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Non-alcoholic fatty liver disease cell model induction liquid and preparation method and application thereof

A fatty liver disease, cell model technology, applied in biochemical equipment and methods, animal cells, tumor/cancer cells, etc., can solve the problems of high requirements, complex methods, long modeling cycle, etc., to achieve a high modeling success rate Effect

Inactive Publication Date: 2019-04-16
AFFILIATED HOSPITAL OF ZUNYI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

At present, the cell models of non-alcoholic steatosis liver disease established by the above induction methods are relatively stable and have good repeatability, but the method is complicated, requires high conditions, and the modeling period is long

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  • Non-alcoholic fatty liver disease cell model induction liquid and preparation method and application thereof
  • Non-alcoholic fatty liver disease cell model induction liquid and preparation method and application thereof

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preparation example Construction

[0020] The invention provides a method for preparing a non-alcoholic fatty liver disease cell model inducing liquid, comprising the following steps:

[0021] 1) Saponification is carried out after mixing stearic acid and sodium hydroxide solution to obtain saponification liquid; the mass volume ratio of the stearic acid and sodium hydroxide solution is 1g: (170~185)mL;

[0022] 2) After mixing the saponification solution with the defatted bovine serum albumin solution with a mass concentration of 35% to 43% in equal volumes, an induction solution mother solution is obtained;

[0023] 3) Diluting the mother solution of the induction solution with fetal bovine serum medium with a volume fraction of 7-12%, to obtain the induction solution.

[0024] In the preparation method of the nonalcoholic fatty liver disease cell model induction solution of the present invention, saponification is carried out after stearic acid and sodium hydroxide solution are mixed, obtain saponification s...

Embodiment 1

[0030] 1) Preparation of 40% fat-free bovine serum albumin: Preheat the PBS solution to 55°C, measure 1.2g of fat-free bovine serum albumin, put it in a 15mL centrifuge tube, add 2mL of PBS solution, do not shake or mix, put directly Centrifuge at 8000rpm in a high-speed centrifuge for 20 minutes to dissolve completely, and dilute to 3mL with PBS. Obtaining 3mL of a mass concentration of 40% defatted bovine serum albumin solution, the appearance is a brownish-yellow clear sample;

[0031] 2) Preparation of 20mM stearic acid solution: Take 0.04g of sodium hydroxide, dissolve it in 10mL of deionized water, make 10mL of 0.1mol / L sodium hydroxide solution, take 3mL. Weigh 0.017g of stearic acid and add it into 3mL of sodium hydroxide solution, place it in a water bath at 100°C for full saponification for 10 minutes, until finally it becomes colorless, transparent and clear, and 20mM sodium stearate saponification solution is obtained.

[0032] 3) Add the insulated stearic acid so...

Embodiment 2

[0034] Dilute the 10mM stearic acid solution by 50, 100, and 200 times with a volume fraction of 10% fetal bovine serum medium to form a stearic acid working solution with a concentration of 200 μM, 100 μM, and 50 μM. HepG2 cells were treated for 24 hours, the above experiment was repeated 3 times, oil red O staining was performed, and the results were as follows figure 1 As shown, it can be seen that 200 μM, 100 μM, and 50 μM stearic acid working solutions can all be successfully modeled.

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Abstract

The invention provides a preparation method of a non-alcoholic fatty liver disease cell model induction liquid and application of the induction liquid and belongs to the technical field of biologicalmodel establishment. The induction liquid comprises a degreased bovine serum albumin solution of stearic acid, wherein the concentration of the stearic acid in the induction liquid is 50-200 micromole / L. According to the induction liquid, liver cell fat hyalinosis is induced through the stearic acid, the modeling success rate is high, the condition requirement is low, the method is simple and convenient, the modeling cycle is show, and the repeatability is high. A targeting tool is offered to research in the occurrence and development of non-alcoholic fatty liver diseases, the liver disease occurrence mechanism is further understood, a new function target point is provided for developing anti-liver injury drugs, and a new thought and a new way for clinically preventing and treating the non-alcoholic fatty liver diseases.

Description

technical field [0001] The invention belongs to the technical field of biological model establishment, and in particular relates to a non-alcoholic fatty liver disease cell model inducing solution and a preparation method and application thereof. Background technique [0002] With the increasing number of obese people in the world, non-alcoholic fatty liver disease has become a common disease, and its incidence is showing an increasing trend year by year. At present, about 20-30% of the adults in the world may suffer from non-alcoholic fatty liver disease, most of them are in the stage of simple fatty liver, and there will be no serious symptoms or liver function damage in the early stage, so it has not received enough attention. And about 20% of people with simple fatty liver may develop into steatohepatitis patients or even liver cirrhosis and liver cancer within a few years. In recent years, a number of studies have shown that patients with non-alcoholic fatty liver are ...

Claims

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Application Information

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IPC IPC(8): C12N5/09
CPCC12N5/0693C12N2500/30C12N2500/12C12N2501/998
Inventor 陈欢何毅怀陈贵梅林世德沈访
Owner AFFILIATED HOSPITAL OF ZUNYI UNIV
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