Cellular internalization of proinsulin C-peptide

Proinsulin C-peptide is known to bind specifically to cell membranes and to exert intracellular effects, but whether it is internalized in target cells is unknown. In this study, using confocal microscopy and immunostained or rhodamine-labeled peptide, we show that C-peptide is internalized and localized to the cytosol of Swiss 3T3 and HEK-293 cells. In addition, transport into nuclei was found using the labeled peptide. The internalization was followed at 37°C for up to 1 h, and was reduced at 4°C and after preincubation with pertussis toxin. Hence, it is concluded to occur via an energy-dependent, pertussis toxin-sensitive mechanism and without detectable degradation within the experimental time course. Surface plasmon resonance measurements demonstrated binding of HEK-293 cell extract components to C-peptide, and subsequent elution of bound material revealed the components to be intracellular proteins. The identification of C-peptide cellular internalization, intracellular binding proteins, absence of rapid subsequent C-peptide degradation and apparent nuclear internalization support a maintained activity similar to that of an intracrine peptide hormone. Hence, the data suggest the possibility of one further C-peptide site of action. Received 31 October 2006; received after revision 27 December 2006; accepted 30 December 2006     

难治性癫痫患者脑组织中SCG10的表达及其意义

目的了解难治性癫痫患者脑组织中SCG10和MAPK的表达,探讨其在难治性癫痫发生、发展中的作用。方法按随机化原则,在我们建立的难治性癫痫患者术后脑组织库中抽取36例患者的脑组织,用免疫组织化学分别检测SCG10和MAPK的表迭,与16例对照组进行比较。并采用免疫荧光双标的方法,检测SCG10和MAPK在病例组中的表达情况。结果免疫组化检测到SCG10在实验组表达（0．4674±0．0258）,高于对照（0．3405±0．0207）,两组比较有显著性差异（P〈0．05）;MAPK在实验组表达（0．4217±0．0141）,同样也高于对照组（0．3189±0．1422,P〈0．05）。免疫荧光双标法,显示SCG10和MAPK表达在同一细胞上。讨论SCG10与MAPK在难治性癫痫患者的脑组织中表达增加,以及它们的共同表达,提示两者的相互作用可能是难治性癫痫发生发展中的一个重要因素。