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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

Changes in expression levels of Trps1 and EMT markers in HIBECs following CPRI.A: RT-PCR results; B–F: semi-quantitative PCR analysis of Trps1, E-cadherin, CK19, Vimentin, and α-SMA mRNA expression. Lane 1: control group; 2: CP for 12 h, RI for 1 h; 3: CP for 24 h, RI for 1 h; 4: CP for 48 h, RI for 1 h. CP, cold preservation. RI, reperfusion injury.
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pone.0123233.g003: Changes in expression levels of Trps1 and EMT markers in HIBECs following CPRI.A: RT-PCR results; B–F: semi-quantitative PCR analysis of Trps1, E-cadherin, CK19, Vimentin, and α-SMA mRNA expression. Lane 1: control group; 2: CP for 12 h, RI for 1 h; 3: CP for 24 h, RI for 1 h; 4: CP for 48 h, RI for 1 h. CP, cold preservation. RI, reperfusion injury.

Mentions: The Trps1 protein was primarily seen in the nucleus, and to a lesser extent in the cytoplasm, of cultured HIBECs (Fig 3). Following 12 h of CP, expression levels of the Trps1 gene and protein were significantly increased (p = 0.018 and 0.031, respectively) in HIBECs compared with those in untreated controls. In contrast, these were significantly decreased in HIBECs subjected to 24- or 48-h CP (p = 0.026 and 0.009, respectively), with a continued decrease observed after 48 h of CP (Fig 4).


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)

Changes in expression levels of Trps1 and EMT markers in HIBECs following CPRI.A: RT-PCR results; B–F: semi-quantitative PCR analysis of Trps1, E-cadherin, CK19, Vimentin, and α-SMA mRNA expression. Lane 1: control group; 2: CP for 12 h, RI for 1 h; 3: CP for 24 h, RI for 1 h; 4: CP for 48 h, RI for 1 h. CP, cold preservation. RI, reperfusion injury.
© Copyright Policy
Related In: Results  -  Collection

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

pone.0123233.g003: Changes in expression levels of Trps1 and EMT markers in HIBECs following CPRI.A: RT-PCR results; B–F: semi-quantitative PCR analysis of Trps1, E-cadherin, CK19, Vimentin, and α-SMA mRNA expression. Lane 1: control group; 2: CP for 12 h, RI for 1 h; 3: CP for 24 h, RI for 1 h; 4: CP for 48 h, RI for 1 h. CP, cold preservation. RI, reperfusion injury.
Mentions: The Trps1 protein was primarily seen in the nucleus, and to a lesser extent in the cytoplasm, of cultured HIBECs (Fig 3). Following 12 h of CP, expression levels of the Trps1 gene and protein were significantly increased (p = 0.018 and 0.031, respectively) in HIBECs compared with those in untreated controls. In contrast, these were significantly decreased in HIBECs subjected to 24- or 48-h CP (p = 0.026 and 0.009, respectively), with a continued decrease observed after 48 h of CP (Fig 4).

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