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Caenorhabditis elegans drunkenness model creation method for rapidly screening alcohol-effect-dispelling plants and application

A Caenorhabditis elegans, model-building technology, applied in the fields of compound screening/testing, pharmaceutical formulations, active ingredients of hydroxyl compounds, etc., can solve the problems of limited sample number in the test group, long test period, high economic cost, and shorten the test period. , reduce the test cost and ensure the effect of accuracy

Active Publication Date: 2018-10-19
HUNAN AGRICULTURAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

This method has the disadvantages of complicated process, limited number of samples in the test group, long test period, and high economic cost.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0015] Example 1 Establishment of Caenorhabditis elegans Drunk Model

[0016] Wild-type Caenorhabditis elegans (N 2 ) as the basis for model establishment, the synchronization and basal culture of Caenorhabditis elegans were carried out with reference to the liquid culture method in the reference (Stiernagle T. Maintenance of C. elegans [J]. Wormbook, 2006: 1-11.). Caenorhabditis elegans feeds on concentrated OP50 with a concentration of 100mg / ml in the bacterial solution, and the concentration of the worm solution is controlled at 2000-2500 worms / mL. At this time, when Caenorhabditis elegans is detected, the head swing frequency is 50-100 times / 30s, the swallowing movement is 50-90 times / 30s, the movement trajectory presents a sinusoidal wavelength coordinated left and right, and the life span is 18-25d. The total antioxidant capacity was 5-12U / mg, the oxygen free radical (ROS) level was 40-60A.U., and no externally ingested alcohol was detected in the nematode.

[0017] Wh...

Embodiment 2

[0018] Example 2 Verification of the Caenorhabditis elegans Drunk Model

[0019] Extraction of medicinal and edible plant extract powder: crush various purchased medicinal and edible plants into powder, and use water extraction method to extract active ingredients repeatedly according to the solid-liquid ratio of 1g:10ml respectively. , filter, and combine the 2 filtrates. Then transfer to a rotary vacuum concentrator to concentrate the filtrate, then place the concentrated solution in a glass dish, seal it, and freeze it in a -80°C refrigerator for at least 2 hours. Finally, the frozen medicine dish is quickly taken out and placed in a freeze dryer to be freeze-dried into powder.

[0020] Method: Set up a blank group (worm solution + water), a control group (worm solution + alcohol solution), and a drug group (worm solution + alcohol solution + medicinal solution), among which:

[0021] Blank group: The synchronization and basal culture of wild-type Caenorhabditis elegans w...

Embodiment 3

[0025] Example 3 Pueraria root water extract to hangover application of drunken nematodes

[0026] Carry out according to the grouping method of embodiment 2, medicinal edible dual-purpose plant adopts kudzu root.

[0027] The pueraria root was purchased from a pharmacy, and the pueraria root extract powder was made according to the extraction method in Example 2. After the pueraria root extract powder was completely dissolved in water, it was sterilized through a 0.22 micron filter membrane, and then added to the control with a volume final concentration of 5 mg / ml In the worm solution of the group, intervene for 24h. Detect various indicators such as head swing frequency, swallowing times, movement trajectory, lifespan, total antioxidant capacity in the nematode, oxygen free radical level and alcohol concentration in the body of Caenorhabditis elegans.

[0028] The results showed that in the drug group, the frequency of head swings was 65 times / 30s, the swallowing movement ...

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Abstract

The invention discloses a caenorhabditis elegans drunkenness model creation method for rapidly screening alcohol-effect-dispelling plants. The caenorhabditis elegans drunkenness model creation methodincludes taking wild caenorhabditis elegans as a base, and cultivating the caenorhabditis elegans in liquid until the content reaches 2000-2500 worm / mL; after the caenorhabditis elegans just grows into the adulthood, adding an alcoholic solution into the worm liquid until the final concentration of the alcoholic solution reaches 1-5%, and performing 24-hour intervention, wherein the caenorhabditiselegans is drunk at the moment. The method has the advantages that a model organism, namely the caenorhabditis elegans, which is convenient and easy to get, is adopted for evaluation, and accordingly, experiment cost can be reduced effectively; the growth cycle of the caenorhabditis elegans is short, and tens of thousands of the caenorhabditis elegans can be reproduced in a short time to be usedfor experiments, so that enough samples can be guaranteed, experiment errors can be reduced, and accuracy of evaluation results can be guaranteed; high-throughput screening experiments can be carriedout, experiment results can be acquired efficiently, and experiment cycle can be shortened greatly.

Description

technical field [0001] The invention belongs to the field of biotechnology, and relates to a method for rapid screening of water extracts of plants (dual-purpose for medicine and food) with hangover effects, in particular to a method for rapidly screening hangover plants for establishment of a Caenorhabditis elegans intoxication model. Background technique [0002] my country has a long history of wine culture. Moderate drinking can improve blood circulation and relieve fatigue. However, when the intake exceeds the limit of the human body, symptoms such as drowsiness, gastric bleeding, liver damage, and nervous system paralysis will occur, which will cause great harm to the human body. Therefore, A large number of anti-alcoholic medicines with anti-alcoholic effects (such as Gehua Tang, Gandele sobering drinks, Xingyue sobering drinks, Dingzun sobering drinks and other plant sobering drugs) have come out. However, the efficacy of most anti-alcoholic drugs is mostly verified b...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K31/045A61K49/00
CPCA61K31/045A61K49/0008
Inventor 李宗军吴中琴李珂戴奕杰
Owner HUNAN AGRICULTURAL UNIV
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