Cheng Jason H, She Hongyun, Han Yuan-Ping, Wang Jiaohong, Xiong Shigang, Asahina Kinji, Tsukamoto Hidekazu
Department of Pathology, Keck School of Medicine of the Univ. of Southern California, Los Angeles, CA 90033-9141, USA.
Am J Physiol Gastrointest Liver Physiol. 2008 Jan;294(1):G39-49. doi: 10.1152/ajpgi.00263.2007. Epub 2007 Nov 15.
Activation of hepatic stellate cells (HSC), a key event in liver fibrosis, is caused by diminished adipogenic transcription. This study investigated whether Wnt signaling contributes to "antiadipogenic" activation of HSC and liver fibrogenesis. Culture-activated HSC from normal rats and HSC from cholestatic rat livers were examined for expression of Wnt, Frizzled (Fz) receptors, and coreceptors by quantitative PCR. Wnt signaling was assessed by nuclear beta-catenin and T cell factor (TCF) promoter activity. Dickkopf-1 (Dkk-1), a Wnt coreceptor antagonist, was transduced by an adenoviral vector to assess the effects of Wnt antagonism on culture activation of HSC and cholestatic liver fibrosis in mice. Messenger RNA for canonical (Wnt3a and 10b) and noncanonical (Wnt4 and 5a) Wnt genes, Fz-1 and 2, and coreceptors [low-density lipoprotein-receptor-related protein (LRP)6 and Ryk] are increased approximately 3-12-fold in culture-activated HSC compared with quiescent HSC. The nuclear beta-catenin level and TCF DNA binding are markedly increased in activated HSC. TCF promoter activity is stimulated with Wnt1 but inhibited by Chibby, a protein that blocks beta-catenin interaction with TCF, and by Dkk-1. Dkk-1 enhances peroxisome proliferator-activated receptor-gamma (PPARgamma)-driven PPAR response element (PPRE) promoter activity, a key adipogenic transcriptional parameter, abrogates agonist-stimulated contraction, and restores HSC quiescence in culture. High expression of Dkk-1 increases apoptosis of cultured HSC. Expression of Wnt and Fz genes is also induced in HSC isolated from experimental cholestatic liver fibrosis, and Dkk-1 expression ameliorates this form of liver fibrosis in mice. These results demonstrate antiadipogenic Wnt signaling in HSC activation and therapeutic potential of Wnt antagonism for liver fibrosis.
肝星状细胞(HSC)的激活是肝纤维化中的关键事件,由脂肪生成转录减少引起。本研究调查了Wnt信号通路是否促成HSC的“抗脂肪生成”激活及肝纤维化。通过定量PCR检测正常大鼠培养激活的HSC和胆汁淤积大鼠肝脏中的HSC中Wnt、卷曲蛋白(Fz)受体和共受体的表达。通过核β-连环蛋白和T细胞因子(TCF)启动子活性评估Wnt信号通路。用腺病毒载体转导Dickkopf-1(Dkk-1),一种Wnt共受体拮抗剂,以评估Wnt拮抗对小鼠HSC培养激活和胆汁淤积性肝纤维化的影响。与静止的HSC相比,培养激活的HSC中经典(Wnt3a和10b)和非经典(Wnt4和5a)Wnt基因、Fz-1和2以及共受体[低密度脂蛋白受体相关蛋白(LRP)6和Ryk] 的信使核糖核酸增加约3至12倍。激活的HSC中核β-连环蛋白水平和TCF DNA结合显著增加。TCF启动子活性被Wnt1刺激,但被Chibby(一种阻断β-连环蛋白与TCF相互作用的蛋白)和Dkk-1抑制。Dkk-1增强过氧化物酶体增殖物激活受体γ(PPARγ)驱动的PPAR反应元件(PPRE)启动子活性,这是一个关键的脂肪生成转录参数,消除激动剂刺激的收缩,并在培养中恢复HSC的静止状态。Dkk-1的高表达增加培养的HSC的凋亡。从实验性胆汁淤积性肝纤维化分离的HSC中也诱导Wnt和Fz基因的表达,并且Dkk-1表达改善小鼠这种形式的肝纤维化。这些结果证明HSC激活中存在抗脂肪生成的Wnt信号通路以及Wnt拮抗对肝纤维化的治疗潜力。