Application of fat globule epidermal growth factor 8 in diagnosis and treatment of heart remodeling and heart failure
A technology for cardiac remodeling and heart failure, applied in disease diagnosis, biochemical equipment and methods, microbial measurement/testing, etc., can solve the problems of low overall survival rate of heart failure, achieve inhibition of myocardial hypertrophy, improvement of heart function, Inhibition and its effect on fibrosis
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Embodiment 1
[0061] Example 1 Correlation between serum MFG-E8 protein level and cardiac remodeling and heart failure severity
[0062] 1. Research objects: Heart failure patients diagnosed with dilated cardiomyopathy (DCM) and hypertrophic cardiomyopathy (HCM) were selected, including 162 patients with DCM heart failure and 43 patients with HCM heart failure. Patients admitted to the hospital were graded according to the New York Heart Association (NYHA) standard. The healthy control group (Control) consisted of 100 healthy subjects. The baseline data of the study subjects are shown in Table 1.
[0063] Table 1
[0064]
[0065] 2. Serum collection: Collect 5mL of venous blood on an empty stomach in the morning, centrifuge at 1500r / min for 5min, collect serum, and freeze at -80°C.
[0066] 3. Determination of serum MFG-E8 level: serum MFG-E8 content was determined by ELISA method, and the kit used was human milk fat globule epidermal growth factor 8 (MFG-E8) ELISA kit provided by R&...
Embodiment 2
[0072] Embodiment 2 experimental animals and raising
[0073] 1. Experimental animals: 8-10 weeks old, 23.5-27.5g body weight, male C57BL / 6 mice (named WT, purchased from Beijing Huafukang Biotechnology Co., Ltd.) with a background of male, systemic MFG-E8 gene Knockout mice (MFG-E8-KO, purchased from RIKEN, Cat. No. 01726). Heart-specific MFG-E8 transgenic mice (MFG-E8-TG) and non-transgenic mice (NTG, same-age littermate control non-transgenic mice) were used as experimental subjects.
[0074] 2. Breeding environment: All experimental mice were raised in the SPF level experimental animal center of Wuhan University. SRF grade mouse feed was purchased from Beijing Huafukang Biotechnology Co., Ltd. Raising conditions: the room temperature is between 22-24° C., the humidity is between 40-70%, the lighting time is 12 hours alternately between light and dark, and free to drink and eat.
Embodiment 3
[0075] Example 3 Construction of heart-specific MFG-E8 transgenic mice (for construction strategies, see Figure 5 A)
[0076] Using the cDNA of the C57BL / 6J mouse MFG-E8 gene as a template, use the following primers to PCR amplify the mouse MFG-E8 gene (NCBI, Gene ID: 17304, CCDS21379.1):
[0077] Upstream primer: 5'-AGCTTTGTTTAAACGCCACCATGCAGGTCTCCCGTGTGC-3',
[0078] Downstream primer: 5'-CTAGCTAGCTTAACAGCCCAGCAGTCCA-3'.
[0079] The amplified product and the pCAG-CAT-LacZ vector digested with restriction endonucleases PmeI and NheI (provided by the laboratory of Teacher Yang Qinglin, School of Basic Science, Peking Union Medical College Medical College, see the references for the preparation process: Kim T, Zhelyabovska O , Liu J, et al.Generation of an Inducible, Cardiomyocyte-Specific Transgenic Mouse Model with PPAR b / d Overexpression [J].PeroxisomeProliferator-Activated Receptors (PPARs), 57.) Ligation to obtain the transgenic vector pCAG-CAT-MFG-E8 -polyA, the expr...
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