Department of Pathology, School of Medicine, Saint Louis University, St. Louis, Missouri 63104, USA.
Endocrinology. 2009 Dec;150(12):5384-94. doi: 10.1210/en.2009-0517. Epub 2009 Oct 6.
Elevated levels of cholesterol/low-density lipoprotein (LDL) are a risk factor for the development of nonalcoholic steatohepatitis and its associated hepatic fibrosis. However, underlying mechanisms remain elusive. We previously reported that curcumin induced gene expression of peroxisome proliferator-activated receptor (PPAR)-gamma and stimulated its activity, leading to the inhibition of the activation of hepatic stellate cells (HSCs), the major effector cells during hepatic fibrogenesis. We recently showed that curcumin suppressed gene expression of LDL receptor in activated HSCs in vitro by repressing gene expression of the transcription factor sterol regulatory element binding protein-2 (SREBP-2), leading to the reduction in the level of intracellular cholesterol in HSCs and to the attenuation of the stimulatory effects of LDL on HSCs activation. The current study aimed at exploring molecular mechanisms by which curcumin inhibits srebp-2 expression in HSCs. Promoter deletion assays, mutagenesis assays, and EMSAs localize a specificity protein-1 (SP-1) binding GC-box in the srebp-2 promoter, which is responsible for enhancing the promoter activity and responding to curcumin in HSCs. Curcumin suppresses gene expression of SP-1 and reduces its trans-activation activity, which are mediated by the activation of PPARgamma. The inhibitory effect of curcumin on SP-1 binding to the GC-box is confirmed by chromatin immuno-precipitation. In summary, our results demonstrate that curcumin inhibits srebp-2 expression in cultured HSCs by activating PPARgamma and reducing the SP-1 activity, leading to the repression of ldlr expression. These results provide novel insights into molecular mechanisms by which curcumin inhibits LDL-induced HSC activation.
胆固醇/低密度脂蛋白(LDL)水平升高是导致非酒精性脂肪性肝炎及其相关肝纤维化的一个危险因素。然而,其潜在机制仍不清楚。我们之前报道姜黄素诱导过氧化物酶体增殖物激活受体(PPAR)-γ基因表达并刺激其活性,从而抑制肝星状细胞(HSCs)的激活,HSCs 是肝纤维化过程中的主要效应细胞。我们最近表明,姜黄素通过抑制转录因子固醇调节元件结合蛋白-2(SREBP-2)的基因表达,抑制激活的 HSCs 中 LDL 受体的基因表达,从而减少 HSCs 内胆固醇的水平,并减弱 LDL 对 HSCs 激活的刺激作用,从而抑制 LDL 受体在体外表达。本研究旨在探讨姜黄素抑制 HSCs 中 SREBP-2 表达的分子机制。启动子缺失试验、突变试验和电泳迁移率变动分析将特异性蛋白-1(SP-1)结合 GC 盒定位于 SREBP-2 启动子,该启动子负责增强启动子活性并对 HSCs 中的姜黄素作出反应。姜黄素抑制 SP-1 的基因表达并降低其转录激活活性,这是由 PPARγ 的激活介导的。姜黄素抑制 SP-1 与 GC 盒结合的抑制作用通过染色质免疫沉淀得到证实。总之,我们的结果表明,姜黄素通过激活 PPARγ和降低 SP-1 活性抑制培养的 HSCs 中 SREBP-2 的表达,从而抑制 LDLR 的表达。这些结果为姜黄素抑制 LDL 诱导的 HSC 激活的分子机制提供了新的见解。