Application of curcumin to preparation of drugs for treating cerebral ischemia/reperfusion injuries
A cerebral ischemia-reperfusion, curcumin technology, used in drug combination, cardiovascular system diseases, digestive system and other directions, can solve problems such as brain tissue reperfusion injury, etc., to improve liver and kidney function damage, neurological function. State-improving, low-cost effects
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Embodiment 1
[0026] Embodiment 1 curcumin injection and mouse focal cerebral ischemia reperfusion (MCAO) model preparation
[0027] (1) Preparation of Curcumin Injection
[0028] ① Povidone (PVP) curcumin solid dispersion:
[0029] Mass ratio (curcumin:PVP=1:8) PVP 24g, dissolved in 40ml absolute ethanol, magnetically stirred
[0030] Dissolve 3g of curcumin in 3ml of absolute ethanol, add it dropwise to the PVP solution, wash the bottle with 10ml of absolute ethanol, stir manually, pour it into a 250ml beaker with a large rotor and stir well, freeze-dry for 4 hours after stirring.
[0031] ② Curcumin solid dispersion solution:
[0032] Weigh 900mg of curcumin solid dispersion, add 5ml of normal saline for injection, that is curcumin 20mg / ml, in a brown bottle, it is transparent orange red, viscous, take 20mg / ml 1ml+ normal saline 7ml is 2.5mg / ml ml of application solution.
[0033] (2) Making mice focal cerebral ischemia-reperfusion injury model: 60 male Kunming mice weighing about 25...
Embodiment 2
[0042] Example 2 Histopathological changes of cerebral ischemia-reperfusion groups in mice
[0043] (1) After 2 hours of HE staining ischemia and 24 hours of reperfusion, the mice were anesthetized with 1% sodium pentobarbital (6mg / 100g), opened the chest and cut the right atrial appendage, and perfused sequentially through the left ventricle (containing 0.4% heparin) with 0.9% physiological Saline 30ml, 4% paraformaldehyde 30ml, the brain was removed immediately, post-fixed in 10% paraformaldehyde solution for 48h. Paraffin embedding was carried out by conventional methods, and 2 mm thick coronal sections were taken at the forehead thickness of 5 mm for HE section staining. The pathological changes of the cerebral cortex on the ischemic side were observed by light microscope.
[0044] (2) HE staining results, such as figure 2 As shown in the light microscope, the number and shape distribution of nerve cells in the sham operation group were normal; the arrangement of cells ...
Embodiment 3
[0045] Example 3 Immunohistochemistry of NSE, Cox-II, and Bcl-2 in brain tissue of mice after cerebral ischemia-reperfusion
[0046] (1) Immunohistochemical detection of NSE, Cox-II, and Bcl-2 expressions According to the results of HE staining, the ischemic infarct was determined to be sectioned, and the paraffin tissue sections were dewaxed, stained according to the immunohistochemical SABC method, and 3% H 2 o 2 Block endogenous peroxidase activity for 30 min, wash with PBS for 5 min x 2 times, and distilled water for 5 min x 2 times. Citrate buffer (pH 6.0) was heat-repaired at 92°C for 5 minutes, allowed to stand for 5 minutes, heat-repaired again for 5 minutes, washed with PBS for 5 minutes×2 times, and washed with distilled water for 5 minutes×2 times. 1.5% goat serum was sealed and incubated in a wet box for 1h (37°C), discarded without washing, and primary antibodies (rabbit anti-mouse Cox-II (1:50) and Bcl-2 (1:200) polyclonal antibodies) were added dropwise 37 Incub...
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