CN109722473A - The application of miRNA marker miR-19b relevant to bone metabolic disease - Google Patents
The application of miRNA marker miR-19b relevant to bone metabolic disease Download PDFInfo
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Abstract
The present invention relates to field of biotechnology.MiR-19b is disclosed for the first time to play an important role in osteoblast differentiation, can be used as marker for screening the drug of prevention or treatment bone metabolism related disease (including osteoporosis, osteoproliferation, Osteoblast Differentiation exception, bone loss etc.).
Description
Technical field
The present invention relates to field of biotechnology.More specifically, it is related to a kind of miRNA mark relevant to bone metabolic disease
The application of will object miR-19b.
Background technique
Osteoporosis (Osteoporosis) is reduced with bone amount, and bone tissue microstructure is impaired, and bone trabecula quantity subtracts
Few, bone brittleness increases and a kind of raised disease of general bone metabolism obstacle of risk of fractures degree.Osteoporosis is also that one kind is moved back
The property changed disease, with age, risk increases.The drug for the treatment of osteoporosis mainly includes anti-bone resorption drug at present
With promoting bone growing drug, such as Diphosphonate, calcitonin, estrogen receptor alternative medicine and parathyroid hormone.But these
Drug all has certain side effect, and the generation that may cause sudden fracture, breast cancer and myeloma is used for a long time, thus is facing
Application on bed all has significant limitation.
The formation of osteoporosis is mainly since the responsible ostosis of osteoblast during bone remoulding and osteoclast are negative
The bone resorption disequilibrium of duty leads to the degeneration of bone density decline and bone microstructure, to easily cause fracture.At present for controlling
Treating osteoporosis mainly includes anti-bone resorption and anti-anabolism class drug.Anti- bone resorption drug, which mainly passes through, inhibits osteoclastic thin
Cytoactive and function to reduce the speed of bone amount reduction, this kind of drug include: diphosphonate, estrogen replacement therapy (HRT),
Selective estrogen receptor modulators (SERMs) and calcitonin, but this kind of drug can reduce the rate of bone remoulding, and clinical evidence is aobvious
Show, diphosphonate, which is used for a long time, can lead to sudden fracture.Osteoporosis in post-menopause women disease incidence is high, mainly thinks at present
It is since a large amount of declines of estrogen level cause bone remoulding to accelerate, bone converts disequilibrium and bone amount is caused to reduce.Therefore, with
Estrogen receptor is that the treatment of target spot includes HRT, SERMs, but this kind of drug has and increases the risk that female patient obtains breast cancer.
The major limitation of anti-bone resorption drug is that it cannot rebuild the bone structure lost, and increases the formation of new bone.
The currently the only anti-anabolism class drug for having obtained U.S. FDA approval is the parathormone of promoting bone growing
(Parathyroidhormone, PTH), PTH are to adjust alcium and phosphor metabolization, maintain the major hormone of body calcium balance, PTH is to bone
On the one hand effect is the number and vigor for increasing osteoclast, promote bone resorption, discharges Ca2+、P3+；On the other hand be promote at
The proliferation and activity of osteocyte simultaneously discharge bone growth factor, thus promoting bone growing.But PTH can increase myeloma incidence,
Clinically only use 2 years, at the same clinical data show it is effective to 60% treatment object.It is another kind of just under study for action
Be Bone formation drug using mature osteocytes as target cell, the Humanized monoclonal of anti-osteosclerosis albumen (sclerostin)
Antibody, currently the sufferers of osteoporosis face of the interim postmenopausal women of clinic Ш verifies its effectively and safely property.In addition, with skeletonization
Cell and osteoclast are target cell, further include using cathepsin K, calcium-sensing receptor and RANK receptor as the inhibitor of target spot
Or the statins that monoclonal antibody and promotion BMP-2 are expressed.But these drugs have its limitation and side effect respectively.
With the fast development of regenerative medicine research field in recent years, stroma stem cell answering in osteanagenesis and Bone Defect Repari
With showing huge potentiality.Stroma stem cell (MSCs) is primarily present a kind of adult stem cell in marrow, has certainly
My updating ability and Multidirectional Differentiation ability, can break up as osteoblast, fat cell, myocyte etc., and have in vitro
Immunoregulation capability becomes research hotspot both domestic and external in recent years.Suffered by opposite embryonic stem cell and inductivity stem cell
Ethics limitation, stroma stem cell can be from the limitations of ethics problem as adult stem cell transplanting.Meanwhile stroma stem cell itself
MHCI and II expression is very low, and does not express CD40, CD80 and the CD86 of activation T cell, has immunoregulation capability, therefore have
Huge potential applicability in clinical practice.Currently, stroma stem cell is a variety of in osteogenesis imperfecta, graft versus host disease(GVH disease), angiocarpy etc.
Disease is clinically applied.However, although recent domestic carries out the cell and molecular mechanism of stroma stem cell
A large amount of research, but some important problem in science are also urgently studied, source, self-renewing including stroma stem cell are (dry
Property maintain) mechanism, raise transfer and its molecular mechanism of directed differentiation etc. between osteocyte-fat cell, therefore study
The cell and molecular mechanism of stroma stem cell self-renewing and differentiation find the novel targets of osteanagenesis and Bone Defect Repari with important
Meaning will provide important theoretical foundation for the application of stroma stem cell clinically.
Currently, self and heteroplastic transplantation MSC regenerative therapy strategy is respectively in nerve degenerative disease, degeneration heart
A large amount of research has been carried out in the loss reparation of disease and bone tissue.Research finds a kind of new compound LLP2A-ALE, can be with
Promote the migration of MSC and MSC is promoted to break up to osteoblast direction, the osteoporosis mouse caused by estrogen deficiency and aging
In model, LLP2A-ALE significantly improves bone amount reduction, it has been found that the both ends of this kind of compound pass through respectively in conjunction in bone structure
4 β 1 of hydroxyapatite and MSCs surface integrin α, promote MSC to bone surface migrate, while increase intracellular Akt swash
Enzyme phosphorylation, promotes it to osteoblast differentiation, and correlative study result is published in Nature Medicine.
No matter any source, MSC has three kinds of fundamental characteristics as stem cell: in undifferentiated state；It can be with
Self-renewing；With multipotency differentiation property, certain types of cell can produce.These characteristics are referred to broadly as " stemness ".With body
The decline of astogeny cell function, the quantity of MSC is significantly reduced, and then reduces Osteoblast Differentiation and new bone.Therefore, researchers
Be actually to stem cell how permanent its stemness of holding it is very interested.LIF ELISA (Leukemia
Inhibitory factor, LIF), fibroblast growth factor (fibroblast growth factor, FGFs), Wnts
The cell factors such as (mammalian homologues of Drosophila), TGF β were once considered the tool of the self-renewing with cell
There is stronger correlation.LIF activates the STAT3 in JAK-STAT access, and the activation of STAT3 be once considered as cell self
The necessary condition of update.FGFs is also proved related to MSC self-renewing.2003, Yamanaka study group find one
The albumen of mouse embryo stem cell and the gene Nanog gene for transplanting single-minded expression in preceding embryo, gene compiling is a kind of tool
There is the homeobox transcription factor for hindering differentiation, for maintaining versatility of mouse embryo stem cell to play critically important work
With.The holding STAT3 approach of stem cell versatility of confirmation is different from early stage, and Nanog's acts on another independent holding
The signal transduction pathway of stem cell versatility.2006, Takahashi and Yamanaka had found four kinds of transcription factors again:
Oct3/4, Sox2, c-Myc and Klf4 can be formed with inducing adult fibroblast has multipotent stem cells (iPSCs).
2007 the study found that OCT4, Sox2 and Nanog also have expression, the base of OCT4 regulation on the MSC of derived from bone marrow
MSCs cell cycle, with increasing for passage number, OCT4 and end is adjusted similar to embryonic stem cell in the target gene of matter stem cell
The expression quantity of granzyme (telomerase) is significantly reduced.There is research to claim recently, height expression MSCs cell membrane surface integral protein
CD49f, which can regulate and control OCT4 and SOX2 by activation PI3K/AKT signal path, to express, and then maintains the stemness of MSCs.Separately grind
Study carefully the expression pattern analysis by MSCs genome, discovery Klf4 is keeping its stemness, maintaining the undifferentiated state of MSCs to have important
Effect.It is another studies have reported that MSCs expands the characteristic for being lost stem cell after several generations.And pass through transduction SOX2 Nanog base
Because after, the proliferation and differentiation potential of MSC can be maintained well.2012 current research report Nanog can overcome MSCs aging,
The ability replying the cell Proliferation of the MSCs of aging and breaking up to myocyte.It can be seen that, MSC similar with embryonic stem cell
In the transcription factor expressions level such as OCT4, Nanog, SOX2, Klf4 and TERT height and undifferentiated state be positively correlated, mention
Show that they can be used as the undifferentiated mark of MSC and target spot.
In recent years, microRNA (miRNA) is gradually known, and miRNA is sent out in cell differentiation, biological development and disease
It plays a great role during hair tonic exhibition, causes more and more to study concern.MiRNA is had found by Lee etc. at first, is a kind of
Containing loop-stem structure, the tiny RNA molecule (21~23 nucleotide) of non-protein coding is similar to siRNA.MiRNA base
Because being similar to many encoding egg white genes, transcribed usually in nucleus by RNA polymerase II, initial product has for big
The Microrna precursor (pri-miRNA) of 5 ' cap sequences and 3 ' poly-A tails.Pri-miRNA in nuclease Drosha and
Under the action of its confactor Pasha, 5 ' cap sequences and 3 ' poly-A tails are sheared, and become about 70 nucleotide compositions
pre-miRNA.Pre-miRNA is transported in cytoplasm by RAN-GTP and transport protein (exportin).Then, another core
Sour enzyme Dicer shears its loop-stem structure, generates the miRNA:miRNA* double-strand of about 22 length of nucleotides.Dicer will be at
After ripe RNA double-strand release, RNA- albumen composition is formed by a histone reaction bonded quickly and is referred to as miRNA induction silencing
Compound.In miRNA induction silencing complex, a mature single-stranded miRNA is retained in this complex, it can be identified
The binding site of 3 ' end noncoding regions on said target mrna.MiRNA with target gene mRNA base pairing by drawing after processing is mature
It leads silencing complex degradation mRNA or hinders its translation, there is the transcription product for destroying desired specificities gene or induction translation
The function of inhibition.MiRNA and RNA- protein complex wide expression in animal and plant.MiRNAs is only at specific group
It knits and is expressed with the stage of development, there is tissue specificity and timing, decide the special adjustment process of the function of tissue and cell
In play an important role.
The development of the embryonic period, embryonic phase of miRNAs and bone, adjusts stem cell osteogenic action and chondrocyte metabolic and osteoclast closes
System is significant.MiRNA adjusts the differentiation of people's bone marrow pluripotent stem cell and the expression of LIF ELISA；When to osteoblast differentiation
When, 19 kinds of miRNA expression up-regulations；When to Adipocyte Differentiation, 20 kinds of miRNA expressions up-regulations；Hsa-mir199a and
The secretion of hsa-mir346 inhibition LIF ELISA.Studies have found that miR-29b can directly lower histone deacetylation
The albumen production quantities such as enzyme, transforming growth factor β 3, II A type activin A receptor, these albumen inhibit as osteoblast differentiation
Object, expression reduction may advantageously facilitate osteoblast differentiation.Meanwhile miR-29b reduces COL1A1, COL5A3, COL4A2
3 end non-coding area sequence activity, to reduce or inhibit the synthesis of collagen.When collagen secretion reaches a stationary phase,
MiR-29b concentration increases, and inhibits the synthesis of collagen, promotes sclerotin mineralising, is conducive to sclerotin and generates.MiR-223 passes through adjusting
NFI-A and macrophage colony-stimulating factor receptor are horizontal, and reach and the differentiation of osteoclast is adjusted.
Studies have reported that miR-194-5p, miR-21, miR-23a, miR-24, miR-25, miR-100 and miR-125b
It is the biomarker closely related with osteoporosis.It is other that there are also much miRNAss related with osteoporosis.Xiao P etc.
MiR-422a and miR-133a in people's discovery osteocyte can also be used as the biomarker of low bone density sufferers of osteoporosis face.
MiR-133a and miR-21 and menopausal women osteoporosis have substantial connection.MiRNAs is by acting on its coherent signal
Access has an impact process of osteoporosis.MiR-29a is proved that, by adjusting Runx2 promotion Osteoblast Differentiation, research is found
MiR-20a also has similar mechanism.Runx2 is the action target spot of a variety of miRNA as participation gene important in bone metabolism,
It further include miR-204, miR-705, miR-103 etc..MiRNA can also participate in bone metabolism mistake by influencing the level of BMP-2
The adjusting of journey, including miR-34c, miR-17, miR-106 etc..In addition, miRNA can also influence the work of osteoclast by RANK
Property, as MiR-503 can inhibit osteoclast to generate.To sum up, miRNA plays an important role during bone metabolism.
And certain miRNAs in serum (or blood plasma) have been studied confirmation and can be used as some diseases, as cancer,
The biomarker of diabetes, osteoporosis and osteoarthritis is for diagnosing and preventing.Serum miRNA can as cancer or its
The serum biomarkers of his disease, significance are to provide a kind of diagnostic means of non-damage.The reality of existing maturation
Room technology is tested, the early detection system based on miRNA can be quickly set up.
Therefore, in conjunction with the advantage of multinomial cutting edge technology, using miRNAs and bone substantial connection and early changes advantage,
MiRNAs is verified to the regulating and controlling effect of Osteoblast Differentiation, the treatment to osteoporosis is of great significance.
Summary of the invention
Therefore, for above-mentioned problems in the prior art, miR-19b is applied the purpose of the present invention is to provide a kind of
Assistant regulating and controlling osteoblast differentiation simultaneously utilizes miR-19b screening prevention or the method for the potential substance for the treatment of bone metabolic disease.
In order to achieve the goal above, one aspect of the present invention provides a kind of miRNA mark relevant to bone metabolic disease
The purposes of object miR-19b, which is characterized in that for screening the drug of prevention or treatment bone metabolic disease.
The bone metabolic disease includes osteoporosis, osteoproliferation, Osteoblast Differentiation exception, bone loss.
According to another aspect of the present invention, the promotor of miR-19b a kind of is provided in preparation prevention or prevention and treatment bone metabolism disease
Purposes in the drug of disease.
According to another aspect of the invention, the side of a kind of screening prevention or the potential substance for treating bone metabolic disease is provided
Method, which comprises
The system of expression miR-19b is handled with candidate substances；With
Detect the expression of miR-19b in the system；
Wherein, if the candidate substances promote the expression of miR-19b, show that the candidate substances are prevention or prevention and treatment bone generation
Thank to the potential substance of disease.
Further, ALP is also expressed in the system, the method also includes: detect the expression of ALP in the system；
Wherein, if the candidate substances increase the expression of ALP, show that the candidate substances are prevention or prevention and treatment bone metabolism disease
The potential substance of disease.
Further, Runx2 is also expressed in the system, the method also includes: detect the table of Runx2 in the system
It reaches；
Wherein, if the candidate substances increase the expression of Runx2, show that the candidate substances are prevention or prevention and treatment bone metabolism
The potential substance of disease.
Further, OCN is also expressed in the system, the method also includes: detect the expression of OCN in the system；
Wherein, if the candidate substances increase the expression of OCN, show that the candidate substances are prevention or prevention and treatment bone metabolism disease
The potential substance of disease.
Beneficial effect
Compared to conventional clinical method, the present invention, by serum/plasma miRNA serum Regulate Osteoblast Differentiation, for screening
The potential substance of prevention or treatment bone metabolic disease, can provide more effectively for the treatment of the bone metabolic diseases such as osteoporosis
Method.Blood plasma miRNA in the circulating cycle can in conjunction with lipoprotein, or with excretion body wrap up form, in soda acid, Frozen-thawed cycled process
Middle holding homeostasis, guarantees the accuracy of result.Blood plasma miRNA is easier to obtain as biomarker, and dosage it is low,
It is also stable and easy to detect simultaneously and quantitative accurate, the potential substance of screening prevention or treatment bone metabolic disease can be greatlyd improve
Validity.
Detailed description of the invention
From detailed description with reference to the accompanying drawing, it will be more clearly understood above-mentioned and other purposes of the invention,
Feature and other advantages, wherein
Fig. 1 shows the signal that mature miRNA is selectively converted to cDNA in miScript HiSpec Buffer
Figure；
Fig. 2 shows miR-19b expressions in hMSC atomization；
Fig. 3 shows miR-19b expression in MC3T3-E1 atomization；
Fig. 4 shows the expression figure that miR-19b adjusts hMSC Osteoblast Differentiation；
Fig. 5 shows the expression figure that miR-19b adjusts MC3T3-E1 Osteoblast Differentiation.
Specific embodiment
In order to make the objectives, technical solutions, and advantages of the present invention clearer, below in conjunction with attached drawing and specific implementation
Example, the present invention will be described in further detail.It should be appreciated that specific embodiment described herein is only to explain this hair
It is bright, it is not intended to limit the present invention.
The present invention utilizes regulation of the miRNAs for osteoblast physiology course, verifies miR-19b to the tune of Osteoblast Differentiation
Control effect, new thinking is provided for osteoporosis treatment.
Embodiment
The expression of 1 MiRNA osteoblast in vitro
Cell culture and transfection
1) hMSC and MC3T3-E1 plating cells (α-MEM complete medium when bed board containing 10%FBS).In 6cm plate
The density normally cultivated about 85-95%.12-18h after bed board, addition 10nM dexamethasone is added in culture medium, and 50 μ g/ml are anti-bad
The beta-glycerophosphate of hematic acid and 5mM are to Osteoblast Differentiation.Then a cell is collected within every 7 days until the 21st day for analyzing spy
Determine the expression of miRNA.
2) it transfects: being used before transfection and add 10nM dexamethasone, the beta-glycerophosphate of 50 μ g/ml ascorbic acid and 5mM
Culture medium changes liquid.It takes two 1.5ml to manage without RNA enzyme EP, is separately added into 120ul 1 × riboFECTTM CP Buffer, 10 μ l
MiRNA mimic/inhibitor, 12 μ l riboFECTTM CP reagent are mixed gently, and are incubated at room temperature 0-15min, are added
Make its final concentration of 200nM of miRNA mimic/inhibitor in orifice plate.In the miRNA mimic/ that third day more renews
Inhibitor and culture medium transfect 6 days, cell are collected, for analyzing the expression quantity for analyzing Alp, OCN and Runx2.
The extraction of 2 MiRNA
Using QIAGEN RNeasy Mini Kit kit, say that cell sample is dissolved in suitable QIAzol Lysis
In Reagent.After chloroform is added, after mixing, it is stored at room temperature 5min.It is added and the isometric chloroform of sample, vortex 15min, room temperature
Stand 2-3min.12000 turns of centrifuge speed of adjustment, is centrifuged 15 minutes by 4 DEG C of operating temperature.It is divided into water phase after lysate centrifugation
And organic phase.RNA is located at upper strata aqueous phase, and DNA is located at interphase, and albumen is located at lower layer's organic phase or interphase.
After extracting upper strata aqueous phase, 100% ethyl alcohol that 1.5 times of volumes are added is mixed, and ethyl alcohol can provide the RNA conjunction of combination
Suitable condition, RNA molecule are greater than 18 nucleotide.Sample adds RNeasy MinElute centrifugal column, and total serum IgE is integrated to film
On, phenol and other pollutants are effectively washed away.High-quality RNA is free of in RNase water in small size and is eluted.Serum and blood plasma
In mainly contain tiny RNA s, there is no need to separated purifying tiny RNAs and big RNA component.
The preparation of 3 cDNA
The RNA extracted is converted into cDNA in case subsequent detection using QIAGEN miScript II RT Kit.Fig. 1
Show the schematic diagram that mature miRNA is selectively converted to cDNA in miScript HiSpec Buffer.It is mature
MiRNA is a kind of non-coding RNA that is natural, being about 22 bases.They regulate and control gene expression after transcription.With
MRNA is different, and miRNA is not polyadenylic acid.Mature miRNA by poly (A) polymerize enzymatic synthesis polyadenylic acid, then by
Oligo-dT primer reverse transcription is at cDNA.Polyadenylic acid synthesis and reverse transcription reaction carry out simultaneously in same test tube.
The base of several degeneracys is contained at 3 ' ends of oligo-dT primer, and 5 ' ends have a common tags, can be used for real-time PCR reaction
When expand maturation miRNA.The perfect combination of polyadenylic acid and common tags, which can ensure that, will not detect genomic DNA.
1) melt RNA template on ice.Melt the water without RNase, 10x miScript Nucleics Mix and 5x in room temperature
miScript HiSpec Buffer。
It is touched with hand and mixes solution, the heavy collection of solution is arrived test tube bottom by of short duration centrifugation, is placed on stand-by on ice.
2) reverse transcription reaction solution is prepared according to following table.
Reaction system is as follows:
Note: miScript Reverse Transcriptase Mix should take out from -20 DEG C of refrigerator before the reaction.
It mixes gently, is placed on stand-by on ice.After being finished, it should put back to immediately in -20 DEG C of refrigerator.The volume of standby reaction solution should exceed
The 10% of required volume.
3) RNA template is added in the test tube of the solution containing reverse transcription reaction.It mixes gently, then of short duration centrifugation is placed on ice
Upper preservation.
4) it is reacted 60 minutes at 37 DEG C.
5) 5 minutes are reacted at 95 DEG C to inhibit reverse transcription reaction Solution Active, then puts and is kept on ice.
6) cDNA is diluted with the pure water of no RNase, carries out real-time PCR reactions immediately
4 use the express spectra of real-time PCR analysis maturation miRNA
1) in room temperature (15-25 DEG C) defrosting 2x QuantiTect SYBR Green PCRMaster Mix, 10x
MiScript Universal Primer, Cdna primer, and the pure water without Rnase, mix each solution.
2) reaction solution is configured according to following table.It mixes gently.
3) PCR program is set according to following table.
Start PCR program.
Fig. 2 and 3 shows hMSC and MC3T3-E1 to during osteoblast differentiation, and the expression of miR-19b is at any time
Between the figure that changes, as shown, with the extension of time, the progress of cell osteogenic differentiation process, miR-19b gradually rise,
After differentiation 14 days, the expression of miR-19b tends towards stability, and changes with time just smaller；Figure 4 and 5 show miR-
19b adjusts the expression figure of hMSC Osteoblast Differentiation and the expression figure of miR-19b adjusting MC3T3-E1 Osteoblast Differentiation.By scheming
4 as can be seen that transfection miR-19b promotor (mimic) can promote skeletonization marker gene in hMSC cell differentiation procedure
The expression of ALP, Runx2 and OCN；Transfection miR-19b inhibitor (inhibitor) can inhibit skeletonization marker gene ALP,
The expression of Runx2 and OCN.As seen from Figure 5, in MC3T3-E1 cell differentiation procedure, miR-19b promotor is transfected
(mimic) expression of skeletonization marker gene ALP, Runx2 and OCN can be promoted；It transfects miR-19b inhibitor (inhibitor)
It can inhibit the expression of skeletonization marker gene ALP, Runx2 and OCN.
Based on the above discovery, miRNA marker miR-19b can be used for screening prevention or treat the medicine of bone metabolic disease
Object.Specifically, the method for the potential substance of screening prevention or treatment bone metabolic disease may include: and be handled to express with candidate substances
The system of miR-19b；With
Detect the expression of miR-19b in the system；
Wherein, if the candidate substances promote the expression of miR-19b, show that the candidate substances are prevention or prevention and treatment bone generation
Thank to the potential substance of disease.
Or also can detecte the expression of skeletonization marker gene ALP, Runx2 and OCN in system, if the candidate substances increase
Add the expression of ALP, Runx2 and OCN, then shows that the candidate substances are the potential substances of prevention or prevention and treatment bone metabolic disease.
The promotor of miR-19b can be used for preparing prevention or prevent and treat the drug of bone metabolic disease.
The foregoing is merely better embodiments of the invention, are not intended to limit the invention, all of the invention
Made any modifications, equivalent replacements, and improvements etc., should all be included in the protection scope of the present invention within spirit and principle.
Claims (7)
1. a kind of purposes of miRNA marker miR-19b relevant to bone metabolic disease, which is characterized in that for screening prevention
Or the drug for the treatment of bone metabolic disease.
2. the purposes of miRNA marker miR-19b relevant to bone metabolic disease as described in claim 1, which is characterized in that
The bone metabolic disease includes osteoporosis, osteoproliferation, Osteoblast Differentiation exception, bone loss.
3. a kind of purposes of promotor of miR-19b in the drug of preparation prevention or prevention and treatment bone metabolic disease.
4. a kind of method of the potential substance of screening prevention or treatment bone metabolic disease, which comprises
The system of expression miR-19b is handled with candidate substances；With
Detect the expression of miR-19b in the system；
Wherein, if the candidate substances promote the expression of miR-19b, show that the candidate substances are prevention or prevention and treatment bone metabolism disease
The potential substance of disease.
5. method as claimed in claim 4, which is characterized in that ALP is also expressed in the system, the method also includes: inspection
Survey the expression of ALP in the system；
Wherein, if the candidate substances increase the expression of ALP, show that the candidate substances are prevention or prevention and treatment bone metabolic disease
Potential substance.
6. method as claimed in claim 4, which is characterized in that Runx2 is also expressed in the system, the method also includes:
Detect the expression of Runx2 in the system；
Wherein, if the candidate substances increase the expression of Runx2, show that the candidate substances are prevention or prevention and treatment bone metabolic disease
Potential substance.
7. method as claimed in claim 4, which is characterized in that OCN is also expressed in the system, the method also includes: inspection
Survey the expression of OCN in the system；
Wherein, if the candidate substances increase the expression of OCN, show that the candidate substances are prevention or prevention and treatment bone metabolic disease
Potential substance.
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110106249A (en) * | 2019-05-09 | 2019-08-09 | 深圳市人民医院 | Application of the long-chain non-coding RNA NONHSAG039951.2 as diagnosing osteoporosis marker |
| CN110157794A (en) * | 2019-05-09 | 2019-08-23 | 深圳市人民医院 | Application of the long-chain non-coding RNA NONHSAT005760.2 as diagnosing osteoporosis marker |
| CN112143697A (en) * | 2020-10-08 | 2020-12-29 | 北京广未生物科技有限公司 | Method for promoting proliferation and differentiation of embryonic stem cells |
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