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Application of sodium orthovanadate in preparation of medicine for resisting myocardial apoptosis

A technology of sodium orthovanadate and myocardial apoptosis, applied in the application field of myocardial apoptosis drugs, to achieve the effect of broad scientific research and clinical development prospects

Inactive Publication Date: 2016-01-20
SHANDONG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the search found that the role and influence of sodium orthovanadate on cardiomyocyte apoptosis induced by excessive lipid accumulation, or whether it can be used to develop anti-myocardial apoptosis drugs, has not been reported at home and abroad.

Method used

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  • Application of sodium orthovanadate in preparation of medicine for resisting myocardial apoptosis
  • Application of sodium orthovanadate in preparation of medicine for resisting myocardial apoptosis
  • Application of sodium orthovanadate in preparation of medicine for resisting myocardial apoptosis

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0030] Preparation of sodium orthovanadate storage solution: Dissolve sodium orthovanadate powder in three-distilled water to make 100mmol / l storage solution, adjust pH to 10.0, boil until colorless, let cool to room temperature, continue to adjust pH repeatedly and boil Until the pH is stable at 10.0, filter, aliquot and store at -20°C.

[0031] Morphological observation of the effect of different concentrations of SOV on the survival of H9c2 cardiomyocytes.

[0032] at 37°C with 5% CO 2 Under the environment of DMEM / 10% fetal bovine serum, the H9c2 cardiomyocytes were cultured for future use. Before morphological observation, the cells in the logarithmic growth phase should be counted with a hemocytometer according to 8×10 4 Cells / ml were inoculated in small dishes, and pre-divided into BSA control group, PA alone treatment group, BSA+SOV, PA+SOV group.

[0033] After the cells grew to a coverage rate of 90%, the drug-dosed group was pretreated with SOV at a concentration...

Embodiment 2

[0036] Western blot detection of different concentrations of SOV in H9c2 cardiomyocytes, the impact on the execution of apoptosis caspases, including caspase3, caspase7 protein precursor cleavage and the impact on the expression of anti-apoptotic protein Bcl-2.

[0037] In the cultured H9c2 cardiomyocytes, 50 μM, 100 μM, and 200 μM SOV were added for pretreatment for 1 h, and after adding PA for 12 h, the total protein was extracted for comparison and statistics.

[0038] The specific method is: wash the treated cells twice with PBS; after discarding the PBS, add the prepared Western and IP cell lysates to the culture plate; The collected protein was centrifuged at 12,000 rpm for 15 minutes at 4°C; the supernatant was transferred to a new eppendorf centrifuge tube and placed on ice; 2 μl of the protein suspension was taken out to measure the protein concentration, and stored at -20°C after aliquoting, and stored at -80°C. ℃ long-term storage.

[0039]Measure the concentration...

Embodiment 3

[0043] The effect of different concentrations of SOV on the apoptosis rate of H9c2 cardiomyocytes induced by PA was detected by flow cytometry.

[0044] In the cultured H9c2 cardiomyocytes, 50μM, 100μM, 200μM SOV were pretreated for 1h, and PA was added for 12h to induce myocardial apoptosis. After AnnexinV / PI staining, the apoptotic cells were detected by flow cytometry, and the apoptosis rate was counted. The specific method is as follows:

[0045] (1) Cell collection: After H9c2 cardiomyocytes were treated with drugs, they were digested with 0.25% trypsin without EDTA, suspended, collected into a 10ml centrifuge tube, and the number of cells per sample was (1-5)×10 6 / mL, centrifuge at 900g / min for 5min, and discard the culture medium.

[0046] (2) Wash once with 1ml stainingbuffer, centrifuge at 900g / min for 5min.

[0047] (3) Resuspend the cells with 50ul of binding buffer, add 5ul of FITC, incubate at room temperature in the dark for 10-15min, flick and mix every 5 mi...

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Abstract

The invention discloses application of sodium orthovanadate in preparation of medicine for resisting myocardial apoptosis caused by excessive lipid accumulation. Experiments prove that the sodium orthovanadate can effectively inhibit the myocardial apoptosis induced by long-chain saturated fatty acid palmitic acid, and it is specifically shown that the sodium orthovanadate inhibits the apoptosis induced by the palmitic acid from executing cutting of a caspase precursor, promotes protein expression of anti-apoptosis protein Bcl-2 and obviously reduces the apotosis rate and pyknosis of nucleus chromatin. On the basis, it is expected that application is supplied to SOV in aspect of preparing the medicine for treating the myocardial apoptosis induced by excessive lipid accumulation along with diseases such as obesity and diabetes in clinical research, and it is indicated that the wide scientific research and clinical development prospects are achieved.

Description

technical field [0001] The present invention relates to a new application of an inorganic vanadium compound sodium orthovanadate (sodium orthovanadate, SOV), in particular to an application of sodium orthovanadate in the preparation of anti-myocardial apoptosis drugs induced by excessive lipid accumulation. Background technique [0002] Since the beginning of the 20th century, the obesity population has increased dramatically, followed by a high incidence of cardiovascular disease. Cardiomyocytes are the structural and functional units of the heart, and even a small amount of apoptosis can lead to very serious consequences. In obesity and related diseases, the excessive accumulation of lipids in cardiomyocytes can lead to myocardial apoptosis, which is considered to play an important role in the development of cardiovascular diseases. Among them, long-chain saturated free fatty acids, especially palmitate (PA), also known as palmitic acid, are the main factors leading to li...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K33/24A61P9/00
Inventor 尹德领赵静张尚立刘静
Owner SHANDONG UNIV
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