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Application of shionone in preparation of medicine for treating cerebral arterial thrombosis

A technology of ischemic stroke and shionone, applied in the field of pharmacotherapeutics, to improve inflammatory response and improve brain function

Pending Publication Date: 2022-07-22
NANJING DRUM TOWER HOSPITAL
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] Shionone (Shionone) is an active ingredient extracted from the dry roots and rhizomes of Aster of the Compositae plant. Studies have reported that Shionone has anti-inflammatory effects on many inflammatory diseases, but it has not been seen in the treatment of ischemic stroke. Report, therefore, the present invention provides Shivanone as the application of preparation treatment ischemic stroke medicine

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  • Application of shionone in preparation of medicine for treating cerebral arterial thrombosis
  • Application of shionone in preparation of medicine for treating cerebral arterial thrombosis
  • Application of shionone in preparation of medicine for treating cerebral arterial thrombosis

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0052] In vitro studies demonstrated that asterone attenuates LPS-induced inflammatory responses in microglia:

[0053] 1) Materials and methods

[0054] Primary microglia were obtained from C57BL / 6 mice and cultured in a humidified incubator at 37°C for 10 days, then primary microglia were isolated and re-cultured in 6-well or 12-well plates with cell culture medium DMEM medium (Thermo Fisher, USA) containing 10% FBS (fetal bovineserum Biological Industries, Israel) and 100 μ / ml penicillin and streptomycin was used.

[0055] The primary microglia were grouped according to the same density: control group, LPS-stimulated group, drug and LPS group. The supernatant of the treated primary microglia was discarded, followed by washing with PBS, and the total RNA of primary microglia and tissues was extracted with Trizol reagent (Invitrogen). Then, cDNA was transcribed using PrimeScript RT kit (Vazyme Biotech). qRT-PCR was performed on an ABI 7500 PCR machine (Applied Biosystems) ...

Embodiment 2

[0060] In vitro studies demonstrate that asterone reduces the production of pro-inflammatory factors at the protein level:

[0061] 1) Materials and methods

[0062] The expression levels of inflammatory factors were determined by Western blot. The supernatant was discarded from the primary microglia treated in different ways in Example 1, and then the cells were washed with PBS buffer, and then RIPA was added for protein lysis. RIPA Lysis Buffer, scraped the protein into a centrifuge tube, centrifuged at 1300rpm for 30min, aspirated the supernatant, used the BCA method to quantify the protein, then dissolved it in 5x Loading buffer and boiled for 5min. After electrophoresis on a 4-20% polyacrylamide gel, the protein was transferred to a PVDF membrane, blocked with 5% nonfat milk for 1 hour at room temperature, and the primary antibody (IL1β, RampD Systems, TNF-α, ab215188, Abcam, USA) was added separately. iNOS, 610328, BD Biosciences, USA), overnight at 4°C, horseradish per...

Embodiment 3

[0066] In vivo studies demonstrate the therapeutic effect of asterone on MCAO model mice:

[0067] The MCAO model was prepared by suture method and administered orally or intraperitoneally at a dose of 20 mg / kg / day. It was proved by behavioral studies that asterone has a protective effect on ischemic brain injury.

[0068] 1) Materials and methods

[0069] Neurological deficit score (NSS): The Modified Neurological Severity Score (mNSS) was used to score the degree of neurological deficit in mice, with a total score of 18 points and 1-6 points as mild injury , 7-12 is divided into moderate injury, 13-18 is severe injury;

[0070] Calculation of cerebral infarction volume: After perfusion, the brain tissue was quickly taken out, placed in a -80 ℃ refrigerator for 5-10 minutes, cut into 2 mm thick slices from the frontal pole to the posterior coronal section, and placed in 2% TTC (2, 3, 5- Triphenyltetrazolium chloride) solution, incubate at 37 °C for 30 min in the dark, the w...

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Abstract

According to the application of the shionone in preparing the medicine for treating the cerebral arterial thrombosis, the functions of the shionone are preliminarily studied by utilizing a modern biological technology, and the fact that the shionone inhibits generation of proinflammatory cytokines in LPS (Lipopolysaccharide) activated microglial cells at a gene level and a protein level is found, symptoms of MCAO model mice can be relieved, and the application of the shionone in preparing the medicine for treating the cerebral arterial thrombosis is realized. The generation of proinflammatory cytokines in the cerebral ischemic penumbra of the MCAO mouse is reduced at the gene level and the protein level, and the shionone can be prepared into the medicine for treating the cerebral arterial thrombosis disease. Therefore, the shionone provided by the invention has a good application prospect in the medicine for treating the cerebral arterial thrombosis.

Description

technical field [0001] The invention relates to the field of drug therapy, in particular to the application of asterone as a medicine for preparing ischemic stroke. Background technique [0002] Cerebrovascular disease has become the number one killer worldwide and can be divided into ischemic stroke and hemorrhagic stroke. Among them, ischemic stroke is extremely common in clinical practice, accounting for about 80%, and belongs to acute cerebrovascular disease. It not only has a very high morbidity, mortality and disability rate, but also has an unsatisfactory prognosis, which seriously threatens the life safety and quality of life of patients. At this stage, no specific drugs or methods have been found for the clinical treatment of this disease. Basic treatments include anticoagulation, thrombolysis, antiplatelet aggregation, thrombectomy, neuroprotective therapy, and gene therapy. In recent years, with the accelerating pace of life and improving living standards, coupl...

Claims

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Application Information

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IPC IPC(8): A61K31/56A61K36/28A61P9/10
CPCA61K31/56A61K36/28A61P9/10
Inventor 徐运徐露珊张智董小红李成刚舒鑫
Owner NANJING DRUM TOWER HOSPITAL
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