Application of medioresinol in preparation of drug for preventing or treating cerebral injury and pharmaceutical composition thereof
A resin alcohol, brain injury technology, applied in the field of medicine to achieve the effect of reducing injury
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Embodiment 1
[0032] Determination of the release of lactate dehydrogenase LDH and the expression of inflammatory factor IL-1β in cells:
[0033] Brain vascular endothelial cells (bEND3) were pretreated with corticosteroids (20 μM) for 12 hours, then the medium was removed, the medium was replaced with sugar-free medium, and placed in an anoxic incubator for 6 hours. The hydrogenase detection kit was used to detect lactate dehydrogenase (LDH) released by vascular endothelial cells (Shanghai Beyond Biotechnology Co., Ltd.), and the adherent cells were collected at the same time, and the mRNA expression of inflammatory factor IL-1β in vascular endothelial cells was detected by RT-PCR .
[0034] (1) Establishment of endothelial cell injury model in vitro by oxygen-glucose deprivation (OGD):
[0035] The mouse brain microvascular endothelial cell line bEND3 was injected at 1×10 5 The density of cells / ml was inoculated on a 24-well plate pre-coated with type I collagen, and after 24 hours of s...
Embodiment 2
[0051] Determination of the permeability of the blood-brain barrier established by the co-culture of primary cerebral microvascular endothelial cells and astrocytes: Firstly, the co-culture model of cerebral microvascular endothelial cells and astrocytes was established. A 24-well cell culture plate and the inside and back of the Millicell chamber were coated with rat tail collagen and allowed to dry. Astrocytes were inoculated on the back side of the Millicell culture chamber, and cultured in the incubator for 6 hours. After the astrocytes were completely attached to the wall, the Millicell culture chamber was placed in a 24-well cell culture plate, and the brain microvascular endothelial cells Inoculated on the inner side of the Millicell culture chamber, cultured with the growth medium of brain microvascular endothelial cells. After the model was successful, the cells were treated with corticoresinol (20μM) for 12h and then treated with OGD for 6h. The effect of corticoresi...
Embodiment 3
[0062] Establishment of mouse middle cerebral artery occlusion (tMCAO) model and measurement of cerebral infarction volume:
[0063] (1) Establishment of mouse tMCAO model
[0064] The mouse tMCAO model refers to the internal carotid artery suture method of Longa et al. The main operation steps are as follows: weigh male C57 / BL6 mice weighing about 25-30g, place them in an anesthesia induction box, give 3%-4% isoflurane to induce anesthesia, and place the mice in a supine position after anesthesia Fixed on the operating table. The mice were put on a breathing mask and given 1.0%-2.0% isoflurane to maintain anesthesia. Wipe the mouse neck skin with 75% alcohol cotton ball, incise the skin in the middle of the neck, and bluntly separate the layers of muscle and soft tissue with curved forceps, expose the right common carotid artery, and put a nylon thread for use. The soft tissue was bluntly dissected from the head side of the mouse along the common carotid artery to expose t...
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