Function and application of thymidylate synthetase CAD (carbamoyl-phosphate synthetase II, aspartate transcarbamylase and dihydroorotase) gene in treating fatty liver and type-II diabetes mellitus

A nucleotide synthetase and fatty liver technology, applied in gene therapy, medical preparations containing active ingredients, metabolic diseases, etc., can solve problems such as liver damage, aggravate heart burden, increase blood volume, etc., and achieve improvement of fatty liver Effect

Active Publication Date: 2014-10-01
武汉惠康基因科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0008] 4) Insulin sensitizers work by activating PPAR-, and their biggest side eff...

Method used

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  • Function and application of thymidylate synthetase CAD (carbamoyl-phosphate synthetase II, aspartate transcarbamylase and dihydroorotase) gene in treating fatty liver and type-II diabetes mellitus
  • Function and application of thymidylate synthetase CAD (carbamoyl-phosphate synthetase II, aspartate transcarbamylase and dihydroorotase) gene in treating fatty liver and type-II diabetes mellitus
  • Function and application of thymidylate synthetase CAD (carbamoyl-phosphate synthetase II, aspartate transcarbamylase and dihydroorotase) gene in treating fatty liver and type-II diabetes mellitus

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0051] [Example 1] Mouse fatty liver and type Ⅱ diabetes model (DIO) obtained

[0052] 1. Grouping of experimental animals: 8-week-old, male, WT mice and CAD-KO mice were selected and given two special diets, D12942 high fat diet (Highfat diet, HFD) and D12450B low fat diet (Normal chow, NC) respectively Breeding, that is, WT NC group, KO NC group, WT HFD group, KO HFD group, a total of 4 groups.

[0053] 2. Operation process of high-fat feed induction model:

[0054] Using WT and KO mice, the DIO model was established, and the phenotype correlation analysis was carried out to clarify that the CAD gene plays an important role in the pathogenesis of metabolic diseases. 8-week-old, male, WT mice and CAD-KO mice were selected and fed with two special diets, D12942 high-fat diet (HFD) and D12450B low-fat diet (Normal chow, NC), respectively, that is, WT NC group , KO NC group, WT HFD group, KO HFD group, a total of 4 groups. The food intake of the mice was recorded in detail ev...

Embodiment 2

[0056] [Example 2] mouse body weight, blood glucose level determination

[0057] (1) Detection of fasting body weight and food intake of mice, and blood collection from orbital sinus

[0058] 1) Weight detection

[0059] ①Fasting: Fast the mice to be tested at 8:00 am (without water), and start the experimental operation at 2:00 pm.

[0060] ② Weighing: Weigh at 0 week, 4 week, 8 week, 12 week, 16 week, 20 week and 24 week respectively, put a small plastic bucket on the dynamic electronic balance, grab the mouse, put it into the weighing In a small bucket, measure the weight and record the data. Feed amount detection: After the weighing operation is completed, add feed to the mice, and record the amount of feed for the mice on the dynamic electronic balance.

[0061] (2) Detection of fasting blood glucose level

[0062] All the mice to be tested were fasted from 8:00 am to 2:00 pm (without water), that is, the experimental operation was started after 6 hours of fasti...

Embodiment 3

[0069] [Example 3] Glucose tolerance test (intraperitoneal glucose tolerance test, IPGTT)

[0070] On the 22nd week of the experiment, the intraperitoneal glucose injection test (IPGTT) was performed to evaluate the glucose tolerance of the mice.

[0071] ① Before measuring blood glucose, measure the fasting body weight of the mouse, and calculate the injection volume of glucose based on 10 μL / g.

[0072] ② First test the fasting blood sugar at 0 minutes before the glucose injection, and inject the glucose solution through the abdominal cavity quickly after the test is completed.

[0073] ③ Operation method of intraperitoneal injection: ①Fix the mouse; grab the mouse, grasp the tail of the mouse with the little finger and ring finger of the left hand, and grasp the neck of the mouse with the other three fingers, so that the head of the mouse is down, and the mouse The abdomen is fully exposed. ②Needle positioning and injection: insert the needle from the side of the abdomen,...

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Abstract

The invention discloses a function and application of thymidylate synthetase CAD (carbamoyl-phosphate synthetase II, aspartate transcarbamylase and dihydroorotase) genes in treating fatty liver and type-II diabetes mellitus, and belongs to the field of gene functions and application. Functions of CAD genes are studied by a high-fat diet (HFD) induced model, and results discover that the levels of body weight and fasting blood-glucose of a CAD gene knock-out mouse in an HFD group are higher than those of a WT (Wild Type) mouse in a control group; glucose tolerance tests by intraperitoneal injection discover that the tolerance of the CAD gene knock-out mouse on glucose is remarkably weakened; the integral liver appearance, liver weight, liver/weight ratio and lipid component pathological staining result of the mouse indicate that the fatty liver disease of the CAD knock-out mouse in the HFD group is remarkably severe, the lipid accumulation is remarkably increased, and these results indicate that CAD gene knock-out can remarkably deteriorate fatty liver and type-II diabetes mellitus. Regarding the effects of CAD, the CAD can be used for preparing medicaments for preventing, relieving and/or treating fatty liver and/or type-II diabetes mellitus.

Description

technical field [0001] The invention belongs to the field of gene function and application, and particularly relates to a nucleotide synthetase CAD: carbamoyl-phosphate synthetase II, aspartate aminotransferase and dihydroorotase (carbamoyl-phosphate synthetase II, aspartate The function and application of transcarbamylase, and dihydroorotase (CAD) in fatty liver and type Ⅱ diabetes, specifically in the application of drugs for the prevention, alleviation and / or treatment of fatty liver and / or type Ⅱ diabetes. Background technique [0002] Diabetes is a chronic metabolic disease that seriously endangers human health. It is estimated that the number of patients will increase to 439 million by 2030, which means that 7.7% of adults aged 20-79 in the world will be diabetics. In the past two or three decades, the number of people with diabetes has doubled, and it is an extremely serious threat to human health around the world. Currently, the incidence of type 2 diabetes and pre-...

Claims

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Application Information

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IPC IPC(8): A61K48/00A61K38/43A61P1/16A61P3/10
Inventor 李红良汪涛张晓东杜成
Owner 武汉惠康基因科技有限公司
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