CN115747145A - Application of RBM22 in preparation of medicine for promoting myocardial regeneration - Google Patents
Application of RBM22 in preparation of medicine for promoting myocardial regeneration Download PDFInfo
- Publication number
- CN115747145A CN115747145A CN202211510049.2A CN202211510049A CN115747145A CN 115747145 A CN115747145 A CN 115747145A CN 202211510049 A CN202211510049 A CN 202211510049A CN 115747145 A CN115747145 A CN 115747145A
- Authority
- CN
- China
- Prior art keywords
- rbm22
- myocardial
- regeneration
- promoting
- medicine
- 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.)
- Pending
Links
Images
Landscapes
- Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
Abstract
The invention belongs to the technical field of biomedicine, and particularly relates to an application of RBM22 in preparation of a medicine for promoting myocardial regeneration. The promotion of the expression of RBM22 promotes the proliferation and regeneration of myocardial cells, thereby improving myocardial ischemic injury. Based on the discovery, the invention provides an application of RBM22 in preparing a medicine for promoting myocardial regeneration. The regeneration of myocardial cell proliferation can be promoted by promoting the expression of the RBM22 by the myocardial regeneration promoting medicine, so that the regeneration and repair of damaged myocardium after ischemic heart disease are realized, the myocardial fibrosis reaction is inhibited, and the effects of protecting the cardiac function and reducing the occurrence of heart failure after ischemic heart disease are achieved.
Description
Technical Field
The invention belongs to the technical field of biomedicine, and particularly relates to application of RBM22 in preparation of a medicine for promoting myocardial regeneration.
Background
Heart failure, as the end-stage course of cardiovascular disease, has a very high mortality rate, with a five-year survival rate of less than 50% for patients. Ischemic myocardial infarction is one of the major factors leading to heart failure. The onset of acute Myocardial Infarction (MI) can lead to ischemic necrosis of cardiomyocytes within a short period of time and cause a decline in cardiac function and the development of heart failure. In the early stages of ischemia, cardiomyocytes are lost on a large scale due to a lack of oxygen and nutrients. The synergistic effect of several processes such as DAMPs released after myocardial cell injury initiates the acute phase inflammatory response and activates cardiac fibroblasts to produce collagen and other extracellular matrix components to maintain the basic structure of the heart. In the late stage of cardiac ischemia, a large amount of myofibroblasts are gathered, and the myocardial contractility is reduced, thereby impairing the cardiac pumping function. Since adult cardiomyocytes have a limited proliferative capacity and cannot repair the damaged area, cardiac rational remodeling and fibrosis are caused, eventually leading to irreversible advanced heart failure.
Except for heart transplantation, clinical means for treating ischemic heart diseases such as ventricular assist and the like cannot replenish a large amount of lost myocardial cells at present. Therefore, the method of enhancing the proliferation ability of cardiomyocytes or stem cell transplantation can replenish lost cardiomyocytes, and thus has become a hot research point for the treatment of heart failure after myocardial infarction.
Previous studies report that RBM22 belongs to SLT11 family, contains a special RNA zinc finger binding domain C-x8-C-x5-C-x3-H, is a component of the catalytic core of spliceosome, and plays an important role in the alternative splicing of pre-mRNA. In addition, RBM22 is involved in gene regulation, and is capable of binding DNA and acting as a true transcription factor on a large number of target genes. It was found that increased expression of RBM22 in glioblastoma, while decreased expression of RBM22 affected cell function: the migration ability of tumor cells is reduced, the secretion of Vascular Endothelial Growth Factor (VEGF) is inhibited, the number of stem/progenitor cells in tumor spheres is reduced, and apoptosis is promoted. However, there is no report on whether RBM22 is involved in regenerative repair after cardiac stress injury.
Disclosure of Invention
The inventors of the present invention have found that the promotion of the expression of RBM22 promotes the proliferation and regeneration of cardiomyocytes, thereby improving ischemic injury of myocardium. Based on the discovery, the invention provides an application of RBM22 in preparing a medicine for promoting myocardial regeneration.
In the application of the RBM22 provided by the invention in preparing the medicine for promoting myocardial regeneration, the RBM can also have the following characteristics: the medicine for promoting myocardial regeneration has the following functions: promoting the proliferation and regeneration of myocardial cells, promoting the regeneration and repair of damaged cardiac muscle after ischemic heart disease, inhibiting myocardial fibrosis reaction, protecting cardiac function and reducing the occurrence of heart failure after ischemic heart disease.
In the application of the RBM22 provided by the invention in preparing the medicine for promoting myocardial regeneration, the RBM can also have the following characteristics: wherein the medicine for promoting myocardial regeneration is used for promoting the expression of RBM22 so as to promote the proliferation and regeneration of myocardial cells.
Action and Effect of the invention
According to the application of the RBM22 in the preparation of the medicine for promoting myocardial regeneration, because the expression of the RBM22 can regulate the proliferation and regeneration level of myocardial cells, the expression of the RBM22 is promoted by the medicine for promoting myocardial regeneration, so that the proliferation and regeneration of the myocardial cells can be promoted, the regeneration and repair of damaged myocardium after ischemic heart disease are realized, the myocardial fibrosis reaction is inhibited, and the effects of protecting the cardiac function and reducing the occurrence of heart failure after the ischemic heart disease are achieved.
Drawings
FIG. 1 is a graph showing a comparison between the expression levels of RBM22 in a sham-operated group and an apex removal model group of wild type newborn mice in examples of the present invention;
FIG. 2 is a graph showing the fluorescence staining results of primary cardiomyocytes of AAV9 control group and wild type suckling mice overexpressing RBM22 group when EDU is used as a cell proliferation marker in an example of the present invention;
FIG. 3 is a graph showing the results of fluorescence staining of primary cardiomyocytes of wild-type suckling mice in a white control group and an interfering RBM22 expression group when EDU is used as a cell proliferation marker in an example of the present invention;
FIG. 4 is a graph of the proliferation of primary cardiomyocytes in a white control group and in a wild-type suckling mouse with RBM22 expression interfering group when EDU is used as a cell proliferation marker in an example of the present invention;
FIG. 5 is a graph showing the fluorescence staining results of primary cardiomyocytes of AAV9 control group and wild type suckling mice overexpressing RBM22 when Ki67 is used as a cell proliferation marker in an example of the present invention;
FIG. 6 is a graph showing the fluorescence staining results of primary cardiomyocytes of wild type suckling mice in a white control group and a group interfering with expression of RBM22 when Ki67 is used as a cell proliferation marker in an example of the present invention;
FIG. 7 is a graph of the proliferation of primary cardiomyocytes in a white control group and in a wild-type suckling mouse interfering with RBM22 expression when Ki67 is used as a cell proliferation marker in an example of the present invention.
Detailed Description
In order to make the technical means, the creation features, the achievement purposes and the effects of the invention easy to understand, the following embodiments are specifically described with reference to the attached drawings.
< example >
In this example, the relationship between RBM22 and the effect of regeneration after myocardial injury was verified by experiments on mice, as follows:
in the embodiment, an apex cordis excision model is constructed for 3-day-old C57 mice, a pseudo-operation group and an apex cordis excision model group are arranged, western-Blot detection is carried out on tissue samples of the apex cordis part after 7 days to detect RBM22 expression change, and the experimental process is as follows:
total tissue protein was extracted using TPER, haltProteaseElhibitor Cocktail, PMSF protease inhibitor (ThermoFisher scientific), westernBlot using transmembrane solution, electrophoresis solution, TBST (Shanghai Yazyme biomedical science Co., ltd.) and using electrophoresis tank and transmembrane apparatus (Bio-Rad). The expression of RBM22 and beta-actin in each myocardial tissue group was analyzed by a full-automatic chemiluminescence fluorescence analysis system using energy and a super-sensitization chemiluminescence developer (ThermoFisher Scientific Co.).
FIG. 1 is a graph showing the comparison between the expression amount of RBM22 in the sham-operated group and the apex cordis removal model group of wild type newborn mice in the examples of the present invention.
As shown in figure 1, RBM22 is normally expressed in the Sham operation group (P3-Sham) without apex excision, and the RBM22 expression level in the myocardial tissue of the newborn mice is obviously increased after myocardial injury after apex excision in the apex excision model group (P3-AR).
In this example, the relationship between RBM22 and cardiomyocyte proliferation and regeneration was further verified by interfering with RBM22 expression and promoting RBM22 expression, and comparing with a control group, specifically as follows:
constructing siRNA which can interfere RBM22 expression in a targeted manner (siRNA-RBM 22, synthesized by Dharmacon, siRNAD-055902-01, siRNAGCAUUAUCAUUUA, siRNAD-055902-02. Separating 1-3 days old mice (C57 strain, purchased from Shanghai Seipple-Bikai laboratory animals Co., ltd.) into a control group (NC) and an interference RBM22 expression group (siRNA-RBM 22), a control group (AAV 9-Ctrl) and an overexpression RBM22 group (AAV 9-RBM 22), co-staining the four groups by using various myocardial cell proliferation indexes such as myocardial cell Marker alpha-actin, ki67, EDU and the like after treating for 48h, and detecting the difference of proliferation levels.
After C57 suckling mouse myocardial cells are cultured in vitro and siRNA-RBM22 and AAV9-RBM22 are transfected, immunofluorescence staining is carried out: after 4% paraformaldehyde (biosystems) was fixed at room temperature for 15 minutes, PBS and PBST prepared by Triton (Amresco) were diluted to prepare 1% bsa (biofrox) and then blocked at room temperature for 1 hour, the antibody against the cardiomyocyte marker α -actin (Abcam) was co-stained with the antibody against the cell growth marker Ki67 (Abcam) and EDU kit (petityagar), incubated overnight at 4 degrees and then rinsed three times with 1XPBS (GeneRay), incubated at room temperature for one hour with Alexa488, 594 secondary antibody (Invitrogen), rinsed three times with 1XPBS again, and after blocking with a DAPI-containing blocking agent (southern biotech), imaged with a fluorescent microscope, and the ratio of double positive cells was counted to reflect the proliferation of cardiomyocytes.
FIG. 2 is a graph showing the fluorescence staining results of primary cardiomyocytes of AAV9 control group and wild type suckling mice overexpressing RBM22 group when EDU is used as a cell proliferation marker in an example of the present invention; FIG. 3 is a graph showing the results of fluorescence staining of primary cardiomyocytes of wild-type suckling mice in a whitespace control group and a group interfering with expression of RBM22 when EDU is used as a cell proliferation marker in an example of the present invention; FIG. 4 is a graph of the proliferation of primary cardiomyocytes in a white control group and in a wild-type suckling mouse interfering with the expression of RBM22 when EDU is used as a marker for cell proliferation in an example of the present invention.
In FIGS. 2 and 3, the results of α -actin staining (cardiomyocytes), EDU staining (proliferating cells), and total fluorescence staining were shown in this order from left to right.
As shown in FIG. 2, the number of proliferating cells in the over-expressed RBM22 group (AAV 9-RBM 22) was significantly greater than that in the AAV9 control group (AAV 9-Ctrl).
As shown in FIGS. 3 and 4, the number of proliferating cells in the group interfering with expression of RBM22 (siRNA-RBM 22) was significantly less than that in the blank control group (NC), and the proportion of proliferating cells in the group interfering with expression of RBM22 was also significantly less than that in the blank control group.
FIG. 5 is a graph showing the fluorescence staining results of primary cardiomyocytes of AAV9 control group and wild type suckling mice overexpressing RBM22 when Ki67 is used as a cell proliferation marker in an example of the present invention; FIG. 6 is a graph showing the fluorescence staining results of primary cardiomyocytes of wild type suckling mice in a white control group and a group interfering with expression of RBM22 when Ki67 is used as a cell proliferation marker in an example of the present invention; FIG. 7 is a graph showing the proportion of proliferation of primary cardiomyocytes in wild type suckling mice in the white control group and in the group interfering with expression of RBM22 when Ki67 is used as a cell proliferation marker in the examples of the present invention.
In FIGS. 5 and 6, the results of α -actin staining (cardiomyocytes), ki67 staining (proliferating cells), and total fluorescence staining were shown from left to right.
As shown in FIG. 5, the number of proliferating cells in the over-expressed RBM22 group (AAV 9-RBM 22) was significantly greater than in the AAV9 control group (AAV 9-Ctrl).
As shown in fig. 6 and 7, the number of proliferating cells in the interfering RBM22 expressing group (siRNA-RBM 22) was smaller than that in the blank control group (NC), and the proportion of proliferating cells in the interfering RBM22 expressing group was also significantly lower than that in the blank control group.
In conclusion, interference with the expression of RBM22 can inhibit the proliferation of myocardial cells, and promotion of the expression of RBM22 can promote the proliferation and regeneration of the myocardial cells. Therefore, the expression of RBM22 is promoted to promote the proliferation and regeneration of myocardial cells, and further, the regeneration and repair of damaged myocardium are promoted, so that the myocardial ischemic injury is improved.
Further, methods for promoting expression of RBM22 include, but are not limited to, AAV9, mRNA technology, and accordingly, drugs for promoting expression of RBM22 are not limited to gene drugs prepared based on AAV9 and/or mRNA technology; any drug that can promote the overexpression of RBM22 can promote the proliferation and regeneration of myocardial cells, promote the regeneration and repair of damaged myocardium after ischemic heart disease, inhibit myocardial fibrosis reaction, protect cardiac function and reduce the occurrence of heart failure after ischemic heart disease.
Effects and effects of the embodiments
According to the application of the RBM22 in the preparation of the medicine for promoting myocardial regeneration, because the expression of the RBM22 can regulate the proliferation and regeneration level of myocardial cells, the expression of the RBM22 is promoted by the medicine for promoting myocardial regeneration, so that the proliferation and regeneration of the myocardial cells can be promoted, the regeneration and repair of damaged myocardium after ischemic heart disease are realized, the myocardial fibrosis reaction is inhibited, and the effects of protecting cardiac function and reducing the occurrence of heart failure after ischemic heart disease are achieved.
The above embodiments are preferred examples of the present invention, and are not intended to limit the scope of the present invention.
Claims (3)
- Application of RBM22 in preparing medicine for promoting myocardial regeneration is provided.
- 2. The use of an RBM22 according to claim 1 in the manufacture of a medicament for promoting myocardial regeneration, wherein:wherein, the medicine for promoting myocardial regeneration has the functions of: promoting proliferation and regeneration of myocardial cells, promoting regeneration and repair of damaged myocardium after ischemic heart disease, inhibiting myocardial fibrosis reaction, protecting cardiac function, and reducing occurrence of heart failure after ischemic heart disease.
- 3. The use of an RBM22 according to claim 1 in the manufacture of a medicament for promoting myocardial regeneration, wherein:wherein the medicine for promoting myocardial regeneration is used for promoting the expression of RBM22 so as to promote the proliferation and regeneration of myocardial cells.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202211510049.2A CN115747145A (en) | 2022-11-29 | 2022-11-29 | Application of RBM22 in preparation of medicine for promoting myocardial regeneration |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202211510049.2A CN115747145A (en) | 2022-11-29 | 2022-11-29 | Application of RBM22 in preparation of medicine for promoting myocardial regeneration |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN115747145A true CN115747145A (en) | 2023-03-07 |
Family
ID=85340082
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202211510049.2A Pending CN115747145A (en) | 2022-11-29 | 2022-11-29 | Application of RBM22 in preparation of medicine for promoting myocardial regeneration |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN115747145A (en) |
-
2022
- 2022-11-29 CN CN202211510049.2A patent/CN115747145A/en active Pending
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Hu et al. | Knockdown of lncRNA MALAT1 attenuates acute myocardial infarction through miR-320-Pten axis | |
| Wang et al. | CircUbe3a from M2 macrophage-derived small extracellular vesicles mediates myocardial fibrosis after acute myocardial infarction | |
| Long et al. | Down-regulation of miR-138 promotes colorectal cancer metastasis via directly targeting TWIST2 | |
| Chen et al. | Mesenchymal stem cells overexpressing MiR-126 enhance ischemic angiogenesis via the AKT/ERK-related pathway | |
| Wang et al. | A long noncoding RNA NR_045363 controls cardiomyocyte proliferation and cardiac repair | |
| Dai et al. | Micro RNA‐155 attenuates activation of hepatic stellate cell by simultaneously preventing EMT process and ERK 1 signalling pathway | |
| JP2021006047A (en) | Functional nucleic acid molecule, and utilization of the same | |
| Liang et al. | miRNA-204 drives cardiomyocyte proliferation via targeting Jarid2 | |
| Xu et al. | Micro RNA‐30c suppresses the pro‐fibrogenic effects of cardiac fibroblasts induced by TGF‐β1 and prevents atrial fibrosis by targeting TGF β RII | |
| Zhang et al. | miR‑29a suppresses IL‑13‑induced cell invasion by inhibiting YY1 in the AKT pathway in lung adenocarcinoma A549 cells | |
| Su et al. | lncRNA MIRF promotes cardiac apoptosis through the miR-26a-Bak1 axis | |
| Gu et al. | LncRNA promoted inflammatory response in ischemic heart failure through regulation of miR-455-3p/TRAF6 axis | |
| Fan et al. | Nuclear miR-665 aggravates heart failure via suppressing phosphatase and tensin homolog transcription | |
| Zhou et al. | MicroRNA-208b alleviates post-infarction myocardial fibrosis in a rat model by inhibiting GATA4 | |
| Yang et al. | miR-30a-5p inhibits the proliferation and collagen formation of cardiac fibroblasts in diabetic cardiomyopathy | |
| CN111718995A (en) | Biomarker for nasopharyngeal carcinoma metastasis diagnosis and/or prognosis evaluation | |
| Wang et al. | Krüppel-like factor 15 reduces ischemia-induced apoptosis involving regulation of p38/MAPK signaling | |
| Chen et al. | MiR-1908 improves cardiac fibrosis after myocardial infarction by targeting TGF-β1. | |
| Ye et al. | Cold-induced Yes-associated-protein expression through miR-429 mediates the browning of white adipose tissue | |
| CN115747145A (en) | Application of RBM22 in preparation of medicine for promoting myocardial regeneration | |
| Zhao et al. | Retracted: Tectonic 1 Is a Key Regulator of Cell Proliferation in Pancreatic Cancer | |
| Wang et al. | miR-409-3p regulated by GATA2 promotes cardiac fibrosis through targeting Gpd1 | |
| CN106390123A (en) | Mir-29 and application of inhibitor thereof in preparing medicine resisting organ transplant rejection | |
| WO2021155825A1 (en) | Use of pax6 gene or expression product thereof in preparation of drug for inhibiting fibrosis | |
| Ji et al. | Downregulation of lncRNA MALAT1 inhibits angiotensin II-induced hypertrophic effects of cardiomyocytes by regulating SIRT4 via miR-93-5p |
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 |