Mouse Resistin (mRetn): cloning, expression and purification in Escherichia coli and the potential regulative effects on murine bone marrow hematopoiesis.

Abstract

BACKGROUND:

Resistin (Retn) is a cytokine which has a controversial physiological role regarding its involvement with obesity and type II diabetes mellitus. Recently, murine Retn was found to be a possibly potential regulator of hematopoiesis in mice shown in the screening results of a set of gene chips which mapped the expression level of murine genes during regeneration of impaired bone marrow (BM) by 5-fluorouracil.

RESULTS:

Recombinant mice Retn was expressed in Escherichia coli and purified using ion exchange chromatography. Totally 11.4 mg rmRetn was obtained from 500 ml culture with endotoxin level less than 1.0 EU/ug. The purity of recombinant murine Resistin reached to at least 97.6% via SDS-PAGE analysis and HPLC. The protein possessed chemotaxis effects in the mouse aortic endothelial cells in vitro in transwell analysis. In vitro, rmRetn could up regulate the CFU number of mice BM and after rmRetn was administered, the cell number of murine bone marrow was significantly increased in vivo after chemotherapy. Finally, rmRetn was found able to protect mice from the chemotoxicity of 5-fluorouracil.

CONCLUSIONS:

The discovery demonstrated a new function of murine Retn and suggested that it could potentially accelerate bone marrow regeneration post chemotherapy.

• Authors: Fangyuan Wang, Jin Gao, Alyssa Malisani, Xiaowei Xi, Wei Han, Xiaoping Wan
• Journal: BMC biotechnology (BMC Biotechnol) Vol. 15 Pg. 105 (Nov 16 2015) ISSN: 1472-6750 [Electronic] England
• PMID: 26572487 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)

Chemical References

• DNA Primers
• Resistin
• Retn protein, mouse
• Fluorouracil

Topics

• Analysis of Variance
• Animals
• Blotting, Western
• Bone Marrow (physiology)
• Chromatography, High Pressure Liquid
• Chromatography, Ion Exchange
• Cloning, Molecular
• DNA Primers (genetics)
• Electrophoresis, Polyacrylamide Gel
• Enzyme-Linked Immunosorbent Assay
• Escherichia coli
• Fluorouracil (toxicity)
• Hematopoiesis (drug effects)
• Mice
• Oligonucleotide Array Sequence Analysis
• Plasmids (genetics)
• Regeneration (drug effects, physiology)
• Resistin (genetics, metabolism, pharmacology)