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Lipid droplet induction and imaging reagent as well as preparation method and application thereof

A reagent and lipid droplet technology, applied in the field of lipid droplet induction and imaging reagents and their preparation, can solve the problems of poor safety, poor water solubility, and inability to trace lipid droplets specifically, and achieve good cytocompatibility, Low toxicity and good water dispersibility

Active Publication Date: 2021-01-22
SOUTHEAST UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0003] Purpose of the invention: In view of the problems existing in the prior art, the present invention provides a highly efficient lipid droplet induction and fluorescent labeling reagent, which can effectively solve the problem of poor water solubility of existing lipid droplet induction reagents and the inability to specifically trace lipid droplets. and disadvantages such as poor security

Method used

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  • Lipid droplet induction and imaging reagent as well as preparation method and application thereof
  • Lipid droplet induction and imaging reagent as well as preparation method and application thereof
  • Lipid droplet induction and imaging reagent as well as preparation method and application thereof

Examples

Experimental program
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Effect test

Embodiment 1

[0034] (1) Weigh 10 mg of dioleoylphosphatidylserine (DOPS, see attached figure 2 ) and dissolved it in 0.5mL chloroform, dried the solvent with nitrogen, and dried it in a vacuum oven at room temperature for 4 hours. After adding 1 mL of phosphate buffer solution to oscillate for hydration and dispersion, the liposomes were prepared using probe ultrasound (163W, 4 minutes), named DOPS liposomes;

[0035] (2) Take 0.2mL fluorescent phospholipid with NBD label on the tail and phosphatidylserine group at the head with a concentration of 1mg / mL (named NBD-PS, see the attached figure 2 ) (solvent is chloroform), and mixed evenly with DOPS (10 mg) dissolved in 0.5 mL chloroform, blowing the solvent with nitrogen gas and drying at room temperature in a vacuum oven for 4 hours. After adding 1mL of phosphate buffer solution for hydration and dispersion, the liposome was prepared by ultrasonic probe (163W, 4 minutes), named as DOPS / NBD-PS liposome (see the attached image 3 ).

Embodiment 2

[0037] Induction of lipid droplets with different saturation and construction of imaging liposomes:

[0038] The preparation method is the same as that of Example 1, except that the DOPS used in (2) of Example 1 is replaced by palmitoyloleoylphosphatidylserine (POPS) of equal mass.

Embodiment 3

[0040] Lipid droplet induction with different head groups and construction of imaging liposomes:

[0041] The preparation method is the same as in Example 1, except that the DOPS used in (2) of Example 1 is replaced by dioleoylphosphatidylglycerol (DOPG), dioleoylphosphatidylinositol (DOPI), bis Oleoylphosphatidic acid (DOPA) or dioleoylphosphatidylglycerol (DOCA).

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Abstract

The invention discloses a lipid droplet induction and imaging reagent as well as a preparation method and application thereof. The reagent is a liposome mainly composed of negatively-charged unsaturated glycerophospholipid and phospholipid marked with fluorescence at the tail of the negatively-charged unsaturated glycerophospholipid. The liposome provided by the invention is good in water dispersibility, can effectively induce cells to generate a large number of lipid droplets under the condition of not influencing the cell activity, and the lipid droplet generation efficiency is greatly improved compared with the existing induction strategy. More importantly, the reagent disclosed by the invention can also be used for carrying out fluorescence labeling on lipid droplets in cells, so thatimaging tracing on the lipid droplets is realized.

Description

technical field [0001] The invention relates to biotechnology, in particular to a lipid droplet induction and imaging reagent and its preparation method and application. Background technique [0002] Lipid droplets (LDs) are the main storage places for neutral lipids in cells, and can store triglycerides, sterol esters, and retinyl esters. They are multifunctional organelles with a single-layer membrane structure and surface-coated proteins. . Recent studies have found that, in addition to being responsible for fat storage, lipid droplets are also essential for many cellular functions, such as lipid metabolism, membrane biogenesis, cell signaling, and inflammation. In addition, accumulating evidence indicates that the development of cancer is related to the abnormality of lipid droplets. These important biological functions of lipid droplets urgently require researchers to develop reagents that can efficiently regulate and monitor cellular lipid droplets. However, current...

Claims

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Application Information

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IPC IPC(8): C09K11/06G01N21/64
CPCC09K11/06G01N21/6456G01N21/6486C09K2211/1048C09K2211/1011C09K2211/1055C09K2211/1007
Inventor 吴富根蒋耀文高歌
Owner SOUTHEAST UNIV
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