Recombinant SFRP5 protein significantly alleviated intrahepatic inflammation of nonalcoholic steatohepatitis

Abstract

BackgroundSecreted frizzled-related protein 5 (SFRP5) is an anti-inflammatory adipokine modulating metabolism dysfunction. This study aims to observe the effect of recombinant SFRP5 protein on nonalcoholic steatohepatitis (NASH).MethodsWe set up a prokaryotic expression system and purified the recombinant SFRP5 protein. Recombinant SFRP5 protein was further identified by SDS-PAGE, western blot, high performance liquid chromatography (HPLC), protein mass spectrometry and in vitro Wnt5a-binding test. NASH mouse model was induced by methionine and choline deficient diet (MCDD) for 2 weeks. SFRP5 treatment group received intraperitoneal injection with a dosage of 10μg/kg SFRP5 twice a day for 2 weeks. Saline was used as control. Inflammation and fatty lesion score of liver tissue pathology and serum transaminase level were compared.ResultsThe purity of recombinant SFRP5 protein is 90% identified by HPLC. Its molecule size is 36,096.08 tested by mass spectrometry. Recombinant SFRP5 can specifically bind with Wnt5a which verifies its activity in vitro. The endotoxin level of this recombinant protein is 0.01EU/μg-0.1EU/μg and is suitable for animal experiment. SFRP5 can significantly improve liver inflammation (SFRP5 vs. control, 1.40 ± 0.70 vs. 2.00 ± 0.47, P < 0.05) as well as fatty lesion scores (SFRP5 vs. control, 1.40 ± 0.97 vs. 2.20 ± 0.63, P < 0.05), and lower ALT and AST levels. The mRNA expression of proinflammatory adipokines (IL-1β, IL-6, TNFα and MCP-1) in liver was down-regulated significantly after SFRP5 intervention. Immunohistochemistry and quantitative PCR revealed a dramatically down-regulation of F4/80 in liver after SFRP5 treatment.ConclusionsRecombinant SFRP5 protein significantly alleviated NASH induced by MCDD.Electronic supplementary materialThe online version of this article (doi:10.1186/s12986-017-0208-0) contains supplementary material, which is available to authorized users.

Newly Discovered Adipokines: Pathophysiological Link Between Obesity and Cardiometabolic Disorders

“…SFRP5 over-expression via adenovirus could alleviate obesity, adipose inflammation, and hepatosteatosis ( Ouchi et al, 2010 ). In an experiment with NASH mouse model, SFRP5 significantly improved liver inflammation and fatty lesion scores ( Chen et al, 2017 ). Moreover, SFRP5 intervention led to the downregulation of pro-inflammatory cytokines, including interleukin (IL)-1β, IL-6, tumor necrosis factor-α (TNF-α), and monocyte chemoattractant protein-1, in the liver.…”

The Wnt antagonist and secreted frizzled-related protein 5: implications on lipid metabolism, inflammation, and type 2 diabetes mellitus

“…SFRP5, is a member of the SFRP family produced from adipose tissue and can act as an inflammatory adipokine. SFRP5 has been shown to act as an antagonist for the Wnt/β-catenin signaling pathway [ 3 ]. It has recently gained much attention, owing to the notion that it is capable of promoting adipocyte differentiation via blockade of the Wnt/β-catenin signaling pathway, thus leading to an increased risk of diet-induced obesity [ 4 , 5 ].…”

PEDF improves cardiac function in rats subjected to myocardial ischemia/reperfusion injury by inhibiting ROS generation via PEDF‑R

“…In brief, 293T cells were transfected with the 20 μ g recombinant vector pGEX 6P-1 (GE Healthcare, Chicago, IL, USA), containing glutathione S-transferase (GST)-tagged PEDF using Lipofectamine ® 3000 according to the manufacturer's protocols. GST-tagged PEDF proteins were purified by high-pressure liquid chromatography purification (>90% purity) and amino-terminal sequence determination ( 23 , 24 ). The resultant proteins were soluble in aqueous solutions.…”