Abstract
Irisin, which was recently identified as a myokine and an adipokine, transforms white adipose tissue to brown adipose tissue and has increasingly caught the attention of the medical and scientific community. However, the signaling pathway of irisin and the molecular mechanisms responsible for the lipolysis effect remain unclear. In this study, we established an efficient system for the expression and purification of GST-irisin in Escherichia coli. The biological activity of GST-irisin was verified using the cell counting kit-8 assay and by detecting the mRNA expression of uncoupling protein 1. Our data showed that GST-irisin regulates mRNA levels of lipolysis-related genes such as adipose triglyceride lipase and hormone-sensitive lipase and proteins such as the fatty acid-binding protein 4, leading to increased secretion of glycerol and decreased lipid accumulation in 3T3-L1 adipocytes. In addition, exogenous GST-irisin can increase its autocrine function in vitro by regulating the expression of fibronectin type III domain-containing protein 5. GST-irisin could regulate glucose uptake in 3T3-L1 adipocytes. Hence, we believe that recombinant GST-irisin could promote lipolysis and its secretion in vitro and can potentially prevent obesity and related metabolic diseases.
Highlights
Obesity, which is characterized by an excess of adipose tissue and associated with various comorbidities such as insulin resistance and type 2 diabetes, has been highlighted as an epidemic in recent decades [1, 2]
The plasmid was transformed into E. coli BL21, and the expression of the Glutathione S-transferase (GST)-irisin fusion protein was induced by isopropyl β-D-1-thiogalactopyranoside (IPTG)
The level of endotoxin in the target protein is less than 5 Eu/ml, and the data is far less than the minimum value of the lipopolysaccharide that stimulated lipolysis in adipocytes [24]
Summary
Obesity, which is characterized by an excess of adipose tissue and associated with various comorbidities such as insulin resistance and type 2 diabetes, has been highlighted as an epidemic in recent decades [1, 2]. This is caused by an increase in the number and/or size of adipocytes [3]. Many studies have focused on increasing energy expenditure of adipose cells to prevent obesity.
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