Anti-aging exosome-superoxide dismutase nano preparation and preparation method thereof

A dismutase nano- and superoxide technology, which is applied in the field of exosome-superoxide dismutase nano-preparation and its preparation, can solve problems such as instability, limited application, and short half-life, and achieve small and low impact on enzyme activity. Immunogenicity, long release effect

Active Publication Date: 2019-09-20
JILIN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the shortcomings of SOD, such as immunogenicity, instability, and short half-life in vivo, limit its clinical and health care applications.

Method used

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  • Anti-aging exosome-superoxide dismutase nano preparation and preparation method thereof
  • Anti-aging exosome-superoxide dismutase nano preparation and preparation method thereof
  • Anti-aging exosome-superoxide dismutase nano preparation and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0024] First, human embryonic kidney epithelial cells (HEK293) were cultured in DMEM medium containing 10% (volume percentage) fetal bovine serum (FBS) from which exosomes were removed, and the culture conditions were 37 ° C, 5% (volume Percentage) carbon dioxide environment, after the cell confluency reaches 90%, obtain 500 mL of HEK293 cell supernatant.

[0025] The above-mentioned HEK293 cell supernatant was centrifuged at 300g for 10min to remove suspended cells, centrifuged at 2000g for 10min to remove cell debris, and centrifuged at 10000g for 30min to remove larger particulate matter; the obtained supernatant was filtered through a 0.22μm filter membrane to remove particles larger than 220nm. Then, the supernatant was concentrated through an ultrafiltration centrifuge tube with a molecular weight cut-off of 100kDa, and the concentrated supernatant from the upper layer of the ultrafiltration centrifuge tube was centrifuged at 100,000g for 1.5h in an ultracentrifuge tube t...

Embodiment 2

[0029]Dilute the empty exosome solution synthesized in Example 1 and the PBS suspensions of three exosome-superoxide dismutase nano-preparations (SOD@EXO) to 1 mg / mL with PBS buffer, and spread on Copper mesh covered with 300-mesh carbon film was air-dried at room temperature. The copper grid coated with the nano-preparation was detected by a transmission electron microscope, and the accelerating voltage was 200kV. Such as figure 1 As shown, it can be clearly observed that the exosomes loaded with SOD molecules are spherical.

Embodiment 3

[0031] Dilute the exosome-loaded solution obtained in Example 1 and the PBS suspensions of the three exosome-superoxide dismutase nano-preparations (SOD@EXO) to 1 mg / mL with PBS buffer, and then dissolve In the PBS buffer filtered through a 0.22 μm filter membrane, a 10 μg / mL solution was finally prepared, and the particle size of the samples was tested with a Zetasizer Nano ZS90 particle size potentiometer. As shown in Table 1, the particle size of exosomes before and after loading SOD was between 100nm and 200nm.

[0032] Table 1: Particle size characterization data of empty exosomes synthesized in Example 1 and three nano-preparations

[0033]

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Abstract

The invention belongs to the technical field of biology and discloses an anti-aging exosome-SOD (superoxide dismutase) nano preparation and a preparation method thereof. According to a strategy of combination of an extrusion method and saponin, SOD enzyme molecules are loaded into exosome, and an action effect and an action mechanism of the nano preparation in senescence resisting are systematically evaluated by taking the prepared SOD@EXO nano preparation as a nano drug and taking N2 wild type caenorhabditis elegans as a model organism. By loading of SOD in the exosome, catalytic activity of natural enzymes is kept, and remarkable advantages such as half-life period prolonging, immunogenicity reducing and membrane permeability improving are achieved as compared with performances of natural SOD. The nano preparation is high in drug-loading capacity and capable of realizing stable drug release, and the low-immunogenicity high-catalytic-activity exosome-superoxide dismutase nano preparation is constructed finally to prolong the half-life period of SOD molecules and improve anti-aging treatment effects remarkably.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to an exosome-superoxide dismutase (SOD) nano preparation with anti-aging effect and a preparation method thereof. Background technique [0002] Senescence is a gradual process in which the functions of cells, organs and tissues of organisms are gradually lost over time. Modern aging theory believes that the aging process of the body is related to the damage of free radicals and induced lipid peroxidation to cells and the body. Free radicals and their derivatives refer to highly active atomic groups with unpaired electrons, which have strong oxidative properties and are important reaction components of redox reactions in the human body. An appropriate amount of free radicals can promote cell proliferation and stimulate phagocytes to kill bacteria. However, excessive free radicals have harmful effects on lipids, DNA and other substances, causing damage to biofilms and putti...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K38/44A61K9/50A61K47/46A61P39/06C12N5/073
CPCA61K38/446C12Y115/01001A61K9/5068A61P39/06C12N5/0603C12N5/0686C12N2509/00
Inventor 李全顺邵新欣李青刘勇梁骁
Owner JILIN UNIV
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