The function and application of sh2b adapter protein 1 (sh2b1) in the treatment of cardiac hypertrophy
A 1. SH2B1, SH2B1 technology, applied in the field of gene function and application, can solve the problem of not fully explaining the mechanism of cardiac hypertrophy, and achieve the effect of worsening cardiac function and promoting cardiac hypertrophy
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Embodiment 1
[0055] The expression of embodiment 1SH2B1 in the heart of normal people and patients with cardiomyopathy
[0056] Normal human hearts (individuals who died from non-cardiac causes), hearts from patients with dilated cardiomyopathy (recipients undergoing heart transplantation, DCM) and hearts from patients with hypertrophic cardiomyopathy (recipients undergoing heart transplantation) were selected. body, HCM), SDS-PAGE-immunoblotting test (Westernblot) was carried out on the protein extracted from the heart, and the specific recognition SH2B1 (R&D, AF6915) protein and cardiomyocyte hypertrophy markers ANP (Millipore, AB2232), β-MHC (santacruz , sc53090) antibody was detected to measure the expression of SH2B1, and GAPDH (Cell Signaling Technology, 2118) was used as an internal reference. Test results such as figure 1 As shown, compared with normal human hearts, the expression of cardiomyocyte hypertrophy markers ANP and β-MHC in patients with cardiomyopathy was significantly ...
Embodiment 2
[0057] The expression of embodiment 2SH2B1 in wild-type mouse Sham group and AB operation 4W, 8W heart
[0058] 1. The myocardial hypertrophy model adopts aortic arch coarctation surgery, and the operation process of the model is as follows:
[0059] 1.1 Preoperative preparation
[0060] (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).
[0061] (2) Preparation of the operation area: the skin of the left chest, left chest and armpit of the left forelimb of the mouse was...
Embodiment 3
[0070] Example 3 Effect of SH2B1 interference (AdshSH2B1) and overexpression (AdSH2B1) adenovirus on the expression of SH2B1 in AngⅡ-stimulated primary cardiomyocytes
[0071] 1. Primary neonatal SD rat cardiomyocyte culture
[0072] (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.
[0073] (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.
[0074] (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 wit...
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