UCD School of Biomolecular and Biomedical Sciences, UCD Conway Institute of Biomolecular and Biomedical Research, University College Dublin, Belfield, Dublin 4, Ireland.
J Lipid Res. 2012 Nov;53(11):2390-404. doi: 10.1194/jlr.M029314. Epub 2012 Sep 11.
Prostacyclin and its prostacyclin receptor, the I Prostanoid (IP), play essential roles in regulating hemostasis and vascular tone and have been implicated in a range cardio-protective effects but through largely unknown mechanisms. In this study, the influence of cholesterol on human IP [(h)IP] gene expression was investigated in cultured vascular endothelial and platelet-progenitor megakaryocytic cells. Cholesterol depletion increased human prostacyclin receptor (hIP) mRNA, hIP promoter-directed reporter gene expression, and hIP-induced cAMP generation in all cell types. Furthermore, the constitutively active sterol-response element binding protein (SREBP)1a, but not SREBP2, increased hIP mRNA and promoter-directed gene expression, and deletional and mutational analysis uncovered an evolutionary conserved sterol-response element (SRE), adjacent to a known functional Sp1 element, within the core hIP promoter. Moreover, chromatin immunoprecipitation assays confirmed direct cholesterol-regulated binding of SREBP1a to this hIP promoter region in vivo, and immunofluorescence microscopy corroborated that cholesterol depletion significantly increases hIP expression levels. In conclusion, the hIP gene is directly regulated by cholesterol depletion, which occurs through binding of SREBP1a to a functional SRE within its core promoter. Mechanistically, these data establish that cholesterol can regulate hIP expression, which may, at least in part, account for the combined cardio-protective actions of low serum cholesterol through its regulation of IP expression within the human vasculature.
前列环素及其前列腺素受体 I 型(IP)在调节止血和血管张力方面发挥着重要作用,并与一系列心脏保护作用有关,但具体机制尚不清楚。在这项研究中,研究了胆固醇对培养的血管内皮细胞和血小板祖细胞巨核细胞中人 IP[(h)IP]基因表达的影响。胆固醇耗竭增加了所有细胞类型中的人前列腺素受体(hIP)mRNA、hIP 启动子指导的报告基因表达和 hIP 诱导的 cAMP 生成。此外,组成型激活固醇反应元件结合蛋白(SREBP)1a,但不是 SREBP2,增加了 hIP mRNA 和启动子指导的基因表达,缺失和突变分析揭示了核心 hIP 启动子内的一个进化保守的固醇反应元件(SRE),紧邻已知的功能 Sp1 元件。此外,染色质免疫沉淀测定证实了 SREBP1a 在体内与该 hIP 启动子区域的直接胆固醇调节结合,免疫荧光显微镜证实胆固醇耗竭显著增加了 hIP 表达水平。总之,hIP 基因受胆固醇耗竭的直接调节,这是通过 SREBP1a 与核心启动子内的功能性 SRE 结合实现的。从机制上讲,这些数据表明胆固醇可以调节 hIP 的表达,这至少部分解释了低血清胆固醇通过调节人血管内 IP 表达的综合心脏保护作用。