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Application of branched alpha ketonic acid serving as antioxidant

A technology of antioxidants and natural antioxidants, applied in the direction of food ingredients as antioxidants, anti-toxic agents, anhydride/acid/halide active ingredients, etc.

Inactive Publication Date: 2018-06-22
WUHAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

So far, the ability of BCKA to scavenge hydroxyl free radicals (HO·) in vitro, and the ability to protect cells at the cellular level, alleviate cell damage, and delay cell aging have not been reported.

Method used

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  • Application of branched alpha ketonic acid serving as antioxidant
  • Application of branched alpha ketonic acid serving as antioxidant
  • Application of branched alpha ketonic acid serving as antioxidant

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0026] [Example 1] BCKA against H 2 o 2 DCF Fluorescence Measurement of Oxidants

[0027] DCF fluorescence measurement is to use the specific fluorescent probe DCF to monitor the content of HO. h 2 o 2 It is very unstable in aqueous solution, and the HO produced by decomposition can oxidize non-fluorescent DCFH to generate fluorescent DCF. The sample can be directly observed by laser confocal microscope (FV1000, Olympus), the excitation wavelength is 488nm, and the emission wavelength is 505nm. Observe the fluorescence intensity values ​​of each group by taking photos with a confocal microscope, and detect the H in each group 2 o 2 HO content in aqueous solution to clarify whether BCKA can reduce H 2 o 2 HO·accumulated in vitro and clarified the important role of BCKA in antioxidation.

[0028] In a 96-well plate, add 50 μl of 5 mM oxidant H 2 o 2 , and 50 μl of different concentrations of BCKA. The concentrations of BCKA were 0mM, 1.5mM, 3mM, 5mM, 7.5mM and 15mM r...

Embodiment 2

[0030] [Example 2] The influence of BCKA on the viability of MEF cells

[0031] 1. MEF cell treatment

[0032] In this experiment, H 2 o 2 Stimulation and calcium ionophore Ca 2+ MEF cells treated with lonophore were used as a model to investigate the mechanism of BCKA on cell protection. This experiment was divided into three groups, namely untreated group (control), H 2 o 2 Treatment group (H 2 o 2 ), and the calcium ionophore Ca 2+ lonophore treatment group (Ca 2+ lonophore). Among them, a PBS treatment group (PBS) and a 5mM BCKA treatment group (BCKA) were set up for each group respectively. 3 parallels for each group. Many functions of cells depend on the existence of extremely high calcium ion concentration difference inside and outside the cell, Ca 2+ As a calcium ionophore, lonophore can allow calcium ions to pass through the cell membrane, reduce the ion concentration difference between the inside and outside of the cell, and then damage the cell and even ...

Embodiment 3

[0041] [Example 3] BCKA against H 2 o 2 DCF Fluorescence Measurement of Oxidants

[0042] To further confirm the respective reducing abilities of KIM, KIV, and KIC, DCF fluorescence generated by the DCF reaction was observed by confocal microscopy. In a 96-well plate, 50 μl of PBS group was added as a control, and 50 μl of 5 mM oxidant H was added to all treatment groups. 2 o 2 , and then add 50 μl of 5 mM KIM, KIV, KIC and BCKA, respectively. After 1 h of reaction, a volume of 1 mL of DCFH (10 mmol / L) was added. The time points of 20min, 30min, 50min and 100mM were taken and observed under a confocal laser microscope. Each set of experiments has three repetitions, and the experiments are repeated three times. Photographs represent typical and average results from triplicate experiments.

[0043] The result is as image 3 As shown, the reducing abilities of KIM, KIV and KIC were compared. 5mM KIM, KIV, KIC and BCKA were mixed with 1.5mM H 2 o 2 Incubate with differe...

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Abstract

The invention discloses application of branched alpha ketonic acid serving as an antioxidant, and belongs to the field of functions and application of metabolic molecules. In-vitro DCF detection research shows that BCKA has a strong capability of removing HO generated by H2O2. The MEF cell injury induced by H2O2 serves as an experimental model on the cell level, BCKA protection cells are researched, the cell injury is reduced, and the effect of delaying cell senescence is achieved. The result shows that the BCKA can effectively enhance the cell activity and has the function of protecting cells against oxidative stress damage. Therefore, the BCKA has a strong antioxidant capacity. According to the functions of the BCKA, the BCKA can be applied to preparation of health care products, medicines, food additives and cosmetics.

Description

technical field [0001] The invention belongs to the field of functions and applications of natural antioxidants, and specifically relates to the application of branched-chain alpha ketoacids as antioxidants in the metabolism of branched-chain amino acids. Branched-chain alpha-ketoacids are α-keto-β-methylpentanoic acid, - one of ketoisocaproic acid, and alpha-ketoisovalerate. Background technique [0002] There are many active oxygen free radicals in the human body. The active oxygen free radicals produced in the human body can cause oxidative damage to cells, leading to various diseases such as human aging, cardiovascular and cerebrovascular diseases, and tumors. Under normal circumstances, there is a scavenging system for active oxygen and free radicals in the body, which keeps active oxygen and free radicals in balance at an extremely low level. Recent studies have confirmed that oxidative stress is involved in multiple links of cell damage. When the active oxygen free ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K31/19A61P39/06A61K8/36A61Q19/08A23L33/10
CPCA23L33/10A23V2002/00A61K8/36A61K31/19A61Q19/08A23V2200/02A23V2250/02
Inventor 王志华李丽莉谢银平何元
Owner WUHAN UNIV