The cAMP effectors Epac and protein kinase a (PKA) are involved in the hepatic cystogenesis of an animal model of autosomal recessive polycystic kidney disease (ARPKD).
The cAMP effectors Epac and protein kinase a (PKA) are involved in the hepatic cystogenesis of an animal model of autosomal recessive polycystic kidney disease (ARPKD).
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营地效应子EPAC和蛋白激酶A（PKA）参与了常染色体隐性多囊肾脏疾病（ARPKD）的动物模型的肝囊肿。
DOI：
10.1002/hep.22636
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发表时间：
2009-01
期刊：
影响因子：
13.5
通讯作者：
LaRusso, Nicholas F.
中科院分区：
文献类型：
--
作者：
Banales, Jesus M.;Masyuk, Tatyana V.;Gradilone, Sergio A.;Masyuk, Anatoliy I.;Medina, Juan F.;LaRusso, Nicholas F.
PCK 大鼠是常染色体隐性多囊肾病 (ARPKD) 的动物模型，会形成与细胞内 3',5'-环单磷酸腺苷 (cAMP) 增加相关的胆管细胞源性肝囊肿，抑制 cAMP 会抑制囊肿生长。我们假设升高的 cAMP 通过两个下游效应器、由 cAMP（Epac1 和 Epac2 亚型）激活的交换蛋白和蛋白激酶 A (PKA) 刺激胆管细胞增殖，并且细胞内钙也参与此过程。在 mRNA 和蛋白质水平上对 Epac 亚型和 PKA 调节亚基 (PKA-R) 进行评估表明，培养的正常大鼠胆管细胞表达 Epac1、Epac2 和所有调节性 PKA 亚基。 Epac 亚型和 PKA RIβ 亚基在培养的 PCK 胆管细胞中过度表达。对 Epac 和 PKA 激活的增殖分析表明，正常胆管细胞和 PCK 胆管细胞在 Epac 特异性刺激下都会增加生长，而 PKA 特异性刺激会产生不同的效果，抑制正常胆管细胞的增殖，但加速 PCK 胆管细胞的增殖过程。另一方面，在 3-D 条件下培养时，正常和 PCK 胆管细胞产生的囊性结构的 PKA 和 Epac 激活导致囊肿生长增加，特别是在 PCK 胆管细胞衍生的囊肿中。药理学抑制剂和 siRNA 介导的基因沉默证明了每种效应器激活的特异性，以及 MEK-ERK1/2 信号传导参与所有观察到的效应器相关增殖变化。 PCK 胆管细胞响应 PKA 刺激而不是 Epac 刺激而过度增殖，被发现与细胞内钙减少有关，钙水平的恢复通过 PI3K/AKT 途径阻断了 PKA 依赖性增殖。我们的数据提供了强有力的证据，表明 cAMP 效应器 Epac 和 PKA 以及细胞内钙水平均参与 ARPKD 的肝囊肿发生。
PCK rats, an animal model of autosomal recessive polycystic kidney disease (ARPKD), develop cholangiocyte-derived liver cysts associated with increased intracellular adenosine 3′,5′-cyclic monophosphate (cAMP), the inhibition of which suppresses cyst growth. We hypothesized that elevated cAMP stimulates cholangiocyte proliferation via two downstream effectors, exchange proteins activated by cAMP (Epac1 and Epac2 isoforms) and protein kinase A (PKA), and that intracellular calcium is also involved in this process. Assessment of Epac isoforms and PKA regulatory subunits (PKA-Rs) at the mRNA and protein level showed that cultured normal rat cholangiocytes express Epac1, Epac2 and all regulatory PKA subunits. Epac isoforms and the PKA RIβ subunit were over-expressed in cultured PCK cholangiocytes. Proliferation analysis in response to Epac and PKA activation indicated that both normal and PCK cholangiocytes increase their growth upon Epac-specific stimulation, while PKA-specific stimulation results in differential effects, suppressing proliferation in normal cholangiocytes but accelerating this process in PCK cholangiocytes. On the other hand, both PKA and Epac activation of cystic structures generated by normal and PCK cholangiocytes when cultured under 3-D conditions resulted in increased cyst growth, particularly in PCK-cholangiocyte derived cysts. Pharmacological inhibitors and siRNA-mediated gene silencing demonstrated the specificity of each effector activation, as well as the involvement of MEK-ERK1/2 signaling in all the observed effector-associated proliferation changes. Hyperproliferation of PCK cholangiocytes in response to PKA stimulation, but not to Epac stimulation, was found to be associated with decreased intracellular calcium, and restoration of calcium levels blocked the PKA-dependent proliferation via PI3K/AKT pathway. our data provide strong evidence that both cAMP effectors, Epac and PKA, and the levels of intracellular calcium are involved in the hepatic cystogenesis of ARPKD.
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