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Application of Brassica napus-Isatis indica e Monomer Addition Line in Inhibition of Influenza Virus

A technology of Brassica napus and influenza virus, applied in the fields of application, antiviral agents, respiratory diseases, etc., can solve the problems of not being able to be applied on a large scale, and achieve the effect of non-toxicity of cells, obvious effect, and antiviral effect

Active Publication Date: 2022-03-08
HUAZHONG AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Huang Luqi's laboratory of China Academy of Chinese Medical Sciences used water as a solvent to carry out 5 monomer addition systems (B monomer, C monomer, D monomer, E monomer, F monomer) and a two-body different addition system (D double body) Extraction, it was found that only the D-dibody showed a certain inhibitory effect on the H1N1 influenza virus, but the D-dibody addition line had male sterile cytoplasm, and had to add a pair of Isatis indigo chromosomes to have antiviral activity, so only tissue Preservation and multiplication of culture methods cannot be applied on a large scale

Method used

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  • Application of Brassica napus-Isatis indica e Monomer Addition Line in Inhibition of Influenza Virus
  • Application of Brassica napus-Isatis indica e Monomer Addition Line in Inhibition of Influenza Virus
  • Application of Brassica napus-Isatis indica e Monomer Addition Line in Inhibition of Influenza Virus

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0047] Preparation of Brassica napus-Isatis indigo line extract:

[0048] The whole plants of different additional lines of Brassica napus-Isatis indica planted in the field were harvested, dried naturally in the shade or at low temperature, crushed, and passed through an 80-mesh sieve, and the collected powder was used for the preparation of crude extraction. The specific extraction steps are as follows:

[0049] 1) Add 100g of different additive powders and 1000mL of methanol into a 2000mL Erlenmeyer flask. Ultrasonic treatment at 55°C for 40-60min, after filtration, collect the supernatant and filter residue respectively.

[0050] 2) Add 1000 mL of methanol to the filter residue in step 1) to continue ultrasonic treatment, repeat step 1 for 5-6 times, and combine all the obtained supernatants.

[0051] 3) Evaporate methanol from the supernatant in step 2) in a rotary evaporator at 50°C.

[0052] 4) Dissolve and evaporate the crude extract of methanol with single distilled...

Embodiment 2

[0057] Effects of extracts of different additional lines of Brassica napus-Isatis indica on the proliferation of influenza virus at the cell level in vitro

[0058] 1. Cell culture

[0059] The A549 cells recovered from cryopreservation were subcultured twice, and expanded with DMEM medium containing 10% fetal bovine serum and double antibodies (penicillin 100 U / ml, streptomycin 100 ug / ml).

[0060] 2. The cells were treated with the extract of Brassica napus-Isatis indigo line and infected with influenza virus

[0061] 1) The A549 cells in good growth state were digested and passaged, and the cell density was adjusted to 1×10 with cell culture medium. 5 / ml, 1ml per well was inoculated in a 12-well plate.

[0062] 2) When the cell growth monolayer density reaches 80%, extracts of different additional lines of Brassica napus-Isatis indigo are added to the A549 cells in the 12-well plate, and the final concentration of the extracts is 200 μg / mL.

[0063] 3) Infect the influe...

Embodiment 3

[0069] Inhibitory Effect of E Monomer Addition Line Extract on Influenza Virus Proliferation at Cellular Level in Vitro

[0070] According to the results of Example 2, it can be seen that the Brassica napus-Isatis indigo E monomer extract has a significant inhibitory effect on avian influenza virus H5N6. effect and its cytotoxicity.

[0071] 1. Toxic effect of E monomer extract on cells

[0072] 1) Take well-grown A549 and MDCK cells for digestion and passage, and adjust the cell density to 2×10 with cell growth medium (DMEM medium + 10% fetal bovine serum + double antibody) 6 / ml, inoculate 96-well plate, inoculate 100μl per well;

[0073] 2) Add 100 μl of E monomer extracts of different concentrations prepared with culture medium (DMEM medium + 10% serum + double antibody) to each well, and mix well. Set 6 concentration gradients, and set 3 replicate wells for each gradient concentration, and the final concentrations are 0 μg / ml, 12.5 μg / ml, 25 μg / ml, 50 μg / ml, 100 μg / ml,...

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Abstract

The invention belongs to the field of biotechnology, and in particular relates to the application of Brassica napus-Isatis indigo E monomer additive system in inhibiting influenza virus. The present invention provides a method for extracting the effective components of Brassica napus-Isatis indigo E monomer additive system. The extract prepared by the method is used for anti-influenza virus experiments, and it is found that in A, D, E, F 4 single Among the three double body addition lines A, B, and C, only the E monomer addition line has a significant anti-influenza virus effect at the cell and animal levels, and has obvious effects on human influenza PR8, swine influenza G4 and avian influenza H5N6. The antiviral effect, the composition not only has no toxicity, but also can promote cell growth, improve cell activity, and can significantly inhibit cell apoptosis. It can be developed and utilized as an antiviral medicinal vegetable, feed, etc., and can be used as a health food for humans and animals to protect the health of animals and humans.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to the application of Brassica napus-Isatis indigo E monomer addition line in inhibiting influenza virus. Background technique [0002] Influenza virus is the main pathogen of humans and animals, belonging to Orthomyxoviridae family Influenzavirus, is a highly contagious disease worldwide. Influenza viruses are divided into Type A, Type B, Type C and Type D. Among them, the antigenicity of Type A influenza virus is prone to variation, which has caused worldwide pandemics many times. In the influenza pandemic from 1918 to 1919, at least 20 million to 40 million people died of influenza in the world; influenza B virus is also highly pathogenic to humans, but it has not been found that influenza B virus has caused worldwide Influenza type C virus only causes inconspicuous or mild upper respiratory tract infection in humans and rarely causes epidemics; influenza type D virus ha...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N5/14A61K36/315A61P31/16A61P11/00A23L19/00A23L33/00A23L33/105A23K10/30A61K36/31
CPCA61K36/31A61K36/315A61P31/16A61P11/00A23L19/00A23L33/00A23L33/105A23K10/30A61K2236/333A61K2236/53A61K2236/51A23V2002/00A23V2200/30A23V2250/21A61K2300/00
Inventor 金梅林杨丽邹忠龚文孝黄坤钟鸣赵雪锦赵丽夏小涵李再云盛锋杜雪竹孙小美康超
Owner HUAZHONG AGRI UNIV
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