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Chinese Herbal Preparation Xuebijing Potently Inhibits Inflammasome Activation in Hepatocytes and Ameliorates Mouse Liver Ischemia-Reperfusion Injury.

Liu X, Hu Z, Zhou B, Li X, Tao R - PLoS ONE (2015)

Bottom Line: The Chinese herb preparation Xuebijing injection (XBJ) has been widely used in the management of various septic disorders or inflammation-related conditions, however the molecular mechanism of its anti-inflammatory effect remains largely elusive.Our data clearly demonstrated that XBJ potently inhibited apoptosis, as well as caspase-1 cleavage and IL-1β production in a time- and dose-dependent manner in isolated hepatocytes, suggesting that in addition to its known modulatory effect on NF-κB-dependent inflammatory gene expression, it also has a direct impact on hepatocyte inflammasome activation.The current study not only deepens our understanding of how XBJ ameliorates inflammation and apoptosis, but also has immediate practical significance in many clinical situations such as partial hepatectomy, liver transplantation, etc.

View Article: PubMed Central - PubMed

Affiliation: Department of Surgery, Ruijin Hospital, Shanghai Jiaotong University School of Medicine, Shanghai, PR China.

ABSTRACT
The Chinese herb preparation Xuebijing injection (XBJ) has been widely used in the management of various septic disorders or inflammation-related conditions, however the molecular mechanism of its anti-inflammatory effect remains largely elusive. In the current study, we found that XBJ treatment potently ameliorated mouse hepatic ischemia-reperfusion (IR) injury, manifested as decreased liver function tests (LDH, ALT, AST), improved inflammation and less hepatocyte apoptosis. Notably, XBJ markedly inhibited inflammasome activation and IL-1 production in mouse livers subjected to IRI, even in the absence of Kupffer cells, suggesting Kupffer cells are not necessary for hepatic inflammasome activation upon Redox-induced sterile inflammation. This finding led us to investigate the role of XBJ on hepatocyte apoptosis and inflammasome activation using an in vitro hydrogen peroxide (H2O2)-triggered hepatocyte injury model. Our data clearly demonstrated that XBJ potently inhibited apoptosis, as well as caspase-1 cleavage and IL-1β production in a time- and dose-dependent manner in isolated hepatocytes, suggesting that in addition to its known modulatory effect on NF-κB-dependent inflammatory gene expression, it also has a direct impact on hepatocyte inflammasome activation. The current study not only deepens our understanding of how XBJ ameliorates inflammation and apoptosis, but also has immediate practical significance in many clinical situations such as partial hepatectomy, liver transplantation, etc.

No MeSH data available.


Related in: MedlinePlus

XBJ significantly ameliorated inflammation in mouse livers that have undergone IRI.B6 mice were either sham operated (sham group) or subjected to 90 minutes of partial warm ischemia, followed by 6 hours of reperfusion. XBJ (2 ml/kg iv) was given either before ischemia (before group) or after initiation of reperfusion (after group), while the same volume of normal saline was administered in the vehicle group. Serum and liver samples were harvested (n = 3-6/group). (A) qRT-PCR detection of CXCL-1, CXCL-10, IL-1β, IL-6 and TNF-a. Data were normalized to HPRT gene expression and expressed as fold increase above the sham group (set as 1). (B) ELISA detection of MCP-1, IL-6 and TNF-α. Accumulation of neutrophils and macrophages after administration of XBJ was analyzed by (C) Ly-6G (neutrophils), (D) CD68 (macrophages) immunohistochemical staining (Original magnifications, 200×magnification) and (E) MPO levels in IRI liver lobes. (F) ROS was detected using the fluorescent probe DCFH-DA. Data were expressed as mean±SEM. *P<0.05, **P<0.01,*** P <0.001.
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pone.0131436.g002: XBJ significantly ameliorated inflammation in mouse livers that have undergone IRI.B6 mice were either sham operated (sham group) or subjected to 90 minutes of partial warm ischemia, followed by 6 hours of reperfusion. XBJ (2 ml/kg iv) was given either before ischemia (before group) or after initiation of reperfusion (after group), while the same volume of normal saline was administered in the vehicle group. Serum and liver samples were harvested (n = 3-6/group). (A) qRT-PCR detection of CXCL-1, CXCL-10, IL-1β, IL-6 and TNF-a. Data were normalized to HPRT gene expression and expressed as fold increase above the sham group (set as 1). (B) ELISA detection of MCP-1, IL-6 and TNF-α. Accumulation of neutrophils and macrophages after administration of XBJ was analyzed by (C) Ly-6G (neutrophils), (D) CD68 (macrophages) immunohistochemical staining (Original magnifications, 200×magnification) and (E) MPO levels in IRI liver lobes. (F) ROS was detected using the fluorescent probe DCFH-DA. Data were expressed as mean±SEM. *P<0.05, **P<0.01,*** P <0.001.

Mentions: IR elicited robust innate immunity within the liver tissues, so we checked the cytokine profile within the IR lobes as well as serum levels of several pro-inflammatory cytokines/chemokines. A previous study has shown that XBJ exerted its effects via inhibition of the NF-κB-dependent transcription of several pro-inflammatory genes, such IL‑6 and TLR4 [12,15]. Consistent with this effects, we found that XBJ treatment either before partial warm ischemia or after reperfusion dramatically decreased the intra-hepatic expression of cytokine IL-1β, IL-6, TNF-α as well as the chemokines C-X-C motif ligand (CXCL)1 and CXCL10 (Fig 2A). The serum levels of MCP-1, IL-6 and TNF-α were also dramatically decreased (Fig 2B). To further prove that the intra-hepatic infiltration of pro-inflammatory cells were decreased after use of XBJ, we used immunohistochemical staining and found the intra-hepatic expression of LY6G which represented neutrophils and CD68 which represented macrophages were also significantly decreased (Fig 2C and 2D). the MPO activity within the IR liver lobes was dramatically decreased after use of XBJ (Fig 2E). Lastly, Hepatic ROS level was measured using DCFH-DA as a probe. The hepatic level of DCFH fluorescence was significantly increased after IR, and the increase of hepatic DCFH fluorescence was prevented in the IR liver lobes treated with XBJ (Fig 2F). Given IL-1β is mainly generated through inflammasome activation, we next examined whether XBJ treatment affected the inflammasome related protein expression. XBJ treatment did not significantly alter the expression of NLRP3 and ASC per se, two main components of the inflammasome protein complex. However, the cleavage of pro-Caspase-1 and pro-IL-1β which was known to be the key step in the processing of mature IL-1β within the inflammasome was significantly decreased (Fig 3). These data clearly indicated that XBJ attenuate liver IRI by minimizing both hepatocyte apoptosis as well as inflammasome activation within the IR liver.


Chinese Herbal Preparation Xuebijing Potently Inhibits Inflammasome Activation in Hepatocytes and Ameliorates Mouse Liver Ischemia-Reperfusion Injury.

Liu X, Hu Z, Zhou B, Li X, Tao R - PLoS ONE (2015)

XBJ significantly ameliorated inflammation in mouse livers that have undergone IRI.B6 mice were either sham operated (sham group) or subjected to 90 minutes of partial warm ischemia, followed by 6 hours of reperfusion. XBJ (2 ml/kg iv) was given either before ischemia (before group) or after initiation of reperfusion (after group), while the same volume of normal saline was administered in the vehicle group. Serum and liver samples were harvested (n = 3-6/group). (A) qRT-PCR detection of CXCL-1, CXCL-10, IL-1β, IL-6 and TNF-a. Data were normalized to HPRT gene expression and expressed as fold increase above the sham group (set as 1). (B) ELISA detection of MCP-1, IL-6 and TNF-α. Accumulation of neutrophils and macrophages after administration of XBJ was analyzed by (C) Ly-6G (neutrophils), (D) CD68 (macrophages) immunohistochemical staining (Original magnifications, 200×magnification) and (E) MPO levels in IRI liver lobes. (F) ROS was detected using the fluorescent probe DCFH-DA. Data were expressed as mean±SEM. *P<0.05, **P<0.01,*** P <0.001.
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Related In: Results  -  Collection

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pone.0131436.g002: XBJ significantly ameliorated inflammation in mouse livers that have undergone IRI.B6 mice were either sham operated (sham group) or subjected to 90 minutes of partial warm ischemia, followed by 6 hours of reperfusion. XBJ (2 ml/kg iv) was given either before ischemia (before group) or after initiation of reperfusion (after group), while the same volume of normal saline was administered in the vehicle group. Serum and liver samples were harvested (n = 3-6/group). (A) qRT-PCR detection of CXCL-1, CXCL-10, IL-1β, IL-6 and TNF-a. Data were normalized to HPRT gene expression and expressed as fold increase above the sham group (set as 1). (B) ELISA detection of MCP-1, IL-6 and TNF-α. Accumulation of neutrophils and macrophages after administration of XBJ was analyzed by (C) Ly-6G (neutrophils), (D) CD68 (macrophages) immunohistochemical staining (Original magnifications, 200×magnification) and (E) MPO levels in IRI liver lobes. (F) ROS was detected using the fluorescent probe DCFH-DA. Data were expressed as mean±SEM. *P<0.05, **P<0.01,*** P <0.001.
Mentions: IR elicited robust innate immunity within the liver tissues, so we checked the cytokine profile within the IR lobes as well as serum levels of several pro-inflammatory cytokines/chemokines. A previous study has shown that XBJ exerted its effects via inhibition of the NF-κB-dependent transcription of several pro-inflammatory genes, such IL‑6 and TLR4 [12,15]. Consistent with this effects, we found that XBJ treatment either before partial warm ischemia or after reperfusion dramatically decreased the intra-hepatic expression of cytokine IL-1β, IL-6, TNF-α as well as the chemokines C-X-C motif ligand (CXCL)1 and CXCL10 (Fig 2A). The serum levels of MCP-1, IL-6 and TNF-α were also dramatically decreased (Fig 2B). To further prove that the intra-hepatic infiltration of pro-inflammatory cells were decreased after use of XBJ, we used immunohistochemical staining and found the intra-hepatic expression of LY6G which represented neutrophils and CD68 which represented macrophages were also significantly decreased (Fig 2C and 2D). the MPO activity within the IR liver lobes was dramatically decreased after use of XBJ (Fig 2E). Lastly, Hepatic ROS level was measured using DCFH-DA as a probe. The hepatic level of DCFH fluorescence was significantly increased after IR, and the increase of hepatic DCFH fluorescence was prevented in the IR liver lobes treated with XBJ (Fig 2F). Given IL-1β is mainly generated through inflammasome activation, we next examined whether XBJ treatment affected the inflammasome related protein expression. XBJ treatment did not significantly alter the expression of NLRP3 and ASC per se, two main components of the inflammasome protein complex. However, the cleavage of pro-Caspase-1 and pro-IL-1β which was known to be the key step in the processing of mature IL-1β within the inflammasome was significantly decreased (Fig 3). These data clearly indicated that XBJ attenuate liver IRI by minimizing both hepatocyte apoptosis as well as inflammasome activation within the IR liver.

Bottom Line: The Chinese herb preparation Xuebijing injection (XBJ) has been widely used in the management of various septic disorders or inflammation-related conditions, however the molecular mechanism of its anti-inflammatory effect remains largely elusive.Our data clearly demonstrated that XBJ potently inhibited apoptosis, as well as caspase-1 cleavage and IL-1β production in a time- and dose-dependent manner in isolated hepatocytes, suggesting that in addition to its known modulatory effect on NF-κB-dependent inflammatory gene expression, it also has a direct impact on hepatocyte inflammasome activation.The current study not only deepens our understanding of how XBJ ameliorates inflammation and apoptosis, but also has immediate practical significance in many clinical situations such as partial hepatectomy, liver transplantation, etc.

View Article: PubMed Central - PubMed

Affiliation: Department of Surgery, Ruijin Hospital, Shanghai Jiaotong University School of Medicine, Shanghai, PR China.

ABSTRACT
The Chinese herb preparation Xuebijing injection (XBJ) has been widely used in the management of various septic disorders or inflammation-related conditions, however the molecular mechanism of its anti-inflammatory effect remains largely elusive. In the current study, we found that XBJ treatment potently ameliorated mouse hepatic ischemia-reperfusion (IR) injury, manifested as decreased liver function tests (LDH, ALT, AST), improved inflammation and less hepatocyte apoptosis. Notably, XBJ markedly inhibited inflammasome activation and IL-1 production in mouse livers subjected to IRI, even in the absence of Kupffer cells, suggesting Kupffer cells are not necessary for hepatic inflammasome activation upon Redox-induced sterile inflammation. This finding led us to investigate the role of XBJ on hepatocyte apoptosis and inflammasome activation using an in vitro hydrogen peroxide (H2O2)-triggered hepatocyte injury model. Our data clearly demonstrated that XBJ potently inhibited apoptosis, as well as caspase-1 cleavage and IL-1β production in a time- and dose-dependent manner in isolated hepatocytes, suggesting that in addition to its known modulatory effect on NF-κB-dependent inflammatory gene expression, it also has a direct impact on hepatocyte inflammasome activation. The current study not only deepens our understanding of how XBJ ameliorates inflammation and apoptosis, but also has immediate practical significance in many clinical situations such as partial hepatectomy, liver transplantation, etc.

No MeSH data available.


Related in: MedlinePlus