Xidakis C, Mastrodimou N, Notas G, Renieri E, Kolios G, Kouroumalis E, Thermos K
Laboratory of Pharmacology, Department Basic Sciences, University of Crete, Faculty of Medicine, Heraklion, Crete, Greece.
Regul Pept. 2007 Oct 4;143(1-3):76-82. doi: 10.1016/j.regpep.2007.03.005. Epub 2007 Mar 30.
The present study investigated the presence of somatostatin receptor subtypes (ssts) and the endogenous peptides somatostatin and cortistatin in rat Kupffer cells, since modulation of these cells by somatostatin may be important for the beneficial effect of somatostatin analogues in a selected group of hepatocellular carcinoma patients. Kupffer cells were isolated from rat liver in agreement with national and EU guidelines. RT-PCR was employed to assess the expression of somatostatin, cortistatin and ssts in Kupffer cells. Western blot analysis and immunocytochemistry were employed to assess the expression and the localization of the receptors, respectively. Quiescent Kupffer cells were found to express sst(1-4) mRNA, while immunocytochemical studies supported the presence of only the sst(3) and sst(4) receptors, which were found to be internalized. However, sst1 and sst(2A) receptors were detected by western blotting. RT-PCR and RIA measurements support the presence of both somatostatin and cortistatin. Stimulation of the cells with LPS activated the expression of the sst(2), sst(3) and sst(4) receptors. The present data provide evidence to support the presence of ssts and the endogenous neuropeptides somatostatin and CST in rat Kupffer cells. Both peptides may act in an autocrine manner to regulate sst receptor distribution. Studies are in progress in order to further characterize the role of ssts in Kupffer cells and in hepatic therapeutics.
本研究调查了大鼠库普弗细胞中生长抑素受体亚型(ssts)以及内源性肽生长抑素和可体松的存在情况,因为生长抑素对这些细胞的调节作用可能对生长抑素类似物在特定组肝细胞癌患者中的有益作用至关重要。按照国家和欧盟指南从大鼠肝脏中分离出库普弗细胞。采用逆转录聚合酶链反应(RT-PCR)评估库普弗细胞中生长抑素、可体松和ssts的表达。分别采用蛋白质印迹分析和免疫细胞化学评估受体的表达和定位。发现静止的库普弗细胞表达sst(1 - 4)mRNA,而免疫细胞化学研究支持仅存在sst(3)和sst(4)受体,且发现这些受体被内化。然而,通过蛋白质印迹检测到了sst1和sst(2A)受体。RT-PCR和放射免疫分析测量结果支持生长抑素和可体松的存在。用脂多糖刺激细胞可激活sst(2)、sst(3)和sst(4)受体的表达。目前的数据提供了证据支持大鼠库普弗细胞中存在ssts以及内源性神经肽生长抑素和CST。这两种肽可能以自分泌方式调节sst受体的分布。为了进一步阐明ssts在库普弗细胞和肝脏治疗中的作用,相关研究正在进行中。