The function and application of nuclear localization protein 1 (nulp1) in the treatment of cardiac hypertrophy
A technology of nuclear localization protein and cardiac fibrosis, which is applied in the field of gene function and application, to achieve the effect of inhibiting cardiac hypertrophy, protecting cardiac function, and resisting cardiac fibrosis and cardiac hypertrophy
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Embodiment 1
[0079] Example 1 Expression of Nulp1 in the hearts of normal people and patients with cardiomyopathy
[0080] SDS-PAGE-Western blot test (Western blot) was performed on proteins extracted from the hearts of normal human hearts (individuals donated by non-cardiac causes of death) and hearts of patients with dilated cardiomyopathy (recipients replaced by patients undergoing heart transplantation, DCM). ), combined with antibodies that specifically recognize Nulp1 protein and cardiomyocyte hypertrophy markers ANP (Millipore, AB2232), β-MHC (santa cruz, sc53090) for detection, and determined the expression of Nulp1 (BETHYL, A303-087A), GAPDH ( Cell Signaling Technology, 2128) as an internal reference. Test results such as figure 1 As shown, the expression of cardiomyocyte hypertrophy markers ANP and β-MHC in the hearts of patients with dilated cardiomyopathy (dilated heart disease) was significantly up-regulated, and the expression of Nulp1 was significantly down-regulated ( fig...
Embodiment 2
[0081] Example 2 Expression of Nulp1 in the heart of wild-type mouse sham operation 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 forelim...
Embodiment 3
[0096] Example 3 Expression of Nulp1 in cardiomyocytes stimulated by control group (PBS) or angiotensin II (Ang II) or phenylephrine (PE)
[0097] Isolate and culture newborn 1-day Sprague-Dawley neonatal rat cardiomyocytes, culture the primary cardiomyocytes for 48 hours and change the medium (see the following example 4 for the specific process of primary neonatal SD rat cardiomyocyte culture), add serum-free DMEM / F12 Starve the cardiomyocytes for 12 hours to synchronize the cells, give PBS, angiotensin II (Ang II, 1 μM) and phenylephrine (PE, 1 μM) stimulation for 48 hours respectively, and perform SDS-PAGE-immunoblotting test on the proteins extracted from cardiomyocytes (Western blot), combined with antibodies that specifically recognize Nulp1 protein and cardiomyocyte hypertrophy markers ANP and β-MHC, were used to detect the expression of Nulp1, and GAPDH was used as an internal reference. Test results such as image 3 As shown, the expressions of ANP and β-MHC in card...
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