Science letters: Proteomic analysis of differentially expressed proteins in mice with concanavalin A-induced hepatitis

Abstract

Objective: To find new protein biomarkers for the detection and evaluation of liver injury and to analyze the relationship between such proteins and disease progression in concanavalin A (Con A)-induced hepatitis.

Methods: Twenty-five mice were randomly divided into five groups: an untreated group, a control group injected with phosphate buffered saline (PBS), and groups with Con A-induced hepatitis evaluated at 1, 3 and 6 h. Two-dimensional gel electrophoresis (2-DE) and mass spectrometry (MS) were used to identify differences in protein expression among groups. Quantitative real-time polymerase chain reaction (qRT-PCR) was performed to verify the results.

Results: In mice with Con A-induced hepatitis, expression levels of four proteins were increased: RIKEN, fructose bisphosphatase 1 (fbp1), ketohexokinase (khk), and Chain A of class pi glutathione S-transferase. Changes in fbp1 and khk were confirmed by qRT-PCR.

Conclusion: Levels of two proteins, fbp1 and khk, are clearly up-regulated in mice with Con A-induced hepatitis.

Fig. 1

Microscopic appearance of the liver in mice representative of each group (a) Untreated mice; (b) PBS-treated mice; (c) 1 h after injecting Con A intravenously; (d) 3 h after injecting Con A intravenously (note a small amount of hepatocyte swelling); (e) 6 h after injecting Con A intravenously (note severe hepatocyte swelling and necrosis)

Fig. 1

Microscopic appearance of the liver in mice representative of each group (a) Untreated mice; (b) PBS-treated mice; (c) 1 h after injecting Con A intravenously; (d) 3 h after injecting Con A intravenously (note a small amount of hepatocyte swelling); (e) 6 h after injecting Con A intravenously (note severe hepatocyte swelling and necrosis)

Fig. 1

Microscopic appearance of the liver in mice representative of each group (a) Untreated mice; (b) PBS-treated mice; (c) 1 h after injecting Con A intravenously; (d) 3 h after injecting Con A intravenously (note a small amount of hepatocyte swelling); (e) 6 h after injecting Con A intravenously (note severe hepatocyte swelling and necrosis)

Fig. 1

Microscopic appearance of the liver in mice representative of each group (a) Untreated mice; (b) PBS-treated mice; (c) 1 h after injecting Con A intravenously; (d) 3 h after injecting Con A intravenously (note a small amount of hepatocyte swelling); (e) 6 h after injecting Con A intravenously (note severe hepatocyte swelling and necrosis)

Fig. 1

Microscopic appearance of the liver in mice representative of each group (a) Untreated mice; (b) PBS-treated mice; (c) 1 h after injecting Con A intravenously; (d) 3 h after injecting Con A intravenously (note a small amount of hepatocyte swelling); (e) 6 h after injecting Con A intravenously (note severe hepatocyte swelling and necrosis)

Fig. 2

Representative 2-DE maps of liver proteins from samples from each group of mice. In the first dimension of electrophoresis, 200 μg protein was loaded on strips (240 mm, pH 3–10 non-linear); the second dimension of electrophoresis was a vertical 12.5% (w/v) SDS-PAGE (a) Untreated controls; (b) PBS controls; (c)–(e) 1, 3, and 6 h after injecting Con A intravenously, respectively

Fig. 2

Representative 2-DE maps of liver proteins from samples from each group of mice. In the first dimension of electrophoresis, 200 μg protein was loaded on strips (240 mm, pH 3–10 non-linear); the second dimension of electrophoresis was a vertical 12.5% (w/v) SDS-PAGE (a) Untreated controls; (b) PBS controls; (c)–(e) 1, 3, and 6 h after injecting Con A intravenously, respectively

Fig. 2

Representative 2-DE maps of liver proteins from samples from each group of mice. In the first dimension of electrophoresis, 200 μg protein was loaded on strips (240 mm, pH 3–10 non-linear); the second dimension of electrophoresis was a vertical 12.5% (w/v) SDS-PAGE (a) Untreated controls; (b) PBS controls; (c)–(e) 1, 3, and 6 h after injecting Con A intravenously, respectively

Fig. 2

Representative 2-DE maps of liver proteins from samples from each group of mice. In the first dimension of electrophoresis, 200 μg protein was loaded on strips (240 mm, pH 3–10 non-linear); the second dimension of electrophoresis was a vertical 12.5% (w/v) SDS-PAGE (a) Untreated controls; (b) PBS controls; (c)–(e) 1, 3, and 6 h after injecting Con A intravenously, respectively

Fig. 2

Representative 2-DE maps of liver proteins from samples from each group of mice. In the first dimension of electrophoresis, 200 μg protein was loaded on strips (240 mm, pH 3–10 non-linear); the second dimension of electrophoresis was a vertical 12.5% (w/v) SDS-PAGE (a) Untreated controls; (b) PBS controls; (c)–(e) 1, 3, and 6 h after injecting Con A intravenously, respectively

Fig. 3

qRT-PCR analysis of selected proteins Relative expression of (a) RIKEN, (b) fbp1, and (c) khk at 1, 3 and 6 h after Con A injection compared with the levels in untreated controls

Fig. 3

qRT-PCR analysis of selected proteins Relative expression of (a) RIKEN, (b) fbp1, and (c) khk at 1, 3 and 6 h after Con A injection compared with the levels in untreated controls

Fig. 3

qRT-PCR analysis of selected proteins Relative expression of (a) RIKEN, (b) fbp1, and (c) khk at 1, 3 and 6 h after Con A injection compared with the levels in untreated controls