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Application of docosahexaenoic acid ethyl ester in preparation of drug for improving excitatory neural toxicity

A technology of ethyl docosahexaenoate and ethyl carbasahexaenoate, which is applied in the field of application of ethyl docosahexaenoate in the preparation of drugs for improving excitatory neurotoxicity, and can solve the protective effect of DHA Few studies, unclear protection mechanism and other issues

Inactive Publication Date: 2016-05-11
GUANGZHOU UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

DHA has physiological effects such as preventing cardiovascular and cerebrovascular infarction, anti-inflammation, lowering blood fat, and anti-cancer. Previous studies have shown that DHA has a protective effect on cerebral ischemia damage, but at the cellular level or by electrophysiological stimulation methods and whole animals. There are few studies on the protective effect of DHA at the same level, and its protective mechanism is still unclear

Method used

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  • Application of docosahexaenoic acid ethyl ester in preparation of drug for improving excitatory neural toxicity
  • Application of docosahexaenoic acid ethyl ester in preparation of drug for improving excitatory neural toxicity
  • Application of docosahexaenoic acid ethyl ester in preparation of drug for improving excitatory neural toxicity

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Embodiment 1

[0026] Example 1 Cell Culture and Treatment

[0027] PC12 cells were cultured in DMEM medium supplemented with 10% (v / v) fetal bovine serum, 100 U / mL penicillin and 100 U / mL streptomycin at 37° C. in humidified air with 5% carbon dioxide. Before the experiment, the cells were divided into plates at a density of 2×105 cells per well. PC12 cells were incubated for 24 h in DMEM medium without or with different concentrations of Et-DHA (final concentration: 10, 30 μM). Then KA (final concentration: 30 mM) was added, and cultured for another 12 h to carry out subsequent experiments. In the above steps, the same amount of DMEM was administered to the control culture group. KA was dissolved in DMEM medium. Et-DHA is dissolved in dimethylsulfoxide (DimethylSulfoxide, DMSO). The final concentration of DMSO was less than 0.1% (v / v). The concentrations of KA and Et-DHA in the above steps are selected through our preliminary experiments. All operations were repeated three times (n=3...

Embodiment 2

[0028] The cytotoxicity of embodiment 2KA

[0029] PC12 cells were plated at a density of 1 x 105 cells per well. Continue to grow for 24h, then add different concentrations of KA (final concentration: 0, 1, 3, 10, 30 and 100mM) in the medium. All assays were started 12 h after KA treatment of cells. Cytotoxicity of KA was determined by the MTT assay (Mosmann, 1983). like figure 1 As shown in B, the results of cell viability detected by MTT showed that the inhibition of KA on cell viability was concentration-dependent and statistically significant (P<0.01). When the concentration of KA was 100mM, the inhibitory rate of PC12 cells also reached about 50%, but there was no significant difference compared with the 30mM treatment group, so KA30mM was selected as the appropriate injury model dose for PC12 cells in subsequent experiments.

Embodiment 3

[0030] Determination of the protective effect of embodiment 3Et-DHA on PC12 cells

[0031] PC12 cells with 1×105 cells per well were treated with different concentrations of Et-DHA (final concentration: 0, 10 and 30 μM) on a 96-well plate for 24 hours, and then added KA (final concentration: 30 mM) for 37 hours. Cultivate at ℃ for 12h. Then MTT (0.5mg / mL) was added to each well, and cultured for another 4h. Then, the MTT was discarded, and the obtained dark blue crystals were dissolved with 100 mL DMSO, and finally read at the absorbance value of 550 nm with a microplate reader (Bio-Rad, Hercules, USA). Depend on figure 2 B It can be seen that the survival rate of PC12 cells treated with KA (30mM) after pretreatment with different concentrations (10 and 30μM) of Et-DHA for 24h was significantly improved, compared with the single KA treatment group, it rose to 55.2±0.5 % and 67.2±0.6%. Statistical analysis showed that the survival rate of PC12 cells pretreated with 10 and ...

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Abstract

The invention provides application of docosahexaenoic acid ethyl ester in preparation of a drug for improving the excitatory neural toxicity, and further provides application of docosahexaenoic acid ethyl ester in preparation of a drug for treating epilepsy. By culturing PC12 cells in vitro, it is shown that docosahexaenoic acid ethyl ester (Et-DHA) prevents the PC12 cells from being subjected to KA-induced oxidative stress damage by reducing LDH releasing and ROS generating, inhibiting the activity of Caspase-3 and regulating and controlling a cell apoptosis pathway on which mitochondria depend, and it is predicted that the protection effect is related to the fact that Et-DHA can change the permeability of cell membranes and mitochondrial membranes. Research results reveal that Et-DHA can be used for preventing and treating diseases such as the epilepsy by serving as a nerve protection ingredient.

Description

technical field [0001] The invention relates to the application of docosahexaenoic acid ethyl ester in the preparation of drugs for improving excitotoxicity. Background technique [0002] Epilepsy is a chronic recurrent short-term brain dysfunction syndrome, and it is one of the common diseases in the Department of Neurology. It is a chronic disease of temporary central nervous system dysfunction caused by recurrent abnormal discharge of neurons. The pathological basis may be the imbalance of excitatory neurotransmitters and inhibitory neurotransmitters, and the neuron death caused by epilepsy may be caused by the excessive production of reactive oxygen species (Reactive Oxygen Species, ROS). The development of functional foods and related drugs that can prevent and treat such diseases has attracted extensive attention in various fields. [0003] Kainic Acid (KA) is a glutamic acid analog, which is a powerful convulsant and neurotoxin in the central nervous system. Rats in...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K31/232A61P25/00A61P25/08
CPCA61K31/232
Inventor 陈鲲曹宽曹丽丽王雪君刘珂莉秦嘉裕陈永顺
Owner GUANGZHOU UNIVERSITY
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