CN113913506A - Application of miR-144-3p in preparation of reagent or medicine for diagnosing or treating myocardial injury - Google Patents
Application of miR-144-3p in preparation of reagent or medicine for diagnosing or treating myocardial injury Download PDFInfo
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Abstract
The invention provides application of miR-144-3p in preparation of a reagent or a medicine for diagnosing or treating myocardial injury, and belongs to the technical field of biological medicines. The invention establishes mouse AngII chronic myocardial injury and inhibits or over-expresses miR-144-3p in vitro to explore a specific mechanism of AngII for inhibiting the myocardial injury caused by Nrf 2. The result shows that AngII can promote miR-144-3p, inhibit the antioxidant function of Nrf2 and aggravate the occurrence of myocardial damage caused by oxidative stress, the invention provides the application of miR-144-3p in the preparation of a reagent or a medicament for diagnosing or treating myocardial damage, can provide a brand new molecular target for the diagnosis and the prevention of myocardial oxidative damage and cardiomyopathy caused by AngII, and lays theoretical and experimental foundation for the prevention and the treatment of clinical cardiomyopathy.
Description
Technical Field
The invention belongs to the technical field of biological medicines, and particularly relates to application of miR-144-3p in preparation of a reagent or a medicine for diagnosing or treating myocardial injury.
Background
Angiotensin ii (angii) has been shown to play an important promoting role in the pathological process of various heart diseases in the local tissues of the heart. AngII can cause oxidative stress by producing excess Reactive Oxygen Species (ROS) and Reactive Nitrogen Species (RNS), thereby promoting the pathological processes of various heart diseases, such as acute myocardial infarction, alcoholic cardiomyopathy and diabetic cardiomyopathy. The Nuclear factor E2-related factor 2(Nuclear factor erythroid2-related factor, Nrf2) is an important regulatory factor of oxidative stress, Nrf2 plays an important protective role in myocardial injury, and according to literature reports, in some diseases such as aging, diabetes, hypertension and chronic inflammation, the risk of cardiovascular diseases is increased by down-regulation of a Nrf2 signal pathway, and heart-specific high-expression Nrf2 can effectively resist myocardial oxidative stress and injury caused by AngII. Furthermore, in the late stage of AngII-induced myocardial injury, exacerbation of myocardial oxidative damage is accompanied by downregulation of myocardial Nrf2 expression. Enhancing the Nrf2 antioxidant signaling pathway may also alleviate AngII-induced myocardial fibrosis and myocardial hypertrophy. The Nrf2 knockout can aggravate heart injury caused by AngII, and promote myocardial fibrosis and myocardial hypertrophy. Although these studies fully confirm the key role of Nrf 2-mediated antioxidant defense system in the development of AngII myocardial injury. However, it is not clear what causes the expression and cardiac function of myocardial Nrf2 are gradually reduced under AngII stimulation, and whether upstream regulatory molecules exist or not. The molecules which specifically act on the myocardial Nrf2 are helpful for more exactly understanding the regulation of myocardial oxidation-reduction balance, and provide a new direction for preventing and treating the cardiomyopathy related to the oxidative stress.
Disclosure of Invention
In view of the above, the invention aims to provide an application of miR-144-3p in preparation of a reagent or a medicament for diagnosing or treating myocardial injury, wherein miR-144-3p realizes a regulation function on myocardial injury by regulating the expression of Nrf 2.
The invention provides application of miR-144-3p as a biomarker in preparation of a reagent for diagnosing myocardial injury.
The invention provides application of miR-144-3p as a molecular target in preparation of a medicament for treating myocardial injury.
The invention provides application of a reagent for inhibiting miR-144-3p expression in preparation of a medicament for treating myocardial injury.
Preferably, the miR-144-3p exacerbates myocardial injury by negatively regulating expression of Nrf 2.
Preferably, the miR-144-3p realizes the Nrf2 gene regulation by complementing with the 3' UTR of Nrf 2.
Preferably, the myocardial damage includes myocardial oxidative damage and cardiomyopathy.
Preferably, the myocardial injury is induced by AngII.
Preferably, the reagent for inhibiting the expression of miR-144-3p comprises miR-144-3p inhibitor.
The invention provides application of a miR-144-3p overexpression reagent in preparation of a myocardial injury aggravation model.
Preferably, the reagent for over-expressing miR-144-3p comprises miR-144-3p imic.
The invention provides application of miR-144-3p as a biomarker in preparation of a reagent for diagnosing myocardial injury. According to the invention, the chronic myocardial injury animal model is constructed by subcutaneously injecting AngII, and the model mouse has the symptoms of left ventricular dilatation, myocardial hypertrophy, cardiac insufficiency, myocardial oxidative stress and injury, and the myocardial injury is more serious along with the prolonging of the treatment time. The application also verifies the expression of miR-144-3p in the myocardial tissues of the mice at different times (2M, 4M and 6M), and the expression of miR-144-3p is obviously reduced at 2M; miR-144-3p expression was significantly upregulated at 4M and 6M. This indicates that the expression level of miR-144-3p can reflect the degree of myocardial injury. Meanwhile, the experimental result also shows that the oxidative damage of the AC16 cardiac muscle cells induced by AngII can be promoted by over-expressing the miR-144-3p, and the oxidative damage of the AC16 cardiac muscle cells induced by AngII can be reduced by inhibiting the miR-144-3 p. Therefore, the occurrence condition of myocardial injury can be accurately diagnosed by detecting the expression quantity of miR-144-3 p. Therefore, the invention provides a new idea for diagnosing myocardial damage.
The invention provides application of miR-144-3p as a molecular target in preparation of a medicament for treating myocardial injury. The chronic myocardial injury animal model successfully constructed by injecting AngII subcutaneously is taken as a material, the expression of Nrf2 is increased in a compensatory way and then gradually decreased when cardiac function is not complete yet in the early stage of myocardial injury, and is obviously lower than a normal level when the cardiac function is abnormal. The application verifies the expression of miR-144-3p in myocardial tissues of 2M, 4M and 6M mice, and shows that the expression of miR-144-3p is obviously reduced at 2M; miR-144-3p expression was significantly upregulated at 4M and 6M. Prediction of a binding site of an Nrf 23 'UTR region is carried out through bioinformatics website TargetScan, and the fact that a complementary binding site exists between miR-144-3p and the 3' UTR of Nrf2 is found, meanwhile, inhibition of miR-144-3p can remarkably increase activation of Nrf2 in AC16 cardiac muscle cells under AngII stimulation, over-expression of miR-144-3p can inhibit activation of Nrf2 in AC16 cardiac muscle cells under AngII stimulation, and miR-144-3p can influence myocardial injury by regulating expression of Nrf 2. Therefore, the invention provides that miR-144-3p influences myocardial cell oxidative damage caused by AngII through reversely regulating the expression of Nrf 2. In addition, because Nrf2 is an important regulator in an oxidative stress pathway, and activation of Nrf2 can protect heart injury and cardiac hypofunction caused by oxidative stress, the purpose of treating myocardial injury is achieved by regulating the expression of miR-144-3 p.
Drawings
FIG. 1A is the results of cardiac ultrasound measurements of control and AngII mice when AngII was treated with 2M;
FIG. 1B is the results of cardiac ultrasound measurements of control and AngII mice when AngII was treated with 4M;
FIG. 1C is the results of cardiac ultrasound measurements of control and AngII mice at 6M treatment with AngII;
FIG. 1D shows the results of the variation of the cardiac ultrasound measurements in the control group of mice and the AngII group of mice at different treatment times;
FIG. 1E is the results of the ratio of heart weight to body weight of control mice and AngII mice at different treatment times;
FIG. 1F shows the results of 4-HNE expression in myocardial tissues of control mice and AngII mice at different treatment times;
FIG. 1G shows the results of Nrf2mRNA expression in hearts of control mice and AngII mice at different treatment times;
FIG. 1H shows the results of miR-144-3p expression in hearts of control mice and AngII mice at different treatment times;
FIG. 2A is the result of prediction of miR-144-3p and Nrf 23' UTR region binding sites using the bioinformatics website TargetScan;
FIG. 2B is a statistical result of the surface area of the cardiomyocytes of the miR-144-3p inhibitor group AC 16;
FIG. 2C is a statistical result of the surface area of the AC16 cardiomyocytes of the miR-144-3p imic group;
FIG. 2D is the expression result of miR-144-3p inhibitor/AngII group 4-HNE;
FIG. 2E is the expression result of miR-144-3p imic/AngII group 4-HNE;
FIG. 2F shows the result of Nrf2 expression in the cardiac myocytes of miR-144-3p inhibitor group AC 16;
FIG. 2G shows the HO-1 expression results of miR-144-3p inhibitor group AC16 cardiomyocytes;
FIG. 2H shows the expression result of Nrf2 in the miR-144-3p mimic group AC16 cardiomyocytes;
FIG. 2I shows the expression result of HO-1 in the cardiac myocytes of miR-144-3p imic group AC 16.
Detailed Description
The invention provides application of miR-144-3p as a biomarker in preparation of a reagent for diagnosing myocardial injury.
The method can reflect whether the myocardial damage occurs or not based on the expression quantity of the miR-144-3p, takes the miR-144-3p as a biomarker, judges whether the myocardial damage occurs or not by detecting the relative expression quantity of the miR-144-3p, and provides a new diagnosis idea for diagnosing the myocardial damage and diseases causing the myocardial damage. The myocardial damage preferably comprises myocardial oxidative damage or cardiomyopathy.
The nucleotide sequence of miR-144-3p is not particularly limited in the invention, and the nucleotide sequence of miR-144-3p known in the field can be adopted. In the embodiment of the invention, the nucleotide sequence of the miR-144-3p is shown as SEQ ID NO 1 (TACAGTATAGATGATGTACT).
In the invention, the reagent is a reagent for detecting the expression quantity of miR-144-3 p. The invention does not specially limit the types of the reagents, and the primers for RT-qPCR detection are designed by adopting a primer design method known in the field and taking miR-144-3p as a template. The method for diagnosing myocardial damage is operated according to a conventional RT-qPCR detection method.
The invention provides application of miR-144-3p as a molecular target in preparation of a medicament for treating myocardial injury.
In the invention, besides the expression quantity of miR-144-3p can reflect the occurrence of myocardial injury, the miR-144-3p can also be used as a molecular target for treating myocardial injury. The results of the examples show that the increase of the surface area of the myocardial cells caused by AngII can be reduced by reducing the expression amount of miR-144-3p, the oxidative damage of AC16 myocardial cells induced by AngII is reduced, the activation of Nrf2 in AC16 myocardial cells under the stimulation of AngII is obviously increased, and the oxidative damage caused by AngII is reduced, while the severity of the myocardial damage is aggravated by over-expressing miR-144-3 p. Therefore, the miR-144-3p is an important molecular target for regulating myocardial damage, and can be used for treating myocardial damage and diseases capable of causing myocardial damage by reducing the expression quantity of the miR-144-3 p. Therefore, the invention provides application of a reagent for inhibiting miR-144-3p expression in preparation of a medicine for treating myocardial injury.
In the present invention, the miR-144-3p preferably exacerbates myocardial injury by negatively regulating expression of Nrf 2. The miR-144-3p is preferably combined with Nrf2 gene through complementation with 3' UTR of Nrf2 so as to inhibit the expression of Nrf2 in myocardial cells, and the miR-144-3p influences oxidative damage caused by AngII by regulating the expression of Nrf2 in view of the effect of protecting myocardial oxidative stress and damage of Nrf 2. The myocardial damage preferably includes myocardial oxidative damage and cardiomyopathy. The myocardial injury is preferably induced by AngII.
In the invention, the reagent for inhibiting the expression of miR-144-3p preferably comprises miR-144-3p inhibitor. In the present example, miR-144-3p inhibitor was purchased from Ruibo, Guangzhou.
The invention provides application of a miR-144-3p overexpression reagent in preparation of a myocardial injury model.
In the invention, the miR-144-3p is taken as a molecular target, and the increase of the expression quantity of the miR-144-3p can aggravate the myocardial damage degree, so that the miR-144-3p overexpression reagent is applied to the preparation of a myocardial damage aggravation model.
In the invention, the reagent for over-expressing miR-144-3p preferably comprises miR-144-3p imic. In the present example, miR-144-3p imic was purchased from Ruibo, Guangzhou. The example results show that the over-expression of miR-144-3p can promote the increase of the surface area of the myocardial cells caused by AngII, promote the oxidative damage of AC16 myocardial cells induced by AngII, reduce the expression of Nrf2 and HO-1, and further inhibit the activation of Nrf 2.
Based on the action relationship between miR-144-3p and Nrf2, the invention also preferably provides application of miR-144-3p in preparation of a reagent for regulating expression of Nrf 2.
The application of miR-144-3p provided by the invention in the preparation of a reagent or medicament for diagnosing or treating myocardial injury is described in detail with reference to the following examples, but the application is not to be construed as limiting the scope of the invention.
Example 1
In vivo experiments, 8-week-old male C57/BL mice were used and grouped as follows:
1. control group: injecting normal saline with corresponding dose every other day subcutaneously for 2 months, and observing for 6 months;
2. AngII group: injecting Ang II (0.5mg/kg) subcutaneously every other day for 2 months, and observing for 6 months;
mice were sacrificed at 2 months (2M), 4 months (4M) and 6 months (6M) after the start of the experiment, and observed for myocardial damage.
Second, index measurement
1. Detecting the structure and the function of the heart by adopting a heart ultrasonic method, and calculating related indexes according to heart ultrasonic image data: cardiac dilation index LVID (left ventricular end-diastolic diameter), myocardial hypertrophy index LVPW (left ventricular posterior wall thickness in diastole), index of systolic function FS% (fractional shortening) and EF% (ejection fraction).
The results show that at 2M, there was no significant change in LVID, LVPW, FS% and EF% in the AngII group mice compared to the control group mice; at 4M, 6M, the LVID and LVPW of the AngII group mice were significantly increased and the FS% and EF% were significantly decreased compared to the control group mice (fig. 1A-1D). This suggests that chronic effects of low doses of AngII cause left ventricular dilatation, myocardial hypertrophy and cardiac insufficiency in mice.
2. The weight of the heart and the body weight of the mouse are measured at different times, and the ratio of the weight of the heart and the body weight of the mouse is calculated.
Compared with the control group of mice, the heart weight/body weight ratio of the mice in the AngII group has no obvious change at 2M; at 4M and 6M, the AngII group mice had significantly increased heart weight/body weight ratios (fig. 1E). The above results indicate that low-dose long-term effects of AngII can cause myocardial hypertrophy in mice.
3. Detecting the expression level of 4-HNE in mouse myocardial tissues at different time, specifically adopting conventional WesternBlot to detect, wherein the internal reference is beta-actin protein.
The expression of 4-HNE in myocardial tissue of the mice of AngII group was significantly higher than that of the control group at 2M, 4M, and 6M (FIG. 1F). The result shows that the low-dose long-term effect of AngII can cause the myocardial oxidative stress and injury of mice.
From the measured cardiac dilation index LVID (left ventricular end diastolic diameter), myocardial hypertrophy index LVPW (left ventricular posterior wall thickness in diastole), indexes of cardiac contractility FS% (fractional shortening) and EF% (ejection fraction), ratio of mouse heart weight and body weight and expression of 4-HNE, the indexes can prove that mouse cardiac muscle is damaged, and the model is successfully constructed.
4. The expression of Nrf2mRNA in mouse hearts at different time is determined by the following specific method:
1) preparing a reverse transcription system: 1000ng total RNA, 4. mu.l reaction mix (5X EasyScript All-in-One SuperMix for qPCR), and 1. mu.l gDNA remover (gDNAROMOVER), the system was made up to 20. mu.l with double distilled water. The reverse transcription reaction temperature was set to: the reaction was carried out at 42 ℃ for 15 minutes and at 85 ℃ for 5 seconds, and the temperature was gradually lowered to 4 ℃ after the reaction was stopped.
2) The real-time fluorescence quantitative PCR is used for detecting the expression of genes, and the primers are synthesized and provided by Shanghai bioengineering GmbH.
Primers for Nrf 2: forward (5 '-3') TCACACGAGATGAGCTTAGGGCAA (SEQ ID NO:2), Reverse (5 '-3') TACAGTTCTGGGCGGCGACTTTAT (SEQ ID NO: 3).
The internal references are beta-actin, Forward (5 '-3') TCATCACTATTGGCAACGAGC (SEQ ID NO:4), Reverse (5 '-3') AACAGTCCGCCTAGAAGCAC (SEQ ID NO: 5).
The reaction system is as follows: mu.l of HiefffqPCR SYBR Green MasterMix (Hight Rox Plus), 0.4. mu.l of Forward Primer (10. mu.M), 0.4. mu.l of Reverse Primer (10. mu.M), 100ng of cDNA, the system was made up to 20. mu.l with double distilled water. The reaction procedure is as follows: performing pre-denaturation reaction at 95 ℃ for 5 min; denaturation reaction at 95 ℃ for 10 seconds, annealing/extension at 60 ℃ for 31 seconds, 40 cycles. The System was subjected to fluorescent quantitative results analysis by Applied Biosystem 7300 Plus Real-time PCR System software and passed 2-△△CtThe method of (3) performs a quantitative statistics of the results.
The results showed that at 2M, Nrf2mRNA expression was significantly elevated in the hearts of AngII mice; whereas at 4M and 6M, Nrf2mRNA expression decreased, and expression downregulation was more pronounced at 6M (fig. 1G). Reflecting that during the early period of myocardial damage caused by AngII, when cardiac insufficiency does not occur, the expression of Nrf2 shows compensatory increase and then gradually declines until the abnormal cardiac function is obviously lower than the normal level.
5. High throughput transcriptome sequencing of myocardial tissues of 2M normal mice and AngII group mice was performed and was committed to Hangzhou Union Biotechnology, Inc.
And simultaneously measuring the miR-144-3p expression quantities at different times, wherein the specific method comprises the following steps:
1) preparing a reverse transcription system: 1000ng total RNA, 1. mu.l 2U/. mu.l Poly A Polymerase, 1. mu.l RTase Mix, 4. mu.l 5XPAP/RT Buffer, and the system was supplemented to 20. mu.l with double disutilled water. The reverse transcription reaction temperature was set to: reacting at 37 ℃ for 60 minutes and 85 ℃ for 5 minutes, and gradually cooling to 4 ℃ after the reaction is stopped.
2) Fluorescent quantitative PCR detection of MiR-144-3 p: the primer of MiR-144-3P is mmu-miR-144-3P (MmirQP0190), and the reference primer is mouse snRNA U6 qPCr primer (MmirQP9002), which are all synthesized and provided by Guangzhou energy recovery company. The reaction system is as follows: mu.l of a 2 Xone-in-One qPCR mixture (All-in-One TM Qpcr Mix), 2. mu.l of Universal adapter PCR Primer (2. mu.M), 2. mu.l of a 2 Xone-in-One microRNA qPCR Primer mixture (All-in-One TM miRNA qPCR Primer) (2. mu.M), 0.4. mu.l of a 50 XRox Reference Dye (ROX Reference Dye), 2. mu.l of cDNA and 3.6. mu.l of double distilled water. The reaction temperature was set as: the pre-denaturation reaction is carried out for 10min at the temperature of 95 ℃, and then the denaturation reaction is carried out for 10 seconds at the temperature of 95 ℃, the renaturation reaction is carried out for 20 seconds at the temperature of 60 ℃ and the extension reaction is carried out for 10 seconds at the temperature of 72 ℃ for 40 cycles in total. The System was subjected to fluorescent quantitative results analysis by Applied Biosystem 7300 Plus Real-time PCR System software and passed 2-△△CtThe method of (3) performs a quantitative statistics of the results.
The result shows that the expression of miR-144-3p is obviously reduced. The expression of miR-144-3p is verified in myocardial tissues of 2M, 4M and 6M mice, and the expression of miR-144-3p is obviously reduced at 2M, which is the same as a sequencing result; miR-144-3p expression was significantly up-regulated at 4M and 6M, in contrast to the sequencing results (fig. 1H). This shows that the expression quantity of miR-144-3p verifies the accuracy of the sequencing result of the high-throughput transcriptome, and the expression trends of miR-144-3p and Nrf2 are opposite in 2M, 4M and 6M, which indicates that miR-144-3p may have a regulation effect on Nrf 2.
Example 2
To further clarify the role relationship between miR-144-3p and Nrf2 and in myocardial injury, in vitro experiments were performed using the AC16 human cardiomyocyte cell line, grouped as follows:
1) control group (NC): transfecting AC16 cardiac muscle cells by using NC with the concentration of 50nM, and culturing for 48 h;
2) NC + AngII group: transfecting AC16 cardiac muscle cells by using NC with the concentration of 50nM, adding AngII with the concentration of 100nmol/L, and culturing for 48 h;
3) inhibition of miR-144-3p group: transfecting AC16 cardiac muscle cells by miRNA-144-3p inhibitor with the concentration of 100nM, and culturing for 48 h;
4) miR-144-3p inhibition group + AngII treatment group: transfecting an AC16 cardiac muscle cell by miRNA-144-3p inhibitor with the concentration of 100nM, adding AngII with the concentration of 100nmol/L, and culturing for 48 h;
5) overexpression of miR-144-3p group: transfecting AC16 cardiac muscle cells by miRNA-144-3p mimic with the concentration of 100nM, and culturing for 48 h;
6) miR-144-3p overexpression group + AngII treatment group: the miRNA-144-3p mimic with the concentration of 100nM is used for transfecting AC16 cardiac muscle cells, 100nmol/LangII is added, and the cells are cultured for 48 h.
Wherein the transfection method comprises the following steps: inoculation before experiment is 1-5 multiplied by 105And (3) continuously culturing the cells in a 24-well plate containing 1ml of culture medium until the cell density is 30-50%. Diluting 2.5. mu.l of 20. mu.M mirNammimic or 2.5. mu.l of 20. mu.M miRNA inhibitor with 30. mu.l of 1 XBOFETTM CP Buffer, gently mixing, adding 3. mu.l of riboFECTTM CP Reagent, gently blowing, mixing, incubating at room temperature for 10min, adding into a culture medium without double antibody, and continuing to culture for 48h for detection. Wherein the MiR-144-3p mimic, miR-144-3p inhibitor and riboFECTTM CP transfection reagents are all purchased from Ruibo Biotechnology, Inc., Guangzhou.
Taking each treatment group as a material, respectively detecting the following indexes by the specific method:
1. prediction of binding sites of the Nrf 23 'UTR region was performed using the bioinformatics website TargetScan, and it was found that miR-144-3p has a complementary binding site with the 3' UTR of Nrf2 (FIG. 2A).
2. And counting the surface area data of each group of cells. The results show that the cell surface area of the AngII treated group is significantly increased compared to the control group; compared with the AngII group, the cell surface area of the miR-144-3p inhibitor/AngII group is obviously reduced, and the cell surface area of the miR-144-3p imic/AngII group is obviously increased (figure 2B and figure 2C). This suggests that AngII can cause the surface area of the myocardial cells to increase, inhibition of miR-144-3p can reduce the surface area increase of the myocardial cells caused by AngII, and overexpression of miR-144-3p can promote the surface area increase of the myocardial cells caused by AngII.
3. Detecting the expression level of 4-HNE in each group of cells, specifically detecting by adopting a conventional Western Blot, wherein the internal reference is beta-actin protein.
Compared with the control group, the AngII treated group has obviously increased AC16 cardiomyocyte 4-HNE expression; compared with the AngII group, the expression of the 4-HNE in the miR-144-3p inhibitor/AngII group is obviously reduced, and the expression of the 4-HNE in the miR-144-3p imic/AngII group is obviously increased (figure 2D and figure 2E). The result shows that AngII stimulation can cause AC16 myocardial cell oxidative damage, inhibition of miR-144-3p can reduce AngII-induced AC16 myocardial cell oxidative damage, and overexpression of miR-144-3p can promote AngII-induced AC16 myocardial cell oxidative damage.
4. The expression levels of Nrf2 and HO-1 in each group of cells are detected, the detection method of Nrf2 is as described in example 1, HO-1 is a downstream antioxidant gene of Nrf2, an upstream primer for detecting HO-1 gene is ATGGCCT CCCTGTACCACATC (SEQ ID NO:6), a downstream primer for detecting HO-1 gene is TGTTGCGCTCAATCTCCTCCT (SEQ ID NO:7), and the detection method is as described in example 1.
Compared with a control group, the expression of Nrf2 and HO-1 in the myocardial cells of the AngII group AC16 is remarkably reduced, the expression of Nrf2 and HO-1 in the myocardial cells of the miR-144-3p inhibitor group AC16 is increased (figure 2F and figure 2G), and the expression of Nrf2 and HO-1 in the myocardial cells of the miR-144-3p imic group AC16 is reduced; compared with the AngII group, the expressions of the miR-144-3p inhibitor/AngII group Nrf2 and HO-1 are obviously increased, and the expressions of the miR-144-3p imic/AngII group Nrf2 and HO-1 are obviously reduced (FIG. 2H and FIG. 2I).
The results show that the activation of Nrf2 in AC16 cardiac muscle cells under the stimulation of AngII can be remarkably increased by inhibiting miR-144-3p, the activation of Nrf2 in AC16 cardiac muscle cells under the stimulation of AngII can be inhibited by over-expressing miR-144-3p, and the oxidative damage caused by AngII can be influenced by regulating the expression of Nrf2 by miR-144-3 p.
The foregoing is only a preferred embodiment of the present invention, and it should be noted that, for those skilled in the art, various modifications and decorations can be made without departing from the principle of the present invention, and these modifications and decorations should also be regarded as the protection scope of the present invention.
Sequence listing
<110> Jilin university
Application of <120> miR-144-3p in preparation of reagent or medicine for diagnosing or treating myocardial injury
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Claims (10)
- Application of miR-144-3p as a biomarker in preparation of a reagent for diagnosing myocardial injury.
- Application of miR-144-3p as a molecular target in preparation of medicines for treating myocardial injury.
- 3. Application of a reagent for inhibiting miR-144-3p expression in preparation of a medicament for treating myocardial injury.
- 4. The use of any one of claims 1 to 3, wherein the miR-144-3p exacerbates myocardial injury by negatively regulating expression of Nrf 2.
- 5. The use of claim 4, wherein the miR-144-3p effects gene regulation of Nrf2 by complementing the 3' UTR of Nrf 2.
- 6. The use of any one of claims 1 to 3, wherein the myocardial damage comprises myocardial oxidative damage and cardiomyopathy.
- 7. The use of any one of claims 1 to 3, wherein the myocardial injury is induced by AngII.
- 8. The use of claim 3, wherein the agent for inhibiting the expression of miR-144-3p comprises miR-144-3p inhibitor.
- 9. Application of a miR-144-3p overexpression reagent in preparation of a myocardial injury aggravation model.
- 10. The use according to claim 9, wherein the agent for over-expressing miR-144-3p comprises miR-144-3p imic.
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