Application of turmeric native
A technology of curcumin and neurotoxin, applied in nervous system diseases, drug combinations, active ingredients of ketones, etc., can solve problems such as unreported effects, achieve great potential value, and reduce the effect of dopaminergic neuron damage
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Embodiment 1
[0029] Example 1 , grouping and injection of animals
[0030] Take 60 healthy 8-week-old male C57BL mice (Shanghai BK Animal Experiment Co., Ltd.), weighing about 20 g, and divide them into 6 groups randomly, with 10 mice in each group, as follows:
[0031] Normal control group: intraperitoneal injection of normal saline for 12 days;
[0032] MPTP group (model group): after continuous intraperitoneal injection of 30mg / kg MPTP for 5 days, it was continuously injected with normal saline for 7 days;
[0033] Treatment group 1: After continuous intraperitoneal injection of 30 mg / kg MPTP for 5 days, 5 mg / kg curcumin was continuously injected for 7 days;
[0034] Treatment group 2: After continuous intraperitoneal injection of 30 mg / kg MPTP for 5 days, 50 mg / kg curcumin was continuously injected for 7 days;
[0035] Treatment group 3: after continuous intraperitoneal injection of 30 mg / kg MPTP for 5 days, 150 mg / kg curcumin was continuously injected for 7 days;
[0036] Solvent...
Embodiment 2
[0037] Example 2 , immunohistochemical analysis
[0038] 2.1. Immunohistochemical method to detect changes in the expression of TH positive cells in the substantia nigra
[0039] Get the 6 groups of mice of embodiment 1, after intraperitoneal injection of 3% pentobarbital (0.15mL per kilogram of body weight) anesthetized, with 0.9% normal saline 100ml (room temperature) and 4% paraformaldehyde 250ml (4 ℃) via The ascending aorta of the left ventricle was perfused to fix the brain tissue, and the midbrain segment of the rat brain was taken and immersed in fixative solution at 4°C for 24 hours. The fixed midbrain segment of the rat brain was dehydrated with graded ethanol, transparentized with xylene, embedded in paraffin, and then serially coronal paraffin sections (thickness: 6 μm) were performed, and the substantia nigra and striatum of the midbrain were selected for immunohistochemical staining Dry the sheet at 60-65°C for 4 hours, dewax with xylene and graded ethanol to ...
Embodiment 3
[0047] Example 3 , Western blot detection
[0048] 3.1. Effect of curcumin on the positive expression (relative value) of TH protein in substantia nigra
[0049] The substantia nigra tissue isolated from (1)-(6) group mice in Example 1 was added in proportion to the lysate (1mmol / L Tris, 50mmol / L sodium chloride, 0.03μmol / L sodium pyrophosphate, 50mmol / L fluorine NaCl, 1% Triton X-100, 1mmol / L phenylmethylsulfonyl fluoride, 1mmol / L sodium vanadate, 20μg / mL aprotinin), made into 50mg / mL homogenate, centrifuged at 15000r / min 4℃ for 15min , take the supernatant and store it at -80°C, and use the bicinchoninic acid (BCA) method to determine the total protein content in the lysate; add 2×sample buffer to the tissue homogenate to dilute in half, boil for 5min, 6000r / Centrifuge for 3 min, take the supernatant and load the sample, take 30 μg of the sample and separate the protein on the gel by 10% sodium dodecylsulfonate polyacrylamide gel electrophoresis (SDS-PAGE); TH polyclona...
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