Activation of Trpv4 reduces the hyperproliferative phenotype of cystic cholangiocytes from an animal model of ARPKD.
Activation of Trpv4 reduces the hyperproliferative phenotype of cystic cholangiocytes from an animal model of ARPKD.
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DOI：
10.1053/j.gastro.2010.04.010
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发表时间：
2010-07
期刊：
影响因子：
29.4
通讯作者：
LaRusso NF
中科院分区：
文献类型：
--
作者：
Gradilone SA;Masyuk TV;Huang BQ;Banales JM;Lehmann GL;Radtke BN;Stroope A;Masyuk AI;Splinter PL;LaRusso NF
在多囊肝病中，囊肿形成涉及胆管细胞过度增殖。在PCK大鼠（常染色体隐性遗传性多囊肾病（ARPKD）的动物模型）中，胆管细胞[Ca 2 +]i降低与过度增殖相关。我们最近发现Trpv 4是一种钙离子进入通道，在正常胆管细胞中表达，其激活导致[Ca 2 +]i增加。因此，我们假设药理学激活Trpv 4可能逆转PCK胆管细胞的过度增殖表型。在正常和PCK大鼠、正常人和常染色体显性PKD（ADPKD）或ARPKD患者的肝脏中检测Trpv 4表达。在来自正常和PCK大鼠的胆管细胞中评估Trpv 4对细胞增殖和囊肿形成的活化作用。在PCK大鼠中分析Trpv 4活化对肾囊肿和肝囊肿的体内作用。Trpv 4在mRNA（8倍）和蛋白质（3倍）水平在CCK-胆管细胞中过表达。共聚焦和免疫金电子显微镜支持Trpv 4在PCK大鼠和ARPKD或ADPKD患者的肝脏中过表达。PCK胆管细胞中的Trpv 4激活使[Ca 2 +]i增加30%，抑制细胞增殖约25-50%，并在3-D培养物中抑制囊肿生长（3倍）。Trpv 4-siRNA沉默阻断了70%的Trpv 4激活剂的作用。Trpv 4激活与Akt磷酸化和β-Raf和Erk 1/2抑制相关。在体内，Trpv 4激活诱导肾囊肿面积显著减少，肝囊肿无显著减少。总之，我们的体外和体内数据表明，通过Trpv 4活化增加细胞内钙可能代表PKD的潜在治疗方法。
In polycystic liver diseases, cyst formation involves cholangiocyte hyperproliferation. In PCK rats, an animal model of autosomal recessive polycystic kidney disease (ARPKD), decreased [Ca2+]i in cholangiocytes is associated with hyperproliferation. We recently showed Trpv4, a calcium-entry channel, is expressed in normal cholangiocytes and its activation leads to [Ca2+]i increase. Thus, we hypothesized that pharmacological activation of Trpv4 might reverse the hyperproliferative phenotype of PCK cholangiocytes. Trpv4 expression was examined in liver of normal and PCK rats, normal humans, and patients with autosomal dominant PKD (ADPKD) or ARPKD. Trpv4 activation effect on cell proliferation and cyst formation was assessed in cholangiocytes derived from normal and PCK rats. The in vivo effects of Trpv4 activation on kidney and liver cysts was analyzed in PCK rats. Trpv4 was overexpressed both at mRNA (8-fold), and protein (3-fold) levels in PCK-cholangiocytes. Confocal and immunogold electron-microscopy supported Trpv4 overexpression in the livers of PCK rats and ARPKD or ADPKD patients. Trpv4 activation in PCK cholangiocytes increased [Ca2+]i by 30% inhibiting cell proliferation by ~25-50% and cyst growth in 3-D-culture (3-fold). Trpv4-siRNA-silencing blocked effects of Trpv4 activators by 70%. Trpv4 activation was associated with Akt phosphorylation and β-Raf and Erk1/2 inhibition. In vivo, Trpv4 activation induced a significant decrease in renal cystic area and a non significant decrease in liver cysts. Taken together, our in vitro and in vivo data, suggest that increasing intracellular calcium by Trpv4 activation may represent a potential therapeutic approach in PKD.
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影响因子：
6
作者：
Banales, Jesus M.;Masyuk, Tatyana V.;LaRusso, Nicholas F.
通讯作者：
LaRusso, Nicholas F.
影响因子：
5.3
作者：
Alessandri-Haber, N;Dina, OA;Levine, JD
通讯作者：
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影响因子：
13.6
作者：
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通讯作者：
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DOI：
10.1073/pnas.0700499104
发表时间：
2007-03-13
影响因子：
11.1
作者：
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通讯作者：
Crews, Craig M.
影响因子：
7.8
作者：
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通讯作者：
Walz, Gerd