Kinase analysis in alcoholic hepatitis identifies p90RSK as a potential mediator of liver fibrogenesis.
Kinase analysis in alcoholic hepatitis identifies p90RSK as a potential mediator of liver fibrogenesis.
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DOI：
10.1136/gutjnl-2014-307979
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发表时间：
2016-05
期刊：
影响因子：
24.5
通讯作者：
Bataller R
中科院分区：
文献类型：
--
作者：
Morales-Ibanez O;Affò S;Rodrigo-Torres D;Blaya D;Millán C;Coll M;Perea L;Odena G;Knorpp T;Templin MF;Moreno M;Altamirano J;Miquel R;Arroyo V;Ginès P;Caballería J;Sancho-Bru P;Bataller R
酒精性肝炎（AH）通常与晚期纤维化相关，这对生存有负面影响。我们的目标是在AH和晚期纤维化患者的肝脏中鉴定激酶的失调，以发现新的分子靶点。在AH （n=12）和正常人肝脏（n=7）中进行了广泛的反相蛋白微阵列分析。采用qPCR、Western blot和免疫组织化学检测肝脏核糖体S6激酶（p90RSK）的表达。山奈酚作为p90RSK通路的选择性药理学抑制剂，通过体内和体外方法评估实验诱导的肝纤维化和损伤的调节。蛋白质组学分析发现p90RSK是AH中最不受调节的激酶之一。慢性肝病患者肝脏p90RSK基因和蛋白表达也上调。免疫组织化学研究显示，肝硬化纤维化活跃区p90RSK染色增加。与给药相比，山奈酚对四氯化碳处理小鼠的治疗性给药导致肝胶原沉积减少，促纤维化和促炎基因的表达减少。山奈酚还能降低肝细胞损伤程度和凋亡程度。在原代肝星状细胞中，山奈酚和小干扰RNA降低了p90RSK的激活，从而调节了关键的促纤维化作用。在原代肝细胞中，山奈酚可减弱促凋亡信号传导。p90RSK在慢性肝病患者中上调，并在体内和体外介导肝纤维化。这些结果表明p90RSK通路可能是晚期纤维化肝脏疾病的一种新的治疗方法。
Alcoholic hepatitis (AH) is often associated with advanced fibrosis, which negatively impacts survival. We aimed at identifying kinases deregulated in livers from patients with AH and advanced fibrosis in order to discover novel molecular targets. Extensive phosphoprotein analysis by reverse phase protein microarrays was performed in AH (n=12) and normal human livers (n=7). Ribosomal S6 kinase (p90RSK) hepatic expression was assessed by qPCR, Western blot and immunohistochemistry. Kaempferol was used as a selective pharmacological inhibitor of the p90RSK pathway to assess the regulation of experimentally-induced liver fibrosis and injury, using in vivo and in vitro approaches. Proteomic analysis identified p90RSK as one of the most deregulated kinases in AH. Hepatic p90RSK gene and protein expression was also upregulated in livers with chronic liver disease. Immunohistochemistry studies showed increased p90RSK staining in areas of active fibrogenesis in cirrhotic livers. Therapeutic administration of kaempferol to carbon tetrachloride-treated mice resulted in decreased hepatic collagen deposition, and expression of profibrogenic and proinflammatory genes, compared to vehicle administration. In addition, kaempferol reduced the extent of hepatocellular injury and degree of apoptosis. In primary hepatic stellate cells, kaempferol and small interfering RNA decreased activation of p90RSK, which in turn regulated key profibrogenic actions. In primary hepatocytes, kaempferol attenuated proapoptotic signalling. p90RSK is upregulated in patients with chronic liver disease and mediates liver fibrogenesis in vivo and in vitro. These results suggest that the p90RSK pathway could be a new therapeutic approach for liver diseases characterised by advanced fibrosis.