Active peptide with antioxidant and antihypertensive effects and application thereof
An active peptide and blood pressure-lowering technology, applied in chemical instruments and methods, peptides, organic chemistry, etc., can solve problems such as reduced nitric oxide bioavailability, vascular endothelial damage, and increased medical costs
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Embodiment 1
[0042] Example 1: Separation and determination of active peptides in Guangdong glutinous rice wine
[0043] 1. Experimental materials
[0044]Guangdong glutinous rice wine is provided by Guangdong Heyuan Lvpure Food Co., Ltd., 1,1-diphenyl-2-picric acid (DPPH), 3-ethylbenzothiazoline-6-sulfonic acid (ABTS), angiotensin conversion Enzyme (ACE, from rabbit lung) was purchased from Sigma (St. Louis, MO, USA). Human hepatoma cells (HepG2) ATCC cell bank; DMEM (dulbecco's modified eaglemedium) medium, fetal bovine serum (FBS) and 0.25% trypsin were purchased from Gibco, USA; DCFH-DA reactive oxygen species ROS fluorescent probe, Phosphate buffer saline (PBS), dimethyl sulfoxide (DMSO), and hydroxyethylpiperazine ethanesulfonic acid (HEPES) were purchased from Sigma-Aldrich, USA; other chemicals and reagents were Analytically pure.
[0045] 2. Determination of polypeptides in Guangdong glutinous rice wine
[0046] Guangdong glutinous rice wine of different years was treated with...
Embodiment 2
[0075] Example 2: Synthesis and Activity Assay of Active Peptides
[0076] 1. Synthesis of Active Peptides
[0077] The active peptide GHVAA was synthesized by Nanjing Synpeptide Co., Ltd. The purity of the synthetic peptide was determined to be 98% by high performance liquid chromatography, and the molecular mass of the synthetic peptide was determined by LC / ESI-MS method. The blood pressure lowering activity and antioxidant activity were verified by blood pressure lowering activity experiment, cellular antioxidant activity experiment and molecular docking.
[0078] 2. Determination of blood pressure lowering activity
[0079] Take 40 μL of the sample solution and mix it with 50 μL of 1.0 mmol / L FAPPG. For the blank group, use 40 μL of 80 mmol / L HEPES buffer to mix with 50 μL of FAPPG. 10μL of 0.1 U / mL ACE was added to both groups at the same time. Immediately after adding ACE, the absorbance values of the two groups were measured at a wavelength of 340 nm, the blank gr...
Embodiment 3
[0105] Example 3: Molecular Mechanism of Computer Simulation of Active Peptides
[0106] 1. Analysis of the inhibition pattern of GHVAA on CD38
[0107]Nicotinamide adenine dinucleotide (NAD+) is a key coenzyme factor in the signaling pathway mediated by SIRT1-7, and is involved in various metabolic pathways and cellular redox reactions. Including assisting and carrying the transfer of hydrogen atoms in metabolic pathways, participating in the partial oxidation reaction in the tricarboxylic acid cycle and the oxidation of fatty acids and amino acids in mitochondria, etc. More importantly, NAD+ can participate in the regulation of apoptosis, cell aging, DNA repair and other processes together with other enzymes to achieve the purpose of anti-oxidation and anti-aging. CD38 is an enzyme that degrades NAD+ in the body. Some of the reported antioxidant peptides can bind to CD38 to inhibit its enzymatic activity and increase the content of NAD+ in the body, thereby exerting anti-ox...
PUM
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