cAMP-mediated regulation of cholesterol accumulation in cystic fibrosis and Niemann-Pick type C cells
cAMP-mediated regulation of cholesterol accumulation in cystic fibrosis and Niemann-Pick type C cells
复制标题
DOI：
10.1152/ajplung.90402.2008
复制
发表时间：
2008-11-01
影响因子：
4.9
通讯作者：
Kelley, Thomas J.
中科院分区：
文献类型：
--
作者：
Manson, Mary E.;Corey, Deborah A.;Kelley, Thomas J.
曼森ME，科里DA，白色NM，凯利TJ。cAMP介导的囊性纤维化和Niemann-Pick C型细胞中胆固醇积累的调节。美国生理学杂志肺细胞分子生理学295：L 809-L 819，2008年。首次发表于2008年9月12日; doi：10.1152/ajplung.90402.2008。本研究的目的是确定囊性纤维化（CF）细胞中胆固醇蓄积的调节机制。CFTR的活化和表达都受cAMP途径的调节，并且假设涉及该途径的反馈反应可能参与胆固醇蓄积的表型。为了检查cAMP途径在胆固醇积累中的作用，我们用腺苷3 '，5'-环单硫代磷酸酯（Rp-cAMPS）的Rp非对映体处理两种CF模型细胞系，并通过菲律宾染色进行可视化。Rp-cAMPS处理消除CF细胞中的胆固醇积累，而8-溴-cAMP处理导致野生型细胞中的胆固醇积累。为了在一个独立的模型系统中证实这些发现，我们还研究了cAMP在调节尼曼-匹克C型（NPC）成纤维细胞中胆固醇积累中的作用。NPC相关蛋白NPC 1的表达也受cAMP直接调控;因此，推测NPC细胞表现出相同的cAMP介导的胆固醇积累控制。Rp-cAMPS的存在也减少了NPC细胞中的胆固醇积累。与相应的对照相比，在CF模型细胞、Cftr（-/-）MNE、从CF受试者鼻刮获得的原发组织和NPC成纤维细胞中，β-抑制蛋白-2（β arr 2）（对cAMP信号传导的细胞应答的标志物）的表达显著升高。
Manson ME, Corey DA, White NM, Kelley TJ. cAMPmediated regulation of cholesterol accumulation in cystic fibrosis and Niemann- Pick type C cells. Am J Physiol Lung Cell Mol Physiol 295: L809-L819, 2008. First published September 12, 2008; doi:10.1152/ajplung.90402.2008.- The goal of this study was to identify a mechanism regulating cholesterol accumulation in cystic fibrosis (CF) cells. Both CFTR activation and expression are regulated by the cAMP pathway, and it is hypothesized that a feedback response involving this pathway may be involved in the phenotype of cholesterol accumulation. To examine the role of the cAMP pathway in cholesterol accumulation, we treated two CF model cell lines with the Rp diastereomer of adenosine 3',5'-cyclic monophosphorothioate (Rp-cAMPS) and visualized by filipin staining. Rp-cAMPS treatment eliminated cholesterol accumulation in CF cells, whereas 8-bromo-cAMP treatment led to cholesterol accumulation in wild-type cells. To confirm these findings in an independent model system, we also examined the role of cAMP in modulating cholesterol accumulation in Niemann-Pick type C (NPC) fibroblasts. Expression of the protein related to NPC, NPC1, is also directly regulated by cAMP; therefore, it is postulated that NPC cells exhibit the same cAMP-mediated control of cholesterol accumulation. Cholesterol accumulation in NPC cells also was reduced by the presence of Rp-cAMPS. Expression of beta-arrestin-2 (beta arr2), a marker of cellular response to cAMP signaling, was significantly elevated in CF model cells, Cftr(-/-) MNE, primary tissue obtained by nasal scrapes from CF subjects, and in NPC fibroblasts compared with respective controls.