CN114990114B - Small molecule RNA for promoting damaged heart myocardial cell proliferation - Google Patents
Small molecule RNA for promoting damaged heart myocardial cell proliferation Download PDFInfo
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- CN114990114B CN114990114B CN202210517123.7A CN202210517123A CN114990114B CN 114990114 B CN114990114 B CN 114990114B CN 202210517123 A CN202210517123 A CN 202210517123A CN 114990114 B CN114990114 B CN 114990114B
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- 230000002107 myocardial effect Effects 0.000 title claims abstract description 20
- 230000001737 promoting effect Effects 0.000 title claims abstract description 15
- 230000004663 cell proliferation Effects 0.000 title claims abstract description 12
- 150000003384 small molecules Chemical class 0.000 title claims description 16
- 210000002216 heart Anatomy 0.000 title abstract description 26
- 239000003814 drug Substances 0.000 claims abstract description 15
- 239000002773 nucleotide Substances 0.000 claims description 6
- 125000003729 nucleotide group Chemical group 0.000 claims description 6
- 108091032973 (ribonucleotides)n+m Proteins 0.000 abstract description 24
- 208000019622 heart disease Diseases 0.000 abstract description 12
- 230000035755 proliferation Effects 0.000 abstract description 11
- 208000010125 myocardial infarction Diseases 0.000 abstract description 7
- 210000004413 cardiac myocyte Anatomy 0.000 abstract description 5
- 108091028043 Nucleic acid sequence Proteins 0.000 abstract description 4
- 241000699670 Mus sp. Species 0.000 description 19
- 241000699666 Mus <mouse, genus> Species 0.000 description 13
- 210000004027 cell Anatomy 0.000 description 8
- 239000012188 paraffin wax Substances 0.000 description 7
- 239000007924 injection Substances 0.000 description 6
- 238000002347 injection Methods 0.000 description 6
- 241000282412 Homo Species 0.000 description 5
- 239000013642 negative control Substances 0.000 description 5
- 210000004165 myocardium Anatomy 0.000 description 4
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 4
- 241000251539 Vertebrata <Metazoa> Species 0.000 description 2
- 210000000038 chest Anatomy 0.000 description 2
- 230000007547 defect Effects 0.000 description 2
- FFYPMLJYZAEMQB-UHFFFAOYSA-N diethyl pyrocarbonate Chemical compound CCOC(=O)OC(=O)OCC FFYPMLJYZAEMQB-UHFFFAOYSA-N 0.000 description 2
- 229940079593 drug Drugs 0.000 description 2
- 230000013020 embryo development Effects 0.000 description 2
- 230000014509 gene expression Effects 0.000 description 2
- 230000003601 intercostal effect Effects 0.000 description 2
- 230000029058 respiratory gaseous exchange Effects 0.000 description 2
- CDEURGJCGCHYFH-DJLDLDEBSA-N 5-ethynyl-2'-deoxyuridine Chemical compound C1[C@H](O)[C@@H](CO)O[C@H]1N1C(=O)NC(=O)C(C#C)=C1 CDEURGJCGCHYFH-DJLDLDEBSA-N 0.000 description 1
- 206010002091 Anaesthesia Diseases 0.000 description 1
- 108091032955 Bacterial small RNA Proteins 0.000 description 1
- 208000032170 Congenital Abnormalities Diseases 0.000 description 1
- 208000002330 Congenital Heart Defects Diseases 0.000 description 1
- 206010010356 Congenital anomaly Diseases 0.000 description 1
- 108020004414 DNA Proteins 0.000 description 1
- WSFSSNUMVMOOMR-UHFFFAOYSA-N Formaldehyde Chemical compound O=C WSFSSNUMVMOOMR-UHFFFAOYSA-N 0.000 description 1
- 206010019280 Heart failures Diseases 0.000 description 1
- 208000013875 Heart injury Diseases 0.000 description 1
- 241001465754 Metazoa Species 0.000 description 1
- 206010028980 Neoplasm Diseases 0.000 description 1
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 1
- 230000037005 anaesthesia Effects 0.000 description 1
- 238000004458 analytical method Methods 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 230000007698 birth defect Effects 0.000 description 1
- 239000008280 blood Substances 0.000 description 1
- 210000004369 blood Anatomy 0.000 description 1
- 230000003683 cardiac damage Effects 0.000 description 1
- 230000000747 cardiac effect Effects 0.000 description 1
- 231100001012 cardiac lesion Toxicity 0.000 description 1
- 238000001516 cell proliferation assay Methods 0.000 description 1
- RNFNDJAIBTYOQL-UHFFFAOYSA-N chloral hydrate Chemical compound OC(O)C(Cl)(Cl)Cl RNFNDJAIBTYOQL-UHFFFAOYSA-N 0.000 description 1
- 229960002327 chloral hydrate Drugs 0.000 description 1
- 210000004351 coronary vessel Anatomy 0.000 description 1
- 206010012601 diabetes mellitus Diseases 0.000 description 1
- 201000010099 disease Diseases 0.000 description 1
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 1
- XACKNLSZYYIACO-DJLDLDEBSA-N edoxudine Chemical compound O=C1NC(=O)C(CC)=CN1[C@@H]1O[C@H](CO)[C@@H](O)C1 XACKNLSZYYIACO-DJLDLDEBSA-N 0.000 description 1
- 239000007850 fluorescent dye Substances 0.000 description 1
- 239000008098 formaldehyde solution Substances 0.000 description 1
- 210000002064 heart cell Anatomy 0.000 description 1
- 230000023560 heart growth Effects 0.000 description 1
- 239000007928 intraperitoneal injection Substances 0.000 description 1
- 210000005240 left ventricle Anatomy 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 108091070501 miRNA Proteins 0.000 description 1
- 239000002679 microRNA Substances 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
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- 230000008506 pathogenesis Effects 0.000 description 1
- 210000003516 pericardium Anatomy 0.000 description 1
- 239000000902 placebo Substances 0.000 description 1
- 229940068196 placebo Drugs 0.000 description 1
- 230000002062 proliferating effect Effects 0.000 description 1
- 108090000623 proteins and genes Proteins 0.000 description 1
- 238000012827 research and development Methods 0.000 description 1
- 229920002477 rna polymer Polymers 0.000 description 1
- 230000007651 self-proliferation Effects 0.000 description 1
- 238000010186 staining Methods 0.000 description 1
- 210000001562 sternum Anatomy 0.000 description 1
- 210000000115 thoracic cavity Anatomy 0.000 description 1
- 210000001519 tissue Anatomy 0.000 description 1
- 210000003437 trachea Anatomy 0.000 description 1
- 238000013518 transcription Methods 0.000 description 1
- 230000035897 transcription Effects 0.000 description 1
- 238000012795 verification Methods 0.000 description 1
- 238000010792 warming Methods 0.000 description 1
Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
- C12N15/09—Recombinant DNA-technology
- C12N15/11—DNA or RNA fragments; Modified forms thereof; Non-coding nucleic acids having a biological activity
- C12N15/113—Non-coding nucleic acids modulating the expression of genes, e.g. antisense oligonucleotides; Antisense DNA or RNA; Triplex- forming oligonucleotides; Catalytic nucleic acids, e.g. ribozymes; Nucleic acids used in co-suppression or gene silencing
-
- 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
- A61K31/70—Carbohydrates; Sugars; Derivatives thereof
- A61K31/7088—Compounds having three or more nucleosides or nucleotides
- A61K31/7105—Natural ribonucleic acids, i.e. containing only riboses attached to adenine, guanine, cytosine or uracil and having 3'-5' phosphodiester links
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P9/00—Drugs for disorders of the cardiovascular system
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P9/00—Drugs for disorders of the cardiovascular system
- A61P9/10—Drugs for disorders of the cardiovascular system for treating ischaemic or atherosclerotic diseases, e.g. antianginal drugs, coronary vasodilators, drugs for myocardial infarction, retinopathy, cerebrovascula insufficiency, renal arteriosclerosis
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N2310/00—Structure or type of the nucleic acid
- C12N2310/10—Type of nucleic acid
- C12N2310/14—Type of nucleic acid interfering N.A.
- C12N2310/141—MicroRNAs, miRNAs
Abstract
The invention relates to the fields of life science and biological medicine, and discloses a small molecular RNA sequence capable of promoting myocardial cell proliferation of a damaged heart, which contains a sequence shown as SEQ ID No. 1. The small molecular RNA has the advantages of promoting the proliferation of damaged cardiac myocytes, being used for preparing medicaments for promoting the proliferation of the cardiac myocytes, medicaments for treating or assisting in treating heart diseases, particularly medicaments for treating or assisting in treating myocardial infarction, and having better application prospects.
Description
Technical Field
The invention relates to the fields of life science and biological medicine, in particular to a small molecular RNA sequence capable of promoting myocardial cell proliferation of a damaged heart and application thereof.
Background
During vertebrate embryo development, the heart is the first tissue organ to begin to form and function during embryo development. Congenital heart diseases, including structural and functional heart diseases, are the most common and serious birth defects in humans, with high morbidity and mortality. How to compensate for the defect of heart growth is a key link for treating heart diseases.
Mice are an ideal vertebrate model for studying human heart pathogenesis and treatment of heart disease. Although the hearts of mice and humans are not exactly the same, the two have quite high similarity. Heart disease in humans can often be reproduced in mice, allowing us to study the occurrence of heart disease in humans and the treatment of heart disease in mice.
Humans cannot repair heart damage by self-proliferation of cardiomyocytes. Similarly, damaged mouse cardiac cardiomyocytes also do not have the ability to self-proliferate, which makes self-repair of damaged human or mouse cardiac lesions difficult. If the adult heart has proliferative heart cells throughout life, it is clear that the repair capacity of the damaged heart can be improved and the heart disease defect can be remedied, which also greatly reduces the incidence of heart disease. How to endow the human myocardial cells with proliferation capability provides thought and direction for realizing the repair of the damaged heart of the human.
The micro RNA is micro ribonucleic acid, is a non-coding single-stranded RNA molecule which is coded by an endogenous gene and has the length of about 22-24 nucleotides, participates in the regulation and control of gene expression after animal and plant transcription, and can inhibit or activate the gene expression. The modification of natural small molecular RNA to obtain small molecular RNA medicine is an important trend in medicine research and development, and the small molecular RNA has been reported to be used for treating diseases such as tumor, diabetes, heart failure and the like.
Disclosure of Invention
The invention aims to provide a small molecular RNA sequence capable of promoting the proliferation of myocardial cells of a damaged heart.
The invention also provides application of the small molecule RNA sequence.
The technical proposal is as follows:
a small molecule RNA has a nucleotide sequence shown in SEQ ID No. 1.
The sequence of SEQ ID No.1 is: 5'-GCAGUACCAUUCAAAGAGCUAU-3'.
Preferably, the nucleotide sequence of the small molecule RNA comprises the nucleotide sequence shown in SEQ ID No. 1.
The small molecule RNA can promote the proliferation of myocardial cells of a damaged heart. Through verification of a mouse heart injury model, myocardial cells in the heart of a damaged mouse show signals of cell proliferation through injection of the small molecule RNA.
Therefore, the small molecular RNA has the function of promoting the proliferation of myocardial cells of the damaged heart, and can be used for preparing medicaments for promoting the proliferation of myocardial cells.
Furthermore, the small molecular RNA can be used for preparing medicines for treating or assisting in treating heart diseases.
Alternatively, the small molecule RNA can be used for preparing medicines for treating or assisting in treating myocardial infarction.
A medicine for promoting myocardial cell proliferation or treating heart disease contains the above small molecule RNA.
The invention has the beneficial effects that the small molecular RNA is provided and used for preparing the myocardial cell proliferation promoting damaged heart. The small molecular RNA can be used for preparing medicaments for promoting myocardial cell proliferation or treating heart diseases, in particular medicaments for treating myocardial infarction, and has good application prospect.
Drawings
FIG. 1 is a paraffin section of a damaged heart of a mouse injected with DEPC water (blank control)
FIG. 2 Paraffin section of damaged heart of mice injected with RNA-NC (negative control)
FIG. 3 Paraffin section of damaged heart of mice injected with RNA-XU1 (experimental group)
Detailed Description
Example 1
(1) Establishing a mouse myocardial infarction model
Six week old female mice of the C57/BL6J strain were anesthetized by intraperitoneal injection of 4% chloral hydrate (200. Mu.L/mouse). After the completion of anesthesia, edU (5-ethyl-2 '-deoxyuridine, 5-ethynyl-2' -deoxyuridine) was intraperitoneally injected (200. Mu.L/min.).
And after fifteen minutes of injection, modeling is started, namely, a mouse myocardial infarction model is built. After dehairing the chest of the mouse by using dehairing paste, fixing the mouse on an operation table, connecting an trachea cannula with a breathing machine, finding the 3 rd and 4 th intercostals beside the sternum of the mouse under a stereoscope, making a parallel incision between the intercostals by using a surgical knife, opening the chest, cutting open the pericardium to expose the heart, ligating the upper 1/3 part of the left coronary artery by using 6-0 suture, and observing that the front wall of the left ventricle of the mouse loses original gloss, the myocardium becomes white, the myocardial activity is weakened and the myocardial infarction model modeling is completed in the ligaturing moment.
The mice were divided into experimental group mice, negative control group mice and blank control group mice.
(2) Mouse myocardial proliferation assay
After successful modeling of the myocardial infarction model, local administration is carried out, and three-point injection is used for injecting small molecular RNA into the myocardium. Mice of the experimental group were injected with small-molecule RNA of SEQ ID No.1 (hereinafter referred to as RNA-XU 1); mice in the negative control group were injected with small-molecule RNA-NC (hereinafter referred to as RNA-NC); mice in the placebo group were injected with DEPC water (diethyl pyrocarbonate, diethyl pyrocarbonate, abbreviated DEPC). Each injection visibly whitens the myocardium, i.e., the myocardium injection administration was successful. After the injection is finished, the thoracic cavity is closed layer by using 3-0 lines, a breathing machine is disconnected, and the mice are observed and awakened by a warming blanket.
The experimental group small molecule RNA and the control group small molecule RNA are synthesized by Ji Ma biological company, and the sequences are as follows:
SEQ ID No.1 laboratory group small molecule RNA (hereinafter referred to as RNA-XU 1): 5'-GCAGUACCAUUCAAAGAGCUAU-3';
control small molecule RNA sequences (RNA-NC): 5'-CAGUACUUUUGUGUAGUACAA-3'.
(3) Observing the materials
After 3 days, the mice are sacrificed, hearts are cut off, blood is washed by normal saline, the mice are soaked in 4% formaldehyde solution and fixed for 48 hours, the mice are transected by ligation points, paraffin block sections are manufactured, and after dewaxing, myocardial cell proliferation is observed under a fluorescence microscope after Apollo fluorescent staining and DNA staining.
Paraffin sections of the mouse-injured heart injected with DEPC water (blank control), paraffin sections of the mouse-injured heart injected with RNA-NC (negative control), and paraffin sections of the mouse-injured heart injected with RNA-XU1 (experimental group) are shown in fig. 1 to 3, respectively.
Analysis of results: a significant positive proliferation signal (white spot in fig. 3) was found in the group injected with RNA-XU1, i.e. a signal that cell proliferation occurred in cardiomyocytes in the heart of injured mice injected with RNA-XU1 sequences.
In contrast, no proliferation signal was found in the RNA-NC negative control group and the DEPC water-injected blank group. This suggests that RNA-XU1 can promote proliferation of myocardial cells in the damaged heart.
Sequence listing
<110> Shanghai university of ocean
<120> a small RNA for promoting proliferation of myocardial cells of a damaged heart
<160> 2
<170> SIPOSequenceListing 1.0
<210> 1
<211> 22
<212> RNA
<213> Artificial sequence (Artificial Sequence)
<400> 1
gcaguaccau ucaaagagcu au 22
<210> 2
<211> 21
<212> RNA
<213> Artificial sequence (Artificial Sequence)
<400> 2
caguacuuuu guguaguaca a 21
Claims (2)
1. The application of the small molecular RNA with the nucleotide sequence shown as SEQ ID No.1 in the aspect of preparing medicaments for promoting myocardial cell proliferation.
2. A medicament for promoting myocardial cell proliferation is characterized by comprising small molecule RNA with a nucleotide sequence shown as SEQ ID No. 1.
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| CN202210517123.7A CN114990114B (en) | 2022-05-12 | 2022-05-12 | Small molecule RNA for promoting damaged heart myocardial cell proliferation |
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| CN114990114B true CN114990114B (en) | 2023-12-01 |
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Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102266570A (en) * | 2011-07-08 | 2011-12-07 | 中国科学院动物研究所 | New application of miRNA-484, pharmaceutical composition containing miRNA-484 and use thereof |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| WO2006047454A2 (en) * | 2004-10-21 | 2006-05-04 | Rutgers, The State University Of New Jersey | Rational probe optimization for detection of micrornas |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102266570A (en) * | 2011-07-08 | 2011-12-07 | 中国科学院动物研究所 | New application of miRNA-484, pharmaceutical composition containing miRNA-484 and use thereof |
Non-Patent Citations (1)
| Title |
|---|
| Rainbow trout exposed to benzo[a]pyrene yields conserved microRNA binding sites in DNA methyltransferases across 500 million years of evolution;Christopher Kuc 等;SCIENTIFIC REPORTS;第7卷(第1期);第9页 表1 * |
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