Inhibition of the hedgehog pathway leads to antifibrotic effects in dermal fibrosis.
Inhibition of the hedgehog pathway leads to antifibrotic effects in dermal fibrosis.
复制标题
刺猬通路的抑制会导致真皮纤维化的抗纤维化作用。
作者：
Cao Hua-Li;Zhou Jiong;Chen Xi-Bei;L;eck Lilla;Yang Jian-Qiang;Chen Jia-Qi;Li Wei;Cai Sui-Qing;Zheng Min;Man Xiao-Yong
真皮纤维化的特征在于产生基质的“阳性”肌成纤维细胞的活化，以及细胞外基质的持续产生和沉积。最近已证明刺猬途径在系统性硬化症（SSc）中以促纤维化方式起作用。Hedgehog通路的负调节因子（Hh），融合的抑制因子（Sufu），被证明参与纤维化疾病的激活。然而，到目前为止，腐乳在纤维化中的确切作用尚未研究。在我们的研究中，我们的目的是确定腐乳在纤维化过程中的作用，使用体外培养的健康供体的真皮成纤维细胞。使用Smo拮抗剂环巴胺和Sufu慢病毒载体处理或转染细胞。Western blot检测纤维化标志物及ERK 1/2、Smad 2、GSK 3 β蛋白水平的表达。通过体外伤口愈合测定来测量成纤维细胞迁移。采用博莱霉素诱导的小鼠皮肤纤维化模型，研究环巴胺对皮肤纤维化的影响。我们发现，环巴胺显着上调表达的Sufu。环巴胺和Sufu lentivector均以统计学显著水平降低人真皮成纤维细胞的迁移和肌成纤维细胞分化。此外，环巴胺逆转TGF-β1诱导的真皮纤维化。Cyclopamine和Sufu过表达可抑制GSK-3β的磷酸化，抑制成纤维细胞的迁移。在硬皮病小鼠模型中，腹膜内注射环巴胺可抑制皮肤纤维化。我们的研究结果表明，cyclopamine和Sufu过表达可以有效地抑制内源性以及TGF-β1诱导的成纤维细胞激活，通过随后的GSK-3β激活。腐乳激动剂可能是一个有前途的方法，在开发抗皮肤纤维化和系统性硬化症的药物。
Dermal fibrosis is characterized by the activation of the matrix-producing 'positive' myofibroblasts, and the relentless production and deposition of extracellular matrix. The hedgehog pathway has recently been demonstrated to work in a pro-fibrotic manner in systemic sclerosis (SSc). A negative regulator of the hedgehog pathway (Hh), the suppressor of fused (Sufu), was shown to be involved in the activation of fibrotic diseases. However, the exact role of Sufu in fibrosis has not been investigated so far. In our study, we aimed to define the role of sufu in the process of fibrosis using dermal fibroblasts of healthy donors that were cultured in vitro. Cyclopamine, a Smo antagonist, and Sufu lentivector were used to treat or transfect cells. The expression of fibrosis markers and ERK1/2, Smad2, and GSK3β at the protein level was determined by Western blot. Fibroblast migration was measured by in vitro wound healing assay. Bleomycin-induced dermal fibrosis mouse model was introduced to assess the effect of cyclopamine on dermal fibrosis in vivo. We found that cyclopamine significantly upregulated the expression of Sufu. Both cyclopamine and Sufu lentivector reduced migration and myofibroblast differentiation of human dermal fibroblasts at a statistically significant level. Furthermore, cyclopamine reversed dermal fibrosis induced by TGF-β1. Cyclopamine and the overexpression of Sufu inhibited the phosphorylation of GSK-3β and restrained the migration of fibroblasts. Dermal fibrosis was inhibited by intraperitoneal injection of cyclopamine in a mouse model of scleroderma. Our findings suggest that cyclopamine and Sufu-overexpression may effectively inhibit the endogenous as well as the TGF-β1-induced activation of fibroblasts through subsequent activation of GSK-3β. Sufu agonists may be a promising approach in the development of antifibrotic medications for dermal fibrosis and systemic sclerosis.