CN114344296B - Application of NAT10 inhibitor in diagnosis and treatment of heart failure caused by pressure overload - Google Patents
Application of NAT10 inhibitor in diagnosis and treatment of heart failure caused by pressure overload Download PDFInfo
- Publication number
- CN114344296B CN114344296B CN202210064755.2A CN202210064755A CN114344296B CN 114344296 B CN114344296 B CN 114344296B CN 202210064755 A CN202210064755 A CN 202210064755A CN 114344296 B CN114344296 B CN 114344296B
- Authority
- CN
- China
- Prior art keywords
- heart failure
- heart
- nat10
- inhibitor
- treatment
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Active
Links
Images
Landscapes
- Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
Abstract
The invention discloses application of a NAT10 inhibitor in diagnosis and treatment of heart failure caused by pressure overload. According to the invention, by establishing a heart failure animal model after aortic arch constriction of a mouse and gastric perfusion administration of the NAT10 inhibitor, the heart function, the ventricular remodeling condition and the expression of related marker molecules are evaluated by utilizing the technologies of heart overload, section staining and molecular biology, the heart function can be improved, the ventricular remodeling is improved, the heart failure related marker is reduced, the deposition of collagen and extracellular matrix is reduced, ac4C is remarkably inhibited after Remodelin intervention, and the TAC-induced heart remodeling and heart function of the mouse can be remarkably improved by the NAT10 inhibitor.
Description
Technical Field
The invention relates to application of a NAT10 inhibitor in diagnosis and treatment of heart failure caused by pressure overload, belonging to the field of biological medicines.
Background
Heart failure, as the terminal stage of progression of various cardiovascular diseases, has a high incidence and a poor prognosis. Although the survival rate of the heart failure patients is improved by the drug treatment of inhibiting the excessive activation of neuroendocrine such as 'golden triangle' and 'new tetrad', the life quality and the long-term prognosis are still poor, and the 1-year death rate of the heart failure patients is still as high as 17%.
Heart failure is classified into different types due to different mechanisms of occurrence. From the pathophysiological point of view, heart failure can be roughly divided into two main categories, i.e., the primary myocardial damage and the long-term volume (or) pressure overload of the heart, which leads to the final development of the pump function from compensation to decompensation. The heart failure is mainly the ventricular remodeling, that is, the pathological remodeling of the ventricles, including the change of the size, the shape, the tissue structure and the functional state of the ventricles, caused by the maladaptive change of the myocardium in the process of the heart failure, is the basic pathological process of the development of the heart failure.
Cardiac remodeling is an important pathophysiological basis for the development and progression of heart failure. At the gross morphological level, myocardial remodeling is mostly manifested by thickening, mass increase of local or most or even the entire ventricular wall or/and atrial wall. At the cellular level, single cardiomyocytes that fail to divide are hypertrophied, interstitial components such as fibroblasts proliferate, collagen synthesis and secretion increase, and apoptosis of the cardiomyocytes is accompanied, causing the myocardial tissue components to change. On the molecular level, the gene expression for promoting cardiac hypertrophy is increased, such as connective tissue growth factor CTGF, transforming growth factor TGF beta, type I and type III collagens, etc., which increase the stiffness of the ventricular wall and reduce the cardiac muscle compliance. Although cardiac remodeling is initially a compensatory mechanism, long-lasting cardiac remodeling can significantly increase the incidence and mortality of heart failure. At present, no effective prevention and treatment method for heart failure caused by cardiac remodeling exists clinically. Therefore, the discovery of specific molecules and molecular pathways for blocking heart remodeling to cause heart failure and the search of drug targets for preventing and treating heart failure have very important theoretical significance and clinical significance.
Epigenetics is increasingly receiving attention from researchers in the field of heart failure as an important means for regulating gene expression. RNA epigenetic post-transcriptional modification is an important link in epigenetics, both the protective role in the progression of various diseases by targeting 6-methyladenosine suggests the potential of RNA epigenetic post-transcriptional modification mechanisms in the treatment of heart failure. 4-acetyl cytidine (ac 4C) is a conserved type of post-transcriptional chemically modified nucleoside present on a variety of RNAs. The formation of ac4C is mainly catalyzed by N-acetyltransferase 10 (N-acetyltransferase 10, nat10), and is involved in the onset and progression of various diseases, including immune inflammatory reaction, metabolic diseases, autoimmune diseases, cancer, and the like. However, there is no report on the role of NAT10 inhibitors in the preparation of a medicament for the treatment of heart failure due to stress overload.
Disclosure of Invention
The technical problem to be solved by the invention is as follows: how to improve the curative effect and prognosis of heart failure caused by pressure overload such as cardiac remodeling.
In order to solve the technical problems, the invention provides application of an inhibitor of NAT10 gene or protein in preparing a medicament for treating heart failure caused by pressure overload.
Preferably, the inhibitor of the NAT10 gene or protein comprises Remodelin, which has the formula C 15 H 14 N 4 S, molecular weight of 282.36, chemical structural formula
The invention also provides a medicament or a pharmaceutical composition for treating the heart failure caused by the pressure overload, which comprises a pharmaceutically acceptable carrier and an effective amount of active ingredients, wherein the active ingredients comprise NAT10 gene or protein inhibitors.
Preferably, the inhibitor of the NAT10 gene or protein comprises Remodelin.
The invention also provides application of the reagent for detecting the expression or activity of the NAT10 gene or protein in preparing a reagent or a kit for diagnosing the heart failure caused by the pressure overload.
Preferably, the reagent for detecting expression or activity of NAT10 gene or protein comprises NAT10 antibody.
The invention also provides the use of an agent for detecting the level of ac4C modification in the manufacture of a reagent or kit for the diagnosis of heart failure due to pressure overload.
Compared with the prior art, the invention has the beneficial effects that:
according to the invention, the specific inhibitor Remodelin of NAT10 is creatively used for interfering TAC-induced pathological myocardial remodeling and heart failure of mice, and the fact that the Remodelin is significantly inhibited in prognosis ac4C is found, so that TAC-induced heart remodeling and heart function of the mice can be significantly improved.
Drawings
FIG. 1 shows ac4C modification and NAT10 expression levels in human heart failure heart tissue and TAC-induced heart failure heart tissue of mice; wherein A is the ac4C modification level in heart failure heart tissue and normal tissue of a human, B is the NAT10 expression level in heart failure heart tissue and normal tissue of the human, C is the ac4C modification level in heart failure heart tissue of a mouse induced by TAC and heart tissue of a sham-operated mouse, and D is the NAT10 expression level in heart failure heart tissue of the mouse induced by TAC and heart tissue of the sham-operated mouse;
fig. 2 is a 2-week intragastric gavage of NAT10 inhibitors based on constructed TAC-induced heart failure mice, and it was found that remodelain intervention significantly reduced the level of modification of ac4C (a), reduced ejection fraction EF (B), and reduced fraction FS (C);
FIG. 3 is a graph of HE staining, masson staining and WGA staining of mouse heart tissue 4 weeks after TAC operation (A), myocardial fibrosis detection (B), myocardial cell hypertrophy level detection (C), RT-qPCR detection of expression level of heart failure-related marker molecules (D), and RT-qPCR detection of expression level of collagen and extracellular matrix deposition-related genes (E);
in the above figures, the symbol indicates that the significant difference between the two groups is shown by statistical analysis, P is less than 0.05; the expression shows that the two groups have significant difference by statistical analysis, and P is less than 0.01; trepresents that the two groups have significant difference through statistical analysis, and P is less than 0.001.
Detailed Description
In order to make the invention more comprehensible, preferred embodiments accompanied with figures are described in detail below.
In a first aspect of the invention, there is provided the use of an inhibitor of the NAT10 gene or protein in the manufacture of a medicament for the treatment of heart failure.
The heart failure is caused by pressure overload.
The inhibitor of NAT10 gene or protein is an inhibitor for instructing the NAT10 protein to lose activity or reduce activity.
The inhibitor of the NAT10 gene or protein is selected from small molecular compounds or biological macromolecules.
The above-mentionedThe small molecule compound of (1) is Remodelin, the molecular formula of which is C 15 H 14 N 4 S, molecular weight of 282.36, chemical structural formula
In a second aspect of the present invention, a pharmaceutical composition for treating heart failure is provided, wherein the pharmaceutical composition comprises an inhibitor of NAT10 gene or protein as an active ingredient, and further comprises a pharmaceutically acceptable carrier.
The heart failure is caused by pressure overload.
The inhibitor of the NAT10 gene or protein is an inhibitor for promoting the loss or reduction of the activity of the NAT10 protein.
The dosage form of the pharmaceutical composition is injection, capsule, granule, tablet, pill or oral liquid, etc.
In a third aspect of the present invention, there is provided a use of NAT10 as a diagnostic marker in the preparation of a reagent or kit for heart failure diagnosis.
In a fourth aspect of the invention, there is provided the use of an agent for detecting the level of ac4C modification in the manufacture of a reagent or kit for the diagnosis of heart failure.
It should be noted that heart failure is classified into different types due to different mechanisms. From the pathophysiological point of view, heart failure can be roughly divided into two main categories, i.e., the primary myocardial damage and the long-term volume (or) pressure overload of the heart, which leads to the final development of the pump function from compensation to decompensation. The heart failure generation mechanism is mainly ventricular remodeling, namely pathological ventricular remodeling caused by maladaptive change of cardiac muscle in the heart failure development process, including change of ventricular size, shape, tissue structure and functional state, and is the basic pathological process of the heart failure generation and development. As used herein, the term "NAT10 inhibitor" includes antagonists, down-regulators, blockers, etc., as long as they are capable of reducing or losing NAT10 protein activity, or down-regulating the expression level of NAT10 or inhibiting its activity. They may be chemical compounds, chemical small molecules, biological molecules. The biomolecule may beAt the nucleic acid level (including DNA, RNA), or a viral product that inhibits NAT10 expression. By way of example, the inhibitors are: compound Remodelin, formula C 15 H 14 N 4 S, molecular weight 282.36. Nucleic acid inhibitors, protein inhibitors, nucleases, nucleic acid binding molecules, as long as they are capable of reducing or abolishing the activity of NAT10 protein, or down-regulating the expression of NAT 10.
The mice referred to in the following examples were purchased from the collective pharmaceutical biotechnology corporation, the NAT10 specific inhibitor Remodelin was purchased from the MCE (MedChemExpress) corporation, and the animal experiments were approved by the ethical committee of the subsidiary zhongshan hospital of the university of counterden.
Examples
This example provides the use of NAT10 inhibitors in the diagnosis of heart failure due to stress overload:
1. constructing a mouse aortic arch constriction (TAC) model:
adult male C57bl/6 mice, 8 weeks old and 20-25g in body weight, were selected, anesthetized, fixed, depilated, prepared, and then the skin was cut from the superior fossa of the sternum and the median line of the sternum to the second intercostal space (approximately 1cm incision). The fascia is separated at the suprasternal fossa in a blunt way, the air outlet pipe is exposed, and then the forceps with small bends enter the suprasternal fossa to push the thymus tissue to two sides. The field of vision is exposed by adopting a retractor, and a No. 5 thread is penetrated into a blood vessel at the head and arm trunk through the front section of a self-made syringe needle (the front section is cut off and made into a right angle) and is knotted in advance. Then pad with a 4 mm 27G needle (made square) and tie off. And (4) slowly pulling out the needle head of 27G with the length of 4 mm after ligation, suturing muscle tissues, placing in a warm environment after skin suturing for revival, and completing model construction. The prescription and method of the pseudo-operation are consistent, and only the threading is needed without ligation.
2. And (3) NAT10 expression and ac4C acetylation modification detection of clinical heart failure heart samples and heart failure mouse heart samples caused by TAC:
through ethical examination and informed consent, 3 cases of left ventricle myocardial tissues of patients who need heart transplantation operation and have final end-stage heart failure caused by primary dilated cardiomyopathy in clinical diagnosis of Zhongshan hospital affiliated to the university of Compound Dan are collected. 3 cases of left ventricular myocardial tissue of a control group of non-heart failure patients were also collected. In addition, 6 cases of the heart tissues of the mice in the sham-operated group and the TAC-induced heart failure group were collected. Respectively extracting total RNA of the samples, detecting the content of ac4C acetylation modification by liquid chromatography-tandem mass spectrometry (LC-MS), and detecting the expression of NAT10 by an RT-qPCR method.
3. NAT10 inhibitor remodelain intervention:
TAC surgery induced stress overload induced heart failure in mice was performed as described above. The treatment group is randomly grouped at 3 days after TAC operation, NAT10 inhibitor Remodelin (100 mg/kg/d) is given to the treatment group in a gastric perfusion mode, DMSO (dimethyl sulfoxide) with the same volume is given to the control group for intragastric perfusion, and the administration is continuously carried out for 2 weeks until 4 weeks after TAC, heart ultrasonic detection on cardiac function, HE (human immunodeficiency Virus) staining detection on myocardial morphological change, masson staining detection on fibrosis, WGA (tissue culture and fibroblast) staining detection on myocardial cell hypertrophy level, and RT-qPCR (reverse transcription-quantitative polymerase chain reaction) detection on heart failure related marker molecules, collagen and extracellular matrix deposition related gene expression.
4. Cardiac ultrasonic detection of cardiac structure and function:
after dehairing, mice were anesthetized with isoflurane and mounted on a heating pad (1% -2%). Isoflurane flow was carefully controlled to maintain the heart rate around 450 beats/min. The geometry and function of the heart were assessed using a two-dimensional (2-D) guided M-mode echocardiogram (VisualSonics Vevo 2100, canada) equipped with a 30mHz linear transducer. A parasternal long axis view was obtained while recording the heart rate. Obtaining the geometric shape and functional index of the heart: ejection Fraction (EF), fractional Shortening (FS). Each measurement indicator is averaged over at least 5 consecutive cardiac cycles.
5. Staining and checking a heart specimen section:
heart material was obtained after TAC 4 weeks, fixed with paraformaldehyde and then embedded in paraffin and sectioned. After the section is made, dewaxing and hydrating are carried out conventionally, and the morphology of the heart tissue, the fibrosis degree and the surface area of the myocardial cells are observed by HE, masson and WGA staining.
6. The RT-qPCR method is used for analyzing the expression of heart failure related markers such as Nppa, nppb, myh7, col1a1, col3a1, ctgf and Lox, collagen and extracellular matrix deposition related genes, and the improvement condition of the NAT10 inhibitor on the TAC-induced heart failure related marker molecule expression is proved.
7. The experimental results are as follows:
(1) In the experiment, ac4C modification and NAT10 expression of heart failure heart tissue which is clinically diagnosed as heart failure caused by dilated cardiomyopathy and heart failure heart tissue of mice induced by TAC are detected by LC-MS and RT-qPCR methods, and the ac4C modification and NAT10 expression are found to be remarkably upregulated in both human heart specimens and heart specimens of mice, as shown in figure 1.
(2) The constructed TAC-induced heart failure mice are subjected to intragastric gavage of the NAT10 inhibitor for 2 weeks, and Remodelin intervention is found to be capable of remarkably reducing the modification level of ac4C and improving the cardiac function index, as shown in figure 2, the NAT10 inhibitor can also improve TAC-induced cardiac insufficiency and ventricular remodeling, reduce the expression of myocardial hypertrophy and heart failure related marker molecules and improve the deposition of collagen and extracellular matrix, as shown in figure 3.
The above-described embodiments are only preferred embodiments of the present invention, and are not intended to limit the present invention in any way and substantially, it should be noted that those skilled in the art may make several modifications and additions without departing from the scope of the present invention, which should also be construed as a protection scope of the present invention.
Claims (1)
- Use of an inhibitor of the NAT10 gene or protein for the manufacture of a medicament for the treatment of heart failure due to stress overload, wherein the inhibitor of the NAT10 gene or protein is Remodelin with the molecular formula C 15 H 14 N 4 S, molecular weight of 282.36, chemical structural formula/>
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202210064755.2A CN114344296B (en) | 2022-01-20 | 2022-01-20 | Application of NAT10 inhibitor in diagnosis and treatment of heart failure caused by pressure overload |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202210064755.2A CN114344296B (en) | 2022-01-20 | 2022-01-20 | Application of NAT10 inhibitor in diagnosis and treatment of heart failure caused by pressure overload |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN114344296A CN114344296A (en) | 2022-04-15 |
| CN114344296B true CN114344296B (en) | 2023-03-31 |
Family
ID=81090615
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202210064755.2A Active CN114344296B (en) | 2022-01-20 | 2022-01-20 | Application of NAT10 inhibitor in diagnosis and treatment of heart failure caused by pressure overload |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN114344296B (en) |
-
2022
- 2022-01-20 CN CN202210064755.2A patent/CN114344296B/en active Active
Also Published As
| Publication number | Publication date |
|---|---|
| CN114344296A (en) | 2022-04-15 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Heather et al. | Isoproterenol induces in vivo functional and metabolic abnormalities; similar to those found in the infarcted rat heart | |
| Stansfield et al. | Periostin is a novel factor in cardiac remodeling after experimental and clinical unloading of the failing heart | |
| US20110196017A1 (en) | Micro-rna that promotes vascular integrity and uses thereof | |
| CN105194651B (en) | CREG albumen is used for the medical usage of Ischemic myocardium reperfusion injury | |
| WO2018028249A1 (en) | Mirna and use thereof in treatment of metabolic disease | |
| CN107693784B (en) | Application of PEDF gene in treatment of diabetic myocardial injury | |
| Zhao et al. | Attenuation of atrial remodeling by aliskiren via affecting oxidative stress, inflammation and PI3K/Akt signaling pathway | |
| Wang et al. | MicroRNA-29b upregulation improves myocardial fibrosis and cardiac function in myocardial infarction rats through targeting SH2B3. | |
| Deng et al. | Distinct phenotypes induced by different degrees of transverse aortic constriction in C57BL/6N mice | |
| Li et al. | Zerumbone, a humulane sesquiterpene from Syringa pinnatifolia, attenuates cardiac fibrosis by inhibiting of the TGF-β1/Smad signaling pathway after myocardial infarction in mice | |
| CN114344296B (en) | Application of NAT10 inhibitor in diagnosis and treatment of heart failure caused by pressure overload | |
| US20240016769A1 (en) | Method for preparing compound or biological drug enhancing CNPase activity for treating heart diseases | |
| CN113456621A (en) | Application of curcumin metabolite in preparation of medicine for treating myocardial ischemia-related injury | |
| WO2021073249A1 (en) | USE OF β-NMN IN PREPARATION OF DRUG FOR TREATING AND PREVENTING SEPSIS-INDUCED ORGAN DAMAGE | |
| Sivakumar et al. | Cardiac mitochondrial dynamics: miR-mediated regulation during cardiac injury | |
| CN114504650B (en) | Application of Lgmn as target in treating ischemic heart disease | |
| Su et al. | Takotsubo-like cardiomyopathy in pheochromocytoma | |
| CN112057624B (en) | Application of LARP7 in diagnosis and treatment of heart failure, atherosclerosis or aging | |
| CN114848622A (en) | Application of SBFI-26 in preparation of medicines for treating pulmonary hypertension | |
| CN107625781B (en) | Application of miRNA inhibitor in preparation of medicine for preventing and treating myocardial infarction | |
| Tai et al. | CREG improves cardiac function by regulating cardiomyocytes’ autophagy in diabetic myocardial infarction rats. | |
| Rahman et al. | Differential effects of doxorubicin on atrial natriuretic peptide expression in vivo and in vitro | |
| Li et al. | The effect of YiQiFuMai on ischemic heart failure by improve myocardial microcirculation and increase eNOS and VEGF expression | |
| CN103243091A (en) | MiRNA-140 inhibitor and applications thereof | |
| Jin | The Therapeutic Effect of miRNA-19a/19b on Heart Failure in Mice and the Mechanism of Myocardial Regeneration and Repair |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |