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Application of daucosterol in preparing medicines for promoting proliferation of neural stem cells
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A technology of neural stem cells and carotene, applied in the field of medicine
Inactive Publication Date: 2012-10-31
NANJING UNIVERSITY OF TRADITIONAL CHINESE MEDICINE +1
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The above-mentioned substances that can promote the proliferation of neural stem cells have limitations in their application: macromolecular substances are difficult to pass through the blood-brain barrier, excessive intake of exogenous hormones can easily lead to disordered release of endogenous hormones, and small molecule drugs have limited sources and are expensive. Long-term use may have certain adverse reactions
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Embodiment 1
[0102] Embodiment 1. In Vitro Experiment
[0103] 1.1 Experimental animals
[0104] Ten pregnant SD rats at 14-16 days of pregnancy were provided by Zhejiang Experimental Animal Center, certificate number: SCXK (Zhejiang) 2008-0033.
[0105] 1.2 Main reagents and reagent preparation:
[0106] Neuralbasal medium (product of GIBCO company, Lot 1094141), B27 medium supplement (product of GIBCO company, Lot 1159693); rat b-FGF and EGF (product of Pepro TECH company, EGF: Lot 0906390-1 H2911); b-FGF: Lot 1305CY12) mouse anti-rat nestin monoclonal antibody, TRITC (rhodamine)-labeled goat anti-mouse IgG antibody, Brdu and its antibody (product of Wuhan Boster Biotechnology Co., Ltd.); CCK-8 kit (Beyond Institute of Biotechnology); Carotin, provided by Jiangsu Provincial Key Laboratory of Prescription Research, Nanjing University of Traditional Chinese Medicine, purity ≥ 98%;
[0107] Dissecting fluid:D-hank , s, (pH7.2-7.4): NaCl8.0g, Na 2 HP0 4 12H 2 O0.132g, KH 2 PO 4...
Embodiment 2
[0143] Embodiment 2. overall experiment
[0144] 2.1 Animals, grouping and handling
[0145] Twenty aged mice (11 months old), half male and half male, were divided into blank control group and carrotin group, 10 mice in each group. The blank control group was administered with normal saline, and the carotin group was administered with 10 mg / kg carotin, once a day for 30 days.
[0146] 2.2 Brdu-nestin double labeling and laser confocal microscope photography
[0147] method: Three days before the animals were sacrificed, the mice were given intraperitoneal injection of Brdu, 50 mg / kg, once every 12 hours, for three consecutive days. 12 hours after the end, the mice were anesthetized by intraperitoneal injection of chloral hydrate, the blood vessels were flushed with normal saline, and the heart was perfused with 4% paraformaldehyde. The brain was quickly fixed and removed, and fixed in 4% frozen paraformaldehyde for 48 hours. Then dehydrate in 20%, 30% sucrose hype...
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Abstract
The invention relates to a new application of daucosterol in medicine and health care product preparation, in particular to application of daucosterol in preparing medicines and health care products for promoting proliferation of neural stem cells. The daucosterol is a phytosterol derivative and is sitosterol-3-O-glucoside which widely exists in plants, and the processes of extracting, separation and content detection are simple and convenient. According to research, the medicines can promote proliferation of osteoblastlike cells, protect endothelial cells of human capillaries and further can reduce blood sugar activity. By means of the application, primarily-cultured neural stem cells and aged mice serve as the materials, and users can observe the effect of daucosterol on the neutral stem cells. In-vitro experiments and integral experiments prove that the daucosterol has the effect of promoting proliferation of the neutral stem cells. In the in-vitro experiments, the daucosterol with the dose ranging from 10 mu mol / L to 100 mu mol / L can promote proliferation of the neutral stem cells, and in the integral experiments, the daucosterol with the dose of 10mg / kg can promote proliferation of the neutral stem cells of the aged mice.
Description
technical field [0001] The invention relates to the application of drugs / health products in the field of medicine, in particular to the application of carotene in the preparation of drugs / health products for promoting the proliferation of neural stem cells. Background technique [0002] 1. The source and physical and chemical properties of carotin Carotin aliases: Sito glucoside, sitosterol-3-O-glucoside; English name: Daucosterol. Molecular weight: 576.85. [0003] Carotin is an important phytosterol derivative, which is widely distributed in nature and exists in the roots, stems, leaves, fruits and seeds of plants. It can be extracted with ethanol under reflux and separated by silica gel column chromatography or macroporous resin column chromatography. This product is a white powder, which can be stored in a dry and low temperature place away from light, soluble in pyridine, DMSO, hot methanol, ethanol, etc., insoluble in petroleum ether, chloroform, water and other so...
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Patent Type & Authority Applications(China)
IPC IPC(8): A61K31/704A61P25/28A23L1/29A23L33/00
Inventor 江励华王明艳杨念云陆大祥郭盛段金厫
Owner NANJING UNIVERSITY OF TRADITIONAL CHINESE MEDICINE