Function and application of SH2B adapter protein 3(SH2B3) for treating cardiac hypertrophy
A technology of SH2B3, 1.SH2B3 is applied in the field of gene function and application to achieve the effect of worsening cardiac function and promoting cardiac hypertrophy
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Embodiment 1
[0052] Example 1 Expression of SH2B3 in the hearts of normal people and patients with cardiomyopathy
[0053]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. SDS-PAGE-Western blotting test (Western blot) for proteins extracted from the heart, combined with antibodies that specifically recognize SH2B3 protein and cardiomyocyte hypertrophy markers ANP (Millipore, AB2232) and Myh7 (santa cruz, sc53090) Detection was performed to measure the expression of SH2B3 (santa cruz, sc7222), and β-Tubulin (Cell Signaling Technology, 2128) was used as an internal reference. Test results such as figure 1 , figure 2 As shown, the expressions of cardiomyocyte hypertrophy markers ANP and Myh7 were significantly up-regulated in the hearts of patients with dil...
Embodiment 2
[0054] Example 2 Expression of SH2B3 in wild-type mouse Sham group and AB operation 4W, 8W hearts
[0055] 1. The myocardial hypertrophy model adopts aortic arch constriction surgery, and the operation process of the model is as follows:
[0056] 1.1 Preoperative preparation
[0057] (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).
[0058] (2) Preparation of the operation area: the skin of the left chest, left chest and armpit of the left forelimb of the mouse was rem...
Embodiment 3
[0067] Example 3 Detection of cardiac hypertrophy and fibrosis in mouse cardiac hypertrophy (AB) model
[0068] 1. The myocardial hypertrophy model adopts aortic arch constriction surgery, and the operation process of the model is as follows:
[0069] Wild-type mice, SH2B3 knockout mice, heart-specific SH2B3 transgenic mice and non-transgenic mice aged 8-10 weeks and weighing 23.5-27.5 g were selected and divided into sham operation group and cardiac hypertrophy model group. Group 10 mice. The modeling method is the same as in Example 2.
[0070] 2. Collect materials
[0071] (1) Preliminary work: prepare in advance a urine cup filled with 20mL of 10% formaldehyde by volume, and label it (mouse number, group, type of operation and date of collection). A petri dish filled with 10% KCl solution was placed at the sampling site. Turn on the analytical balance and set it to zero for later use. Mice were then weighed and sacrificed.
[0072] (2) Material collection: Ophthalmic...
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