Urechis unicinctus polypeptide with antioxidant activity and application thereof
A technology of anti-oxidative activity and single-ring thorn, which is applied in the field of biomedicine to achieve the effect of reducing the release of LDH, reducing the increase of ROS, and ensuring the integrity
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Embodiment 1
[0042] Example 1 Effect of the monocyclic echinococcoid polypeptide of the present invention on blood pressure regulatory factors in vascular endothelial cells
[0043] 1. Experimental materials and instruments
[0044] 1.1 Experimental materials
[0045] F4: Trp-Pro-Gly-Lys-Ala; F5: Gly-Leu-Thr-Lys-Pro;
[0046] Human umbilical vein endothelial cells (HUVEC) were provided by the Cell Bank of Shanghai Institute of Biological Sciences, Chinese Academy of Sciences.
[0047] 1.2 Main reagents see Table 1
[0048] Table 1 Main Reagents
[0049]
[0050] 1.3 Experimental equipment see Table 2
[0051] Table 2 Experimental Instruments
[0052]
[0053] 2 Experimental methods
[0054] 2.1 Peptide inhibition mode
[0055] According to the methods reported in previous studies (see [Forghani B, Zarei M, Ebrahimpour A, et al. Purification and characterization of angiotensin converting enzyme-inhibitory peptides derived from Stichopus horrens: Stability study against the ACE ...
Embodiment 2
[0119] Example 2 The repairing effect of the monocyclic spinypin polypeptide of the present invention on HUVEC cell damage caused by oxidative stress
[0120] The experimental materials, reagents and instruments used in this example are the same as those in Example 1.
[0121] 1. Experimental method
[0122] 1.1 Determination of superoxide anion scavenging ability
[0123] Prepare F4 and F5 into a series of concentration gradients, add reagents according to Table 6, bathe in water at 25°C for 5 minutes, and measure absorbance at 560nm.
[0124] Table 6 Determination of superoxide anion scavenging rate
[0125]
[0126] The formula for determining the clearance rate is as follows:
[0127] 1.2 Construction of oxidative damage model
[0128] Human umbilical vein endothelial cells (HUVEC) in the logarithmic growth phase were seeded in 96-well culture plates. 180 μL per well, cell number 1.5×10 4 pcs / well, placed at 37°C, 5% CO 2 Incubate for 24 h in an incubator. Div...
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