论文部分内容阅读
正常朊蛋白 (Pr PC)也可以在外周组织包括人的血液中表达。血液中大部分 Pr PC在血浆中被发现。我们提出了血管内皮细胞是血液中朊蛋白来源泉的假说。反转录 PCR证实人的脐静脉血管内皮细胞 (HU VEC)和微血管内皮细胞 (HMEC- 1)可表达朊蛋白的 m RNA;流式血细胞计数法证实 ,在细胞活性没有一定的特殊规律时 ,HUVECS 和HMEC- 1S(两种细胞所含的 Pr PC分子分别为每细胞中12 0 90 0± 15 0 5 8和 5 832 7± 4 5 77)能表达 Pr PC;免疫荧光共聚焦显微镜检测证实 ,HMEC- 1S和 HU VECS膜表面呈现 Pr PC阳性 ,采用时控强化解离免疫荧光分析法 (DEL FIA )对细胞培养基质进行分析 ,表明这两种细胞可缓慢释放可溶性Pr PC位与其活性无关。与 Von Willebrand致病因子抗原相反 ,当病人患有 Von Willebrand疾病时 ,采用去氨加压素治疗时 ,病人体内血浆 Pr PC水平下降。采用不同数量的血液样品测定病人体内血浆的 Pr PC证明 ,Pr PC的浓度和细胞的数量没有相关性。但是 ,当病人处于肾功能衰竭时 ,血浆中Pr PC的水平较高。总之 ,不论是大血管还是小血管的内皮细胞都可以表达高水平的 Pr PC,这些 Pr PC能释放入细胞培养基中。
Normal prion protein (Pr PC) can also be expressed in peripheral blood, including human blood. Most of the blood Pr PC was found in plasma. We hypothesize that vascular endothelial cells are a source of prion protein in the blood. Reverse transcriptase PCR confirmed the expression of prion protein m RNA in human umbilical vein endothelial cells (HU VECs) and microvascular endothelial cells (HMEC-1). Flow cytometry confirmed that there was no specific regulation of cell activity, Pr PC was expressed in both HUVECs and HMEC-1S (Pr PC molecules contained in both cells were 12 0 90 0 ± 15 0 58 and 5 832 7 ± 4 5 77 per cell, respectively). Immunofluorescence confocal microscopy confirmed , PrEC positive on HMEC-1S and HU VECS membrane surfaces, and analysis of cell culture media using time-resolved enhanced dissociation immunofluorescence assay (DEL FIA) showed that these two cells can slowly release soluble Pr PC sites independently of their activity . In contrast to Von Willebrand’s Pathogenic Factor Antigen, plasma Pr PC levels are reduced in patients with desmopressin when patients have Von Willebrand disease. Using different numbers of blood samples to determine PrPc in patients’ plasma proves that there is no correlation between the PrPc concentration and the number of cells. However, Pr PC levels in plasma are higher when the patient is in renal failure. In conclusion, endothelial cells of both large and small blood vessels can express high levels of Pr PC, which can be released into the cell culture medium.