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Cordyceps sobolifera active site and application thereof in preparing drugs for nerve protection and aging resistance

A technology for active parts and neuroprotection, applied in the field of medicine, can solve the problems such as the lack of refined Cicada flower extract and the lack of very clear active functional components.

Active Publication Date: 2014-05-21
JIANGSU UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] In view of the fact that cicadae and the anamorph of the famous and precious Chinese medicine Cordyceps sinensis belong to fungi (Cordyceps genus), and Paecilomyces cicadae has various pharmacological effects such as immunity, and has the advantages of low toxicity and easy cultivation, it is hopeful that it can be used as a substitute for Cordyceps sinensis. Its corresponding active functional components are not very clear, and most of the pharmacological experiments only stay on the basis of the original medicinal materials or crude extracts, and there is no report that the extract of Cicada japonica is further refined to the active site and used to prepare neuroprotective and anti-aging drugs

Method used

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  • Cordyceps sobolifera active site and application thereof in preparing drugs for nerve protection and aging resistance
  • Cordyceps sobolifera active site and application thereof in preparing drugs for nerve protection and aging resistance
  • Cordyceps sobolifera active site and application thereof in preparing drugs for nerve protection and aging resistance

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0016] The n-butanol extraction part (JCH n-BuOH ) preparation:

[0017] Take 80g of dried cicadae powder that has been crushed through the No. 2 sieve of Pharmacopoeia, and extract twice with 800mL water under reflux at 90°C for 2 hours each time. After the extract is concentrated under reduced pressure, add 4 times the volume of absolute ethanol and place in Precipitate overnight in refrigerator at ℃; centrifuge at 3000r / min for 15min; concentrate the supernatant, transfer the concentrated solution into a separatory funnel, and use equal volumes of petroleum ether, ethyl acetate, and n-butanol for fractional extraction, a total of 3 extractions, and combine , concentrated under reduced pressure to recover the solvent, and respectively obtained the petroleum ether part, the ethyl acetate part, and the n-butanol part (JCH n-BuOH ), water part; freeze-dried to get JCH n-BuOH 1.92g.

Embodiment 2

[0018] Example 2JCH n-BuOH Protective effect on glutamate-induced senescence of PC12 cells

[0019] (1) Determination of cell viability by MTT method and LDH method:

[0020] Take PC12 cells in the logarithmic growth phase (provided by Jiangsu University School of Medicine), and use 2 × 10 4 Inoculate in a 96-well culture plate at 200 μl / L per well. At 37°C, 5% CO 2 After culturing overnight under the condition, the cells were divided into control group, model group and drug treatment group. That is, the control group (complete medium without Glu); model group (complete medium + Glu); drug treatment group (complete medium + JCH n-BuOH +Glu, the concentrations were 200μg / mL, 100μg / mL, 50μg / mL, diluted with culture medium. )JCH n-BuOH After pretreatment for 1 h, add Glu and incubate for 24 h. Set up 4 parallel wells in each group. After culturing for 24 hours, add 20 μl of 5g / L MTT to each well, continue culturing for 4 hours, then terminate the culture, carefully suck th...

Embodiment 3

[0037] Example 3 JCH n-BuOH In vitro antioxidant activity assay:

[0038] (1) Determination of DPPH free radical scavenging ability:

[0039] Add 1.0mL 200μmol / DPPH ethanol solution to 1mL different concentrations of Cicada flower extract solution, then add 2.0mL80% ethanol and mix evenly. After 30min in a dark place, measure the absorbance A at 517nm with a UV spectrophotometer 样品 , and simultaneously measure the absorbance value A of the mixture of 1.0mL DPPH ethanol solution and 3.omL ethanol solution 空白 and the absorbance value A of the mixture of 3mL ethanol and 1.0mL sample 对照 , clearance rate formula: DPPH clearance rate=[A 空白 -(A 样品 -A 对照 ) / A 空白 ]×100%. The data show that JCH n-BuOH It has the strongest ability to clear DPPH. The results are shown in Table 5

[0040] (2) Superoxide anion (O 2·- ) Determination of free radical scavenging ability:

[0041] Add 3ml Tris-HCl (PH8.2) to 1mL different concentrations of Cicada flower extract solution, after 20min ...

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Abstract

The invention relates to a Cordyceps sobolifera active site and application thereof in preparing drugs for nerve protection and aging resistance. The active site is an n-butanol site of a Cordyceps sobolifera water extract. The active site is prepared by the following steps: pulverizing Cordyceps sobolifera, extracting with water under reflux, concentrating the extracting solution under reduced pressure, adding anhydrous ethanol, and precipitating over night; centrifuging, and concentrating; and carrying out fractional extraction on the concentrated solution with petroleum ether, ethyl acetate and n-butanol, concentrating under reduced pressure to recover the solvent, and carrying out freeze-drying to respectively obtain a petroleum ether site, an ethyl acetate site, an n-butanol site and a water site. The n-butanol site can obviously inhibit aging and damage of the glutamic-acid-induced PC12 cells, prevent the cell LDH from release, enhance the survival rate of the cells, lower the intracellular oxygen free radical level, enhance the activity of the glutathione reductase (GSH-Px) and superoxide dismutase (SOD), and have favorable in-vitro oxidation resistance activity in the experiment of clearing DPPH. and superoxide anion (O<2.->) free radicals.

Description

technical field [0001] The invention belongs to the technical field of medicine, and in particular relates to the preparation of parts with neuroprotective and anti-aging activities from Cicada japonica, a Chinese medicinal material, and its application in the preparation of neuroprotective and anti-aging drugs. Background technique [0002] With the aging population in our country, the incidence of neurodegenerative diseases is increasing day by day, and the research on aging and neurodegenerative diseases has become a neuroscience hotspot. The aging of the human body often leads to an accelerated reduction in the number of nerve cells, resulting in some neurodegenerative diseases represented by Alzheimer's Disease (AD). At present, the etiology of neurodegenerative diseases is unknown, and there is no effective treatment. Only a few drugs can be used to treat neurodegenerative diseases. [0003] Cicada is a dry complex formed by the fungus Cordyceps cicadae Shing of the f...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K36/066A61P39/06A61P25/00A61P25/28A61K35/64
Inventor 欧阳臻张魏琬麒赵明王吉标尚磊王璠汪愿
Owner JIANGSU UNIV
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