Application of oligopeptide or derivative thereof in preparation of drug or food for improving neural tube defect
A neural tube and drug technology, applied in the application field of oligopeptides and their derivatives in the preparation of drugs or food for improving neural tube defects, to achieve the effects of reversing the decrease in expression levels, reducing adverse effects, and improving neural tube defects
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Embodiment 1
[0044] Example 1: Preparation of leucopenzopeptide (LPF).
[0045] The oligopeptide of the present invention is derived from the active peptide released from the zearase process, the specific preparation method comprising the following steps:
[0046] (1) Corn protein concentration and purification: references (Chinese Journal of Cereals and Oils, 2007, 22(5): 44-48) reported method to purify corn protein in corn yellow powder, reduce the interference of non-protein substances on the preparation process of corn peptides;
[0047]The specific steps are as follows: 500g of corn yellow powder is added to a 7.5L alkali alcohol solution (0.1mol / L NaOH solution and 95% ethanol are prepared according to the volume ratio of 45:55), soaked in a 55 °C water bath for 3 hours, during which 500W, 50kHz ultrasound assisted extraction is given, and 4000 rpm centrifugation is centrifuged for 10 minutes to collect the extract; Add an equal volume of ultrapure water to the extract and adjust the pH...
Embodiment 2
[0051] Example 2: Leucoprosyl peptide (LPF) in vitro antioxidant capacity
[0052] The antioxidant capacity of leucopenzopeptide in vitro was determined by the Oxygen Radical Absorbance Capacity (ORAC) experiment.
[0053] ORAC experimental process: Add different concentrations of 20 μl of leucopenzopeptide aqueous solution in 96-well plates, add 40 μl of AAPH and 20 μl of phosphate buffer, and finally add 20 μl of fluorescein sodium and quickly place the 96-well plate in a luminesce microplate reader at a set temperature of 37 °C to start the assay. One point is measured every 2 min for a total of 2 h. The ORAC values are calculated as the protection integration area corresponding to the 1 μmol / l Trollox on the fluorescence attenuation curve as the standard control.
[0054] The results are as follows Figure 2 As shown, the in vitro antioxidant capacity of leucoprotein increases dose-dependently, indicating that the small molecule peptide has direct free radical scavenging activit...
Embodiment 3
[0055] Example 3: Effect of leuproline (LPF) in vitro on the viability of chicken embryonic fibroblasts
[0056] This implementation case is used to detect cell viability by classical MTT experiments. MTT is a yellow powder with the full name thiazolilam; The detection principle is that MTT can enter the cell through the cell membrane, and the amber dehydrogenase reduced by the amber dehydrogenase in the mitochondrials of the living cell is difficult to dissolve in water and deposited in the cell, while the dead cell has no function; The crystals formed by living cells can be dissolved by dimethyl sulfoxide (DMSO), and the light absorption value detected by it can indirectly reflect the survival and growth of cells, thereby evaluating the toxic side effects of drugs.
[0057] The specific experimental process is as follows: Chicken fibroblasts DF-1 in a good state, in the logarithmic growth phase, and growing density of about 80% in the petri dish are laid in 96-well plates, 10 pe...
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