CN107982537A - Curative drug and its application for microRNA-155 - Google Patents
Curative drug and its application for microRNA-155 Download PDFInfo
- Publication number
- CN107982537A CN107982537A CN201711149052.5A CN201711149052A CN107982537A CN 107982537 A CN107982537 A CN 107982537A CN 201711149052 A CN201711149052 A CN 201711149052A CN 107982537 A CN107982537 A CN 107982537A
- Authority
- CN
- China
- Prior art keywords
- microrna
- epilepsy
- medicine
- mouse
- microglia
- 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
- A61K45/00—Medicinal preparations containing active ingredients not provided for in groups A61K31/00 - A61K41/00
Landscapes
- Health & Medical Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Medicinal Chemistry (AREA)
- Pharmacology & Pharmacy (AREA)
- Epidemiology (AREA)
- Life Sciences & Earth Sciences (AREA)
- Animal Behavior & Ethology (AREA)
- General Health & Medical Sciences (AREA)
- Public Health (AREA)
- Veterinary Medicine (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
Abstract
Curative drug and its application for microRNA 155, are related to microRNA 155.The antiepileptic is prevention or treatment epilepsy.The medicine is made of effective dose comprising the nucleic acid of sequence and pharmaceutically acceptable carrier or auxiliary material shown in sequence table.Ejection preparation, oral formulations, spray agent, ointment formulation or patch etc. can be made according to the common method of medicine preparation in the medicine.Purposes of 155 antagonists of microRNA by microglia in prevention, treatment epileptic condition is provided.
Description
Technical field
The present invention relates to microRNA-155, more particularly, to the curative drug for microRNA-155 and its answers
With.
Background technology
Epilepsy is one of current most common nervous disorders, and the whole world about 65,000,000 people are diseased[1,2].There is research table
Bright, spongiocyte is by increasing the excitability of nerve cell and promoting inflammation to promote epilepsy, inflammation and neuroglia mistake
Degree activation causes the excitability of neuron excessive, eventually epileptogaenic generation of recurrent repeatedly etc.[3].It is it has recently been demonstrated that small
RNA (miRNA) plays a crucial role in the pathogenesis of the nervous system disease such as epilepsy[4~7].MiRNA is by with reference to target
The 3' non-translational regions (3'UTR) of mRNA are translated to reduce the stability of said target mrna and reduce, so as to adjust the small of gene expression
(~22nt) non-coding RNA[8,9].About 60% human protein is by miRNA direct regulations and controls[10].Under different pathological conditions,
Their differential expressions in brain, it is thus possible to as the therapy target of the nervous system disease including epilepsy and diagnosis, pre-
Biological markers afterwards[11~14]。
MicroRNA-155 (miR-155) is a typical multi-functional miRNA molecule.Nearest experimental evidence shows,
MiR-155 plays a significant role in inflammation, immune response and tumour generation[15~18].In nervous system, miR-155 quilts
It is considered inflammation modulators main in immune response[19,20], and in Macrophage Inflamatory reaction and multiple sclerosis lesion
Period highly raises[21,22].In addition, there is the core that studies have shown that microglia is probably the neurological disorder including epilepsy
The heart[3,23,24].Foregoing background shows that miR-155 may participate in the generation of epilepsy by adjusting microglia activity.
Bibliography：
[1]de Boer HM,Mula M,Sander JW.The global burden and stigma of
epilepsy.Epilepsy&behavior:E&B.2008；12:540-6.
[2]Moshe SL,Perucca E,Ryvlin P,Tomson T.Epilepsy:new
advances.Lancet.2015；385:884-98.
[3]Devinsky O,Vezzani A,Najjar S,De Lanerolle NC,Rogawski MA.Glia and
epilepsy:excitability and inflammation.Trends in neurosciences.2013；36:174-
84.
[4]Li MM,Li XM,Zheng XP,Yu JT,Tan L.MicroRNAs dysregulation in
epilepsy.Brain research.2014；1584:94-104.
[5]Tan CL,Plotkin JL,Veno MT,von Schimmelmann M,Feinberg P,Mann S,et
al.MicroRNA-128 governs neuronal excitability and motor behavior in
mice.Science.2013；342:1254-8.
[6]Jimenez-Mateos EM,Engel T,Merino-Serrais P,McKiernan RC,Tanaka K,
Mouri G,et al.Silencing microRNA-134 produces neuroprotective and prolonged
seizure-suppressive effects.Nature medicine.2012；18:1087-94.
[7]Rajman M,Metge F,Fiore R,Khudayberdiev S,Aksoy-Aksel A,Bicker S,et
al.A microRNA-129-5p/Rbfox crosstalk coordinates homeostatic downscaling of
excitatory synapses.The EMBO journal.2017；36:1770-87.
[8]Kosik KS.The neuronal microRNA system.Nature reviews
Neuroscience.2006；7:911-20.
[9]Lagos-Quintana M,Rauhut R,Lendeckel W,Tuschl T.Identification of
novel genes coding for small expressed RNAs.Science.2001；294:853-8.
[10]Friedman RC,Farh KK,Burge CB,Bartel DP.Most mammalian mRNAs are
conserved targets of microRNAs.Genome research.2009；19:92-105.
[11]Dogini DB,Avansini SH,Vieira AS,Lopes-Cendes I.MicroRNA
regulation and dysregulation in epilepsy.Frontiers in cellular
neuroscience.2013；7:172.
[12]Eacker SM,Dawson TM,Dawson VL.Understanding microRNAs in
neurodegeneration.Nature reviews Neuroscience.2009；10:837-41.
[13]Henshall DC.Antagomirs and microRNA in status
epilepticus.Epilepsia.2013；54 Suppl6:17-9.
[14]Bartel DP.MicroRNAs:genomics,biogenesis,mechanism,and
function.Cell.2004；116:281-97.
[15]Sheedy FJ,O'Neill LA.Adding fuel to fire:microRNAs as a new class
of mediators of inflammation.Annals of the rheumatic diseases.2008；67 Suppl
3:iii50-5.
[16]O'Connell RM,Zhao JL,Rao DS.MicroRNA function in myeloid
biology.Blood.2011；118:2960-9.
[17]Vigorito E,Kohlhaas S,Lu D,Leyland R.miR-155:an ancient regulator
of the immune system.Immunological reviews.2013；253:146-57.
[18]Kohanbash G,Okada H.MicroRNAs and STAT interplay.Seminars in
cancer biology.2012；22:70-5.
[19]Ponomarev ED,Veremeyko T,Weiner HL.MicroRNAs are universal
regulators of differentiation,activation,and polarization of microglia and
macrophages in normal and diseased CNS.Glia.2013；61:91-103.
[20]Su W,Aloi MS,Garden GA.MicroRNAs mediating CNS inflammation:Small
regulators with powerful potential.Brain,behavior,and immunity.2016；52:1-8.
[21]Thamilarasan M,Koczan D,Hecker M,Paap B,Zettl UK.MicroRNAs in
multiple sclerosis and experimental autoimmune encephalomyelitis.Autoimmunity
reviews.2012；11:174-9.
[22]Murugaiyan G,Beynon V,Mittal A,Joller N,Weiner HL.Silencing
microRNA-155ameliorates experimental autoimmune encephalomyelitis.Journal of
immunology.2011；187:2213-21.
[23]Mosser CA,Baptista S,Arnoux I,Audinat E.Microglia in CNS
development:Shaping the brain for the future.Progress in neurobiology.2017；
149-150:1-20.
[24]Luo C,Koyama R,Ikegaya Y.Microglia engulf viable newborn cells in
the epileptic dentate gyrus.Glia.2016；64:1508-17.
The content of the invention
It is an object of the invention to provide the curative drug for microRNA-155 and its application.
The curative drug for microRNA-155 is prevention or the medicine or pharmaceutical composition for the treatment of epilepsy.Institute
Medicine is stated to be made of comprising the nucleic acid of sequence and pharmaceutically acceptable carrier or auxiliary material shown in sequence table effective dose.
Ejection preparation, oral formulations, spray can be made according to the common method of medicine preparation in the medicine or pharmaceutical composition
Mist preparation, ointment formulation or patch etc..
The nucleotide sequence design anti sense nucleotide sequence and its trim of the microRNA-155, the modification include appointing
Anticipate nucleotide ribose modification, base modification and phosphate backbones modification in one or more of combinations or any nucleotide increasing
Subtract, replace, as long as the nucleotide sequence after modification still has the activity with microRNA-155 reverse complementals.
The invention discloses microRNA-155 antagonists in prevention, treatment epilepsy and the nerve characterized by epilepsy outbreak
Purposes in systemic disease.By substantial amounts of it is experimentally confirmed that in the mouse model for inducing epileptic attack, Cerebral Cortex and
The horizontal significantly rises of microRNA-155 in hippocampal tissue.Give the antagonist of mouse intracerebroventricular injection microRNA-155
(antagomir) mouse epileptic attack symptom can be significantly inhibited, it is meant that microRNA-155 rises in the pathogenesis of epilepsy
Key effect.It is contemplated that the purposes of the biological agent of open antagonist or inhibitor based on microRNA-155,
To treat and prevent the generation of temporal epilepsy.
Brief description of the drawings
Fig. 1 is the horizontal up-regulations of miR-155 in the epileptic attack mouse model brain tissue that KA induces.
Fig. 2 is the horizontal up-regulations of miR-155 in the brain sample of TLE patient.
Fig. 3 can significantly improve epileptic attack for miR-155 antagonists.
Fig. 4 can significantly improve the death of epilepsy Mouse Neuron for miR-155 antagonists.
Fig. 5 can significantly improve the expression of epilepsy mouse inflammatory factor TNF-α for miR-155 antagonists.
Fig. 6 can significantly improve epilepsy mouse inflammatory factor IL-1 β expression for miR-155 antagonists.
Fig. 7 can significantly improve epilepsy mouse inflammatory factor IL-6 expression for miR-155 antagonists.
Fig. 8 is primarily targeted for intracerebral spongiocyte for miR-155.
Fig. 9 is the microglia miR-155 up-regulated expressions of KA processing.
Figure 10 is that the microglia specific proteins CSF1R expression of KA processing is lowered.
Figure 11 has found the selectively targeted CSF1R of miR-155 for the processing of miR-155 activators.
Figure 12 can significantly raise CSF1R expression for miR-155 antagonists.
Figure 13 can significantly reverse Activated Microglia state for miR-155 antagonists.
Figure 14 is that the microglia conditioned medium of miR-155 antagonists processing can significantly reduce the neuron of KA inductions
The granting of action potential.
Figure 15 is that the microglia conditioned medium of miR-155 antagonists processing can significantly reduce the neuron of KA inductions
The frequency of excitatory postsynaptic potential (EPSP) (EPSC).
Figure 16 is that the microglia conditioned medium of miR-155 antagonists processing can significantly reduce the neuron of KA inductions
The frequency of inhibitory postsynaptic potential (IPSC).
Embodiment
The present invention is further described with reference to specific embodiment, all embodiments are only used for the illustration present invention, never limit
Protection scope of the present invention processed.
With bull C57BL/6 mouse (18~22g), be injected intraperitoneally it is classical cause epilepsy agent digenic acid (Kainic acid,
KA) 15mg/kg induces mouse epileptic attack, establishes mouse animal epileptic model.Mouse shows eyes and coagulates after injection digenic acid
Depending on, dynamic must increase, be short of breath, wash one's face action repeatedly, twitches with Head And Face, gradually develop into standing and fall down complete
The epilepsy syndromes of body tonic, breaking-out grade reach Racine and are classified IV~V grade.After 24h, broken end takes brain, takes out
Bilateral hippocampus, extracts RNA, and fluorescent quantitative PCR experiment is shown, the more normal control group mice of KA group mouse, microRNA- in hippocampus
155 horizontal significantly rises are (referring to Fig. 1).The brain tissue sample of clinical treatment of intractable epilepsy is collected, detection shows epileptic
The more non-epileptic of microRNA-155 levels is also significantly raised (referring to Fig. 2) in brain tissue.This means microRNA-155
Expression change it is closely related with the occurrence and development of epilepsy.
Further give the specific antagonists of bull C57BL/6 mouse intracerebroventricular injections microRNA-155
(antagomir-155), the epileptic attack symptom of mouse significantly improves, and breaking-out grade reaches Racine and is classified II~III grade, brain
Electrograph is shown as abnormal spike frequency and substantially reduces (referring to Fig. 3).FJB immunofluorescence experiments are shown, inject microRNA-155
After antagonist, hippocampus of mice neurodegeneration significantly improves (referring to Fig. 4), TNF-α (referring to Fig. 5), IL-1 β (referring to Fig. 6) and IL-
The neuroinflamations such as 6 (referring to Fig. 7) significantly improve, it is meant that antagonism microRNA-155 can improve mouse epileptic attack, mitigate god
Through degenerative conditions, improve neuroinflamation.
Because the main cell in brain is neuron, astroglia and microglia, by analyzing cerebral tissue
Expressions of the middle microRNA-155 in different cells, it is found that it is mainly expressed in spongiocyte (referring to Fig. 8).We connect
And shown in original cuiture microglia with KA processing, fluorescent quantitative PCR experiment, its microRNA-155 level is also notable
Rise further finds specific receptor PROTEIN C sf1r tables in microglia (referring to Fig. 9) by microRNA target prediction and detection
Lowered up to notable (referring to Figure 10).Further detected by luciferase reporter gene and find that Csf1r is microRNA-155's
Target gene (referring to Figure 11), and its protein level is significantly raised (referring to Figure 12) after the processing of microRNA-155 antagonists.This
Outside, microRNA-155 significantly improves the Activated Microglia state of KA processing (referring to Figure 13).We further collect
The microglia conditioned medium of microRNA-155 processing, is acted on Primary cultured neurons, finds microRNA-
The microglia CMC model liquid energy of 155 processing significantly reduces the formation (referring to Figure 14) of neuron action potential, reduces god
Frequency through first excitatory postsynaptic potential (EPSP) (referring to Figure 15) and inhibitory postsynaptic potential (referring to Figure 16).Excitement through member
Property.The studies above is prompted, and microRNA-155 antagonists significantly improve neuronal excitability, is that epilepsy and its relevant disease are fallen ill
Important protectiveness small molecule nucleic acid.Finally, and the blood of 40 clinical epileptic patients and 40 normal healthy controls is collected, detects it
The content of middle microRNA-155, it is found that its content in epileptic also significantly raises.Prompting, it is expected to become clinical insane
The biomarker of epilepsy patient.
By substantial amounts of it is experimentally confirmed that the horizontal significantly rises of microRNA-155 in small Epiletic model and epileptic.
Mouse epilepsy outbreak can be improved by giving the antagonist (antagomir) of mouse intracerebroventricular injection microRNA-155, be prompted
MicroRNA-155 antagonists are antagonism epilepsy and its important protectiveness small molecule nucleic acid of relevant disease morbidity.
The present invention is on the basis of the experiment of a large amount of animals, cell and clinical sample, it is intended to develops and is directed to microRNA-
155 treating and preventing the biological agent of epilepsy.It is specific as follows：
MicroRNA-155 antagonists or inhibitor biological agent：The medicine of preparation can be containing only for microRNA-155
Antisense base sequences or its antagonist/inhibitor and its trim, may be made as a kind of pharmaceutical composition, it is characterised in that
The nucleotide sequence and pharmaceutically acceptable carrier of microRNA-155 antagonists or inhibitor comprising effective dose；Its
In, the carrier can be nano particle, liposome, cholesterol, chitosan, virus etc..Synthesis is for microRNA-155's
Antisense base sequences or its antagonist/inhibitor, and be connected to form medicine with above-mentioned carrier, it can be made into ejection preparation or oral
Preparation, by way of vein or intramuscular injection or oral or snuffing, for treating epilepsy.
The detection of microRNA-155 expressions in 1. epilepsy model hippocampus of mice of embodiment
1) this experiment uses 6~8 week old male C57BL/6 mouse, is specifically divided into 2 groups：Epilepsy model mouse group and control
Group.Epilepsy model mouse group mouse gives intraperitoneal injection digenic acid (KA) 15mg/kg, induces mouse epileptic attack, manufactures mouse
Epilepsy model.
2) after 24hr, traditional extraction mouse bilateral hippocampus RNA：
1ml TRIZOL reagents, electric homogenizer homogenate are added in tissue per 100mg.After placing 5min at room temperature, add
0.2ml chloroforms, room temperature, which is transferred, after mixing puts 5min, and 4 DEG C of 12,000r centrifuge 15min.Upper strata aqueous phase liquid is transferred to new EP pipes
In, isometric (0.3ml) isopropanol is added, room temperature, which is transferred, after mixing puts 10min, and 4 DEG C of 12,000r centrifuge 10min.Abandon supernatant,
75% ethanol of 1ml is added, 4 DEG C of 12,000r centrifugations 10min after mixing.Supernatant is abandoned, precipitation 10min is air-dried, adds 10 μ l
DEPC handles water, dissolves RNA in 55~60 DEG C of water-bath 10min.The RNA solution of acquisition is stored in -80 DEG C.
3) ultraviolet absorption method measure RNA concentration and purity：
Operated according to microplate reader operation instruction, first returned to zero before measurement with the DEPC water of dissolving RNA.According to 260nm
Locate readings and obtain RNA concentration, RNA purity is judged with the ratio of A260/A280.
4) reverse transcription is carried out using the miRNA tailing methods Reverse Transcriptase kit of TIANGEN (Beijing, China)：
Reverse transcription reaction system (10 μ l)：
2×miRNA Buffer Mix 5μl
MiRNA RT Enzyme Mix 1μl
Total RNA (10pg/ μ l~1 μ g/ μ l) 4 μ l
After above system mixes, brief centrifugation, after 42 DEG C are incubated 60min, 95 DEG C are incubated 3min (enzyme inactivation reaction).Stop
Afterwards 4 DEG C it is spare, product is stored in -20 DEG C.
MiRNAs real-time PCR：(PCR reagent derives from ROCHE companies, and the primer such as miR-155, U6 is sharp by Guangzhou
Rich biology Co., Ltd synthesis)
Reaction system (20 μ l)：
Response procedures：
95℃30sec--(95℃3sec--65℃30sec)×40cycles—Melting Curve
The results show：There is bright the normal group mouse of microRNA-155 contents relatively control in epilepsy model mouse cortex and hippocampus
Aobvious rise, illustrates may there is rush epilepsy effect (Fig. 1) in epilepsy mouse microRNA-155.
The expression of 2. Brain Tissue of Patients with Epilepsy microRNA-155 of embodiment
1) 12 epileptics are collected and 11 normal control's Brain Tissue of Patients with Epilepsy samples (is derived from 2013 to 2016
The attached success hospital Inpatients in Neurosurgical ward of year Xiamen University, each case are made a definite diagnosis point by two or more clinicians
Class).RNA is extracted according to 2 the method for embodiment, and detects the expression of microRNA-155.
2) the results show：MicroRNA-155 expressions are significantly raised (Fig. 2) in Brain Tissue of Patients with Epilepsy.Prompting
The expression change of microRNA-155 is closely related with the occurrence and development of epilepsy.
3. mouse electroencephalographic record of embodiment shows specific antagonists (antagomir-155) energy of microRNA-155
Improve mouse epileptic attack
Traced using Nicolet Buddhist nun's high-tensile strength digitlization electroencephalograph, mouse electroencephalogram is traced with scalp electrode, is used
6 leads, respectively place a pin electrode, reference electrode puts a left side on mouse or so volume, left and right temporo and the opposite scalp of left and right top
Ear is simultaneously grounded.Sensitivity：20 μ V/mm, chart speed：20mm/sec, High frequency filter：70HZ, low frequency filtering：0.53Hz.
1) this experiment uses 6~8 week old male C57BL/6 mouse, is specifically divided into 3 groups：KA groups, at antagomir-219
Manage intervention group and negative control group.KA group mouse give intraperitoneal injection KA 15mg/kg and Injected into Cerebral Ventricle antagomir-
155 3 μ l of negative control agent antagomir-NC.Antagomir-155 processing intervention group mouse give intraperitoneal injection KA
15mg/kg, when breaking-out reaches the IV grades of above, by mouse anesthesia, placement is fixed on mouse stereotaxic apparatus, gives side brain
The 3 μ l (200nM) of antagonist (antagomir-155) of room micro-injection microRNA-155.Control group mice gives homonymy side
The 3 μ l of negative control agent antagomir-NC of ventricles of the brain micro-injection antagomir-155.
2) behavior expression of mouse is observed：KA processing mouse show as obvious epileptic attack symptom, and Racine classifications reach
To IV grade.Electroencephalogram shows that intervention group mouse shows spike frequency and substantially reduces (Fig. 3).Control group mice is shown as normally
Behavior, electroencephalogram also epilepsy waveform (Fig. 3) without exception.
It is small that embodiment 4.FJB dyeing shows that the specific antagonists (antagomir-155) of microRNA-155 can improve
Mouse neuronal death
Above-mentioned mouse model makes frozen section, FJB dyeing displays, antagomir-155 processing intervention group hippocampus of mice
Interior neural death situation is obviously improved (Fig. 4) compared with KA groups.
Embodiment 5-7.qRT-PCR shows that the specific antagonists (antagomir-155) of microRNA-155 can improve
The inflammatory reaction such as Mice brain tissues TNF-α, IL-1 β, IL-6
After above-mentioned mouse model 2hr, two groups of Mice brain tissues RNA of traditional extraction, as the step of embodiment 3 detects three respectively
The inflammatory Cytokines Expressions such as the TNF-α in group Mice brain tissues, IL-1 β, IL-6 are horizontal.The results show antagomir-155 processing
Inflammatory Cytokines Expression level substantially reduces (Fig. 5~7) compared with KA groups in intervention group Mice brain tissues.These are the result shows that antagonism mouse
Antagomir-155 expressions in hippocampus can improve mouse epileptic attack, explanation by suppressing inflammatory reaction
MicroRNA-155 plays key effect in the pathogenesis of epilepsy.
Embodiment 8.qRT-PCR shows that miR-155 is mainly expressed in microglia and astroglia
We are separately cultured the main cell in brain tissue, neuron, astroglia and microglia first, lead to
The expression of the different intracellular miR-155 of detection is crossed, it is found that it is mainly expressed in microglia and astroglia (figure
8)。
Embodiment 9.qRT-PCR significantly raises miR-155 expression in original cuiture microglia after showing KA processing
By detect 50 μM of KA handle microglias 2 it is small when, 12 it is small when, it is found that its miR-155 is significantly raised (Fig. 9).
Embodiment 10.qRT-PCR shows that KA handles primary microglia specific proteins CSF1R expression and significantly lowers
After above-mentioned cell model 2hr, 12hr, traditional extraction each group RNA, such as the step of embodiment 1, to detect each group respectively thin
The expression of the miR-155 target genes of intracellular, microglia specific proteins CSF1R expression is notable after finding KA processing
Lower (Figure 10).
Embodiment 11.Luciferase assay show miR-155 Targeted-controls CSF1R
By building 3 ' the UTR wild types and its mutant of CSF1R, transfection microglia cell line BV2Cell, with
The processing of miR-155 activators, finds the selectively targeted regulation and control CSF1R (Figure 11) of miR-155.
Embodiment 12.Western Blot show the small colloid of original cuiture that miR-155 antagonists significantly reverse KA to handle
The downward of CSF1R in cell
1) this experiment carries out microglia original cuiture, point such as the following group using newborn 1-3 days mouse：Normal group
(addition physiological saline)；Epilepsy cell model group (add KA, final concentration of 50 μM)；Epilepsy cell model antagomir-155
Processing intervention group (after addition antagomir-155 makes final concentration of 100nM half an hour, adding KA makes final concentration of 50 μM)；
2) after 12hr, the RNA of traditional extraction each group cell, as the step of embodiment 1 detects the CSF1R of each group cell respectively
Expression.
By detecting the pretreated microglia of miR-155 antagonists, it is found that miR-155 antagonists significantly reverse KA
The downward (Figure 12) of CSF1R in the original cuiture microglia of processing.
Small glue caused by KA is significantly inhibited after the antagonist of microRNA-155 is added in 13. epilepsy cell model of embodiment
The activation of cell plastid
As the step of embodiment 12 handles cell.After 2hr, immunofluorescence dyeing observation Activated Microglia state, hair
The activation (Figure 13) of microglia caused by KA is significantly inhibited after the antagonist of existing microRNA-155.
The microglia conditioned medium of embodiment 14-16.microRNA-155 antagonists processing significantly inhibits nerve
The excitability of member
1. an experiment is carried out microglia original cuiture, is divided such as the following group using newborn 1-3 days mouse：Normal group
(addition physiological saline)；Epilepsy cell model group (add KA, final concentration of 50 μM)；Epilepsy cell model antagomir-155
Processing intervention group (after addition antagomir-155 makes final concentration of 100nM half an hour, adding KA makes final concentration of 50 μM)；
After 2.24hr, the conditioned medium of each group cell is collected, Primary cultured neurons is handled, neuron is detected after 8hr
Action potential and IPSC/EPSC.As a result as shown in Figure 14~16, the microglia bar of antagomir-155 antagonists processing
Part nutrient solution significantly inhibits the granting (Figure 14) of neuron action potential, excitatory postsynaptic potential (EPSP) (Figure 15) and inhibitory synapse
The frequency of current potential (Figure 16) afterwards, prompts antagomir-155 antagonists to influence the excitability of neuron by microglia.
Sequence table
<110>Xiamen University
<120>Curative drug and its application for microRNA-155
<160> 1
<170> SIPOSequenceListing 1.0
<210> 1
<211> 65
<212> RNA
<213>People (Homo sapiens)
<400> 1
cuguuaaugc uaaucgugau agggguuuuu gccuccaacu gacuccuaca uauuagcauu 60
aacag 65
Claims (5)
1. for the curative drug of microRNA-155, it is characterised in that to prevent or treating the medicine or drug regimen of epilepsy
Thing；The medicine is made of effective dose comprising the nucleic acid of sequence and pharmaceutically acceptable carrier or auxiliary material shown in sequence table.
2. the curative drug of microRNA-155 is directed to as claimed in claim 1, it is characterised in that the medicine or medicine group
One in ejection preparation, oral formulations, spray agent, ointment formulation, patch is made according to the common method of medicine preparation for compound
Kind.
3. the curative drug of microRNA-155 is directed to as claimed in claim 1, it is characterised in that the microRNA-155
Nucleotide sequence design anti sense nucleotide sequence and its trim, it is described modify include any nucleotide ribose modify, base
Modification and one or more of combinations or the increase and decrease of any nucleotide, replacement in phosphate backbones modification, as long as the core after modification
Nucleotide sequence still has the activity with microRNA-155 reverse complementals.
4. the curative drug of microRNA-155 is directed to as claimed in claim 1, it is characterised in that the medicine is
MicroRNA-155 antagonists or inhibitor.
5. the purposes of temporal epilepsy is being treated and prevented for the curative drug of microRNA-155 as claimed in claim 1.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201711149052.5A CN107982537A (en) | 2017-11-17 | 2017-11-17 | Curative drug and its application for microRNA-155 |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201711149052.5A CN107982537A (en) | 2017-11-17 | 2017-11-17 | Curative drug and its application for microRNA-155 |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN107982537A true CN107982537A (en) | 2018-05-04 |
Family
ID=62031542
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201711149052.5A Pending CN107982537A (en) | 2017-11-17 | 2017-11-17 | Curative drug and its application for microRNA-155 |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN107982537A (en) |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2013124816A2 (en) * | 2012-02-22 | 2013-08-29 | Brainstem Biotec Ltd. | Generation of neural stem cells and motor neurons |
| WO2015025995A1 (en) * | 2013-08-21 | 2015-02-26 | 서울대학교산학협력단 | Pharmaceutical composition and method for preventing or treating diseases associated with epilepsy or convulsions by targeting microrna |
| CN105163721A (en) * | 2012-05-23 | 2015-12-16 | 俄亥俄州立大学 | Lipid nanoparticle compositions and methods of making and methods of using the same |
-
2017
- 2017-11-17 CN CN201711149052.5A patent/CN107982537A/en active Pending
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2013124816A2 (en) * | 2012-02-22 | 2013-08-29 | Brainstem Biotec Ltd. | Generation of neural stem cells and motor neurons |
| CN105163721A (en) * | 2012-05-23 | 2015-12-16 | 俄亥俄州立大学 | Lipid nanoparticle compositions and methods of making and methods of using the same |
| WO2015025995A1 (en) * | 2013-08-21 | 2015-02-26 | 서울대학교산학협력단 | Pharmaceutical composition and method for preventing or treating diseases associated with epilepsy or convulsions by targeting microrna |
Non-Patent Citations (4)
| Title |
|---|
| HUA TAO ET AL.: "Association of Tag SNPs and Rare CNVs of the MIR155HG/miR-155 Gene with Epilepsy in the Chinese Han Population", 《BIOMED RESEARCH INTERNATIONAL》 * |
| HUAJUN FU ET AL.: "Silencing MicroRNA-155 Attenuates Kainic Acid-Induced Seizure by Inhibiting Microglia Activation", 《NEUROIMMUNOMODULATION》 * |
| MUHAMMAD USMAN ASHHAB ET AL.: "Expressions of Tumor Necrosis Factor Alpha and MicroRNA-155 in Immature Rat Model of Status Epilepticus and Children with Mesial Temporal Lobe Epilepsy", 《J MOL NEUROSCI》 * |
| ZHENGXU CAI ET AL.: "Antagonist Targeting microRNA-155 Protects against Lithium-Pilocarpine-Induced Status Epilepticus in C57BL/6 Mice by Activating Brain-Derived Neurotrophic Factor", 《FRONTIERS IN PHARMACOLOGY》 * |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Strickland et al. | MicroRNA dysregulation following spinal cord contusion: implications for neural plasticity and repair | |
| Ning et al. | microRNAs in spinal cord injury: potential roles and therapeutic implications | |
| Chu-Tan et al. | MicroRNA-124 dysregulation is associated with retinal inflammation and photoreceptor death in the degenerating retina | |
| Chen et al. | MiR‐92b‐3p promotes neurite growth and functional recovery via the PTEN/AKT pathway in acute spinal cord injury | |
| US20090082297A1 (en) | Compositions and Methods for Regulating Gene Expression | |
| Li et al. | Inhibition of miR-21 ameliorates excessive astrocyte activation and promotes axon regeneration following optic nerve crush | |
| US9315812B2 (en) | Methods of using microRNA 195 in providing neuroprotection | |
| CN111035648B (en) | Application of long-chain non-coding RNAGAS5 in preparation of medicine for promoting nerve regeneration and repairing nerve injury | |
| Li et al. | miR‐22‐3p enhances the intrinsic regenerative abilities of primary sensory neurons via the CBL/p‐EGFR/p‐STAT3/GAP43/p‐GAP43 axis | |
| CA2850258A1 (en) | Inhibition of microrna-134 for the treatment of seizure-related disorders and neurologic injuries | |
| CN112716971B (en) | Application of lncRNA XR _595534.2 in preparation of medicines for treating or preventing chronic pain | |
| Campbell et al. | AntimiR targeting of microRNA-134 reduces seizures in a mouse model of Angelman syndrome | |
| Hu et al. | MicroRNA-132 regulates total protein of Nav1. 1 and Nav1. 2 in the hippocampus and cortex of rat with chronic cerebral hypoperfusion | |
| US11497762B2 (en) | MiRNA molecule, equivalent, antagomir, or source thereof for treating and/or diagnosing a condition and/or a disease associated with neuronal deficiency or for neuronal (re)generation | |
| Sierra et al. | The lncRNA Snhg11, a new candidate contributing to neurogenesis, plasticity, and memory deficits in Down syndrome | |
| Ling et al. | A causal relationship between the neurotherapeutic effects of miR182/7a and decreased expression of PRDM5 | |
| CN107982537A (en) | Curative drug and its application for microRNA-155 | |
| CN103656685B (en) | MicroRNA-219 is preparing the application in antiepileptic | |
| Wang et al. | Morphine induces the apoptosis of mouse hippocampal neurons HT-22 through upregulating miR-181-5p | |
| Jeelani | miRNAs in epilepsy: A review from molecular signatures to therapeutic intervention | |
| CN111593114B (en) | Application of miR-122 and inhibitor thereof in prevention/treatment of radioactive brain injury | |
| CN110229879B (en) | PiRNA-500 nucleotide analogue, application of antisense nucleotide thereof and product using same | |
| CN113827729B (en) | Application of PD-1/PD-L1 antagonist in anti-epilepsy | |
| Zhang | Research progress of miRNA in different depression model samples | |
| Tian et al. | Alterations of microRNAs expression profiles in small extracellular vesicle after traumatic brain injury in mice |
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: 20180504 |
|
| RJ01 | Rejection of invention patent application after publication |