CN104645352B - A kind of piRNA GEM 132s medical composition and its use - Google Patents
A kind of piRNA GEM 132s medical composition and its use Download PDFInfo
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Abstract
The present invention relates to a kind of piRNA GEM 132s medical composition and its use, pharmaceutical composition includes piRNA GEM 132s and carrier；PiRNA antisense base sequences are：5’‑CCUUGGCACAUGCGCAGAUUAUUUGUUUA‑3’；The carrier is the one or more in chitosan, cholesterol, nano particle and liposome；The pharmaceutical composition is used for myocardial infarction, treating myocardial ischemia damage and the treatment of myocardial fibrosis and preventing and treating；Its raw material is scientific and reasonable, and preparation technology is simple, and medicine effect is obvious, and use range is wide, and securely and reliably, application environment is friendly.
Description
Technical field：
The invention belongs to biomedicine technical field, is related to a kind of endogenic non-coding tiny RNA s new drug and purposes,
The pharmaceutical composition of especially a kind of GEM 132 containing piRNA and its purposes in preventing or treating heart disease.
Background technology：
Angiocardiopathy is the chief threat of human health and life, is the number one killer of human health, and the whole world is annual
There are more than 1,700 ten thousand people to die from angiocardiopathy, its death rate is close to the summation of all cancer mortalities.As the mankind give birth to
Running water benefit raising on ordinary days and the change of dietary structure, the angiocardiopathy death rate is in obvious ascendant trend, is turned into more than cancer
The first big cause of death.Recent statistics result according to announcing in the recent period shows, 18 years old and above prevalence of hypertension rate are
18.8%, estimate national number of patients more than 1.6 hundred million, myocardial hypertrophy, myocardial infarction, coronary heart disease and the heart failure thereby resulted in
The patient for the apoptosis related cardiac conditions such as exhausting is up to several ten million.It is not fully understood at present on heart disease pathogenesis, heart
Prevention, diagnosis and the treatment of disease can not still reach satisfactory effect, are badly in need of the new medicine of exploitation and are used for cardiopathic examine
Disconnected and preventing and treating.
Apoptosis (apoptosis) is also known as apoptosis, refers to cell in certain physiology or pathological conditions
Under, it then follows the program of itself, the process of oneself end lives, be an active, high-sequential, by gene control and a system
The process that row enzyme participates in, to normal embryo development, maintain to play an important role during cell colony and malignant change, ensureing
The role of key is play in the healthy survival processes of multicellular organism.Under many myocardial damages or heart pathological state, such as
Apoptosis can occur for myocardial ischemia-reperfusion injury, myocardial hypertrophy and heart failure etc., cardiac muscle cell.Due to the cardiac muscle cell of maturation
Division growth is unable to, cardiac muscle cell's excessive Apoptosis will necessarily reduce cardiac muscle cell's number, and this is probably what heart disease was fallen ill
Mechanism.
PiRNA (piwi-interacting RNA) is a kind of non-coding small RNA molecular newfound in recent years, and sum is super
40,000 kinds, length about 26~3l nt are crossed, the end of sequence 5 ' has uracil skewed popularity (about 86%), and piRNA clusters form locus
(loci) it is distributed on genome, specifically with the Piwi subfamily protein bindings in Argonuat protein families, and therefore
And gain the name.2006 are isolated in mouse sperm cell first, are originally found piRNA and are mainly expressed in germ cell line,
To maintaining reproduction DNA complete, the transcription of suppression transposons, the formation for suppressing translation, participating in heterochromatin, execution epigenetic regulation
With reproduction cell occur etc. play an important role.With going deep into for research, it has been found that piRNA can be in many somatic tissues
In extensive or specific expressed, including heart, brain, liver, spleen, kidney, lung, stomach, small intestine, colon, ovary, uterus, testis
Ball.
Research for the piRNA of heart has very extensive practice significance and application value.First, to piRNA's
Functional study is advantageous to find the mechanism of new regulation and control myocardial infarction, deepens understanding of the people to existing signal pathway.Secondly, it is right
Heartspecific piRNA research is advantageous to find new drug target and medicine.Now, Many researchers all utilize people
The piRNA inhibitor of work synthesis carries out functional study to piRNA.This process such as methylates, the inhibitor of cholesterol modification
Time length is metabolized in vivo, is specifically acted on target piRNA, can effectively be suppressed piRNA expression, thus with turning into
The possibility of curative drug.This inhibitor structure is simple, will not produce immune response in vivo, has very high medicinal open
Make an offer value.Present many non-coding RNAs, such as our expression regulations and function of the piRNA that studies emphatically in heart is still
And it is not known, we want to carry out this kind of piRNA expressed in heart the research of expression regulation and function, so as to
Deepen understanding of the people to myocardial infarction molecular mechanism, there is provided new drug target.This is for developing using piRNA as strategy
Heart disease diagnosis and medicine has and its important meaning and application prospect.
The content of the invention：
The shortcomings that it is an object of the invention to overcome prior art to exist, there is provided a kind of new pharmaceutical composition, it is determined that adjusting
Control the piRNA of the heartspecific expression of cardiac muscle cell apoptosis and its in myocardial ischemia-reperfusion, myocardial infarction, coronary heart disease, cardiac muscle
Purposes in the heart diseases such as loose and myocardial fibrosis.
In order to realize foregoing invention purpose, it is anti-that piRNA GEM 132s pharmaceutical composition of the present invention includes piRNA
Adopted nucleotides and pharmaceutically acceptable carrier；SEQ ID NO in piRNA antisense base sequences used such as sequence table:1 institute
Show：5’-CCUUGGCACAUGCGCAGAUUAUUUGUUUA-3’；The carrier is chitosan, cholesterol, nano particle and lipid
One or more in body, it is therefore preferable to liposome nano granule.
The content of piRNA GEM 132s is 0.5-1 grams in pharmaceutical composition of the present invention.
Described pharmaceutical composition is administered in a manner of oral or injection；Wherein, drug administration by injection mode is selected from intravenous injection, flesh
Meat injection, intracoronary injection or myocardial injection.
The piRNA GEM 132s pharmaceutical composition of the present invention is used for myocardial infarction, treating myocardial ischemia damage and cardiac muscle fibre
The treatment and preventing and treating of change.
The pharmaceutical composition of the present invention can be made into the kit for preventing or treating heart disease, and the piRNA contained is anti-
Adopted nucleotides forms medicine with carrier and imported in vivo, and the carrier is one in chitosan, cholesterol, nano particle and liposome
Kind is a variety of, it is therefore preferable to liposome nano granule.PiRNA antisense nucleosides acid content is 0.5-1 grams in kit, is obstructed for cardiac muscle
Extremely, treating myocardial ischemia damage and the prevention and treatment of myocardial fibrosis.
PiRNA tables during the cardiac muscle cell apoptosis for the treatment of myocardial ischemia damage and hypoxia inducible are found through experiments that in the present invention
Up to notable up-regulation；By transfect and inject piRNA GEM 132s suppress piRNA expression can suppress cardiac muscle cell apoptosis,
Treating myocardial ischemia damage and myocardial infarction area are reduced；There is protective effect to heart by suppressing cardiac muscle cell apoptosis.
Compared with prior art, the raw material that it is selected is scientific and reasonable, and preparation technology is simple by the present invention, and medicine effect is obvious,
Use range is wide, safe and reliable, and application environment is friendly.
Brief description of the drawings：
Fig. 1 is piRNA expressions during treating myocardial ischemia damage and the cardiac muscle cell apoptosis of anoxic treatment (Anoxia)
Variation diagram；
Fig. 2 is to suppress inhibitory action figures of the endogenic piRNA to cardiac muscle cell apoptosis；
Fig. 3 is inhibitory action figure of the piRNA GEM 132s to treating myocardial ischemia damage.
Embodiment：
The present invention is expanded on further below by specific embodiment and with reference to accompanying drawing.
Material, experimental method and the operating procedure being related in following embodiment, including primary myocardial cell culture, cell
Transfection, myocardial ischemia and reperfusion operating procedure, the double dyes of Evans blue/TTC and TUNEL detections are referring to documents below：Wang
JX,etal,miR-499 regulates mitochondrial dynamics by targeting calcineurin and
dynamin-related protein-1.2011 January 17:71-78；Experimental animal wherein used is mouse, and its is small
Mouse kind is C57/BL6, purchased from Beijing Medical University；Enzyme used such as restriction enzyme, amplification enzyme and reverse transcriptase etc.,
Purchased from the extensive and profound in meaning biological Co., Ltd in Beijing；Reagent used is analytical grade reagent, and commercially available from regular channel.
Embodiment 1,
PiRNA GEM 132s pharmaceutical composition includes piRNA GEM 132s and pharmaceutically acceptable carrier；It is used
PiRNA antisense base sequences such as sequence table in SEQ ID NO:Shown in 1：5’-CCUUGGCACAUGCGCAGAUUAUUUGUU
UA-3’；The carrier is the one or more in chitosan, cholesterol, nano particle and liposome, it is therefore preferable to which liposome is received
The grain of rice.The content of piRNA GEM 132s is 0.5-1 grams in the pharmaceutical composition.
PiRNA expressions are examined during the cardiac muscle cell apoptosis of embodiment 2, myocardial ischemia and Myocytes Anoxia induction
Survey
The present embodiment is thin using the primary cardiac muscle of the method culture rat suckling mouse of routine to cardiac muscle cell apoptosis experimental model
Born of the same parents, hypoxemia incubator (oxygen concentration is less than 1%) culture different time, extract RNA, Real-Time Fluorescent Quantitative PCR Technique detection piRNA
Expression, Fig. 1 be treating myocardial ischemia damage and anoxic treatment (Anoxia) cardiac muscle cell apoptosis during piRNA express water
Flat variation diagram, wherein Figure 1A show primary cardiomyocytes anoxic treatment piRNA expressions；Figure 1B shows mouse cardiac muscle ischemic
Damage piRNA expressions；Significantly (Figure 1A) is raised in hypoxemia processing 4h-8h；The heart is established using ligation mouse coronary artery
Myocardial ischemia model, ischemic different time are cored dirty ischemic region and non-ischemic region cardiac muscular tissue, are extracted total serum IgE, are passed through real-time fluorescence
Quantitative PCR technique detects piRNA expression, as a result shows the more non-ischemia group of ischemic 30min-120min myocardial ischemia tissues
Knit and the piRNA expressions of non-ischemia model group significantly rise (Figure 1B), wherein with sham-operation group (sham) as a control group.
Embodiment 3, piRNA GEM 132s suppress the experiment of cardiac muscle cell apoptosis
The cardiac muscle cell that original cuiture is used in the present embodiment is model, transfects piRNA GEM 132s, (piRNA antisenses
Nucleotide sequence is to be completely reversed complementary sequence, 5 '-CCUUGGCACAUGCGCAGAUUAUUUGUUUA- with piRNA sequences
3 ', combined with piRNA to suppress piRNA expression), transfection assay is referring to aforementioned documents, after transfecting 24 hours, in hypoxemia
Culture processing cell 3 hours in incubator, inducing cell apoptosis, platform expect blue colouring method detection cardiac muscle cell apoptosis situation, knot
Fruit sees that Fig. 2, wherein Fig. 2A represent that primary cardiomyocytes transfection piRNA GEM 132s (anta-piRNA) suppress endogenic
PiRNA expression；Fig. 2 B represent the influence to the cardiac muscle cell apoptosis of hypoxia inducible, and NC is as negative control for transfection.Fig. 2 shows
Show, the cardiac muscle cell apoptosis of hypoxia inducible can be significantly inhibited by suppressing endogenous piRNA.
Embodiment 4, piRNA GEM 132s suppress treating myocardial ischemia damage and apoptosis experiment
The present embodiment is using C57/BL6 mouse as experimental subjects, every group of 8 mouse, the result one-way that will be measured
ANOVO statistical softwares carry out statistical analysis, inject 30mg/kg piRNA GEM 132s (anta- as follows
PiRNA) and it negative control (NC) (the piRNA GEM 132s be 2 ' end carried out methoxy modification, i.e. 2 ' in nucleic acid
The hydroxyl at end is by methoxy substitution to strengthen the stability of nucleic acid.The modification is modified by Shanghai Ji Ma companies) (with purchase
It is negative control sequence from one section of unrelated sequences of Shanghai Ji Ma companies, it does not suppress the expression of existing known any gene,
Its sequence is：5′-CAGUACUUUUGUGUAGUACAA-3′)；Mouse is weighed and anaesthetized, and dorsal position is fixed, and cropping simultaneously uses the tincture of iodine
Sterile surgical area.Neck midsection tracheae is simultaneously intubated, and even animal respirator carries out positive airway pressure, at left border of sternum and heart
Longitudinal incision skin about 3cm at beating, successively blunt separation hypodermis, muscle, open chest, carefully lift pericardium and cut off, fill
Divide exposure heart；Dilute piRNA GEM 132s in phosphate buffer (PBS), final volume is 200 μ L, No. 26 conduits from
The apex of the heart enters artery root, and folder closes sustainer and pulmonary artery simultaneously when injecting piRNA GEM 132s, and lasting folder closes 20 seconds, supervises
5min is surveyed, treats that heart recovers Dou Lvhou, hematocele in thoracic cavity is removed and spends needle applicator intake-gas closing thoracic cavity, wait mouse
After clear-headed, depart from lung ventilator, put back in cage；The whole surgical procedure of electrocardiogram monitoring；It is laggard to inject piRNA GEM 132s 3 days
Row heart ischemia reperfusion is performed the operation, and concrete operations are as follows：After mouse anesthesia, back of the body position is fixed on experiment plank, and carries out the heart
Electrograph monitors.Neck midsection tracheae is simultaneously intubated, and is interlocked thing breathing apparatus, is opened chest in the 4th intercostal, carefully lift pericardium
And cut off, fully expose heart, blood vessel；Between pulmonary conus and left auricle of heart, using great cardiac vein trunk as mark, Yu Zuo
Inserting needle at the 2mm of auricle root lower section, depth of needle 0.5mm；Myocardium top layer is passed through with 6/0 band pin suture, in pulmonary artery cone
Branch pin；After electrocardiogram recovers stable 10min, ramus descendens anterior arteriae coronariae sinistrae (LAD) is ligatured；With I, aVL ST-Segment
It is hunchbacked to raise upwards more than 0.1mv and continue more than 0.5h as the successful mark of ligation, after ischemic 45min, unclamp ligation
Line starts Reperfu- sion, and mark is dropped to ST sections；Remove hematocele in thoracic cavity and spend needle applicator intake-gas closing thoracic cavity；
Reperfu- sion determines myocardial infarction areas in 24 hours with the double dyes of Evans blue/TTC；
PiRNA GEM 132s are as shown in Figure 3 to the resistant function for the treatment of myocardial ischemia damage；Wherein Fig. 3 A are mouse core intramuscular injection
Implement the result of the test that myocardial ischemia-reperfusion is performed the operation after penetrating piRNA GEM 132s, ARR/LV represents the hazardous area gross area/left side
Ventricle area, INF/AAR represent Infarct area/hazardous area gross area, and INF/LV represents Infarct area/left ventricular area,
Wherein every group is from left to right followed successively by the control group not plus handled, the control group (Con) through ischemia-reperfusion, through Ischemia Reperfusion
The negative control treatment group (NC) of note and the piRNA GEM 132s treatment group (anta-piRNA) through ischemia-reperfusion；Fig. 3 B
The result of cardiomyocyte apoptosis after piRNA GEM 132s is injected for mouse cardiac muscle.
As a result show：The mouse I/R groups myocardial infarction area for injecting piRNA GEM 132s is brighter than wild mouse I/R groups
It is aobvious to reduce, there is significant difference (p<0.01) Fig. 3 A, are seen.
The myocardial infarct size that the present embodiment is related to calculates as follows：Dangerous area (AAR/LV, %)=(hazardous area is total
Area/left ventricular area) × 100%, myocardial infarct size (INF/AAR, %)=(Infarct area/hazardous area gross area) ×
100%；It has detected influence of the injection piRNA GEM 132s to I/R apoptosis again simultaneously, detect (Fig. 3 B) with TUNEL, as a result
Display piRNA GEM 132s can significantly inhibit the generation of cardiac muscle cell apoptosis caused by myocardial ischemia-reperfusion, wherein with vacation
Operation group is as a control group；Result above illustrates that mouse underwent coronary injection piRNA GEM 132s can significantly inhibit the heart
Myocardial ischemia reperfusion injury.
Sequence table
SEQ ID NO:1
5’- CCUUGGCACAUGCGCAGAUUAUUUGUUUA-3’
Claims (1)
1. a kind of piRNA GEM 132s pharmaceutical composition is preparing treatment myocardial infarction, treating myocardial ischemia damage and cardiac muscle fibre
Application in the medicine of change, it is characterised in that the piRNA GEM 132s pharmaceutical composition includes piRNA GEM 132s
And carrier；The piRNA antisense base sequences are：
5’-CCUUGGCACAUGCGCAGAUUAUUUGUUUA-3’；The carrier is chitosan, cholesterol, nano particle and fat
One or more in plastid；The content of piRNA GEM 132s is 0.5- in the piRNA GEM 132s pharmaceutical composition
1 gram；The piRNA GEM 132s pharmaceutical composition is administered in a manner of oral or injection.
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| CN107151695B (en) * | 2016-12-08 | 2021-11-09 | 青岛大学 | PiRNA combination for detecting acute myocardial ischemia diseases and detection method and application thereof |
| CN106916885B (en) * | 2017-01-13 | 2021-11-12 | 青岛大学 | PiRNA combination for detecting heart disease and application thereof |
| CN107604058B (en) * | 2017-09-22 | 2020-12-18 | 青岛大学 | PiRNA-514 nucleotide analogue and application of antisense nucleotide thereof and product using same |
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| CN116694761B (en) * | 2023-04-29 | 2024-06-11 | 中国人民解放军总医院第二医学中心 | PiRNA marker for diagnosing breast cancer chemotherapy heart injury, kit and application thereof |
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Non-Patent Citations (3)
| Title |
|---|
| A germline-specific class of small RNAs binds mammalian Piwi proteins;A. Girard 等;《Nature》;20060713;第442卷(第7099期);摘要，第202页右栏最后一段 * |
| Cloning and expression profiling of testis-expressed piRNA-like RNAs;Seungil Ro 等;《RNA-a Publication of the RNA Society》;20071031;第13卷(第10期);第1693-1702页 * |
| 长链非编码RNA在心脏发育中的作用;李婷婷 等;《上海交通大学学报 医学版》;20141228;第34卷(第12期);第1834-1838页 * |
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