Knockout of secretin receptor reduces large cholangiocyte hyperplasia in mice with extrahepatic cholestasis induced by bile duct ligation.
Knockout of secretin receptor reduces large cholangiocyte hyperplasia in mice with extrahepatic cholestasis induced by bile duct ligation.
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DOI：
10.1002/hep.23657
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发表时间：
2010-07
期刊：
影响因子：
13.5
通讯作者：
Alpini, Gianfranco
中科院分区：
文献类型：
--
作者：
Glaser, Shannon;Lam, Ian P.;Franchitto, Antonio;Gaudio, Eugenio;Onori, Paolo;Chow, Billy K.;Wise, Candace;Kopriva, Shelley;Venter, Julie;White, Mellanie;Ueno, Yoshiyuki;Dostal, David;Carpino, Guido;Mancinelli, Romina;Butler, Wendy;Chiasson, Valorie;DeMorrow, Sharon;Francis, Heather;Alpini, Gianfranco
在胆管结扎（BDL）过程中，大胆管细胞的生长受到cAMP/ERK 1/2通路的调节，并与分泌素受体（SR）表达增加密切相关。尽管有人认为SR调节胆管细胞的生长，但缺乏分泌素依赖性增殖的直接证据。SR+/+（野生型，WT）或SR敲除（SR-/-）小鼠进行假手术或BDL 3或7天。我们评估了：（i）肝脏切片中的SR表达、胆管细胞增殖和凋亡;（ii）WT和SR−/− BDL小鼠纯化的大胆管细胞中的PCNA蛋白表达和ERK 1/2磷酸化。正常WT小鼠用促胰液素处理（2.5纳摩尔/Kg BW/天，通过渗透微型泵处理1周），并评价胆汁质量。使用小型和大型胆管细胞来评估促胰液素（100 nM）对增殖、PKA活性和ERK 1/2磷酸化的体外作用。SR表达也被稳定敲低的shRNA，和基础和促分泌素刺激的cAMP水平（胆汁生长的功能指数）和增殖进行了测定。大胆管细胞表达SR。SR基因敲除显著降低BDL诱导的大胆管细胞生长，这与增强的凋亡有关。与WT BDL小鼠相比，SR−/− BDL小鼠的大胆管细胞中PCNA表达和ERK 1/2磷酸化水平降低。对正常WT小鼠体内给予促胰液素增加了导管质量。在体外，促胰液素增加大胆管细胞的增殖，PKA活性和ERK 1/2磷酸化，这被PKA和MEK抑制剂阻断。SR表达的稳定敲低降低了基底胆管细胞增殖。SR是维持胆汁生长的重要营养调节因子。目前的研究为促胰液素治疗胆管减少性肝病的潜在用途提供了强有力的支持。
During bile duct ligation (BDL), the growth of large cholangiocytes is regulated by the cAMP/ERK1/2 pathway and is closely associated with increased secretin receptor (SR) expression. Although it has been suggested that the SR modulates cholangiocyte growth, direct evidence for secretin-dependent proliferation is lacking. SR+/+ (wild-type, WT) or SR knockout (SR−/−) mice underwent sham surgery or BDL for 3 or 7 days. We evaluated: (i) SR expression, cholangiocyte proliferation, and apoptosis in liver sections; and (ii) PCNA protein expression and ERK1/2 phosphorylation in purified large cholangiocytes from WT and SR−/− BDL mice. Normal WT mice were treated with secretin (2.5 nmoles/Kg BW/day by osmotic minipumps for 1 week) and biliary mass was evaluated. Small and large cholangiocytes were used to evaluate the in vitro effect of secretin (100 nM) on proliferation, PKA activity and ERK1/2 phosphorylation. SR expression was also stably knocked down by shRNA, and basal and secretin-stimulated cAMP levels (a functional index of biliary growth) and proliferation were determined. SR was expressed by large cholangiocytes. Knockout of SR significantly decreased large cholangiocyte growth induced by BDL, which was associated with enhanced apoptosis. PCNA expression and ERK1/2 phosphorylation were decreased in large cholangiocytes from SR−/− BDL compared to WT BDL mice. In vivo administration of secretin to normal WT mice increased ductal mass. In vitro, secretin increased proliferation, PKA activity and ERK1/2 phosphorylation of large cholangiocytes that was blocked by PKA and MEK inhibitors. Stable knockdown of SR expression reduced basal cholangiocyte proliferation. SR is an important trophic regulator sustaining biliary growth. The current study provides strong support for the potential use of secretin as a therapy for ductopenic liver diseases.
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影响因子：
25.7
作者：
Korner, Meike;Hayes, Gregory M.;Reubi, Jean Claude
通讯作者：
Reubi, Jean Claude
DOI：
10.1038/labinvest.2009.6
发表时间：
2009-04
期刊：
Laboratory investigation; a journal of technical methods and pathology
影响因子：
--
作者：
通讯作者：
--
DOI：
10.1152/ajpgi.1999.276.5.g1289
发表时间：
1999-05-01
影响因子：
4.5
作者：
LeSage, GD;Glaser, SS;Alpini, G
通讯作者：
Alpini, G
影响因子：
29.4
作者：
Miyoshi, H;Rust, C;Gores, GJ
通讯作者：
Gores, GJ
影响因子：
29.4
作者：
Alpini, G;Roberts, S;LaRusso, NF
通讯作者：
LaRusso, NF