Application of maribateol a as preparation of medicine for treating ischemic brain injury
A technology of ischemic brain injury and butyrol, which is applied in the application field of maributol A as a drug for the treatment of ischemic brain injury, can solve the problems of increased bleeding risk, many contraindications, time window restrictions, etc., and achieve improvement Effects on brain function after ischemia
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Embodiment 1
[0065] In vivo studies demonstrated that MA has a therapeutic effect on MCAO mice:
[0066] The middle cerebral artery infarction model on one side of C57L / B6 mice was established by suture method, that is, the MCAO model, and treated with MA tail vein injection. MA has been shown to have a protective effect against ischemic brain injury through behavioral studies.
[0067] 1) Materials and methods
[0068] The MCAO mouse model was established by thread embolization method, and the experiment was randomly divided into 3 groups: sham group, MCAO group, MCAO+MA (Nanjing University, Nanjing, China) 10 mg / kg group, 12 mice in each group, and tail vein injection .
[0069] Neurological Severity Severity (NSS) Scoring: The Modified Neurological Severity Severity Score (mNSS) was used to score the neurological deficit of mice. The total score of mNSS is 18 points, 1-6 is divided into mild injury, 7-12 is divided into moderate injury, and 13-18 is divided into severe injury.
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Embodiment 2
[0077] In vitro studies demonstrated that MA improves cell viability after OGD and promotes the differentiation of BV2 cells into M2 type:
[0078] 1) Materials and methods
[0079] Microglioma cell culture: Microglioma cells, namely BV2 cells, were purchased from ATCC in the United States and grown in a medium containing 10% fetal bovine serum, 2 mmol / L L-glutamine, penicillin (100 U / ml) and streptomycin (100 μg / ml) DMEM high glucose medium, 37°C, 5% CO2 incubator.
[0080] Viability detection of BV2 cells: BV2 cells were treated with OGD, and cells were treated with different concentrations of MA. Cell viability was detected by the tetramethylazolium salt colorimetric method (MTT method). Add MTT to 100 μl of culture solution / well, continue culturing for 4 hours in an incubator at 37°C, 5% CO2, then pour out all the culture solution, add 100 μl of DMSO solution to each well, shake on a shaker until the blue-purple crystals are completely dissolved, and the enzyme The sta...
Embodiment 3
[0088] In vivo studies have demonstrated that MA promotes the differentiation of microglia towards the replacement type and attenuates the post-ischemic inflammatory response:
[0089] 1) Materials and methods
[0090] Detection of mRNA expression levels of inflammatory factors: After MCAO in mice, MA was injected into the tail vein at the time of ischemia for 2 hours and reperfusion. The brains were removed at 6, 24, 48 and 96 hours after reperfusion, and the expression levels of inflammatory factors were measured by RT-PCR, the method was the same as before.
[0091] Detection of M2 type markers: After MCAO of mice, cortical tissues of mice in each group were taken, and total RNA was extracted in each group.
[0092] The specific method is the same as above.
[0093] Determination of the activation state of microglial cells in the mouse cortex tissue: 24 hours after the mouse MCAO was reperfused, the whole brain tissue of each group of mice was collected after perfusion wi...
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