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Application of thymoquinone (TQ) to preparation of drugs for preventing vascular stent restenosis

A vascular stent, thymoquinone technology, applied in the field of biomedicine, can solve problems such as unclear role

Pending Publication Date: 2019-11-08
WENZHOU PEOPLES HOSPITAL
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

But the role of TQ in cardiovascular disease is still unclear
The effects of TQ on the proliferation, migration and apoptosis of vascular smooth muscle cells in vitro and the formation of neointima in vivo have yet to be confirmed

Method used

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  • Application of thymoquinone (TQ) to preparation of drugs for preventing vascular stent restenosis
  • Application of thymoquinone (TQ) to preparation of drugs for preventing vascular stent restenosis
  • Application of thymoquinone (TQ) to preparation of drugs for preventing vascular stent restenosis

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0027] Embodiment 1 TQ measures cytotoxicity, cell viability and proliferation

[0028] The cell viability and proliferation rate were determined by MTT assay. VSMCs were inoculated at 1×10 per well in a 96-well culture plate. 4 Cells were cultured for 24 hours and incubated with 1% FBS serum for 48 hours. In the viability assay, cells were pretreated with 40 ng / ml PDGF-BB and various concentrations (0, 5, 10, 12.5, 15, 20, and 40 μM) of TQ for 24 hours, and then treated with 40 ng / ml PDGF-BB and TQ (15 μM ) Stimulate VSMCs for 6h, 12h, 24h, 48h and each group was treated with different concentrations of TQ (5-15μΜ) for 24h. After stimulation, 5mg / ml MTT solution was added to each well for 4h, and then 150μl DMSO was used to replace the MTT solution. Absorbance was measured at 570 nm with a microplate reader.

[0029] Such as figure 1 As shown in a, each group of TQ (5-15 μ M) has no toxic effect on VSMCs after 24 hours of treatment. Therefore, TQ concentrations of 5 μM, ...

Embodiment 2

[0033] Example 2 Effect of TQ on the Migration of Vascular Smooth Muscle Cells

[0034] 1. Scratch test and detection of cell migration

[0035] VSMCs were seeded in 6-well culture plates for 48 hours, and the cells in the culture plates reached 90-100% confluency. VSMCs were cultured with starvation medium (1% FBS) for 48 hours. After drawing a straight line in the middle of 200μl tip cells, they were divided into control group (no PDGF-BB treatment), model group (40ng / ml PDGF-BB treatment) and drug group (40ng / ml PDGF-BB treatment + TQ5, 10, 12.5, 15 μM and other groups treated with different concentrations) to treat VSMCs. Images were acquired using Leica Application Suite software, and cell migration was determined by using Image J software to determine the percentage of scratch closure area. PDGF-BB treated cells for 24 hours ( figure 2 a and figure 2 b) and 48 hours ( figure 2 c and figure 2 After d), the scratch narrowing increases significantly, while TQ can...

Embodiment 3

[0039] Example 3 Cell Apoptosis Detection

[0040] VSMCs were inoculated on coverslips, starved for 48 hours in serum-free medium, and divided into control group (no PDGF-BB treatment), model group (40ng / ml PDGF-BB treatment) and drug group (40ng / ml PDGF-BB treatment+TQ5, 10, 12.5, 15 μ M and other groups treated with different concentrations) were used to treat VSMCs. After deparaffinization and rehydration with xylene and ethanol, VSMCs cells were fixed in 4% paraformaldehyde solution (pH7.4). Incubate for 1 hour at 25°C in 3% H 2 o 2 Block for 10 minutes, infiltrate with 0.1% Triton X-100 sodium citrate solution for 3 minutes, label apoptotic cells with dUTP nick end labeling (TUNEL) mediated by terminal deoxyribonucleic acid transferase, and label cell nuclei with DAPI. Images were acquired with a fluorescence microscope (BX53, Olympus), and apoptotic cells were analyzed with Image J software. The result is as Figure 5 As shown, PDGF-BB treatment can significantly re...

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Abstract

The invention discloses application of thymoquinone (TQ) to preparation of drugs for preventing vascular stent restenosis. By verifying the inhibiting effect of the TQ on proliferation and migration,induced by platelet-derived growth factors (PDGF-BB) in vitro, of vascular smooth muscle cells (VSMCs) and in vivo neointima formation, according to the action mechanism of the TQ, the TQ inhibits proliferation, induced by the PDGF-BB, of the VSMCs, the TQ further induces apoptosis through the mitochondrial dependence apoptotic pathway and p38, the TQ prevents migration of the VSMCs by inhibitingMMP2 activity and expression, and finally, the TQ has a significant treating effect on ligation-induced rat neointima formation. The TQ is a potential candidate drug for preventing and treating occlusive vascular diseases such as in-stent restenosis, a strong molecular biology basis is provided for treating vascular stent restenosis, and far-reaching clinical significance and popularity are achieved.

Description

technical field [0001] The invention relates to the technical field of biomedicine, in particular to the application of thymoquinone in the preparation of drugs for preventing restenosis of vascular stents. Background technique [0002] Vascular remodeling is a major cause of various vascular diseases, such as atherosclerosis and pulmonary hypertension, and is characterized by changes in the structure and function of the vessel wall. In response to vascular injury or changes in local environmental factors, vascular remodeling involves a series of pathological processes, including endothelial dysfunction, proliferation and migration of vascular smooth muscle cells (VSMCs), arterial calcification, and extracellular matrix remodeling. This injury-induced vascular remodeling is mainly due to the excessive proliferation and migration of VSMCs and the invasion of VSMCs into the intimal space, which together lead to neointimal formation and restenosis after angioplasty. Therefore,...

Claims

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Application Information

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IPC IPC(8): A61K31/122A61P9/10
CPCA61K31/122A61P9/10
Inventor 朱宁向贻佳赵旭勇蔡昌宏陈皓姜文兵王毅曾春来
Owner WENZHOU PEOPLES HOSPITAL
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