Application of miRNA inhibitor in the preparation of drugs for preventing and treating myocardial infarction
A myocardial infarction and inhibitor technology, applied in the field of biomedicine, can solve problems such as the unclear role of genetics
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Embodiment 1
[0033] Example 1: Myocardial injection of miR-9 inhibitor preparations effectively reduces cardiac miR-9 content
[0034] 1. Inject the miR-9 inhibitor into the myocardium in vivo and verify the expression level of cardiac miR-9
[0035]About 25 g of C57BL / 6J male mice were selected as the experimental subjects, and the mixed preparation of miR-9 inhibitor and Dharmafect Duo was injected at two sites in the left ventricle, and antagomir NC was used as a negative control. The mixed preparation injected per mouse included 60 μg miR-9 inhibitor and 8 μl Dharmafect Duo;
[0036] 2. Detection of cardiac miR-9 expression level by QPCR
[0037] According to the routine cardiac total RNA extraction, inversion and QPCR, 4 replicate wells were set up for each sample, the reaction system was 10 μl, and U6 was used as an internal reference;
[0038] See attached figure 1 , is the expression level map of myocardial miR-9 detected by QPCR. It can be seen that the administration of miR-9 ...
Embodiment 2
[0039] Example 2: miR-9 inhibitor effectively improves left ventricular ejection fraction (ejection fraction, EF, %) and left ventricular fractional shortening (fractional shortening, FS, %) after myocardial infarction
[0040] 1. Establishment of myocardial infarction model in mice
[0041] About 25 g C57BL / 6J male mice were selected as the experimental subjects, and the myocardial infarction model was established by ligation of the left anterior descending coronary artery. After intraperitoneal injection of anesthesia, orotracheal intubation, connected to an air ventilator, respiratory rate 110 times / min, tidal volume 3ml, breath-to-breath ratio 1:1.3. In the right decubitus position, the outer skin was incised through the left thoracic longitudinal incision, the pectoralis major muscle was peeled off, the chest was opened through the third and fourth intercostal transverse incisions, the heart was exposed, and the pericardium was torn off with tweezers. The left coronary a...
Embodiment 3
[0045] Example 3: miR-9 inhibitor significantly reduces myocardial infarction size
[0046] 1. Establishment of mouse myocardial infarction model: same as Example 2;
[0047] 2. In vivo myocardial injection of miR-9 inhibitor: same as Example 1;
[0048] 3. Masson staining: the mice were killed 7 days after the operation, and the left ventricle tissue was taken for cardiac tissue staining to observe the therapeutic effect. Follow the routine Masson staining procedure, observe and take pictures under an ordinary light microscope. The image analysis software Image J was used to analyze the area of each part, and the area of myocardial infarction / heart area was calculated;
[0049] See attached Figure 4 , attached Figure 5 , the infarct area of each group was observed 7 days after myocardial infarction by Masson staining ( Figure 4 A), it was found that the size of myocardial infarction in the miR-9 suppressed subgroup was significantly lower than that in the contro...
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