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Applications of inhibition of Dcf1 gene expression in preparing drugs used for treating obesity

A dcf1, inhibition technology, applied in the application field of inhibiting the expression of Dcf1 gene in the preparation of obesity treatment drugs, can solve the problems of fat-soluble vitamin deficiency and other problems

Active Publication Date: 2017-07-04
SHANGHAI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, there are few weight-loss drugs approved by the FDA on the market, mainly targeting lipase, but long-term use can cause steatorrhea, which can cause fat-soluble vitamin deficiency

Method used

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  • Applications of inhibition of Dcf1 gene expression in preparing drugs used for treating obesity
  • Applications of inhibition of Dcf1 gene expression in preparing drugs used for treating obesity
  • Applications of inhibition of Dcf1 gene expression in preparing drugs used for treating obesity

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0012] Example 1: All experiments were performed in male mice (maintained on a C57BL / 6J background). After the animal is born, dcf1 - / - and dcf1 + / + mice were reared individually in the third week and maintained 12 hours of light (06:00-18:00) and 12 hours of darkness (18:00-06:00), and controlled temperature (24±1°C). Food and water were available ad libitum and sufficient. All animal handling was performed in accordance with Society for Neuroscience guidelines. Three-month-old size comparison see ( figure 1 A), the results of body weight and diet are shown in ( figure 1 B, 1C). (n=28, *P<0.05; **P<0.01; *P<0.001).

Embodiment 2

[0013] Example 2: Take out the target area, hypothalamus, hippocampus and cortex from the pre-cooled PBS. Then homogenize in a homogenizer with Western and IP cell lysate until there is no obvious tissue block and it becomes a transparent homogenate. Continue to lyse on ice for 30 min, and then collect the lysate in a 1.5ml EP tube. The supernatant was collected by centrifugation at 8000 rpm for 10 minutes at 4°C. Continue to centrifuge the supernatant at 4°C, 15,000rpm for 20min, collect the supernatant again to be the tissue protein extract, add the corresponding amount of Loading buffer, heat and denature it for later use. Afterwards, Western blotting was performed. The electrophoresis conditions are stacking gel 80V, 30 minutes; separating gel 120V, 90 minutes. The transfer condition was 25V for 40 minutes. The primary antibody dilution ratio is 1:1000, and the secondary antibody dilution ratio is 1:10000. ( figure 2 , B, C, D, , E, F are the statistical analysis of...

Embodiment 3

[0014] Example 3: Immunohistochemical detection

[0015] Firstly, the experimental mice were perfused and fixed with 4% PFA, dehydrated with sucrose, and embedded. Then cryosection (20um) was performed. Finally, immunohistochemistry was performed. Wash 3 times with 0.01M PBS, 5 minutes each; Permeabilize with 0.1%TritonX-100-PBS for 30 minutes; Wash 3 times with 0.01MPBS, 5 minutes each; Block with 2% BSA-PBS for 1 hour at room temperature; Blot dry with 2% BSA - After PBS, directly drop the primary antibody (MBP, 1:500) diluted in PBS. Incubate at 37°C for 2 hours, or overnight at 4°C; blot dry the primary antibody, wash with 0.01M PBS 3 times, 5 minutes each time. Under dark conditions, add secondary antibody diluted in PBS (1:500 dilution ratio). Incubate at 37°C for 2 hours; blot dry the secondary antibody, add DAPI (1:1000) diluted in PBS, and incubate at room temperature for 10 minutes; blot dry DAPI, wash 3 times with 0.01M PBS, 5 minutes each time; seal with fluore...

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Abstract

The invention relates to applications of inhibition of Dcf1 gene expression in preparing drugs used for treating obesity. According to the invention, behavior tests of wild type mice (as a reference) and dcf1 knock-out mice are carried out, it is detected that the weight and food intake of mice are reduced because of dcf1 deletion, and NPY protein down-regulation is detected in Westernblotting and immunohistochemical detection, so that it is confirmed that dcf1 is at the upstream of NPY. ATP kit is used for detecting that after dcf1 knockout, mice hypothalamus tissue and primary culture cell ATP level increases. Expression of dcf1 and expression of NPY in 293T cells are both capable of reducing cell ATP level, so that dcf1 possesses an important effect in energy balance, and a novel method used for treating obesity is provided.

Description

technical field [0001] The present invention relates to an inhibitory dcf1 Application of gene expression in preparation of medicine for treating obesity. Background technique [0002] Obesity has significant and widespread impacts on human health, and it is currently the top healthcare priority and challenge of this century. Genetic studies suggest that energy balance disorders are the result of key regulators, such as neuropeptide Y (NPY) or proopiomelanocortin gene (POMC). Current research shows that NPY can increase food intake and reduce thermogenic effects, while POMC has the opposite effect, it can suppress appetite and increase energy consumption. Obesity is caused by energy intake greater than expenditure, which is stored and accumulated in the form of fat. Currently, there are few FDA-approved weight-loss drugs on the market, which mainly work on lipase, but long-term use can cause steatorrhea and cause fat-soluble vitamin deficiency. It has also recently been ...

Claims

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Application Information

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IPC IPC(8): A61K45/00A61K49/00G01N33/68A61P3/04
CPCA61K45/00A61K49/0008G01N33/68
Inventor 文铁桥陈瑜冯瑞丽王娇
Owner SHANGHAI UNIV
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