Dexamethasone-entrapped macrophage-derived microvesicle as well as preparation method and application thereof

A technology of macrophages and dexamethasone, applied in the field of biomedicine, to achieve the effect of reducing dosage, wide sources, and improving adverse reactions

Active Publication Date: 2018-10-09
SOUTHEAST UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, there is no report in the prior art that macrophage microvesicles are use

Method used

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  • Dexamethasone-entrapped macrophage-derived microvesicle as well as preparation method and application thereof
  • Dexamethasone-entrapped macrophage-derived microvesicle as well as preparation method and application thereof
  • Dexamethasone-entrapped macrophage-derived microvesicle as well as preparation method and application thereof

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Experimental program
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Effect test

Embodiment 1

[0036] Dexamethasone-loaded macrophage-derived microvesicles Extraction, identification and drug concentration determination

[0037] 1, preparation of

[0038]The mouse macrophage cell line RAW 264.7 (purchased from the Cell Bank of the Chinese Academy of Sciences) was cultured in vitro, and the RAW264.7 cells were cultured in RIPM 1640 (Gibco, USA) medium containing 10% fetal bovine serum (Gibco, USA) by volume fraction, wherein The fetal bovine serum used was centrifuged to remove microvesicles. When the confluence of RAW 264.7 cells in the culture flask reaches 70%-80%, wash the cells twice with RIPM 1640 medium, and replace it with FBS-free RIPM 1640 medium, add DEX (Sigma, USA) to make the final concentration 30 μmol / L, treated for 16h. After 16 hours of treatment, aseptically collect the culture supernatant in a 50ml sterile centrifuge tube and centrifuge at 4°C, 2000×g for 20 minutes to remove dead cells and large debris; carefully transfer the supernatant to a ...

Embodiment 2

[0047] Dexamethasone-loaded macrophage-derived microvesicles Therapeutic effect on LPS-induced glomerular endothelial cell inflammation

[0048] 1. Uptake by glomerular endothelial cells detection

[0049] Mouse primary glomerular endothelial cells (GECs) were extracted in vitro, spread in six-well plates or confocal small dishes, and when the cell fusion reached 80%, add 2×10 7 PKH26 (US Sigma) labeled Add 5×10 to the confocal cuvette 6 PKH26-tagged Observe the uptake of GECs by LPS (final concentration 10 μg / ml) (Sigma, USA) at different time points effect, in order to observe GEC phagocytosis The amount of the average fluorescence intensity of GECs in the six-well plate was detected by flow cytometry, and the results were as follows figure 2 As shown in A, the average fluorescence intensity of GECs in the LPS intervention group (final concentration 10 μg / ml) was higher (the control group was without LPS). Observation of GEC uptake by laser confocal microscopy...

Embodiment 3

[0056] Dexamethasone-loaded macrophage-derived microvesicles Therapeutic effect on LPS-induced sepsis model in mice

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Abstract

The invention discloses a dexamethasone-entrapped macrophage-derived microvesicle as well as a preparation method and application thereof. The dexamethasone-entrapped macrophage-derived microvesicle is formed by entrapping dexamethasone by a microvesicle from a cell RAW 264.7 of a mouse macrophage system. The dexamethasone-entrapped macrophage-derived microvesicle disclosed by the invention can beuptaken by an injured cell more effectively, and fulfills the aim of improving the kidney inflammation by inhibiting the activation of a proinflammatory signal pathway and infiltration of inflammatory cells. Meanwhile, the preparation method disclosed by the invention is simple, convenient and efficient; the prepared microvesicle is from the cell RAW 264.7, so that sufficient microvesicles can beprovided, and the microvesicles are wide in source; large-scale production can be implemented. The dexamethasone-entrapped macrophage-derived microvesicle prepared through the preparation method disclosed by the invention can be applied to preparation of a drug or preparation for treating kidney diseases and can also be applied to preparation of an anti-inflammatory or immunosuppression drug or preparation.

Description

technical field [0001] The invention belongs to the field of biomedicine and relates to a microvesicle-based drug delivery system, in particular to a method for preparing macrophage-derived microvesicles loaded with dexamethasone and its application in the anti-inflammatory treatment of kidney diseases. Background technique [0002] Glucocorticoids such as dexamethasone have been widely used in the treatment of inflammatory and immune diseases because of their significant anti-inflammatory and immune-suppressing effects. syndrome, IgA nephropathy, etc. However, long-term heavy use of glucocorticoids can lead to serious adverse reactions and glucocorticoid resistance, which seriously hinders clinical use. Therefore, exploring ways to reduce the adverse reactions of glucocorticoids and improve the sensitivity of glucocorticoids has become a research hotspot. [0003] Microvesicles are extracellular vesicles with a diameter of about 100-1000nm secreted by a variety of living ...

Claims

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Application Information

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IPC IPC(8): A61K9/50A61K9/127A61K47/46A61K31/573A61P13/12A61P29/00A61P37/06
CPCA61P13/12A61P29/00A61P37/06A61K9/1278A61K9/5068A61K31/573A61K49/0091
Inventor 刘必成汤涛涛吕林莉
Owner SOUTHEAST UNIV
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