Long non-coding RNA and application thereof in diagnosis and/ or treatment of T2DM

A non-coding, therapeutic target technology, applied in the field of genetic engineering, can solve problems such as beta cell developmental defects and impaired glucose tolerance in adult mice

Active Publication Date: 2017-02-15
江苏省省级机关医院
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The same research group recently discovered that lncRNAβlinc1 maintains islet function by regulating islet-specific transcription factors, and knockout can cause defects in beta cell development and impaired glucose tolerance in adult mice

Method used

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  • Long non-coding RNA and application thereof in diagnosis and/ or treatment of T2DM
  • Long non-coding RNA and application thereof in diagnosis and/ or treatment of T2DM
  • Long non-coding RNA and application thereof in diagnosis and/ or treatment of T2DM

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0077] Example 1 detects the expression of Linpint in normal Balb / c mice

[0078] Extraction of primary islet cells, with HBSS solution (KCl 0.4g, KH2PO 40.06g, MgCl2.6H2O 0.1g, MgSO4.7H2O 0.1g, CaCl2 0.1g, NaCl 8g, NaHCO3 0.35g, NaHPO4.7H2O 0.09g deionized water To IL, PH7.4; prepare collagenase (collagenase V prepared with HBSS solution, 1mg / ml); mice fasted for more than 12h, prepare pentobarbital sodium, and anesthetize mice. , ligated with a thread; cut the heart to bleed, wipe off the blood, find the common pancreatic duct, and intubate; push the syringe first slowly and then quickly. Pancreas filling generally takes 2ml. Peel off the pancreas, put it in a sterilized 50ml centrifuge tube, and then Put it on ice; add collagenase V to 5ml centrifuge tube to 5ml. Bath in 37°C water for 25min; take out the centrifuge tube and put it on ice; vortex the centrifuge tube to vortex the tissue into sand. Add HBSS with serum 2 times the volume of the solution, mix well to stop dig...

Embodiment 2

[0098] Example 2 In db / db mouse islets, the expression of Linpint is down-regulated

[0099] The db / db (diabetes) mouse is a mouse model of T2DM. There is a recessive mutation in the db gene of the mouse autosome 4. It is a spontaneous type 2 diabetic mouse caused by leptin receptor deficiency. It is a good way to study T2DM. animal models.

[0100] The db / db mice began to develop hyperglycemia, hyperinsulinemia, and insulin resistance in about 2 weeks, and then became obese. Generally, after 4 weeks, the blood sugar increased significantly, and the islet beta cells were obviously damaged. After eight weeks, the random blood sugar reached Above 20mmol / L, typical symptoms of diabetes appear. 8-week-old and 12-week-old control mice and db / db mice were simultaneously detected for random blood glucose changes (Fig.2.A): Compared with control mice, the random blood glucose of db / db mice was elevated (P <0.05). The islets of control mice and db / db mice were extracted, and the exp...

Embodiment 3

[0102] Example 3 In mouse cell lines, down-regulation of Linpint inhibits the secretion of insulin

[0103] In order to detect the effect of Linpint on the function of islet cells, Invitrogen Company designed and synthesized three interference sequences of mouse Linpint, and synthesized the corresponding scrambled sequences as a control. We transfected the three interference sequences of Linpint in MIN6 and NIT-1 cells, and detected their interference efficiency by real-time quantitative PCR. The results showed (Fig.4A-B): Compared with the control, the interference efficiency of Linpint siRNA#1 Up to 80±3% and 68±3% (P<0.01).

[0104] The MIN6 cells were planted in a 48-well culture plate, and when the cell density reached 60-70%, the medium was discarded and transfected. After 48h, the GSIS experiment was carried out. Prepare glucose-free Krebs-Ringer bicarbonate (Krb) (115mM NaCl, 4.7mM KCl, 1.2mM MgSO4.7H2O, 1.2mM KH2PO4, 20mM NaHCO3, 16mM HEPES, 2.56mM CaCl2, 0.2% BSA) ...

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Abstract

The invention belongs to the field of gene engineering, and in particular relates to application of Lincpint in preparation of T2DM prediction and targeted treatment drugs, down-regulation of the Lincpint in T2DM mice is relate to function injuries of pancreatic beta cells, the Lincpint of a low expression level may participate the pathogenesis of the T2DM, and by change of the expression of the Lincpint, the apoptosis of the pancreatic beta cells and synthesis and secretion of insulin can be influenced.

Description

technical field [0001] The invention belongs to the field of genetic engineering, in particular to a long non-coding RNA and its application in diagnosis / treatment of T2DM. Background technique [0002] Diabetes is a chronic disease that seriously affects the health of the population. The incidence rate is increasing year by year. It not only brings physical and mental damage to individuals, but also brings economic burdens to individuals and the country. The complex organizational structure of the pancreas and the physiological function of the islets, as well as the high incidence of diabetes in clinical practice, make the study of islet function particularly important. The maintenance of islet function is one of the current research focuses on diabetes. [0003] More and more reports have shown that lncRNA is involved in the development and function maintenance of islet cells, and the occurrence and development of diabetes and its complications. A research group found by ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/113C12Q1/68
CPCC12N15/113C12N2310/10C12Q1/6883C12Q2600/136C12Q2600/158C12Q2600/178
Inventor 唐伟金飞燕德伟朱亚男王宁
Owner 江苏省省级机关医院
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