Liver kinase B1 regulates hepatocellular tight junction distribution and function in vivo.
Liver kinase B1 regulates hepatocellular tight junction distribution and function in vivo.
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DOI：
10.1002/hep.28724
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发表时间：
2016-10
期刊：
影响因子：
13.5
通讯作者：
Arias, Irwin M.
中科院分区：
文献类型：
--
作者：
Porat-Shliom, Natalie;Tietgens, Amber J.;Van Itallie, Christina M.;Vitale-Cross, Lynn;Jarnik, Michal;Harding, Olivia J.;Anderson, James M.;Gutkind, J. Silvio;Weigert, Roberto;Arias, Irwin M.
肝激酶B1（LKB 1）及其下游效应子AMP活化蛋白激酶（AMPK）通过调节膜运输和能量代谢在极性建立中发挥关键作用。在胶原夹心培养的肝细胞中，LKB 1或AMPK的缺失损害了顶端ABCB 11（Bsep）的运输和胆小管的形成。在本研究中，我们使用肝脏特异性（白蛋白-Cre）LKB 1基因敲除小鼠（LKB 1 −/−）来研究LKB 1在体内维持功能性紧密连接（TJ）中的作用。透射电子显微镜检查显示，具有微绒毛的肝细胞顶膜基本上延伸到LKB 1 −/−肝脏的基底外侧域。免疫荧光研究表明，LKB 1的损失导致更长和更宽的小管结构与错误定位的连接蛋白，扣带蛋白。为了检测连接功能，我们使用活体显微镜对6-羧基荧光素二乙酸酯（6-CFDA）的转运动力学进行了定量，6-CFDA在肝细胞中被加工成其荧光衍生物6-羧基荧光素（6-CF）并分泌到小管中。在LKB 1 −/−小鼠中，6-CF主要保留在肝细胞中，小管分泌延迟，6-CF出现在血液中。为了测试6-CF是否通过渗透性TJ转运，我们静脉注射低分子量（3 kDa）葡聚糖与6-CFDA联合。在野生型小鼠中，3 kDa葡聚糖保留在血管系统中，而它迅速出现在LKB 1 −/−小鼠的异常胆小管中，证实了连接中断导致血流和胆小管之间的细胞旁交换。结论：LKB 1在调节TJ和细胞旁通透性的维持中起着关键作用，这可以解释抑制LKB 1/AMPK通路的各种药物、化学物质和代谢状态如何导致胆汁淤积。（肝病学2016;64：1317 - 1329）
Liver kinase B1 (LKB1) and its downstream effector AMP‐activated protein kinase (AMPK) play critical roles in polarity establishment by regulating membrane trafficking and energy metabolism. In collagen sandwich‐cultured hepatocytes, loss of LKB1 or AMPK impaired apical ABCB11 (Bsep) trafficking and bile canalicular formation. In the present study, we used liver‐specific (albumin‐Cre) LKB1 knockout mice (LKB1−/−) to investigate the role of LKB1 in the maintenance of functional tight junction (TJ) in vivo. Transmission electron microscopy examination revealed that hepatocyte apical membrane with microvilli substantially extended into the basolateral domain of LKB1−/− livers. Immunofluorescence studies revealed that loss of LKB1 led to longer and wider canalicular structures correlating with mislocalization of the junctional protein, cingulin. To test junctional function, we used intravital microscopy to quantify the transport kinetics of 6‐carboxyfluorescein diacetate (6‐CFDA), which is processed in hepatocytes into its fluorescent derivative 6‐carboxyfluorescein (6‐CF) and secreted into the canaliculi. In LKB1−/− mice, 6‐CF remained largely in hepatocytes, canalicular secretion was delayed, and 6‐CF appeared in the blood. To test whether 6‐CF was transported through permeable TJ, we intravenously injected low molecular weight (3 kDa) dextran in combination with 6‐CFDA. In wild‐type mice, 3 kDa dextran remained in the vasculature, whereas it rapidly appeared in the abnormal bile canaliculi in LKB1−/− mice, confirming that junctional disruption resulted in paracellular exchange between the blood stream and the bile canaliculus. Conclusion: LKB1 plays a critical role in regulating the maintenance of TJ and paracellular permeability, which may explain how various drugs, chemicals, and metabolic states that inhibit the LKB1/AMPK pathway result in cholestasis. (Hepatology 2016;64:1317‐1329)
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影响因子：
25.7
作者：
Gissen P;Arias IM
通讯作者：
Arias IM
影响因子：
11.8
作者：
Aijaz, S;D'Atri, F;Matter, K
通讯作者：
Matter, K
影响因子：
3.7
作者：
Homolya L;Fu D;Sengupta P;Jarnik M;Gillet JP;Vitale-Cross L;Gutkind JS;Lippincott-Schwartz J;Arias IM
通讯作者：
Arias IM
影响因子：
3.4
作者：
Derangeon, Mickael;Spray, David C.;Bourmeyster, Nicolas;Sarrouilhe, Denis;Herve, Jean-Claude
通讯作者：
Herve, Jean-Claude
DOI：
10.1073/pnas.1018376108
发表时间：
2011-01-25
影响因子：
11.1
作者：
Fu, Dong;Wakabayashi, Yoshiyuki;Arias, Irwin M.
通讯作者：
Arias, Irwin M.