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Nucleotide molecule SR4B and application of the same in producing antidiabetic medicament

A nucleic acid molecule and anti-diabetic technology, applied in the field of biomedicine, can solve problems such as anti-diabetic gene drugs that have not been reported yet

Inactive Publication Date: 2011-06-15
FUDAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Rely on insulin, but when diet and oral hypoglycemic drugs are not effective, or because of complications and concomitant diseases, insulin is sometimes required to control hyperglycemia
[0005] However, so far, no one has reported gene medicine for antidiabetic

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0033] Example 1 Effect of siRNA interference on endogenous BRSK2 expression on insulin secretion

[0034] (1) Dilute NIT cells at 1×10 5 Cells / well were seeded on a 24-well plate, and the cells were placed in CO 2 Cultivate in an incubator at 37° C. for 18-24 hours to make the cell abundance reach 70-80%.

[0035](2) Invitrogen lipofectamine 2000 eukaryotic transfection kit was used for transfection. 5 μl of Nonsilence (negative control for siRNA interference), 1 μl, 3 μl and 5 μl of siRNA (dTdTAAAGUCUGCGAUGCGGAUG and CAUCCGCAUCGCAGACUUUUUdTdT)-5 (concentration of 20 μM per μl) were transfected respectively. Four wells were replicated for each transfection.

[0036] (3) 16 hours after transfection, replace with complete medium (DMEM high glucose medium + 15% fetal calf serum) and continue to culture for 24 hours.

[0037] (4) The complete medium (sugar concentration: 25 mM) was replaced again, and the culture supernatant was collected after culturing for 20 minutes.

[0...

Embodiment 2

[0043] Example 2 Effect of BRSK2 overexpression on insulin secretion

[0044] (1) Dilute NIT cells at 1×10 5 Cells / well were seeded on a 24-well plate, and the cells were placed in CO 2 Cultivate in an incubator at 37° C. for 18-24 hours to make the cell abundance reach 70-80%.

[0045] (2) Invitrogen lipofectamine 2000 eukaryotic transfection kit was used for transfection. Two plasmids, Pcmv-Myc empty vector and Pcmv-Myc-BRSK2, were transfected respectively, and each plasmid was transfected into 4 wells at a concentration of 200ng / well.

[0046] (3) 16 hours after transfection, replace with complete medium (DMEM high glucose medium + 15% fetal calf serum) and continue to culture for 24 hours.

[0047] (4) The complete medium (sugar concentration: 25 mM) was replaced again, and the culture supernatant was collected after culturing for 20 minutes.

[0048] (5) The concentration of insulin in the culture supernatant was measured by radioimmunoassay. The detection kit was pro...

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PUM

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Abstract

The invention discloses a nucleic acid molecule and the application to prepare antidiabetic in the biological medical domain, wherein the diabetes mellitus is a group of clinical syndrome interacted by genetic and environmental factor, which bases on hyperglycemia as main mark to lead the harm of multiple systems due to lingering illness; the nucleic acid molecule SR4B is added into Langerhans' islet beta-cell to increase the content of insulin in the cell culture supernatant. The invention can accelerate the secretory function of insulin as new antidiabetic, which provides a new path and method to release and treat diabetes.

Description

technical field [0001] The invention belongs to the field of biomedicine and relates to a nucleotide molecule and its application in preparing antidiabetic drugs. Background technique [0002] Diabetes mellitus is a group of clinical syndromes caused by the interaction of genetic and environmental factors. Absolute or relative insufficiency of insulin secretion and decreased sensitivity of target tissue cells to insulin lead to a series of metabolic disorders such as sugar, protein, fat, water and electrolytes. Clinically, hyperglycemia is the main sign, and prolonged illness can cause damage to multiple systems. Acute metabolic disorders such as ketoacidosis can occur in severe illness or stress. [0003] The main clinical type of diabetes Insulin-dependent diabetes mellitus (IDDM, type I) can occur at any age, but mostly occurs in young adults. The clinical features are acute onset, obvious symptoms such as polyphagia, polyuria, polydipsia, and weight loss, with a tende...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/12C07H21/02C12N15/63A61K48/00A61P3/10
Inventor 余龙汤文文郭泽坤袁健唐丽莎
Owner FUDAN UNIV
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