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CN-121975519-A - Luteolin carbon dot for guiding lysosomes as well as preparation method and application thereof

CN121975519ACN 121975519 ACN121975519 ACN 121975519ACN-121975519-A

Abstract

The invention discloses a luteolin carbon dot of a guide lysosome and a preparation method and application thereof in the technical field of carbon dot application, wherein luteolin and 2, 5-dihydroxyterephthalic acid are used as carbon source precursors, the luteolin carbon dot is prepared by a hydrothermal method, the particle size of the prepared luteolin carbon dot is 0.5-4 nm, the fluorescence excitation wavelength is 360-440 nm, the maximum emission wavelength is 540 nm, the zeta potential is-19 mv, the luteolin carbon dot can be guided to macrophages and fat cell lysosomes through research, the polarization of M2 type macrophages is promoted, the heat production of beige fat cells and the expression of brown marker gene Ucp are upward regulated, and furthermore, the M2 type macrophages loaded with the luteolin carbon dot can promote the expression of the beige fat cells Ucp through releasing the carbon dot and the paracrine effect, so that the heat production and the energy consumption are increased, and the intervention scheme for the development of the basis of obesity and related metabolic diseases is laid for future based on the carbon dot.

Inventors

  • WANG LU
  • CHEN WENBIN
  • LIU JIAN
  • ZHANG ZHIXIN

Assignees

  • 合肥工业大学

Dates

Publication Date
20260505
Application Date
20260407

Claims (9)

  1. 1. The luteolin carbon dot for guiding the lysosomes is characterized in that the luteolin carbon dot is prepared by taking luteolin and 2, 5-dihydroxyterephthalic acid as carbon source precursors through hydrothermal reaction, and the luteolin carbon dot is specifically guided to the lysosomes of macrophages and adipocytes.
  2. 2. The lysosome-oriented luteolin carbon dot of claim 1, wherein the luteolin carbon dot is spherical particles with a particle size of 0.5-4 nm, the luteolin carbon dot has a fluorescence excitation wavelength of 360-440 nm, a maximum emission wavelength of 540 nm and a Zeta potential of-19 mv.
  3. 3. A method for preparing a lysosome-oriented luteolin carbon dot according to any one of claims 1 to 2, wherein the method comprises the steps of dispersing luteolin and 2, 5-dihydroxyterephthalic acid in deionized water to obtain a mixed solution, performing a hydrothermal reaction on the mixed solution, filtering and dialyzing the mixed solution to obtain a carbon dot solution, and freeze-drying the carbon dot solution to obtain the lysosome-oriented luteolin carbon dot.
  4. 4. The method for preparing a lysosome-oriented luteolin carbon dot according to claim 3, wherein the molar ratio of luteolin to 2, 5-dihydroxyterephthalic acid is 1:1-3, the hydrothermal reaction temperature is 160-260 ℃, the reaction time is 2-6 h, the pore diameter of a filtering membrane of the filtering treatment is 0.22 μm, and the dialysis bag size of the dialysis treatment is 1000 Da.
  5. 5. Use of a carbon dot of luteolin according to any of claims 1-2 for cell lysosome targeted fluorescence imaging, wherein the carbon dot of luteolin is used as a green fluorescent probe for directing lysosomes enriched in macrophages and adipocytes for fluorescent imaging of lysosomes of macrophages and adipocytes.
  6. 6. Use of luteolin carbon dots according to any one of claims 1-2 for the preparation of a formulation for promoting macrophage M2 type polarization, wherein the luteolin carbon dots are used to treat macrophages, up-regulate the mRNA expression level of M2 type macrophage marker gene Arg1, and increase the M2 subpopulation ratio of macrophages to promote macrophage M2 type polarization.
  7. 7. Use of a carbon dot of luteolin according to any one of claims 1-2 for the preparation of a formulation for promoting browning of beige adipocytes, wherein the beige adipocytes are treated with the carbon dot of luteolin to up-regulate the mRNA expression level of the beige adipocyte thermogenesis and browning marker gene Ucp1 for promoting browning of the beige adipocytes.
  8. 8. Use of M2 type macrophages loaded with luteolin in the preparation of a preparation for promoting brown adipose cells, characterized in that the M2 type macrophages are pretreated with the carbon dots of the luteolin according to any one of claims 1-2, induced to polarization towards the M2 type, the pretreated M2 type macrophages are co-cultured with the brown adipose cells, and the brown adipose cells Ucp gene expression is regulated indirectly by releasing the loaded carbon dots of the luteolin and paracrine effects, thereby promoting brown adipose cells.
  9. 9. The use according to claim 8, wherein the concentration of luteolin carbon dot pretreatment macrophages is 200 μg/mL, the treatment time is 24h and the co-cultivation time is 24 h.

Description

Luteolin carbon dot for guiding lysosomes as well as preparation method and application thereof Technical Field The invention relates to the technical field of carbon dot application, in particular to a luteolin carbon dot guiding lysosomes, and a preparation method and application thereof. Background Obesity is a global high-rise metabolic disease, the core of which is excessive accumulation of body fat, white fat cells are responsible for storing energy in white adipose tissue, abnormal accumulation of the white fat cells is a main cause of obesity, beige fat cells are also scattered in the white adipose tissue, and high-expression of a heat-generating and browning marker gene Ucp1 can convert chemical energy into heat energy for consumption, and after activation, browning and heat generation can be further increased. In addition, in adipose tissues, M2 type macrophages can promote brown of beige adipocytes by paracrine catecholamine molecules, slit3 proteins and other substances, so that M2 type polarization of the macrophages is induced, and the M2 type macrophages drive brown of the adipocytes, so that the macrophages can be developed into an intervention treatment scheme aiming at obesity and related metabolic diseases. Luteolin is a common dietary flavonoid compound, can regulate and control lysosome activity, induces moderate elevation of intracellular ROS and lysosome-mitochondrial interaction so as to promote macrophage M2 type polarization, can regulate and control lysosome to maintain adipocyte mitochondrial steady state, activate AMPK/PGC1 alpha signal path, up-regulate heat production of beige adipocyte and expression of brown marker gene Ucp1, but has poor self water solubility, is difficult to sufficiently dissolve in aqueous solution, and cannot be effectively dispersed and dissolved in gastrointestinal tract, so that oral absorption efficiency is greatly influenced. In addition, luteolin has strong biological toxicity, and can easily cause irritation and damage to organs such as gastrointestinal tract, liver and the like when used at high dosage, thereby increasing cytotoxicity. The luteolin cell membrane penetration capability is also weak, and is difficult to enter target cells such as adipocytes, macrophages and the like efficiently, so that the practical application of the luteolin in improving obesity is greatly limited. Carbon Dots (CDs) are novel carbon-based nano materials with the size smaller than 10 nm, have the advantages of good water solubility, biocompatibility, high stability and easy functional modification, have wide application prospects in the aspects of guiding delivery, drug carriers and the like in the field of biological medicine, and can effectively enhance the bioavailability and the guiding property of natural active substances. In conclusion, if luteolin is prepared into carbon dots and is guided to a lysosome, the defects of poor water solubility, low bioavailability and the like of the luteolin can be overcome, and the brown of adipocytes driven by M2 type macrophages can be realized, so that the heat production and the energy consumption are increased, and a foundation is laid for developing an intervention treatment scheme aiming at obesity and related metabolic diseases. Through searching, in the prior literature and published patents, the luteolin carbon dots of the guided lysosomes and the application of the luteolin carbon dots are not shown yet. Disclosure of Invention The invention aims to solve the problems, and provides a lysosome-oriented luteolin carbon dot, a preparation method and application thereof, in particular to application in M2 type macrophage-driven beige adipocyte browning. The invention realizes the above purpose through the following technical scheme: as a first aspect of the present invention, there is provided a lysosome-directed luteolin carbon dot prepared by hydrothermal reaction using luteolin and 2, 5-dihydroxyterephthalic acid as carbon source precursors, the luteolin carbon dot being specifically directed to lysosomes of macrophages and adipocytes. As a further optimization scheme of the invention, the luteolin carbon dots are spherical particles with the particle size of 0.5-4 nm, the fluorescence excitation wavelength of the luteolin carbon dots is 360-440 nm, the maximum emission wavelength is 540-nm and the Zeta potential is-19 mv. According to the preparation method, the luteolin and the 2, 5-dihydroxyterephthalic acid are dispersed in deionized water in a molar ratio to obtain a mixed solution, the mixed solution is subjected to a hydrothermal reaction, a carbon dot solution is obtained after the hydrothermal reaction and the filtration and the dialysis treatment, and the carbon dot solution is subjected to freeze drying treatment to obtain the luteolin carbon dot of the guided lysosome. As a further optimization scheme of the invention, the molar ratio of luteolin to 2, 5-dihydroxyterephthalic acid is 1:1-3, the hydrot