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Application of danshinolic acid B in inhibition of islet amyloid polypeptide aggregation and treatment of diabetes

A technology of amyloid polypeptide and salvianolic acid, applied in the field of medicine, can solve problems such as aggravation of diabetes and β-cell function damage

Inactive Publication Date: 2013-03-13
HUAZHONG UNIV OF SCI & TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In addition, the polymer of hIAPP leads to the damage of β-cell function in patients with advanced type 2 diabetes by breaking the coupling between β-cells and inducing the apoptosis of β-cells, which aggravates the condition of diabetes.

Method used

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  • Application of danshinolic acid B in inhibition of islet amyloid polypeptide aggregation and treatment of diabetes
  • Application of danshinolic acid B in inhibition of islet amyloid polypeptide aggregation and treatment of diabetes
  • Application of danshinolic acid B in inhibition of islet amyloid polypeptide aggregation and treatment of diabetes

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0023] Example 1 Thioflavin-T(ThT) fluorescence assays

[0024] Specific operation: IAPP was dissolved in HFIP and sonicated for 2 minutes, then dissolved in 25 mM PBS (50 mM NaCl), pH 7.4, and diluted to an IAPP solution containing 1% HFIP with a final concentration of 15 μM. At 25°C, they were bathed together with salvianolic acid B at 1, 2, and 5 times the molar ratio (compound / IAPP), and analyzed with a Hitachi 2700 fluorescence analyzer. The detection system contained 25mM PBS, 50mM NaCl, pH 7.4 and 20μM thioflavin- T, excitation and emission wavelengths are 450nm and 482nm.

[0025] The result is as figure 2 As shown, IAPP gradually begins to aggregate as the incubation time increases, and the fluorescence signal intensity increases with the aggregation of IAPP. Then, after adding salvianolic acid B, the fluorescence signal intensity of IAPP decreased significantly, while showing a stable dose-dependent manner. This shows that salvianolic acid B has a significant inh...

Embodiment 2

[0026] Example 2 Electron microscope experiment Transmission electronic microscopy (TEM)

[0027] Specific operation: the samples incubated for 24 hours in Example 1 were used to observe the morphology of the fibers after IAPP aggregation from a morphological point of view by using a scanning electron microscope.

[0028] The result is as image 3 As shown, the IAPP after 24h of incubation alone showed a typical fibrous morphology under the scanning electron microscope (see image 3 A). After co-incubating with salvianolic acid B, observed under the electron microscope, the fibrous morphology gradually decreased in a dose-dependent manner (see image 3 B-D) This shows that salvianolic acid B has a significant inhibitory effect on the aggregation of IAPP.

Embodiment 3

[0029] Example 3 Photocatalytic Cross-Linking of Unmodified Proteins (Photo-induced Cross-Linking of Unmodified Proteins (PICUP) assay)

[0030] Specific operation: IAPP was dissolved in HFIP and ultrasonicated for 2 minutes, dissolved with 10mM PBS (pH7.4) and diluted to a final concentration of 1% HFIP, which was 82 μM of IAPP solution, respectively with 1 and 5 times the molar ratio (compound / hIAPP) of salvianolic acid B was irradiated with incandescent light for 5 s in a dark box under the action of a cross-linking agent. Then, 7.5 μl of loading buffer was added to the reaction solution, denatured at 97° C. for 10 minutes, then electrophoresed on 20% Tricine-Urea gel, and stained with visualized silver staining (Beyotime rapid silver staining kit).

[0031] The result is as Figure 4 As a result, IAPP reacted alone formed more multimers (the second band) after irradiating for 5 seconds; while IAPP reacted with different doses of salvianolic acid B after irradiating for 5...

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Abstract

The invention belongs to the field of medicines. Danshinolic acid B has functions of inhibition of islet amyloid polypeptide aggregation and treatment of diabetes. The in-vivo islet amyloid polypeptide aggregation can destroy a beta-cell membrane structure and a normal form function of mitochondria to cause beta-cell apoptosis and damage a beta-cell function, thereby being taken as an important nosogenesis of type-2 diabetes. By danshinolic acid B, the aggregation of the islet amyloid polypeptide can be inhibited, and the process of damaging the beta-cell membrane structure and the normal form function of mitochondria during aggregation is changed over to protect the beta-cells, so that danshinolic acid B can be used for preventing and treating the diabetes.

Description

technical field [0001] The invention belongs to the field of medicine and relates to a medicine for treating diabetes. Background technique [0002] Diabetes is an endocrine disease caused by the body's own nutritional and metabolic disorders. Its basic biochemical characteristics are persistent high blood sugar. It has become the fifth leading cause of death in the world. disease. [0003] Diabetes is divided into type 1 and type 2, and the number of type 2 diabetes (T2DM) accounts for more than 90% of the total number of diabetic patients. The basic feature of type 2 diabetes is: on the basis of insulin resistance, the function of β cells is damaged. The pathogenesis of type 2 diabetes has not yet been elucidated. Islet amyloid polypeptide (Islet amyloidpolypeptide, IAPP) is co-secreted by pancreatic β cells together with insulin, and consists of 37 amino acid polypeptides. Under physiological conditions, IAPP can cooperate with blood sugar regulating hormones such as i...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K31/343A61P3/10
Inventor 黄昆程彪龚皓李晓超孙粤张鑫陈红吴谦
Owner HUAZHONG UNIV OF SCI & TECH
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