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Hepatic SATB1 induces paracrine activation of hepatic stellate cells and is upregulated by HBx

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ABSTRACT

Chronic hepatitis B virus (HBV) infection is a major cause of chronic liver diseases, but its involvement in hepatic fibrogenesis remains unclear. Special AT-rich binding protein 1 (SATB1) has been implicated in reprogramming chromatin organization and transcription profiles in many cancers and non-cancer-related conditions. We found that hepatic SATB1 expression was significantly up-regulated in fibrotic tissues from chronic hepatitis B virus (HBV)-infected patients and HBV transgenic (HBV-Tg) mouse model. Knockdown of SATB1 in the liver significantly alleviated CCl4-induced fibrosis in HBV-Tg mouse model. Moreover, we suggested HBV encoded x protein (HBx) induced SATB1 expression through activation of JNK and ERK pathways. Enforced expression of SATB1 in hepatocytes promoted the activation and proliferation of hepatic stellate cells (HSCs) by secretion of connective tissue growth factor (CTGF), Interleukin-6 (IL-6) and platelet derived growth factor-A (PDGF-AA). Our findings demonstrated that HBx upregulated hepatic SATB1 which exerted pro-fibrotic effects by paracrine activation of stellate cells in HBV-related fibrosis.

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Knockdown of SATB1 ameliorates CCl4-induced fibrosis in HBV-Tg mouse model.(a) Adenovirus carrying shRNA against SATB1 (AdshSATB1) or negative control (AdshNC) was injected into HBV-Tg mouse model. Inhibition efficiency of SATB1 expression was examined by real-time PCR in the fibrotic livers from AdshSATB1 or AdshNC-treated mice (n = 6 in each group). **P < 0.01 vs. AdshNC group. (b) Western blot was used to detect the protein levels of SATB1 and α-SMA in liver tissues (n = 3 in each group). Full-length blots are included in the Supplementary Fig. S6. (c) Representative IHC staining was used to determine the expression of α-SMA in the fibrotic liver. H&E and Masson’s trichrome staining were used to show pathological conditions and collagen deposition (100×). (d) Plasmid levels of ALT and AST were detected by elisa Kit (n = 6). *P < 0.05 vs. AdshNC group. (e) Semi-quantitative analysis of Masson’s trichrome staining in the fibrotic livers from AdshSATB1 or AdshNC-treated mice (n = 6). *P < 0.05 vs. AdshNC group. (f) Real-time PCR was used to detect the expression of SATB1, α-SMA, COL1A1, IL-6, CTGF in liver tissues from HBV-Tg mice. (n = 6). *P < 0.05, **P < 0.01 vs. AdshNC group.
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f2: Knockdown of SATB1 ameliorates CCl4-induced fibrosis in HBV-Tg mouse model.(a) Adenovirus carrying shRNA against SATB1 (AdshSATB1) or negative control (AdshNC) was injected into HBV-Tg mouse model. Inhibition efficiency of SATB1 expression was examined by real-time PCR in the fibrotic livers from AdshSATB1 or AdshNC-treated mice (n = 6 in each group). **P < 0.01 vs. AdshNC group. (b) Western blot was used to detect the protein levels of SATB1 and α-SMA in liver tissues (n = 3 in each group). Full-length blots are included in the Supplementary Fig. S6. (c) Representative IHC staining was used to determine the expression of α-SMA in the fibrotic liver. H&E and Masson’s trichrome staining were used to show pathological conditions and collagen deposition (100×). (d) Plasmid levels of ALT and AST were detected by elisa Kit (n = 6). *P < 0.05 vs. AdshNC group. (e) Semi-quantitative analysis of Masson’s trichrome staining in the fibrotic livers from AdshSATB1 or AdshNC-treated mice (n = 6). *P < 0.05 vs. AdshNC group. (f) Real-time PCR was used to detect the expression of SATB1, α-SMA, COL1A1, IL-6, CTGF in liver tissues from HBV-Tg mice. (n = 6). *P < 0.05, **P < 0.01 vs. AdshNC group.

Mentions: To confirm the role of SATB1 in HBV-related liver fibrosis, we silenced SATB1 in vivo in a CCl4-induced fibrosis model of HBV-Tg mice. Adenovirus carrying a SATB1-specific (AdshSATB1) or control shRNA (AdshNC) was given through tail vein injection once a week after CCl4 treatment. Mice were sacrificed 2 weeks after adenovirus administration. Inhibition efficiency of SATB1 was confirmed by real-time PCR and western blot in whole liver extracts (Fig. 2a,b). Our results showed that knockdown of SATB1 led to reduced histological liver damage, as seen by H&E staining (Fig. 2c) and decreased ALT, AST serum levels (Fig. 2d). Furthermore, fibrosis development was attenuated after AdshSATB1 administration, as shown by Masson’s trichrome staining and decreased expression of the fibrotic marker a-SMA (Fig. 2c). Masson staining indicated that liver treated with AdshSATB1 had less ECM deposition (5.1% ± 0.013) compared with AdshNC controls (10.4% ± 0.025) as indicated by reduced ECM area by 50% in the CCl4-induced fibrosis model (Fig. 2e). Besides, the expression of fibrotic markers, including a-SMA, COL1A1 and CTGF as well as proinflammatory cytokines IL-6 were also decreased in AdshSATB1-treated group (Fig. 2f). These data collectively confirm that down-regulation of SATB1 inhibits hepatic fibrogenesis in HBV transgenic mice.


Hepatic SATB1 induces paracrine activation of hepatic stellate cells and is upregulated by HBx
Knockdown of SATB1 ameliorates CCl4-induced fibrosis in HBV-Tg mouse model.(a) Adenovirus carrying shRNA against SATB1 (AdshSATB1) or negative control (AdshNC) was injected into HBV-Tg mouse model. Inhibition efficiency of SATB1 expression was examined by real-time PCR in the fibrotic livers from AdshSATB1 or AdshNC-treated mice (n = 6 in each group). **P < 0.01 vs. AdshNC group. (b) Western blot was used to detect the protein levels of SATB1 and α-SMA in liver tissues (n = 3 in each group). Full-length blots are included in the Supplementary Fig. S6. (c) Representative IHC staining was used to determine the expression of α-SMA in the fibrotic liver. H&E and Masson’s trichrome staining were used to show pathological conditions and collagen deposition (100×). (d) Plasmid levels of ALT and AST were detected by elisa Kit (n = 6). *P < 0.05 vs. AdshNC group. (e) Semi-quantitative analysis of Masson’s trichrome staining in the fibrotic livers from AdshSATB1 or AdshNC-treated mice (n = 6). *P < 0.05 vs. AdshNC group. (f) Real-time PCR was used to detect the expression of SATB1, α-SMA, COL1A1, IL-6, CTGF in liver tissues from HBV-Tg mice. (n = 6). *P < 0.05, **P < 0.01 vs. AdshNC group.
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f2: Knockdown of SATB1 ameliorates CCl4-induced fibrosis in HBV-Tg mouse model.(a) Adenovirus carrying shRNA against SATB1 (AdshSATB1) or negative control (AdshNC) was injected into HBV-Tg mouse model. Inhibition efficiency of SATB1 expression was examined by real-time PCR in the fibrotic livers from AdshSATB1 or AdshNC-treated mice (n = 6 in each group). **P < 0.01 vs. AdshNC group. (b) Western blot was used to detect the protein levels of SATB1 and α-SMA in liver tissues (n = 3 in each group). Full-length blots are included in the Supplementary Fig. S6. (c) Representative IHC staining was used to determine the expression of α-SMA in the fibrotic liver. H&E and Masson’s trichrome staining were used to show pathological conditions and collagen deposition (100×). (d) Plasmid levels of ALT and AST were detected by elisa Kit (n = 6). *P < 0.05 vs. AdshNC group. (e) Semi-quantitative analysis of Masson’s trichrome staining in the fibrotic livers from AdshSATB1 or AdshNC-treated mice (n = 6). *P < 0.05 vs. AdshNC group. (f) Real-time PCR was used to detect the expression of SATB1, α-SMA, COL1A1, IL-6, CTGF in liver tissues from HBV-Tg mice. (n = 6). *P < 0.05, **P < 0.01 vs. AdshNC group.
Mentions: To confirm the role of SATB1 in HBV-related liver fibrosis, we silenced SATB1 in vivo in a CCl4-induced fibrosis model of HBV-Tg mice. Adenovirus carrying a SATB1-specific (AdshSATB1) or control shRNA (AdshNC) was given through tail vein injection once a week after CCl4 treatment. Mice were sacrificed 2 weeks after adenovirus administration. Inhibition efficiency of SATB1 was confirmed by real-time PCR and western blot in whole liver extracts (Fig. 2a,b). Our results showed that knockdown of SATB1 led to reduced histological liver damage, as seen by H&E staining (Fig. 2c) and decreased ALT, AST serum levels (Fig. 2d). Furthermore, fibrosis development was attenuated after AdshSATB1 administration, as shown by Masson’s trichrome staining and decreased expression of the fibrotic marker a-SMA (Fig. 2c). Masson staining indicated that liver treated with AdshSATB1 had less ECM deposition (5.1% ± 0.013) compared with AdshNC controls (10.4% ± 0.025) as indicated by reduced ECM area by 50% in the CCl4-induced fibrosis model (Fig. 2e). Besides, the expression of fibrotic markers, including a-SMA, COL1A1 and CTGF as well as proinflammatory cytokines IL-6 were also decreased in AdshSATB1-treated group (Fig. 2f). These data collectively confirm that down-regulation of SATB1 inhibits hepatic fibrogenesis in HBV transgenic mice.

View Article: PubMed Central - PubMed

ABSTRACT

Chronic hepatitis B virus (HBV) infection is a major cause of chronic liver diseases, but its involvement in hepatic fibrogenesis remains unclear. Special AT-rich binding protein 1 (SATB1) has been implicated in reprogramming chromatin organization and transcription profiles in many cancers and non-cancer-related conditions. We found that hepatic SATB1 expression was significantly up-regulated in fibrotic tissues from chronic hepatitis B virus (HBV)-infected patients and HBV transgenic (HBV-Tg) mouse model. Knockdown of SATB1 in the liver significantly alleviated CCl4-induced fibrosis in HBV-Tg mouse model. Moreover, we suggested HBV encoded x protein (HBx) induced SATB1 expression through activation of JNK and ERK pathways. Enforced expression of SATB1 in hepatocytes promoted the activation and proliferation of hepatic stellate cells (HSCs) by secretion of connective tissue growth factor (CTGF), Interleukin-6 (IL-6) and platelet derived growth factor-A (PDGF-AA). Our findings demonstrated that HBx upregulated hepatic SATB1 which exerted pro-fibrotic effects by paracrine activation of stellate cells in HBV-related fibrosis.

No MeSH data available.


Related in: MedlinePlus