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Method for building diabetes model through inducing human liver cancer cells HepG2 by glucose and application

A technology of liver cancer cells and glucose, applied in metabolic diseases, active ingredients of heterocyclic compounds, drug combinations, etc., can solve the problems of unpublished diabetes models and unresearched effects of HepG2 cells.

Inactive Publication Date: 2019-03-26
JIANGSU ACADEMY OF AGRICULTURAL SCIENCES
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Problems solved by technology

[0005] Previously, although there was a patent (CA 104357397A) disclosing "a method for establishing a drug-resistant liver cancer HepG2 cell model", there was no research on the effect of drugs on the growth status of HepG2 cells, and the human liver cancer cell HepG2 was only It has been applied in the research of anti-tumor and anti-oxidation
In addition, the patent for establishing a diabetes model with HepG2 cells as the research object has not yet been published.

Method used

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  • Method for building diabetes model through inducing human liver cancer cells HepG2 by glucose and application
  • Method for building diabetes model through inducing human liver cancer cells HepG2 by glucose and application

Examples

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Embodiment example 1

[0019] HepG2 cells were placed in complete medium (1% double antibody, 1% Hepes 1M, 10% FBS, 5.5mM glucose) at 37°C, 5% CO 2 Cultivate them in an incubator until they are completely adhered to the wall. When the HepG2 cells adhere to the wall to about 80%, add 2 mL of trypsin and place in the incubator to digest for 2 minutes, prepare a suspension, drop it into the cell counting plate, count under the microscope, and adjust the cell concentration to 1×10 5 pieces / ml. Set up the control group and the model group, inoculate 100 μL per well in a 96-well plate, and culture in a 37°C incubator for 24 hours until the cells are 70% full. The supernatant was discarded, and 100 μL of incomplete medium (1% double antibody, 1% Hepes 1M, 5.5 mM glucose) was added for starvation culture for 4 hours to synchronize all HepG2 cells. The model group was stimulated by adding glucose solution with a final concentration of 30mM for 24 hours. According to the MTT assay, cell viability was measu...

Embodiment example 2

[0021] HepG2 cells were placed in complete medium (1% double antibody, 1% Hepes 1M, 10% FBS, 5.5mM glucose) at 37°C, 5% CO 2 Cultivate them in an incubator until they are completely adhered to the wall. When the HepG2 cells adhere to the wall to about 80%, add 2 mL of trypsin and place in the incubator to digest for 2 minutes, prepare a suspension, drop it into the cell counting plate, count under the microscope, and adjust the cell concentration to 1×10 5 pieces / ml. Set up the control group, model group, and experimental group, inoculate 100 μL per well in a 96-well plate, and culture in a 37°C incubator for 24 hours until the cells are 70% full. The supernatant was discarded, and 100 μL of incomplete medium (1% double antibody, 1% Hepes 1M, 5.5 mM glucose) was added for starvation culture for 4 hours to synchronize all HepG2 cells. Firstly, blueberry anthocyanin extract with a final concentration of 5 μg / mL was added to the experimental group for protection for 24 hours, a...

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Abstract

The invention relates to a method for preparing a diabetes model through inducing human liver cancer cells HepG2 by glucose and application, and belongs to the technical field of biology. By using themethod, at 37 DEG C and in a 5-percent CO2 constant temperature culture box, the HepG2 cells are cultured until the wall attachment growth reaches 80 to 90 percent; the cells are inoculated to a 96-hole plate according to the cell concentration being 1*10<5> / mL; when the cells grow to 50 to 70 percent of the bottom of the 96-hole plate, after performing starved culture for 4 h by an incomplete culture medium, glucose with the final concentration being 30 to 120 mM is added for induction for 24 to 48 h to build a diabetes cell model. The glucose induced in vitro diabetes cell model is built; through high-concentration glucose stimulation, the survival rate of the HepG2 cells is obviously reduced; the survival rate of model group cells is 35 to 75 percent of a control group. The method is simple; the implementation is easy; the method can be used for studying the influence of medicine on glycometabolism; the screening speed on the hypoglycemic medicine is improved; the far-reaching significance is realized on the diabetes study.

Description

1. Technical field [0001] The invention relates to a preparation method and application of a glucose-induced human liver cancer cell HepG2 diabetes model, belonging to the field of biotechnology. 2. Background technology [0002] Diabetes is a metabolic disease characterized by hyperglycemia, a series of clinical syndromes caused by absolute or relative lack of insulin in the body, and is closely related to genetics. Nowadays, the incidence of diabetes and its complications has been increasing and has become a global problem. According to the statistics of the International Diabetes Federation, there were 150 million people with diabetes in the world in 2000, and now it has reached 285 million. According to the current growth rate, it is estimated that 500 million people in the world will suffer from this disease in 2030, and China has become the hardest hit area for diabetes. . [0003] Diabetes is mainly divided into type 1, type 2 and gestational diabetes, with type 2 d...

Claims

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Application Information

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IPC IPC(8): A61K31/35A61P3/10
CPCA61K31/35A61P3/10
Inventor 黄午阳吴寒宝琳柴智李大婧刘春泉徐亚平
Owner JIANGSU ACADEMY OF AGRICULTURAL SCIENCES
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