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Composition for preventing or treating tissue fibrosis using mfg-e8

A MFG-E8, fiberization technology, applied in the field of MFG-E8

Active Publication Date: 2021-07-20
NEXELL CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

But the reality is that there are no reports of what effect it has on liver fibrosis in liver tissue or other stages of fibrosis in tissue other than

Method used

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  • Composition for preventing or treating tissue fibrosis using mfg-e8
  • Composition for preventing or treating tissue fibrosis using mfg-e8
  • Composition for preventing or treating tissue fibrosis using mfg-e8

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0078] Culture of hepatocytes obtained from between mesenchymal stem cells

[0079] Methylactic stem cells used in this experiment are derived from cord blood, in order to differentiate to hepatocytes, 3 × 10 4 Cell / cm 2 The form is attached to a culture dish coated with collagen.

[0080] The MESENPRO RSTM medium (GIBCO) was used as the basic medium. If the culture dish is filled with 70-80%, epidermal growth factor (EGF; PEPROTECH EC LTD, London, England, 20 ng / ml) and basic fibrous fibrous factor (BFGF) PEPROTECH EC LTD, 10 ng / ml) is placed in the first differentiated medium, iscove's modified Dulbecco medium (IMDM; Invitrogen, Carlsbad, CA, USA) and cultivating two day.

[0081] Thereafter, hepatocyte growth factor (HGF; PEPROTECH EC Ltd., 20 ng / ml), BFGF (10 ng / ml), nicotinamide (Sigma, 0.61 g / l), 1% insulin - transfer INSULIN-TRANSFERRIN-SELENIUM (ITS) Premix (INVITROGEN) is added to the basal medium IMDM as a second differentiation medium and cultured for ten d...

Embodiment 2

[0084] Effect of MFG-E8 in chronic liver disease model and hepatic fibrosis model

[0085]In order to induce chronic liver disease, thioacetamide (TAA) was diluted in a 200 mg / kg body weight, and a mouse (C57BL / C) of 5 to 6 weeks was permutable. Abdominal injection was repeated for 3 times in 1 week. After 8 weeks, the protein of the concentrated secreted protein group was quantified by Bradford Assay, and then the amount of 500 μg was injected in the abdominal cavity. After injection of the secreted protein group, after 3 and 7, the degree of liver fibrosis of the mouse was analyzed. The MFG-E8 protein (R & D Systems, kit) was found to be found in the MFG-E8 protein (R & D Systems, kit) in order to confirm the effect of different concentrations, and 160 μg / kg body weight is set to an intermediate concentration. The 800 μg / kg body weight was set to a high concentration and inserted into the abdominal cavity. After investing in the MFG-E8 protein, after 3 days, the degree ...

Embodiment 3

[0095] Effect of MFG-E8 in HTERT-HSCS and PrimaryHSCS in the PRIMARYHSCS

[0096] Experiments were carried out using an in-Vitro effect of the secreted protein group using hepatic star cell lines and initial cultured hepatar cells. To this end, hepatic star cell lines were cultured in a 10% FBS and 100 units of Penicillin, 100 μg of streptomycin culture solutions in the pHICS DMEM (Dulbecco's Modifiedeagle Medium). In the experiment, in order to produce deactivated hepatar cell lines, a 0.2% FBS was cultured in a culture solution of 0.2% FBS in a culture medium of 0.2% FBS, TGF [beta] 1 protein in the same medium in the same medium. The treatment is performed so that hepatic star cell line is activated. The MFG-E8 protein used in the experiment was treated with TGF [beta] 1 protein in a form of substantially 500 ng / ml and cultured for 48 hours. When processed at different concentrations, cultivation was carried out in the form of 100 ng / ml, 250 ng / ml, 500 ng / ml, 1 μg / ml...

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Abstract

The present invention relates to a composition for preventing or treating tissue fibrosis using milk fat globule epidermal growth factor 8 protein (MFG‑E8). In more detail, MFG‑E8 inhibits TGFβ / Smad signaling induced The expression of collagen, and inhibit the activity of hepatic stellate cells, thereby playing a role in improving liver fibrosis, and has the following effects: in the mouse model of liver fibrosis, the degree of fibrosis is reduced, if cultured in vitro The treatment on the hepatic stellate cells can inhibit the activation, so it can be used as a preventive or therapeutic agent for tissue fibrosis.

Description

Technical field [0001] The present invention relates to the use of MFG-E8 for preventing or treating tissue fibrosis by fibrosis by milk fat ball skin growth factor 8 protein (MFG-E8, MILK FATGLOBULE-EGF FACTOR 8) to prevent or treat tissue fibrosis MFG-E8, the milk The fat gloss skin growth factor 8 protein (MFG-E8) is a protein that is secreted from a mesenchymal stem cell. Background technique [0002] In the case of the study of the differentiation of mesenchymal steel to hepatocytes to hepatocytes, although the intermodulation intermium stem cells exceeding the endoderm in the early stage and has an inner embryonic hepatocyte differentiation. The possibility of transdifferentiation, but recently proposed as follows: According to the accumulated results, it is not the result of differentiation transformation, but through the active factors secreted after being transplanted with mesenchymal stem cells to inhibit the liver within the host tissue. The death of cells, the regener...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K38/18A61P43/00A61P1/16A61P11/00A61P13/12A61P25/00A61P9/00G01N33/15
CPCA61K38/1808G01N33/15A61K35/28A61K35/407A61K38/18
Inventor 金钟埙安姝妍张有真
Owner NEXELL CO LTD
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