidant genes downstream
(HO1) and cell detoxification and the expression for preventing pre- gene (NQO1)；C: white reed alcohol reduces mitochondria free radical caused by hyperglycemia
It is horizontal；D: white reed alcohol significantly reduces programmed cell death caused by hyperglycemia；
Note: Conc.: control；Res:(resveratrol) white reed alcohol；Glucose: glucose；MitoSox: mitochondria
Free radical tracer, it is red；Hoechest: nucleus dyestuff, it is blue；Nrf2: transcription regulatory factor；NQO1: cell it is anti-pre- and
Detoxification genes；HO1: cellular anti-oxidant albumen；Caspase 3/7Glo: the programmed cell death albumen 3/7 of luminescent marking；
Apoptosis: programmed cell death；
Fig. 2 shows that (1-20 μM) processing human umbilical cord mesenchymal stem cells (hUMSC) of white reed alcohol can increase cell colony and be formed
Rate, the expression of real-time PCR research discovery stem cell stemness mark OCT4, Nanog and SOX2 increased after white reed alcohol processing with
And (1-20 μM) of white reed alcohol enhance the survival of class pancreas islet corpusculum induce from hUMSC and function and can increase class pancreas islet corpusculum and divide
Secrete the function of insulin；Wherein, A: white reed alcohol increases human umbilical cord mesenchymal stem cells (hUMSC) cell colony formation rate；B:
Real-time PCR: the expression of white reed alcohol increase stem cell stemness gene；C: immunofluorescence: white reed alcohol increases the class pancreas islet of induction
The secretion of corpusculum insulin；
Note: Conc.: control Res:(resveratrol) white reed alcohol；Nanog, Sox2, OCT4 are to represent stem cell to do
The gene of property；Insulin: insulin, it is red；DAPI: nucleus dyestuff, it is blue；
Fig. 3 shows the diabetes model that the mouse that Nrf2 gene knocks out induces STZ, and neurological susceptibility increases compared with control mice
By force, blood glucose rise is fast, and the duration is long, and symptom is more serious；Wherein, the glycosuria that the mouse that A:Nrf2 gene knocks out induces STZ
The influence of disease model, the mouse that STZ processing Nrf2 gene knocks out keep blood glucose rise fast, and the duration is long, and hyperglycemia is more serious；B:
Insulin level is lower in the mouse blood that Nrf2 gene knocks out；C: mice pancreatic histochemistry and immunohistochemical staining: STZ is serious
Normal islets structure is destroyed, STZ is more serious to the pancreas islet destruction for the mouse that Nrf2 gene knocks out, and insulin secretion is substantially reduced；
Note: Conc.: control；Blood glucose: blood glucose；Insulin: insulin；H&E staining: histochemistry
Dyeing；WT (wild type, WT): wild type or normal genotype；Nrf2KO:Nrf2 gene knocks out type；STZ
(Streptozotocin), streptozotocin, to inducing mouse and rat diabetes；
Fig. 4 shows the diabetes mice of the hUMSC treatment STZ induction with hUMSC and white reed alcohol processing, can reduce STZ and lure
The hyperglycemia of hair reduces islet damage caused by STZ, increases insulin secretion, significantly improves the general signs of mouse；White reed Chenopodiaceae
The hUMSC therapeutic effect of alcohol processing is better than hUMSC group；Wherein, the hUMSC treatment STZ induction of A:hUMSC and white reed alcohol processing
Diabetes mice, hUMSC and white reed alcohol processing hUMSC can obviously reduce STZ induction hyperglycemia, white reed alcohol processing
HUMSC hypoglycemic effect it is more preferable；B: the mice pancreatic immunohistochemical staining prompt hUMSC and hUMSC of white reed alcohol processing can hinder
Only and the destruction of Pancreas Islet Structure caused by STZ is repaired, increases insulin secretion obviously；What C:hUMSC and white reed alcohol were handled
HUMSC reduces programmed cell death caused by STZ, and the hUMSC hypoglycemic effect of white reed alcohol processing is more preferable；
Note: Conc.: control；Blood glucose: blood glucose；Insulin: insulin；Glucagon glucagon；
STZ (Streptozotocin), streptozotocin, to inducing mouse and rat diabetes.
Specific embodiment
The invention discloses the application of Nrf2 agonist, those skilled in the art can use for reference present disclosure, be suitably modified
Realization of process parameters.In particular, it should be pointed out that all similar substitutions and modifications be for a person skilled in the art it is aobvious and
It is clear to, they are considered as being included in the present invention.Method and application of the invention is retouched by preferred embodiment
It states, related personnel can obviously not depart from the content of present invention, change in spirit and scope to method described herein and application
Dynamic or appropriate changes and combinations, carry out implementation and application the technology of the present invention.
Largely research shows that transcription regulatory factor Nrf2 is internal anti-oxidant, cell detoxification, anti-aging and protection cell
The master switch of activity.The inactivation of Nrf2 and the generation of many chronic metabolic diseases of human body such as diabetes, development have close pass
System, and the generation of disease, development and pathogenesis can be prevented and be slowed down to the activity for enhancing Nrf2.The present invention is activated with white reed alcohol
The characteristic of Nrf2 handles the class islet cells corpusculum of stem cell and induction, discovery white reed alcohol protection cell activity and promotion point
Secrete the function of insulin.The present invention is of great significance to the clinical therapeutic efficacy for improving mescenchymal stem cell.
Raw materials used, auxiliary material and reagent are available on the market in the application of Nrf2 agonist provided by the invention.
Relational language is explained:
Mescenchymal stem cell (mesenchymal stem cell, MSC): being a kind of with the of self-replication capacity and multidirectional
The adult stem cell of differentiation potential, this stem cell are present in a variety of organs of human body, in tissue, can induce differentiation into os osseum,
Cartilage, fat and other kinds of cell.Mescenchymal stem cell has low immunogenicity, can receive transplanting.
Human umbilical cord mesenchymal stem cells (human umbilical cord mesenchymal stem cell, hUMSC):
It is the mescenchymal stem cell that sub-argument comes out from fetal cord China Tong Shi glue (Wharton ' s jelly).
Nrf2 (nuclear factor erythroid 2-related factor 2): transcription regulatory factor Nrf2 is
The key factor for regulating and controlling internal cellular oxidation emergency reaction, belongs to the nuclear factor of CNC-bZiP transcription factor family.It is logical
Regulation a series of anti-oxidant albumen and removing toxic substances Protecting gene in vivo are crossed, the survival ability of cell is improved.
White reed alcohol (resveratrol): white reed alcohol (resveratrol) also known as resvertrol, 3,4', 5- trihydroxy
Stilbene, (E) -5-2- (4- hydroxyphenyl)-vinyl-Resorcinol, chemical formula (molecular formula): C14H12O3 is a kind of with weight
The polyphenols for wanting physiological activity, is grape, and main composition in the various plants body such as uranium skin has anti-oxidant, anti-
Only organism aging process and other effects.
Induction differentiation (induced differentiation): made with specific reagent and method a kind of more original thin
The phenomenon that born of the same parents break up to next stage or terminal direction.
Human pancreatic islet β cell: there are two main classes for the cell in human pancreatic islet: α and β, β cells secrete insulin, main function
It can be adjusting blood glucose.
Class pancreas islet corpusculum: stem cell is induced into vitro similar to the pancreas islet in Human Pancreas with the method for induction differentiation
Corpusculum.
STZ (Streptozotocin), streptozotocin, to inducing mouse and rat diabetes.
Below with reference to embodiment, the present invention is further explained:
Embodiment 1
1): the separation of umbilical cord Wharton ' s jelly mescenchymal stem cell (hUMSC), β cell induction and white reed alcohol at
Reason: 10cm umbilical cord is taken, artery and vein are isolated, rest part is cut into 2-3mm3 fritter, is transferred to the 10%FBS's containing DMEM
T75 Tissue Culture Flask.Small fusiform mescenchymal stem cell climbs out of tissue fritter, and rapid growth when cultivating 7-14 days.β cell lures
Lead and white reed alcohol processing: take it is well-grown, fluidic cell examine and determine CD105, the cell of CD90, CD70 positive set adds or be not added
1xITS is added in DMEM10%FBS culture medium and cultivates 4 days for 1-20 μM of white reed alcohol processing group when β cell induces, then plus
0.3mM turine cultivate 4 days, then plus 100mM Buddhist nun's amide (nicotinamide) cultivate 10 days, class islet cells corpusculum will
It is formed.
2): cell culture and colony-forming efficiency: isolated hUMSC is cultivated in 10% fetal calf serum DMEM, 5%CO2,37 DEG C
In constant incubator.It counts 100 cell kinds to enter in 60mm Tissue Culture Dish, adds DMEM, 10% fetal calf serum, in 5%CO2,
It is cultivated 14 days in 37 DEG C of constant incubators, with violet staining, counts bluish violet cell colony.Every group parallel three.Experiment weight
Again three times.
3): real-time fluorescence PCR (Real-time PCR): with 2 μ gRNA of well-grown thin extraction RNA (QIAGEN)
Reverse transcription is used at cDNA.cDNA using reverse transcriptase III (Invitrogen) .cDNA sample
ABsolute Blue qPCR SYBR Green Rox Master Mix (Thermo Scientific) kit carries out real-time
Fluorescent PCR (ABI 7900HT (SABiosciences).Program: 95 DEG C of for 15min as initial denaturing,
followed by 40cycles of 95℃for 15sec,60℃for 30sec,and 72℃for 30sec.
4): protein electrophoresis and immune chromogenic assay (Immunoblotting): cell cell pyrolysis liquid (modified
RIPA Buffer) cracking, SDS_PAGE electrophoresis is added in 40 μ g albumen, transfer albumen to pvdf membrane is sealed with 5% skimmed milk power
It closes 1 hour, primary antibody is added, 4 DEG C overnight, rinsing, secondary antibody is added, room temperature 1 hour, ECL developed the color.
5): programmed cell death: the cell kind of culture is entered in 12 porocyte culture plates, is cultivated 16h., is then handled
24 hours.Cell is washed twice with PBS, is then cracked with cell pyrolysis liquid, Caspas3/7 activity Caspas3/7-Glo reagent
It is quantitative that box (Promega) measures GloMax 20/20.
6): ROS detection: the cell kind of culture, which is entered 8 hole chamber slide culture 16h. cell culture, terminates first 30 points
Clock is added mitosox deep red (Invitroge) 5 μM of cells and is washed twice with PBS, and 4%paraformaldehyde is fixed,
With DAPI dye nucleus cell image fluorescence microscope and quantitatively.
7): animal model: STZ is injected intraperitoneally in 6-8 weeks C57/B6 mouse and Nrf2 gene knockout type mouse, 18-22g
75mg/kg, while injecting hUMSC or huMSC+Res cell 1x106/ only, mouse was surveyed with blood glucose meter at 0,4,7,9,13,15 day
Tail vein blood glucose.After 21 days, each group mouse is handled, weighing extracts the internal organs such as pancreas islet, and paraffin embedding carries out histotomy, H&E
Dyeing and immunohistochemistry.
8): statistical disposition: M ± SD is used in all experiments, and one way ANOVA analysis and t are examined.P < 0.05.
The effective dose of Nrf2 agonist (such as white reed alcohol) is 1~20 μM/(1~20 μM/5x105-1x106Cell)
Or 20~50 μ g/kg the weight of animals (rat or mouse)/d, also there is close effect to the mankind.
The above is only a preferred embodiment of the present invention, it is noted that for the ordinary skill people of the art
For member, various improvements and modifications may be made without departing from the principle of the present invention, these improvements and modifications are also answered
It is considered as protection scope of the present invention.
Claims (10)
- Application of the 1.Nrf2 agonist in the drug that preparation improves the bioactivity of mescenchymal stem cell.
- 2.Nrf2 agonist improves the application in the anti-drug at programmed cell death ability of mescenchymal stem cell in preparation.
- Application of the 3.Nrf2 agonist in the drug that preparation improves mescenchymal stem cell anti-aging ability.
- Application of the 4.Nrf2 agonist in the drug that preparation extends the service life of mescenchymal stem cell.
- Application of the 5.Nrf2 agonist in the protective agent of p-Isletβ cell for preparing mescenchymal stem cell induction.
- Application of the 6.Nrf2 agonist in the survival of preparation enhancing class pancreas islet corpusculum and/or the drug of function.
- Application of the 7.Nrf2 agonist in the drug that preparation improves the function of class pancreas islet corpusculum excreting insulin.
- Application of the 8.Nrf2 agonist in the drug of preparation treatment diabetes.
- 9. application as claimed in any one of claims 1 to 8, the Nrf2 agonist includes tea polyphenols, curcumin, radish sulphur Mixture more than one or both of alkane, resveratrol, tanshinone or feverfew.
- 10. it is a kind of improve mescenchymal stem cell bioactivity and/or it is anti-at the ability of programmed cell death and/or anti-aging and/ Or extend the service life of mescenchymal stem cell and/or increase the method for cell therapy effect, which is characterized in that with Nrf2 agonist work For human or animal body.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201810375101.5A CN110393799A (en) | 2018-04-24 | 2018-04-24 | The application of Nrf2 agonist |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201810375101.5A CN110393799A (en) | 2018-04-24 | 2018-04-24 | The application of Nrf2 agonist |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN110393799A true CN110393799A (en) | 2019-11-01 |
Family
ID=68322529
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201810375101.5A Pending CN110393799A (en) | 2018-04-24 | 2018-04-24 | The application of Nrf2 agonist |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN110393799A (en) |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111035632A (en) * | 2019-12-27 | 2020-04-21 | 天津市肿瘤医院 | Activator of Nrf2 and application of activator in preparation of drugs for preventing and treating environmental toxic substances |
| CN113694200A (en) * | 2021-07-05 | 2021-11-26 | 温州医科大学附属第二医院（温州医科大学附属育英儿童医院） | Targeted medicine for treating cognitive function damage caused by nerve cell damage and application |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101413012A (en) * | 2008-11-28 | 2009-04-22 | 暨南大学 | Method for inducing differentiation of mesenchyma stem cell into islet beta-like cells, and use thereof |
| CN102433302A (en) * | 2011-12-15 | 2012-05-02 | 成都清科生物科技有限公司 | Serum-free culture medium for mesenchymal stem cells |
-
2018
- 2018-04-24 CN CN201810375101.5A patent/CN110393799A/en active Pending
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101413012A (en) * | 2008-11-28 | 2009-04-22 | 暨南大学 | Method for inducing differentiation of mesenchyma stem cell into islet beta-like cells, and use thereof |
| CN102433302A (en) * | 2011-12-15 | 2012-05-02 | 成都清科生物科技有限公司 | Serum-free culture medium for mesenchymal stem cells |
Non-Patent Citations (6)
| Title |
|---|
| L.-T. LEI，等: "Resveratrol attenuates senescence of adipose-derived mesenchymal stem cells and restores their paracrine effects on promoting insulin secretion of INS-1 cells through Pim-1", 《EUROPEAN REVIEW FOR MEDICAL AND PHARMACOLOGICAL SCIENCES》 * |
| PEZZOLLA，等: "Resveratrol Ameliorates the Maturation Process of β-Cell-Like Cells Obtained from an Optimized Differentiation Protocol of Human Embryonic Stem Cells", 《PLOS ONE》 * |
| 余元勋: "《中国分子支气管哮喘病学》", 31 August 2017, 合肥：安徽科学技术出版社 * |
| 尹磊，等: "白藜芦醇对干细胞生物特性和功能的调控", 《江苏大学学报（医学版）》 * |
| 王欣欣: "人脐带间充质干细胞联合白藜芦醇对阿尔茨海默鼠的干预及 SIRT1 分子信号的调控", 《郑州大学 博士学位论文》 * |
| 马晓龙: "间充质干细胞联合白藜芦醇对 1 型糖尿病小鼠胰岛功能的保护作用", 《中国优秀硕士学位论文全文数据库 医药卫生科技辑》 * |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111035632A (en) * | 2019-12-27 | 2020-04-21 | 天津市肿瘤医院 | Activator of Nrf2 and application of activator in preparation of drugs for preventing and treating environmental toxic substances |
| CN113694200A (en) * | 2021-07-05 | 2021-11-26 | 温州医科大学附属第二医院（温州医科大学附属育英儿童医院） | Targeted medicine for treating cognitive function damage caused by nerve cell damage and application |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| US10251824B2 (en) | Method for inducing pluripotent stem cells and pluripotent stem cells prepared by said method | |
| Yang et al. | Curcumin inhibits microglia inflammation and confers neuroprotection in intracerebral hemorrhage | |
| EP3549589A1 (en) | Pharmaceutical composition containing mitochondria | |
| CN108938618A (en) | Delay compound and its application of human mesenchymal stem cell aging | |
| CN109312303A (en) | Express stem cell, its composition and preparation method thereof of mesenchyma and neuronal marker | |
| CN110393799A (en) | The application of Nrf2 agonist | |
| Zhang et al. | Erythropoietin Modification Enhances the Protection of Mesenchymal Stem Cells on Diabetic Rat‐Derived Schwann Cells: Implications for Diabetic Neuropathy | |
| Ho et al. | Artemisia argyi exhibits anti-aging effects through decreasing the senescence in aging stem cells | |
| Kumar et al. | Pharmacological potential of zonisamide and Nigella sativa per se and combination in high‐impact trauma device‐induced traumatic brain injury in Drosophila melanogaster | |
| Huang et al. | Maintaining blood retinal barrier homeostasis to attenuate retinal ischemia-reperfusion injury by targeting the KEAP1/NRF2/ARE pathway with lycopene | |
| Prijanti et al. | Java Sea Spirulina platensis chemical analysis and its protective ability against H2O2-exposed umbilical cord mesenchymal stem cells according to CD73, CD90, and CD105, viability, and HIF-1 alpha docking | |
| KR102159332B1 (en) | Composition for inducing selective apoptosis undifferentiated pluripotent stem cells comprising magnolia extracts and the Method for inducing selective apoptosis using the same | |
| Celik et al. | The antioxidant role of Sideritis caesarea infusion against TCA toxicity in rats | |
| Finocchio et al. | Recent advances of adipose-tissue-derived mesenchymal stem cell-based therapy for retinal diseases | |
| JP5496951B2 (en) | Platelet-derived growth factor (PDGF) -BB production enhancer and stem cell stabilizer containing the same | |
| RU2727540C1 (en) | Application of membrane vesicles of multipotent stromal cells induced by b cytochalasin, for restoration and increase of mitochondrial function | |
| KR101802668B1 (en) | Treatment use of liver failure and recovery use of liver function by liver toxin-induced adipose tissue-derived stem cell secretome | |
| CN105969725B (en) | The purposes of fructus lycii red pigment | |
| Alipanah-Moghadam et al. | Andrographolide induced heme oxygenase-1 expression in MSC-like cells isolated from rat bone marrow exposed to environmental stress | |
| CN104491496B (en) | Application of the Gastrodin/Rhizoma Gastrodiae powder in anti-hepatic fibrosis medicines are prepared | |
| Shukry et al. | Bergenin Ameliorates Glutamate-Induced Death of Hippocampal Neuronal Cells through Modulation of the Antioxidant Activity and reducing the redox oxygen Species Creation. | |
| KR102323817B1 (en) | Composition comprising the extract of winged spindle for improving memory and cognitive function | |
| TWI483742B (en) | Method of treating cells in vitro with composition containing extract of peony root cortex and application thereof | |
| Poor et al. | The neuroprotective effects of dental pulp stem cells and erythropoietin in mice hippocampus after ischemia-reperfusion | |
| CN115120600A (en) | Application of diosgenin and analogues thereof in preparation of medicines for preventing or treating diabetes |
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 | ||
| RJ01 | Rejection of invention patent application after publication |
Application publication date: 20191101 |
|
| RJ01 | Rejection of invention patent application after publication |