CN107604058A - The application of the nucleotide analogs of piRNA 514 and its GEM 132 and apply its product - Google Patents
The application of the nucleotide analogs of piRNA 514 and its GEM 132 and apply its product Download PDFInfo
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- CN107604058A CN107604058A CN201710866272.3A CN201710866272A CN107604058A CN 107604058 A CN107604058 A CN 107604058A CN 201710866272 A CN201710866272 A CN 201710866272A CN 107604058 A CN107604058 A CN 107604058A
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Abstract
The invention provides the application of a kind of nucleotide analogs of piRNA 514 and its GEM 132 and its product is applied, is related to the technical field of engineering in medicine.The application of the nucleotide analogs of piRNA 514 provided by the invention or its bioactive functions fragment or variant in preparing for the product of diagnosis and/or prognosis evaluation heart disease, by detecting the expression of the nucleotides of piRNA 514 in peripheral blood blood plasma, there is the potential value of diagnosis and/or prognosis evaluation to many heart diseases；Also, the application in being used to prevent and/or treat the product of heart disease is being prepared by the GEM 132s of piRNA 514 provided by the invention, the prevention and/or treatment to many heart diseases provide basis.
Description
Technical field
The present invention relates to engineering in medicine technical field, more particularly, to a kind of piRNA-514 nucleotide analogs and its instead
The application of adopted nucleotides and apply its product.
Background technology
World Health Organization's issue《Global disease burden》Report points out that it is dead that angiocardiopathy has become global human
The main reason for dying.According to China's statistics of 2014, the number of angiocardiopathy is suffered from more than 2.7 hundred million by China, and annual 300
Ten thousand people die from angiocardiopathy.Angiocardiopathy significantly raises in the morbidity and mortality of mid-aged population, therefore, in face of me
The present situation of state's aging population, carrying out pathogenesis and clinical research to heart disease has important theory and practice meaning.
Myocardial hypertrophy is that cardiac muscle cell is directed to a kind of increased responsing reaction of Hemodynamics, is usually expressed as tissue level
Thicken and the cellular level of upper cardiac muscular tissue set one's heart the increase of myocyte's volume, are many angiocardiopathies (hypertension, acute hearts
Muscle infarction, congenital heart disease etc.) pathological change that shares during occurrence and development.Induce the stimulus master of myocardial hypertrophy
To include mechanical tension and a variety of neurohumor factors, such as Angiotensin II (Angiotensin II, Ang II), isopropyl kidney
Upper parathyrine (Isoproteronol, Iso), insulin-like growth factor-i (IGF-1), il-1 (IL-1) etc..
PiRNA (piwi-interacting RNA) is after siRNA and the class tiny RNAs of miRNA two, is existed first within 2006
A kind of new non-coding tiny RNA found in male sex-cell, length are generally phonetic with strong urine at 26~31nt, 5 ' ends
Pyridine skewed popularity, species is more than 50,000.
Past research was once once thinking that piRNA was that reproduction cell institute is peculiar, therefore focused primarily upon in reproduction cell
PiRNA functional study, existing result of study show that piRNA is primarily involved in the development of reproduction cell, the silence, different of transposons
The processes such as chromatinic formation and the maintenance of reproduction cell DNA integralities.2012, in the research hair that Cell magazines are delivered
Show the piRNA expressed in sea hare brain, regulate and control the memory of nerve cell by regulating and controlling the DNA methylation of CREB albumen
Function.Thus, researcher has been attracted to carry out the interest of functional study in non-germ cells for piRNA.
At present, the crucial piRNA in the heart disease such as cardiac hypertrophy and myocardial fibrosis there is no report now, and it is sent out
The function of waving is the challenge to field scientific research personnel.
In view of this, it is special to propose the present invention.
The content of the invention
First purpose of the present invention is to provide miRNA-485-5p nucleotide analogs or its bioactive functions piece
The application of section or variant in preparing for the product of diagnosis and/or prognosis evaluation heart disease；Second object of the present invention
It is to provide a kind of medicine for being used for diagnosis and/or prognosis evaluation heart disease；Third object of the present invention is to provide one
Kind is for diagnosis and/or the kit of prognosis evaluation heart disease；Fourth object of the present invention is to provide miRNA-485-
5p GEM 132s are preparing the application in being used to prevent and/or treat the product of heart disease；The 5th purpose of the present invention
It is to provide a kind of medicine for being used to preventing and/or treating heart disease, to alleviate present in prior art in heart fertilizer
Crucial piRNA researchs in the heart diseases such as thick and myocardial fibrosis still belong to the technical problem of blank.
The invention provides piRNA-514 nucleotide analogs or its bioactive functions fragment or variant to be used in preparation
Application in the product of diagnosis and/or prognosis evaluation heart disease；
The piRNA-514 nucleotide analogs contain the sequence shown in SEQ ID NO.1.
Further, the product is medicine or kit.
Further, each base has carried out chemistry in the nucleotide sequence of the piRNA-514 nucleotide analogs
Modification.
Present invention also offers a kind of medicine for being used for diagnosis and/or prognosis evaluation heart disease, the medicine includes
Imitate the piRNA-514 nucleotide analogs or its bioactive functions fragment or variant of dosage.
Present invention also offers a kind of kit for being used for diagnosis and/or prognosis evaluation heart disease, the kit bag
Include the primer for specific detection piRNA-514 nucleotide analogs.
Present invention also offers piRNA-514 GEM 132s to prepare the production for preventing and/or treating heart disease
Application in product；
The piRNA-514 GEM 132s contain the sequence shown in SEQ ID NO.2.
Further, the product is medicine.
In addition, present invention also offers a kind of medicine for being used to preventing and/or treating heart disease, the medicine includes
Imitate the piRNA-514 GEM 132s of dosage.
Further, the administering mode of the medicine includes oral administration or drug administration by injection.
Further, the heart disease includes myocardial hypertrophy, myocardial fibrosis, coronary heart disease, myocarditis, heart valve disease
The one or more of disease, hypertension and heart failure.
PiRNA-514 nucleotide analogs provided by the invention are found through experiments that, are expressed in the cardiac muscle cell of hypertrophy
Significantly lower, the myocardial cell surface product increase induced by Iso can be significantly inhibited by being overexpressed piRNA-514.It is described above,
PiRNA-514 can suppress the generation of myocardial hypertrophy.This means piRNA-514 can be used as a kind of early diagnosis and early prevention
The biomarker of the heart diseases such as myocardial hypertrophy, myocardial fibrosis.Therefore, by detecting piRNA-514 in peripheral blood blood plasma
The expression of nucleotides, for having the potential value of diagnosis and/or prognosis evaluation to many heart diseases.
Brief description of the drawings
Fig. 1 is that the myocardial hypertrophy disease model using isoprel (Iso) induction that the embodiment of the present invention 1 provides is small
In the heart of mouse and control mice, the result of variations figure of piRNA-514 expressions；
Fig. 2 is the cardiac muscle cell that the cardiomegaly that the embodiment of the present invention 2 provides stimulates and isoprel (Iso) induces
The result of variations figure of loose process piRNA-514 expressions；
To cardiac muscle after the primary cardiomyocytes transfection piRNA-514 nucleotide analogs that Fig. 3 provides for the embodiment of the present invention 3
The influence result figure of cell surface product.
Embodiment
Technical scheme is clearly and completely described below in conjunction with accompanying drawing, it is clear that described implementation
Example is part of the embodiment of the present invention, rather than whole embodiments.Based on the embodiment in the present invention, ordinary skill
The every other embodiment that personnel are obtained under the premise of creative work is not made, belongs to the scope of protection of the invention.
The invention provides piRNA-514 nucleotide analogs or its bioactive functions fragment or variant to be used in preparation
Application in the product of diagnosis and/or prognosis evaluation heart disease, following (the SEQ ID of sequence of piRNA-514 analog thuja acids
NO.1)：
5’-AAUCAGAUUUGCACCACUCAUCUUGGUGA-3’。
Wherein, the bioactive functions fragment or variant of piRNA-514 nucleotide analogs, such as can be, but it is unlimited
In following (1)-(3)：
(1) recombinant vector of the encoding gene containing piRNA-514 nucleotide analogs；
(2) recombinant virus of the encoding gene containing piRNA-514 nucleotide analogs；
(3) recombinant viral vector of the encoding gene containing piRNA-514 nucleotide analogs.
In the present invention, product is medicine or kit.
In the present invention, each base has carried out chemistry and repaiied in the nucleotide sequence of piRNA-514 nucleotide analogs
Decorations.
Wherein, chemical modification for example can be, but be not limited to ribose modification, base modification or phosphate backbones modification.
In one preferred embodiment, each base is equal in the nucleotide sequence of piRNA-514 nucleotide analogs
The modification of 2 '-methoxyl group is carried out.
It is used to diagnose and/or the medicine of prognosis evaluation heart disease present invention also offers a kind of, including effective dose
PiRNA-514 nucleotide analogs or its bioactive functions fragment or variant.
Present invention also offers a kind of kit for being used for diagnosis and/or prognosis evaluation heart disease, including for special
Property detection piRNA-514 nucleotide analogs probe or primer.
Wherein, the primer sequence for specific detection piRNA-514 nucleotide analogs is：
Reverse transcription primer：
5’-CTCAACTGGTGTCGTGGAGTCGGCAATTCAGTTGAGTCACCAA G-3’(SEQ ID NO.3)；
Sense primer：5’-CGGGCAATCAGATTTGCACCACTCATCT-3’(SEQ ID NO.4)；
Anti-sense primer：5’-TGGTGTCGTGGAGTCG-3’(SEQ ID NO.5).
To be prepared in addition, present invention also offers piRNA-514 GEM 132s for preventing and/or treating heart disease
Product in application, the sequences of piRNA-514 GEM 132s is following (SEQ ID NO.2)：
5’-UCACCAAGAUGAGUGGUGCAAAUCUGAUU-3’。
In the present invention, product is medicine.
It is used to preventing and/or treat the medicine of heart disease present invention also offers a kind of, including effective dose is above-mentioned
PiRNA-514 GEM 132s.
In the present invention, for prevent and/or treat the medicine of heart disease also include pharmaceutically acceptable supporting agent or
Auxiliary material.
Wherein, supporting agent or auxiliary material for example can be, but be not limited to chitosan, cholesterol, liposome, cyclodextrin, microballoon, micro-
One or more in capsule.
In the present invention, for the administering mode for the medicine for preventing and/or treating heart disease, including it is administered orally or notes
Penetrate administration.
Wherein, the mode of drug administration by injection for example can be, but be not limited to intravenous injection, intramuscular injection, the interior note of coronary artery
Penetrate and myocardial injection.
In the present invention, heart disease include myocardial hypertrophy, myocardial fibrosis, coronary heart disease, myocarditis, heart valve disease,
The one or more of hypertension and heart failure.
In order to contribute to it is clearer understand present disclosure, be described in detail as follows in conjunction with specific embodiment.Such as
Not yet explicitly point out, the Examination on experimental operation being related in following examples is the conventional molecular biology manipulations method in this area, is related to
And reagent be analytical grade reagent, the reagent or instrument being related to can be commercially available from regular channel.
Unless otherwise indicated, the various experimental methods being related in following embodiment and operation, including cell culture, RNA
Extraction, RCR amplifications, quantitative fluorescent PCR, cell dyeing etc., can be found in documents below：Wang K,etal,miR-361-
regulated prohibitin inhibits mitochondrial fission and apoptosis and
protects heart from ischemia injury.Cell Death Differ. 2015Jun；22(6):1058-
68.W.-Q.Tan,etal,Foxo3a Inhibits Cardiomyocyte Hypertrophy through
Transactivating Catalase J Biol Chem.2008October 31； 283(44):29730-29739；Wang
K,etal,miR-9and NFATc3regulate myocardin in cardiac hypertrophy,J Biol
Chem.2010Apr16；285(16):11903-12；Lin Z,etal, miR-23a functions downstream of
NFATc3toregulate cardiac hypertrophy, PNAS,2009,106(29):12103-12108) the above-mentioned documents of
Which is hereby incorporated by reference.
Embodiment 1 uses the myocardial hypertrophy disease model mouse of isoprel (Iso) induction and the heart of control mice
In dirty, the detection of piRNA-514 expressions change
C57BL/6J public affairs mouse 20 is chosen, is divided into control group and model group, is permeated respectively by implanted microcapsule permeable pump
Iso and physiological saline, build the mouse model of myocardial hypertrophy.The total serum IgE of two groups of mouse hearts, real time fluorescent quantitative are extracted respectively
Round pcr detects piRNA-514 expression, as a result sees Fig. 1.
From figure 1 it appears that in the myocardial hypertrophy model group mouse heart of Iso inductions, piRNA-514 expression is lowered,
Illustrate that the change by detecting piRNA-514 expressions can be used as the foundation of myocardial hypertrophy clinical diagnosis.
The cardiac myocyte hypertrophy process piRNA-514 expressions change of the isoprel of embodiment 2 (Iso) induction
Detection
For cardiac myocyte hypertrophy model, the primary cardiac muscle of method culture rat suckling mouse established using this laboratory is thin
Born of the same parents, cardiac muscle cell is handled with 10 μM of isoprels, in the different time of culture, extract the total serum IgE of cell, in real time
Fluorescent quantitative PCR technique detects piRNA-514 expression.The present embodiment uses and carries out isopropyl respectively to primary cardiomyocytes
After adrenaline is handled 0 hour, 8 hours, 16 hours, 24 hours, the table of piRNA-514 different time points in heart is detected
Up to level.The total serum IgE (using TRIZOL kits) of heart is extracted, real-time fluorescence quantitative PCR detects piRNA-514 expression water
It is flat, as a result see Fig. 2.
From figure 2 it can be seen that after Iso is handled 0,8,16,24 hour, piRNA-514 level is with time dependent
Mode is gradually reduced, and further illustrates that piRNA-514 can be as the potential mark of myocardial hypertrophy clinical diagnosis.
Embodiment 3 is overexpressed the experiment that piRNA-514 suppresses primary cardiomyocytes cellular mast
The primary cardiomyocytes that embodiment 2 provides are divided into four groups, respectively blank group, Iso processing in the present embodiment
Group, control group and experimental group, (cell concentration is about 1 × 10 for every group of three repetitions6)。
Wherein, blank group primary cardiomyocytes are only cultivated using normal cell nutrient solution.
Iso treatment groups primary cardiomyocytes are handled with 10 μM of isoprels.
After experimental group is handled with 10 μM of isoprels primary cardiomyocytes, piRNA-514 nucleotides is carried out
(its sequence is 5 '-AAUCAGAUUUGCACCACUCAUCUUGGUGA-3 ', SEQ ID NO.1 to analog, entrusts Shanghai Ji agate
Pharmaceutical Technology Inc. synthesize) transfection.
Control group is identical with the processing method of experimental group, and piRNA-514 nucleotide analogs only are replaced with into meaningless sequence
The negative control RNA (agomir-control) of row.
From figure 3, it can be seen that the piRNA-514 of piRNA-514agomir inductions, which is overexpressed, can reduce Iso induction increasings
The cell surface product added.
In summary, piRNA-514 can suppress the generation of myocardial hypertrophy, can be used as a kind of early diagnosis and early prevention
The biomarker of the heart diseases such as myocardial hypertrophy, myocardial fibrosis.Therefore, by detecting piRNA-514 in peripheral blood blood plasma
Expression, for many heart diseases have diagnosis and/or prognosis evaluation potential value；Suppressed by detecting
PiRNA-514 expression, for having the potential value of prevention and/or treatment to many heart diseases.
Finally it should be noted that：Various embodiments above is merely illustrative of the technical solution of the present invention, rather than its limitations；To the greatest extent
The present invention is described in detail with reference to foregoing embodiments for pipe, it will be understood by those within the art that：Its according to
The technical scheme described in foregoing embodiments can so be modified, either which part or all technical characteristic are entered
Row equivalent substitution；And these modifications or replacement, the essence of appropriate technical solution is departed from various embodiments of the present invention technology
The scope of scheme.
SEQUENCE LISTING
<110>University Of Qingdao
<120>The application of piRNA-514 nucleotide analogs and its GEM 132 and apply its product
<160> 5
<170> PatentIn version 3.5
<210> 1
<211> 29
<212> DNA
<213>Artificial sequence
<400> 1
aaucagauuu gcaccacuca ucuugguga 29
<210> 2
<211> 29
<212> DNA
<213>Artificial sequence
<400> 2
ucaccaagau gaguggugca aaucugauu 29
<210> 3
<211> 44
<212> DNA
<213>Artificial sequence
<400> 3
ctcaactggt gtcgtggagt cggcaattca gttgagtcac caag 44
<210> 4
<211> 28
<212> DNA
<213>Artificial sequence
<400> 4
cgggcaatca gatttgcacc actcatct 28
<210> 5
<211> 16
<212> DNA
<213>Artificial sequence
<400> 5
tggtgtcgtg gagtcg 16
Claims (10)
1.piRNA-514 nucleotide analogs or its bioactive functions fragment or variant are being prepared for diagnosis and/or prognosis
Assess the application in the product of heart disease；
The piRNA-514 nucleotide analogs contain the sequence shown in SEQ ID NO.1.
2. application according to claim 1, it is characterised in that the product is medicine or kit.
3. application according to claim 1, it is characterised in that the nucleotides sequence of the piRNA-514 nucleotide analogs
Each base has carried out chemical modification in row.
4. a kind of medicine for being used for diagnosis and/or prognosis evaluation heart disease, it is characterised in that the medicine includes effective dose
PiRNA-514 nucleotide analogs or its bioactive functions fragment or variant.
5. a kind of kit for being used for diagnosis and/or prognosis evaluation heart disease, it is characterised in that the kit includes being used for
The primer of specific detection piRNA-514 nucleotide analogs.
6.piRNA-514 GEM 132s are preparing the application in being used to prevent and/or treat the product of heart disease；
The piRNA-514 GEM 132s contain the sequence shown in SEQ ID NO.2.
7. application according to claim 6, it is characterised in that the product is medicine.
8. a kind of medicine for being used to preventing and/or treating heart disease, it is characterised in that the medicine includes effective dose
PiRNA-514 GEM 132s.
9. the medicine according to claim 4 or 8, it is characterised in that the administering mode of the medicine include be administered orally or
Drug administration by injection.
10. the application according to claim 1 or 6, the reagent described in medicine or claim 5 described in claim 4 or 8
Box, it is characterised in that the heart disease includes myocardial hypertrophy, myocardial fibrosis, coronary heart disease, myocarditis, heart valve disease, height
The one or more of blood pressure and heart failure.
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| CN201710866272.3A CN107604058B (en) | 2017-09-22 | 2017-09-22 | PiRNA-514 nucleotide analogue and application of antisense nucleotide thereof and product using same |
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110229879A (en) * | 2019-07-01 | 2019-09-13 | 青岛大学 | Its product of the application and application of a kind of piRNA-500 nucleotide analog and its GEM 132 |
| CN111705061A (en) * | 2020-07-22 | 2020-09-25 | 青岛大学 | Antisense nucleotide of piRNA-P1 and piRNA-P1 related to heart disease, application and medicament |
| CN114736956A (en) * | 2022-04-07 | 2022-07-12 | 山东第一医科大学(山东省医学科学院) | Application of piRNA as diagnostic marker in forensic identification of acute myocardial infarction and/or inference of acute myocardial infarction duration |
| CN114965645A (en) * | 2022-06-10 | 2022-08-30 | 山东大学 | In-vitro piRNA detection method based on photoelectrochemistry and double-strand specific nuclease |
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| CN104548136A (en) * | 2015-01-27 | 2015-04-29 | 青岛大学 | PiRNA medical composition and application thereof |
| CN104645352A (en) * | 2015-01-27 | 2015-05-27 | 青岛大学 | piRNA antisense nucleotide pharmaceutical composition and application thereof |
| CN106916885A (en) * | 2017-01-13 | 2017-07-04 | 青岛大学 | For detecting cardiopathic piRNA combinations and its applying |
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| CN104548136A (en) * | 2015-01-27 | 2015-04-29 | 青岛大学 | PiRNA medical composition and application thereof |
| CN104645352A (en) * | 2015-01-27 | 2015-05-27 | 青岛大学 | piRNA antisense nucleotide pharmaceutical composition and application thereof |
| CN106916885A (en) * | 2017-01-13 | 2017-07-04 | 青岛大学 | For detecting cardiopathic piRNA combinations and its applying |
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Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110229879A (en) * | 2019-07-01 | 2019-09-13 | 青岛大学 | Its product of the application and application of a kind of piRNA-500 nucleotide analog and its GEM 132 |
| CN110229879B (en) * | 2019-07-01 | 2022-11-18 | 青岛大学 | PiRNA-500 nucleotide analogue, application of antisense nucleotide thereof and product using same |
| CN111705061A (en) * | 2020-07-22 | 2020-09-25 | 青岛大学 | Antisense nucleotide of piRNA-P1 and piRNA-P1 related to heart disease, application and medicament |
| CN114736956A (en) * | 2022-04-07 | 2022-07-12 | 山东第一医科大学(山东省医学科学院) | Application of piRNA as diagnostic marker in forensic identification of acute myocardial infarction and/or inference of acute myocardial infarction duration |
| CN114736956B (en) * | 2022-04-07 | 2024-03-08 | 山东第一医科大学(山东省医学科学院) | Application of piRNA as diagnostic marker in forensic identification of acute myocardial infarction and/or inference of duration of acute myocardial infarction |
| CN114965645A (en) * | 2022-06-10 | 2022-08-30 | 山东大学 | In-vitro piRNA detection method based on photoelectrochemistry and double-strand specific nuclease |
| CN114965645B (en) * | 2022-06-10 | 2022-12-09 | 山东大学 | In-vitro piRNA detection method based on photoelectrochemistry and double-strand specific nuclease |
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