Application of HucMSC-sEV loaded with miR13896 to acute liver injury caused by sepsis

A technology of acute liver injury and hucmsc-sev, applied in the medical field, can solve the problem of acute liver injury without effective treatment measures and other problems

Pending Publication Date: 2022-07-29
JIANGSU UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0004] At present, there is still no effective treatment for acute liver injury caused by sepsis (hereinafter referred to as acute liver injury caused by sepsis), and it is of great significance to seek effective ways to prevent and treat acute liver injury caused by sepsis

Method used

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  • Application of HucMSC-sEV loaded with miR13896 to acute liver injury caused by sepsis
  • Application of HucMSC-sEV loaded with miR13896 to acute liver injury caused by sepsis
  • Application of HucMSC-sEV loaded with miR13896 to acute liver injury caused by sepsis

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preparation example Construction

[0048] In the present invention, the preparation method of the miR13896-loaded hucMSC-sEV preferably includes the following steps:

[0049] The miR13896 mimics solution was mixed with the hucMSC-sEV solution before electroporation;

[0050] The electroporation solution obtained by electroporation was incubated at 37° C. for 30 min to obtain the hucMSC-sEV loaded with miR13896.

[0051] In the present invention, the concentration of the miR13896 mimics solution is preferably 1 μg / μL; the miR13896 mimics solution preferably includes a DEPC aqueous solution of miR13896 mimics. The present invention has no special limitation on the miR13896 mimics solution.

[0052] In the present invention, the concentration of the hucMSC-sEV solution is preferably 20 μg / μL; the hucMSC-sEV solution preferably includes a PBS solution of hucMSC-sEV. The present invention does not have a special limitation on the hucMSC-sEV solution, and the prepared hucMSC-sEV can be dissolved in a PBS solution. ...

Embodiment 1

[0069] hucMSC-sEV isolation and identification

[0070] according to figure 1 The steps shown are to isolate hucMSC-sEV, specifically:

[0071] (1) Isolation, culture and identification of hucMSC: The method of Qiao Chun et al. was used to isolate and culture human umbilical cord mesenchymal stem cells (hucMSC, Qiao Chun et al. Human mesenchymal stem cells isolated fromtheumbilical cord. Cell Biol Int. 2008; 32(1): 8-15.), the isolated hucMSCs were stored at 37°C, 5% CO 2 Cultured in a humidified incubator; primary culture of hucMSCs was performed with 15% α-MEM. After the hucMSCs climbed out, they were digested and cultured to the P3 generation with 10% α-MEM. Adipogenic and osteogenic differentiation of hucMSC was conducted, and the surface markers of hucMSC were detected by flow cytometry. When the fusion degree of hucMSC in P3 generation reaches 50%-60%, discard the supernatant, wash with PBS for 3 times, change to serum-free medium for 48 hours, collect the supernatan...

Embodiment 2

[0075] miR13896 Molecular Screening

[0076]Shanghai Ouyi Biomedical Technology Co., Ltd. performed miRNA sequencing on human embryonic lung fibroblast exosomes (HFL-sEV) and human umbilical cord mesenchymal stem cell exosomes (hucMSC-sEV). Differentially expressed miRNA molecules were found, and the results were as follows figure 2 shown, in which miR13896 was highly expressed in hucMSC-sEVs.

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Abstract

The invention belongs to the technical field of medical treatment, and particularly relates to application of miR13896-loaded engineered hucMSC-sEV in the aspect of acute liver injury caused by sepsis. The nucleotide sequence of the miR13896 is shown as SEQ ID NO.1. The miR13896-loaded engineered hucMSC-sEV can induce polarization from M1 type to M2 type of macrophages, inhibit generation of IL-1beta, IL-6, TNF-alpha and iNOS, promote expression of IL-10 and Arg1, reduce the content of glutamic-pyruvic transaminase and glutamic oxalacetic transaminase, relieve changes of liver functions and prevent and treat acute liver injury of sepsis.

Description

technical field [0001] The invention belongs to the field of medical technology, in particular to the application of miR13896-loaded hucMSC-sEV in acute liver injury in sepsis. Background technique [0002] Sepsis is a syndrome of invasive infection and host systemic inflammatory response caused by bacteria, fungi, parasites, etc., which can be accompanied by multiple organ dysfunction and failure. Sepsis is one of the common complications after severe infection, trauma, burn, shock and surgery, and it has become one of the important causes of death in patients with clinical hazards. The incidence of sepsis is high, with more than 18 million severe sepsis cases worldwide each year. In recent years, despite great progress in anti-infective therapy and organ function support technology, the mortality rate of sepsis is still as high as 30% to 70%. The high cost of sepsis treatment and the large consumption of medical resources have seriously affected the quality of human life...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K31/7105A61K47/46A61P1/16A61P29/00
CPCA61K31/7105A61K47/46A61P1/16A61P29/00
Inventor 钱晖金灿吴佩佩尹思琪许文荣纪成杨大凯
Owner JIANGSU UNIV
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