Application of trihydroxyflavone in preparing medicament used for preventing and treating abdominal aortic aneurysm
A technology of trihydroxyflavonoids and abdominal aortic aneurysm, applied in the direction of drug combination, antineoplastic drugs, antipyretics, etc., can solve problems such as uncertain conclusions, no literature reports, and influence on the formation of abdominal aortic aneurysm
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Embodiment 1
[0056] Embodiment 1. Establishment of the model
[0057] 1. Identification of Apoe- / - mice
[0058] Following the methods well known to those skilled in the art, the mice were identified by PCR and the Apoe- / - mice were passaged. Three primers oIMR180, oIMR181 and oIMR182 recommended by Jackson Laboratory (Bar Harbor, ME) were used (primer sequences are available from the supplier).
[0059] 2. Identification of AngII-induced Apoe- / - mice to produce an abdominal aortic aneurysm model
[0060] Angiotensin II (angiotensin II, Ang II) is an important member of the renin-angiotensin system, and activates downstream signaling pathways such as MAPKs (p38, ERK, JNK) by binding to its receptor AT1R, thereby inducing the generation of oxygen free radicals, cell Apoptosis, inflammatory cytokine expression, and matrix metalloproteinase synthesis. Angiotensin II stimulates Apoe- / - mice, which can simulate almost all the pathogenesis of abdominal aortic aneurysm formation, with an incid...
Embodiment 2
[0063] Example 2. Therapeutic regimen
[0064] 1. BAI intraperitoneal injection: BAI (Sigma, 465119) powder was dissolved in sterile saline (saline, vehicle) to form a suspension with a concentration of 7.5 mg / mL. BAI concentration can be between 20-200mg / kg / d. In this embodiment, two concentrations and doses are selected: 30 mg / kg / d and 100 mg / kg / d.
[0065] 2. Experimental animals are divided into 6 groups:
[0066] The AngII experimental group included:
[0067] Control group (VehicleApoe- / -, VehApoe- / -),
[0068] BAI low dose group (BAI-30Apoe- / -),
[0069] BAI high dose group (BAI-100Apoe- / -);
[0070] Saline control groups included:
[0071] Control group (VehicleApoe- / -, VehApoe- / -),
[0072] BAI low dose group (BAI-30Apoe- / -),
[0073] BAI high dose group (BAI-100Apoe- / -).
[0074] BAI pre-stimulation was started one week before AngII stimulation, the dose was 30mg / kg / d, 100mg / kg / d, once a day at the same time. Grab the mouse, disinfect the left groin area wi...
Embodiment 3
[0080] Example 3.Western detection
[0081] 1. Total Protein Extraction from Mouse Aorta
[0082] The extraction of mouse aortic tissue protein was done by completely stripping off the perivascular tissue and adventitia after perfusion, weighing first, and then adding RIPA lysate to 3 times the volume of the tissue block, fully homogenized with a glass homogenizer, and iced. Bath 30min. Ultrasonic instrument 7W power ultrasonic 10sec, interval 1min, a total of 3 times, and then ice bath for 30min. Centrifuge at 12500rpm for 30min at 4°C. Take the supernatant and measure the protein concentration by BCA method, add it to a new EP tube, add 5× loading buffer, mark it after mixing, boil in boiling water for 10 minutes, store at -20°C for short-term use, and store in long-term Standby at -80°C.
[0083] 2. Protein Concentration Determination
[0084] Protein concentration BCA determination method: BCA (BicinchoninicAcid) is a pair of monovalent copper ions (Cu + ) sensitive,...
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