Function and application of dual-specificity phosphatase 14 (DUSP 14) in curing cardiac hypertrophy
A bispecific, myocardial hypertrophy technology, applied in the field of gene function and application, can solve problems such as malignant arrhythmia, myocardial cell apoptosis, sudden death, etc., to protect cardiac function, resist cardiac fibrosis, and inhibit cardiac hypertrophy. Effect
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Embodiment 2
[0081] Example 2 Expression of DUSP14 in the hearts of wild-type mouse sham-operated group and cardiac hypertrophy model group
[0082] 1. Aortic arch constriction (AB) was used to establish a mouse model of myocardial hypertrophy. The model operation process:
[0083] 1.1 Preoperative preparation
[0084] (1) Anesthesia: First weigh the mice, calculate the required amount of anesthetic (3% pentobarbital sodium) according to 90 mg / kg body weight, inject intraperitoneally, and record the injection time point. There is no obvious reaction between tail and toe pinching and the mouse is in good condition. This is the standard for successful anesthesia (generally there is no obvious reaction about 10 minutes after injection, and the mouse has a reaction to pinch toe about 50 minutes after anesthesia, and about 30 minutes after anesthesia is the best operation time).
[0085] (2) Preparation of the operation area: the skin of the left chest, left chest and armpit of the left forelimb...
Embodiment 3D
[0096] Example 3 Effect of DUSP14 interference (AdshDUSP14) and overexpression (AdDUSP14) adenovirus on AngII-stimulated primary cardiomyocyte hypertrophy
[0097] 1. Primary neonatal SD rat cardiomyocyte culture
[0098] (1) Eight newborn Sprague-Dawley suckling mice were disinfected with 75% alcohol below the neck, and the heart was removed with ophthalmic scissors and micro forceps, and placed in a glass plate filled with 10mL DMEM / F12 solution. Take another one and repeat the above process.
[0099] (2) Wash the heart with DMEM / F12 medium, and cut the heart into 1-2mm 3 fragments. Transfer to a serum bottle with a rotor, suck off DMEM / F12, and add trypsin digestion solution. Rotate at 120r / min, digest for 15min, rest for a few seconds, and discard the supernatant.
[0100] (3) Add trypsin digestion solution, the speed is 120r / min, and digest for 15min. Stand still for a few seconds, aspirate the supernatant, terminate the digestion with DMEM / F12 medium with 20% calf s...
Embodiment 4
[0106] Example 4 Myocardial hypertrophy and fibrosis detection and cardiac function detection in myocardial hypertrophy model mice
[0107] 1. Using aortic arch constriction (AB) to establish a mouse model of myocardial hypertrophy
[0108] Wild-type mice (WT), DUSP14 knockout mice (DUSP14-KO), heart-specific DUSP14 transgenic mice (DUSP14-TG) and non-transgenic mice aged 8-10 weeks and weighing 23.5-27.5 g were selected. Rats (NTG) were divided into sham operation group (Sham) and AB myocardial hypertrophy model group, that is, WTSham group, WTAB group, DUSP14-KOSham group, DUSP14-KOAB group, DUSP14-TGSham group, DUSP14-TGAB group, NTGSham group, NTGAB group, 10 mice in each group. The modeling method is the same as in Example 2. The detection of myocardial hypertrophy and fibrosis and the detection of cardiac function were carried out 4 weeks after AB.
[0109] 2. Take materials
[0110] (1) Preliminary work: prepare in advance a urine cup filled with 20mL of 10% formald...
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