Limits...
Tissue Transglutaminase-Regulated Transformed Growth Factor-β1 in the Parasite Links Schistosoma japonicum Infection with Liver Fibrosis.

Tang J, Zhu X, Zhao J, Fung M, Li Y, Gao Z, Yan S, Li X, Ji X, Su F, Li Z - Mediators Inflamm. (2015)

Bottom Line: During tTG inhibition, TGF-β1 level decreased in sera and liver of infected mice.The TGF-β1 mature peptide cDNA sequence and its extended sequence were amplified through RT-PCR and RACE-PCR using adult worms as template, and sequence is analyzed and loaded to NCBI GenBank (number GQ338152.1).TGF-β1 transcript in Sj eggs was higher than in adult worms.

View Article: PubMed Central - PubMed

Affiliation: Guangzhou Hoffmann Institute of Immunology, School of Basic Sciences, Guangzhou Medical University, Guangzhou 510182, China.

ABSTRACT
Transforming growth factor (TGF-β1) is among the strongest factors of liver fibrogenesis, but its association with Schistosoma-caused liver fibrosis is controversial. Tissue transglutaminase (tTG) is the principal enzyme controlling TGF-β1 maturation and contributes to Sj-infected liver fibrosis. Here we aim to explore the consistency between tTG and TGF-β1 and TGF-β1 source and its correlation with liver fibrosis after Sj-infection. TGF-β1 was upregulated at weeks 6 and 8 upon liver fibrosis induction. During tTG inhibition, TGF-β1 level decreased in sera and liver of infected mice. TGF-β1 showed positive staining in liver containing Sj adult worms and eggs. TGF-β1 was also detected in Sj adult worm sections, soluble egg antigen and Sj adult worm antigen, and adult worms' culture medium. The TGF-β1 mature peptide cDNA sequence and its extended sequence were amplified through RT-PCR and RACE-PCR using adult worms as template, and sequence is analyzed and loaded to NCBI GenBank (number GQ338152.1). TGF-β1 transcript in Sj eggs was higher than in adult worms. In Sj-infected liver, transcriptional level of TGF-β1 from Sj, but not mouse liver, correlated with liver fibrosis extent. This study provides evidence that tTG regulates TGF-β1 and illustrates the importance of targeting tTG in treating Sj infection-induced fibrosis.

No MeSH data available.


Related in: MedlinePlus

TGF-β1 level increased gradually in mice liver during the courses of Sj infection. (a) TGF-β1 level in serum was determined by ELISA in BALB/c mice with 20 ± 3 infective Sj cercariae for 5, 6, 8, and 12 weeks, and uninfected mice were used as control. Data are shown as means ± SD of 10 mice/group. Experiment was performed four times (∗P < 0.05; and ∗∗P < 0.01 compared with uninfected group). (b) Equal amounts of proteins of mouse liver tissue lysates at indicated time points were used in the Western blot assay to detect protein expression levels of TGF-β1, p-Smad2, and Smad2. GAPDH was used as loading control. (c) Mouse livers at indicated time points were fixed in paraformaldehyde, embedded in paraffin, and then sliced and immunohistochemically stained for TGF-β1. Representative staining for TGF-β1 is shown at ×400 magnification. Top: TGF-β1 expression in egg, Sj egg granuloma, and the surrounding tissue. Bottom: TGF-β1 expression in hepatic cell and liver tissue around the liver sinusoid.
© Copyright Policy - open-access
Related In: Results  -  Collection


getmorefigures.php?uid=PMC4493306&req=5

fig1: TGF-β1 level increased gradually in mice liver during the courses of Sj infection. (a) TGF-β1 level in serum was determined by ELISA in BALB/c mice with 20 ± 3 infective Sj cercariae for 5, 6, 8, and 12 weeks, and uninfected mice were used as control. Data are shown as means ± SD of 10 mice/group. Experiment was performed four times (∗P < 0.05; and ∗∗P < 0.01 compared with uninfected group). (b) Equal amounts of proteins of mouse liver tissue lysates at indicated time points were used in the Western blot assay to detect protein expression levels of TGF-β1, p-Smad2, and Smad2. GAPDH was used as loading control. (c) Mouse livers at indicated time points were fixed in paraformaldehyde, embedded in paraffin, and then sliced and immunohistochemically stained for TGF-β1. Representative staining for TGF-β1 is shown at ×400 magnification. Top: TGF-β1 expression in egg, Sj egg granuloma, and the surrounding tissue. Bottom: TGF-β1 expression in hepatic cell and liver tissue around the liver sinusoid.

Mentions: We previously reported a high extent of post-Sj-infection hepatic fibrosis in mice. Liver granuloma began at week 5, and fibrosis progressed most seriously at week 8, whereas chronic liver fibrosis appeared at week 12 [34]. TGF-β1 was usually the key factor in inducing liver fibrosis compared with other causes [2, 3]. TGF-β1 concentration in mouse serum increased 5, 6, and 8 weeks after Sj infection, and the highest level was observed at week 6 (Figure 1(a)). Western blot analysis and IHC assay revealed that TGF-β1 protein level in Sj-infected mouse liver also increased (Figures 1(b) and 2(c)). In addition, Smad2, the downstream signaling protein of TGF-β1 pathway, was also activated in mice liver after Sj infection (Figure 1(b)). TGF-β1 could be involved in liver fibrosis in a Smad2-dependent manner during Sj infection. TGF-β1 was localized either in the cells of blood vessels where Sj adult worms reside or in the eggs of Sj and in the cells of liver tissue where eggs are deposited (Figure 1(c)). The results suggested that TGF-β1 likely promoted hepatic fibrosis after Sj infection.


Tissue Transglutaminase-Regulated Transformed Growth Factor-β1 in the Parasite Links Schistosoma japonicum Infection with Liver Fibrosis.

Tang J, Zhu X, Zhao J, Fung M, Li Y, Gao Z, Yan S, Li X, Ji X, Su F, Li Z - Mediators Inflamm. (2015)

TGF-β1 level increased gradually in mice liver during the courses of Sj infection. (a) TGF-β1 level in serum was determined by ELISA in BALB/c mice with 20 ± 3 infective Sj cercariae for 5, 6, 8, and 12 weeks, and uninfected mice were used as control. Data are shown as means ± SD of 10 mice/group. Experiment was performed four times (∗P < 0.05; and ∗∗P < 0.01 compared with uninfected group). (b) Equal amounts of proteins of mouse liver tissue lysates at indicated time points were used in the Western blot assay to detect protein expression levels of TGF-β1, p-Smad2, and Smad2. GAPDH was used as loading control. (c) Mouse livers at indicated time points were fixed in paraformaldehyde, embedded in paraffin, and then sliced and immunohistochemically stained for TGF-β1. Representative staining for TGF-β1 is shown at ×400 magnification. Top: TGF-β1 expression in egg, Sj egg granuloma, and the surrounding tissue. Bottom: TGF-β1 expression in hepatic cell and liver tissue around the liver sinusoid.
© Copyright Policy - open-access
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC4493306&req=5

fig1: TGF-β1 level increased gradually in mice liver during the courses of Sj infection. (a) TGF-β1 level in serum was determined by ELISA in BALB/c mice with 20 ± 3 infective Sj cercariae for 5, 6, 8, and 12 weeks, and uninfected mice were used as control. Data are shown as means ± SD of 10 mice/group. Experiment was performed four times (∗P < 0.05; and ∗∗P < 0.01 compared with uninfected group). (b) Equal amounts of proteins of mouse liver tissue lysates at indicated time points were used in the Western blot assay to detect protein expression levels of TGF-β1, p-Smad2, and Smad2. GAPDH was used as loading control. (c) Mouse livers at indicated time points were fixed in paraformaldehyde, embedded in paraffin, and then sliced and immunohistochemically stained for TGF-β1. Representative staining for TGF-β1 is shown at ×400 magnification. Top: TGF-β1 expression in egg, Sj egg granuloma, and the surrounding tissue. Bottom: TGF-β1 expression in hepatic cell and liver tissue around the liver sinusoid.
Mentions: We previously reported a high extent of post-Sj-infection hepatic fibrosis in mice. Liver granuloma began at week 5, and fibrosis progressed most seriously at week 8, whereas chronic liver fibrosis appeared at week 12 [34]. TGF-β1 was usually the key factor in inducing liver fibrosis compared with other causes [2, 3]. TGF-β1 concentration in mouse serum increased 5, 6, and 8 weeks after Sj infection, and the highest level was observed at week 6 (Figure 1(a)). Western blot analysis and IHC assay revealed that TGF-β1 protein level in Sj-infected mouse liver also increased (Figures 1(b) and 2(c)). In addition, Smad2, the downstream signaling protein of TGF-β1 pathway, was also activated in mice liver after Sj infection (Figure 1(b)). TGF-β1 could be involved in liver fibrosis in a Smad2-dependent manner during Sj infection. TGF-β1 was localized either in the cells of blood vessels where Sj adult worms reside or in the eggs of Sj and in the cells of liver tissue where eggs are deposited (Figure 1(c)). The results suggested that TGF-β1 likely promoted hepatic fibrosis after Sj infection.

Bottom Line: During tTG inhibition, TGF-β1 level decreased in sera and liver of infected mice.The TGF-β1 mature peptide cDNA sequence and its extended sequence were amplified through RT-PCR and RACE-PCR using adult worms as template, and sequence is analyzed and loaded to NCBI GenBank (number GQ338152.1).TGF-β1 transcript in Sj eggs was higher than in adult worms.

View Article: PubMed Central - PubMed

Affiliation: Guangzhou Hoffmann Institute of Immunology, School of Basic Sciences, Guangzhou Medical University, Guangzhou 510182, China.

ABSTRACT
Transforming growth factor (TGF-β1) is among the strongest factors of liver fibrogenesis, but its association with Schistosoma-caused liver fibrosis is controversial. Tissue transglutaminase (tTG) is the principal enzyme controlling TGF-β1 maturation and contributes to Sj-infected liver fibrosis. Here we aim to explore the consistency between tTG and TGF-β1 and TGF-β1 source and its correlation with liver fibrosis after Sj-infection. TGF-β1 was upregulated at weeks 6 and 8 upon liver fibrosis induction. During tTG inhibition, TGF-β1 level decreased in sera and liver of infected mice. TGF-β1 showed positive staining in liver containing Sj adult worms and eggs. TGF-β1 was also detected in Sj adult worm sections, soluble egg antigen and Sj adult worm antigen, and adult worms' culture medium. The TGF-β1 mature peptide cDNA sequence and its extended sequence were amplified through RT-PCR and RACE-PCR using adult worms as template, and sequence is analyzed and loaded to NCBI GenBank (number GQ338152.1). TGF-β1 transcript in Sj eggs was higher than in adult worms. In Sj-infected liver, transcriptional level of TGF-β1 from Sj, but not mouse liver, correlated with liver fibrosis extent. This study provides evidence that tTG regulates TGF-β1 and illustrates the importance of targeting tTG in treating Sj infection-induced fibrosis.

No MeSH data available.


Related in: MedlinePlus