Application of inhibition of SOCS3 expression through lentivirus-mediated RNA interference in preventing and controlling insulin resistance in diabetics

A technology for insulin resistance and diabetes, applied in the direction of DNA / RNA fragments, recombinant DNA technology, metabolic diseases, etc., to achieve the effect of improving insulin resistance, broad application background and economic value

Inactive Publication Date: 2017-11-24
张旭
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

T2D is a disease caused by multiple factors. At present, its etiology and pathogenesis have not been fully elucidated. At home and abroad, lifestyle intervention and drug intervention are mainly

Method used

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  • Application of inhibition of SOCS3 expression through lentivirus-mediated RNA interference in preventing and controlling insulin resistance in diabetics
  • Application of inhibition of SOCS3 expression through lentivirus-mediated RNA interference in preventing and controlling insulin resistance in diabetics
  • Application of inhibition of SOCS3 expression through lentivirus-mediated RNA interference in preventing and controlling insulin resistance in diabetics

Examples

Experimental program
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Effect test

Embodiment 1

[0030] In this example, C6 rat glioma cells were transfected with SOCS3shRNA, the SOCS3shRNA with the highest efficiency of interfering with SOCS3 gene was screened out, and SOCS3RNAi lentivirus was packaged with the lentiviral packaging plasmid.

[0031] 1. Construction of SOCS3 short hairpin RNA (SOCS3 shRNA)

[0032] Search the rat SOCS3 gene nucleotide sequence (Gene ID: 89829) in NCBI GenBank, design four siRNA sequences of 21 bases according to the design principles of small interfering RNA (siRNA), and use BLAST search to confirm the specificity of the siRNA sequence sex. At the same time, a non-interfering sequence siRNA-Negtive of the same length was randomly designed as a control. The designed siRNA sequences were synthesized into shRNA double-stranded DNA, and the sequence TTCAAGAGA was used as a circular structure to avoid the formation of a termination signal, and the sequence TTTTTT was added at the 3' end to terminate transcription. The shRNA double-stranded D...

Embodiment 2

[0042] This example is an example of the application of SOCS3RNAi lentivirus to treat a T2D rat model in vivo. In Example 2, the shRNA that inhibits the expression of SOCS3 of the present invention was applied to treat the T2D rat model in vivo, FBG, FINS and IRI were reduced, but ISI was significantly increased. Compared with the control group, the difference was statistically significant.

[0043] 1. Model preparation

[0044] 40 SPF-grade male SD rats with a body weight of 180g-220g (purchased from Slack Experimental Animal Center, license number: SCXK (Shanghai) 2012-0002) were fed with normal diet for 1 week and then given high-fat and high-sugar Feed feeding. After 4 weeks, rats were fasted at night for 12 hours, and received a single intraperitoneal injection of freshly prepared 1% streptozotocin (STZ) (30mg / kg). After 5 days, random blood glucose was measured through tail vein blood collection for 3 consecutive times, and those with blood glucose value >16.7mmol / L we...

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Abstract

The invention discloses an application of inhibition of SOCS3 expression through lentivirus-mediated RNA interference in preventing and controlling insulin resistance in diabetics. According to the invention, on the basis of a lentiviral vector-mediated RNAi technique, an SOCS3 gene silenced T2D rat model is established and experimental detection is implemented, and it is discovered that by inhibiting an SOCS3 gene, expression of IRS1 (insulin receptor substrate 1) can be improved, serine phosphorylation of the IRS1 can be reduced, insulin sensitivity of a T2D rat can be improved and an insulin resistance degree can be relieved. Therefore, a new idea is provided for prevention and control of human diabetes and experimental basis are offered for searching of effective treatment measures.

Description

technical field [0001] The invention belongs to the field of prevention and treatment of diabetes, and in particular relates to the application of a shRNA interference lentivirus capable of inhibiting the expression of SOCS3 in the prevention and treatment of insulin resistance in diabetes. Background technique [0002] Diabetes is one of the major diseases that endanger human life and health. With the change of lifestyle and the increase of obese people, the prevalence of diabetes worldwide is increasing year by year. Type 2 diabetes (T2D) accounts for about 90% of diabetic patients. . T2D is a disease caused by multiple factors. At present, its etiology and pathogenesis have not been fully elucidated. At home and abroad, lifestyle intervention and drug intervention are mainly used to treat T2D. Although a variety of new drugs for T2D have been developed in recent years, but Failure to fundamentally treat diabetes thoroughly to prevent other complications from occurring. ...

Claims

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Application Information

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IPC IPC(8): C12N15/113A61K31/713A61P3/10
CPCC12N15/113C12N2310/14C12N2320/30
Inventor 张旭吴鸣陈俊国陆燕章爱莲王琼平明芳
Owner 张旭
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