Application of elecampane inula root and radix bupleuri composition in preparation of antidepressant drug
A composition, the technology of inulin, which is applied in the direction of drug combination, medical formula, plant raw materials, etc., can solve the problem that the antidepressant effect of the composition of inula bupleuri has not been reported, and achieve the goal of improving depression-like behavior and improving brain function Effect
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Embodiment 1
[0023] Effects of Inula radix Bupleurum composition on chronic restraint stress-induced depression-like behavior in mice
[0024] 1. Experimental method
[0025] 1.1 Preparation of Inulina Bupleurum composition water decoction
[0026] Weigh 75g of Inulin and 25g of Bupleurum, rinse with distilled water, add 500ml of double-distilled water to soak for 20min. Boil with an electric pottery stove and keep it boiling for 30 minutes, simmer for another 30 minutes, and stop heating. After cooling, filter with multi-layer gauze to obtain the first decoction. Collect the filter residues of medicinal materials, add 250ml of double-distilled water and continue to boil with an electric pottery stove for 30min, and stop heating. After cooling, the filtrate was collected with multiple layers of gauze again, and the second decoction was obtained. Combine the decoctions collected twice, and use a rotary evaporator to concentrate the filtrate to 100 ml, and refrigerate for later use.
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Embodiment 2
[0043] Effects of Inula Radix Bupleuri composition on neurotransmitters in the brain of chronic restraint stress mice
[0044] 1. Experimental method
[0045] 1.1 Collection of samples
[0046] After the behavioral study, the mice were sacrificed by dislocating their cervical vertebrae, and the prefrontal cortex tissue was quickly taken out and snap-frozen in liquid nitrogen. Before the test, the prefrontal cortex tissue was taken out, added with normal saline, and mixed with normal saline at a weight ratio of 1:9 to homogenize. After 12000r / min refrigerated centrifugation for 10min, the supernatant was taken and stored in a 4°C refrigerator for later use.
[0047] 1.2 ELISA experiment
[0048] ELISA kits were used to detect the contents of 5-HT, DA and NE in the prefrontal cortex of mice. The operation steps were carried out according to the kit instructions, the OD value of each well was measured at a wavelength of 532 nm, and then the expression content of each neurotrans...
Embodiment 3
[0052] Effects of Inula Radix Bupleuri composition on BDNF / TrkB / CREB signaling pathway in the brain of chronic restraint stress mice
[0053] 1. Experimental method
[0054] 1.1 Total protein extraction
[0055] The mouse prefrontal cortex tissue was taken, and RIPA Buffer containing 1% PMSF was added at a weight-to-volume ratio of 1:5, and the homogenate was frozen for 5 minutes. After that, the homogenate was transferred to a centrifuge tube for further lysing for 40 min, and then the samples were refrigerated and centrifuged for 30 min at a speed of 12000 r / min. The supernatant was collected, and the protein concentration was first determined by BCA method, and then 5×SDS-PAGE Loading Buffer was added with a volume of 1 / 2 and mixed well, and heated at 100 °C for 10 min with a metal heater. Finally, protein samples were collected and stored at -20°C for future use.
[0056] 2. Western blot experiment
[0057] Protein samples were separated by SDS-PAGE gels, transferred t...
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