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Trps1 regulates biliary epithelial-mesenchymal transition and has roles during biliary fibrosis in liver grafts: a preliminary study.

Zhe C, Yu F, Tian J, Zheng S - PLoS ONE (2015)

Bottom Line: Mesenchymal markers were seen in biliary epithelial cells (BECs), with collagen deposited around the bile duct.Expression of epithelial marker mRNAs and proteins in HIBECs decreased with prolonged cold preservation (CP), while mesenchymal marker expression increased.Expression of E-cadherin was increased in HIBECs following Trps1 adenovirus infection and CP/reperfusion injury (CPRI), with vimentin expression levels reduced and CPRI-mediated epithelial-mesenchymal transition (EMT) inhibited.

View Article: PubMed Central - PubMed

Affiliation: Institute of Hepatobiliary Surgery, Southwest Hospital, Third Military Medical University, No. 29 Gaotanyan Road, Shapingba District, Chongqing, 400038, China.

ABSTRACT

Objective: To investigate the role(s) of Trps1 in non-anastomotic biliary stricture (NABS) following liver transplantation.

Methods: Immunohistochemical and histological techniques were used to detect Trps1, E-cadherin, CK19, vimentin, α-SMA, and collagen deposition. Human intrahepatic biliary epithelial cells (HIBECs) were infected with a Trps1 adenovirus, or transfected with Trps1 short-interfering RNAs (siRNAs). Reverse transcription polymerase chain reaction (RT-PCR) assays and western blotting were used to determine expression levels of epithelial and mesenchymal markers, and Trps1 in HIBECs.

Results: Expression of Trps1 and epithelial markers was down-regulated or absent in NABS liver samples. Mesenchymal markers were seen in biliary epithelial cells (BECs), with collagen deposited around the bile duct. Trps1 expression positively correlated with epithelial markers. Expression of epithelial marker mRNAs and proteins in HIBECs decreased with prolonged cold preservation (CP), while mesenchymal marker expression increased. A 12-h CP period led to increased Trps1 mRNA and protein levels. Expression of E-cadherin was increased in HIBECs following Trps1 adenovirus infection and CP/reperfusion injury (CPRI), with vimentin expression levels reduced and CPRI-mediated epithelial-mesenchymal transition (EMT) inhibited. Transfection of HIBECs with Trps1 siRNAs in conjunction with CPRI revealed that E-cadherin expression was decreased, vimentin expression was increased, and CPRI-mediated EMT was promoted.

Conclusion: Trps1 is involved in NABS pathogenesis following liver transplantation and negatively correlates with BEC EMT and biliary fibrosis in liver grafts. Trps1 demonstrates antagonistic effects that could reverse EMT.

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Related in: MedlinePlus

Trps1 expression in cultured HIBECs.Left: cultured HIBECs; Right: immunohistochemical staining of Trps1 in cultured HIBECs.
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pone.0123233.g002: Trps1 expression in cultured HIBECs.Left: cultured HIBECs; Right: immunohistochemical staining of Trps1 in cultured HIBECs.

Mentions: Positive staining for E-cadherin and CK19 was seen on the cellular membrane and in the cytoplasm. In control groups, epithelial markers were normally expressed in all BECs In the NABS group, expression of some of the epithelial markers was absent Vimentin and α-SMA were expressed at variable levels in the NABS group and not detected in the control group Trps1 was mainly expressed in the nuclei of BECs, with a small fraction expressed in the cytoplasm. In the control group, Trps1s was normally expressed in almost all BECs however in the NABS group, biliary epithelial Trps1 expression was less extensive (Fig 2). In the NABS group, bile duct hyperplasia was distinct, and a large amount of collagen was deposited around the bile ducts. There appeared to be severe damage to BECs as distinguished by altered cell morphologies. The BECs in the control group had normal morphologies and very little collagen deposited around the bile duct. Samples from NABS patients exhibited significantly reduced expression levels of E-cadherin, CK-19, Vimentin, α-SMA, and Trps1 (P < 0.01; Table 3) in comparison with those for the control group. Significant differences in biliary fibrosis were also seen between the two groups (P < 0.01), with NABS samples having more severe bile duct fibrosis (Table 4). In NABS samples, Trps1 expression levels positively correlated with epithelial marker expression, and negatively correlated with mesenchymal marker expression, and degree of biliary fibrosis in the liver graft (Table 5).


Trps1 regulates biliary epithelial-mesenchymal transition and has roles during biliary fibrosis in liver grafts: a preliminary study.

Zhe C, Yu F, Tian J, Zheng S - PLoS ONE (2015)

Trps1 expression in cultured HIBECs.Left: cultured HIBECs; Right: immunohistochemical staining of Trps1 in cultured HIBECs.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4401436&req=5

pone.0123233.g002: Trps1 expression in cultured HIBECs.Left: cultured HIBECs; Right: immunohistochemical staining of Trps1 in cultured HIBECs.
Mentions: Positive staining for E-cadherin and CK19 was seen on the cellular membrane and in the cytoplasm. In control groups, epithelial markers were normally expressed in all BECs In the NABS group, expression of some of the epithelial markers was absent Vimentin and α-SMA were expressed at variable levels in the NABS group and not detected in the control group Trps1 was mainly expressed in the nuclei of BECs, with a small fraction expressed in the cytoplasm. In the control group, Trps1s was normally expressed in almost all BECs however in the NABS group, biliary epithelial Trps1 expression was less extensive (Fig 2). In the NABS group, bile duct hyperplasia was distinct, and a large amount of collagen was deposited around the bile ducts. There appeared to be severe damage to BECs as distinguished by altered cell morphologies. The BECs in the control group had normal morphologies and very little collagen deposited around the bile duct. Samples from NABS patients exhibited significantly reduced expression levels of E-cadherin, CK-19, Vimentin, α-SMA, and Trps1 (P < 0.01; Table 3) in comparison with those for the control group. Significant differences in biliary fibrosis were also seen between the two groups (P < 0.01), with NABS samples having more severe bile duct fibrosis (Table 4). In NABS samples, Trps1 expression levels positively correlated with epithelial marker expression, and negatively correlated with mesenchymal marker expression, and degree of biliary fibrosis in the liver graft (Table 5).

Bottom Line: Mesenchymal markers were seen in biliary epithelial cells (BECs), with collagen deposited around the bile duct.Expression of epithelial marker mRNAs and proteins in HIBECs decreased with prolonged cold preservation (CP), while mesenchymal marker expression increased.Expression of E-cadherin was increased in HIBECs following Trps1 adenovirus infection and CP/reperfusion injury (CPRI), with vimentin expression levels reduced and CPRI-mediated epithelial-mesenchymal transition (EMT) inhibited.

View Article: PubMed Central - PubMed

Affiliation: Institute of Hepatobiliary Surgery, Southwest Hospital, Third Military Medical University, No. 29 Gaotanyan Road, Shapingba District, Chongqing, 400038, China.

ABSTRACT

Objective: To investigate the role(s) of Trps1 in non-anastomotic biliary stricture (NABS) following liver transplantation.

Methods: Immunohistochemical and histological techniques were used to detect Trps1, E-cadherin, CK19, vimentin, α-SMA, and collagen deposition. Human intrahepatic biliary epithelial cells (HIBECs) were infected with a Trps1 adenovirus, or transfected with Trps1 short-interfering RNAs (siRNAs). Reverse transcription polymerase chain reaction (RT-PCR) assays and western blotting were used to determine expression levels of epithelial and mesenchymal markers, and Trps1 in HIBECs.

Results: Expression of Trps1 and epithelial markers was down-regulated or absent in NABS liver samples. Mesenchymal markers were seen in biliary epithelial cells (BECs), with collagen deposited around the bile duct. Trps1 expression positively correlated with epithelial markers. Expression of epithelial marker mRNAs and proteins in HIBECs decreased with prolonged cold preservation (CP), while mesenchymal marker expression increased. A 12-h CP period led to increased Trps1 mRNA and protein levels. Expression of E-cadherin was increased in HIBECs following Trps1 adenovirus infection and CP/reperfusion injury (CPRI), with vimentin expression levels reduced and CPRI-mediated epithelial-mesenchymal transition (EMT) inhibited. Transfection of HIBECs with Trps1 siRNAs in conjunction with CPRI revealed that E-cadherin expression was decreased, vimentin expression was increased, and CPRI-mediated EMT was promoted.

Conclusion: Trps1 is involved in NABS pathogenesis following liver transplantation and negatively correlates with BEC EMT and biliary fibrosis in liver grafts. Trps1 demonstrates antagonistic effects that could reverse EMT.

Show MeSH
Related in: MedlinePlus