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A method to increase the expression level of gonadotropin after adding vitamin E to the pituitary cells of semi-smooth tongue sole

A technology of gonadotropin and pituitary cells, applied in biochemical equipment and methods, animal cells, vertebrate cells, etc., can solve problems that have not yet been reported on gonadotropin research

Active Publication Date: 2019-11-26
YELLOW SEA FISHERIES RES INST CHINESE ACAD OF FISHERIES SCI
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Problems solved by technology

At present, in fish, there is no research report on the relationship between vitamin E and gonadotropins secreted by the pituitary gland.

Method used

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  • A method to increase the expression level of gonadotropin after adding vitamin E to the pituitary cells of semi-smooth tongue sole
  • A method to increase the expression level of gonadotropin after adding vitamin E to the pituitary cells of semi-smooth tongue sole
  • A method to increase the expression level of gonadotropin after adding vitamin E to the pituitary cells of semi-smooth tongue sole

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Embodiment Construction

[0043] The method of the present invention will be further described below in conjunction with the accompanying drawings. However, this embodiment is only limited to explain the steps of the present invention, and is not limited thereto. The specific experimental conditions not indicated in the following examples can generally be operated under conventional conditions.

[0044] (1) Isolation and culture of pituitary cells of semi-smooth tongue sole

[0045] 1) Preparation of cell culture medium: Take 9.6 grams of L-15 medium and 4.76 grams of Hepes, fully dissolve and mix for 4 hours, adjust the pH to 7.4 with NaOH, then filter with suction, and subpackage, which is the L-15 basal medium , stored at 4°C; L-15 basal medium was added with a final volume of 5% fetal bovine serum, a final concentration of 100 U / ml of penicillin, and a final concentration of 100 μg / ml of streptomycin, which was L-15 complete culture base, stored at 4°C.

[0046] 2) Primary culture: Ovary extract...

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Abstract

The present invention relates to a method for increasing the expression level of gonadotropin after adding vitamin E to pituitary cells of semi-smooth tongue sole. Carry out the cultivation of primary cells: (2) add vitamin E to semi-smooth tongue sole pituitary cells: prepare vitamin E solution, add vitamin E to the cells cultured in step (1), and collect samples on the fourth day after adding; (3) Quantitative PCR analysis of FSH and LH after vitamin E was added to the pituitary cells of semi-smooth tongue sole; the cell pellet in step (2) was taken for RNA extraction and reverse transcription, and then real-time fluorescent quantitative PCR reaction; (4) ELISA analysis of FSH and LH in the pituitary cells of half-smooth tongue sole after adding vitamin E; the supernatant collected in step (2) was used as the material to carry out the dilution and sample addition, sample addition, incubation, liquid preparation, washing, and addition of the standard in sequence Enzyme, Incubation, Washing, Color Development, Termination, Assay.

Description

technical field [0001] The invention belongs to the field of aquaculture in aquatic biology technology, and in particular relates to a method for increasing the expression level of gonadotropin after adding vitamin E to pituitary cells of semi-smooth tongue sole. Background technique [0002] Vitamin E (Vitamin E) includes two types of tocopherols and tocotrienols, a total of 8 compounds, namely α, β, γ, δ tocopherols and α, β, γ, δ tocotrienols. α-tocopherol is the most widely distributed, most abundant and most active form of vitamin E in nature. Vitamin E can participate in the mediation of tissue respiration and oxidative phosphorylation, and has the functions of promoting reproduction, anti-oxidation, and immune regulation (Maconson, Aquatic Animal Nutrition and Feed Science, China Agricultural Press, 2011). [0003] Semi-smooth tongue sole (Cynoglossμs semilaevis) is a warm-temperate large and medium-sized demersal fish unique to the coastal waters of my country. It i...

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N5/071
CPCC12N5/0616C12N2500/38
Inventor 王娜王蔚芳黄滨史宝王若青马佳璐陈松林
Owner YELLOW SEA FISHERIES RES INST CHINESE ACAD OF FISHERIES SCI