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Application of LncRNA-266 in preparation of drug for inducing brown adipose cell differentiation

A technology of lncRNA-266, 1.lncRNA-266 is applied in the preparation of drugs for inducing brown adipocyte differentiation and promoting energy metabolism in the body, and can solve problems such as developmental defects and various diseases

Active Publication Date: 2021-02-02
NANTONG UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, recent studies have found that many lncRNAs have very important physiological functions, and their abnormal levels can lead to developmental defects and the occurrence and development of various diseases

Method used

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  • Application of LncRNA-266 in preparation of drug for inducing brown adipose cell differentiation
  • Application of LncRNA-266 in preparation of drug for inducing brown adipose cell differentiation
  • Application of LncRNA-266 in preparation of drug for inducing brown adipose cell differentiation

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0024] Example 1 LncRNA-266 overexpression and interference virus construction

[0025] Preparation of overexpression virus: In this study, the overexpression virus was constructed using Invitrogen’s Gateway series kits. Firstly, PCR is used to amplify the complete sequence of LncRNA, and then it is connected into the intermediate vector pENTR3C-Entry Vector and then sent to the company for sequencing. The sequenced correct vector is connected into the target vector (pAd-CMV-DESTVector) by recombination method (LR). , after the sequencing detection was correct, the positive plasmids were extracted and single-cut by the endonuclease Pac I, and then transfected into 293A cells with Lipofectamine 2000 liposome from Invitrogen Company. Observe the formation of viral plaques for about a week. After most of the viral plaques appear, collect 293A cells and perform repeated freezing and thawing at -80°C and 37°C to break the cells, and collect the supernatant by centrifugation to obta...

Embodiment 2

[0027] Example 2 LncRNA-266 induces differentiation of preadipocytes

[0028] Primary preadipocyte culture: 8-week-old male C57BL / 6J mice, take groin white adipose tissue in a centrifuge tube, collagenase digestion solution (collagenase 1mg / ml dissolved in separation buffer ----- including 0.123 M NaCl, 5mM KCl, 1.3mM CaCl 2 , 5nM glucose, 100mM Hepes, 4% BSA) 37°C water bath for 30min, shake once every 5min, filter with 100μm nylon mesh, centrifuge at 200g×5min. Remove the supernatant, add 2ml separation buffer to fully suspend the cells, and centrifuge at 200g×5min. Remove the supernatant, add 2ml of culture medium (DMEM high-glucose medium, 20% fetal bovine serum, 20mM Hepes, 100U / ml penicillin / streptomycin) to fully suspend the cells, plant in a culture dish, place at 37°C, 5 %CO 2 cultured in an incubator.

[0029] Directed differentiation of brown adipocytes: In order to detect the transformation of lncRNA-266 from preadipocytes to brown adipocytes, before adding ind...

Embodiment 3

[0039] Example 3 Effect of LncRNA-266 on Glucose Metabolism in Type Ⅱ Diabetic Mice

[0040] 6-week-old male C57BL / 6J mice were fed with a high-fat diet (Research Diet, 45% calories from fat) for 90 days to induce obesity model mice (ie, type II diabetes model mice). The adenovirus expressing LncRNA-266 (Ad-LncRNA-266) was directly injected into the inguinal beige adipose tissue of type Ⅱ diabetes model mice, and the dose used was to inject 40 μl of virus (1×10 12 PFU). Mice in the control group were injected with the same dose of control virus (Ad-LacZ). Three weeks after virus injection, various physiological indicators were detected.

[0041] After the mice were fasted for 12 hours, blood was taken from the tail vein, and the blood glucose level was quantitatively detected with a blood glucose meter (Bayer, Mishawaka, IN). The results are as follows: image 3 shown. image 3 The results showed that LncRNA-266 treatment could reduce blood glucose levels in type 2 diabete...

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Abstract

The invention relates to medical application of long-chain non-coding RNA (LncRNA-266), in particular to application of LncRNA-266 in preparation of a drug for inducing brown adipose cell differentiation, which can promote body energy consumption and resist obesity. Obese mice are treated by the LncRNA-266, and a result shows that the LncRNA-266 can promote differentiation of beige fat cells of the obese mice to brown fat cells and consume body energy, so that weight gain is inhibited. It is further found that the LncRNA-266 can reduce the blood glucose level of obese mice and improve the insulin sensitivity.

Description

technical field [0001] The invention belongs to the field of medicine, and in particular relates to the application of LncRNA-266 in the preparation of medicines for inducing brown fat cell differentiation and promoting body energy metabolism. Background technique [0002] Obesity is a physiological state of energy metabolism disorder in the body, which is characterized by a large amount of excess energy accumulated in the body in the form of fat. Obesity is a serious threat to human health, it can induce a variety of metabolic diseases, such as fatty liver, type 2 diabetes, cardiovascular disease, certain types of cancer, etc. There are two main types of adipose tissue in the body, namely white adipose tissue and brown adipose tissue. In terms of energy metabolism, white adipocytes contain a large amount of triglycerides, so white adipose tissue is mainly used as an organ for storing body energy; in contrast, brown adipocytes contain a large number of mitochondria and high...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K31/7105A61P3/04A61P3/10
CPCA61K31/7105A61P3/04A61P3/10
Inventor 孙诚汤欣马谨瑜刘晓宇
Owner NANTONG UNIVERSITY
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