Hepatic stellate cells require a stiff environment for myofibroblastic differentiation
Hepatic stellate cells require a stiff environment for myofibroblastic differentiation
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DOI：
10.1152/ajpgi.00412.2010
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发表时间：
2011-07-01
影响因子：
4.5
通讯作者：
Wells, Rebecca G.
中科院分区：
文献类型：
--
作者：
Olsen, Abby L.;Bloomer, Steven A.;Wells, Rebecca G.
Olsen AL，布卢默SA，Chan EP，Gaca MDA，Georges PC，Sackey B，Uemura M，Janmey PA，威尔斯RG.肝星状细胞需要一个严格的环境，肌纤维母细胞分化。美国生理学胃肠和肝脏生理学杂志301：G110-G118，2011年。首次发表于2011年4月28日; doi：10.1152/ajpgi.00412.2010.-肝星状细胞（HSC）的肌纤维母细胞分化是肝纤维化的关键事件，是所有原因引起的慢性肝病中最终共同途径的一部分。驱动HSC分化的分子机制尚未完全了解。由于宏观组织硬化是纤维化疾病的一个特征，我们假设底层基质的机械性质是HSC活化的主要决定因素。原代大鼠HSC培养的惰性聚丙烯酰胺支持可变的，但精确定义的剪切模量（刚度）涂有不同的细胞外基质蛋白或聚-L-赖氨酸。通过细胞形态学、免疫荧光染色和基因表达来确定HSC分化。随着所有涂层基质（包括Matrigel）的基质硬度增加，HSC逐渐变得肌纤维母细胞化。HSC活化的程度而不是速度与基板刚度，与细胞培养的支持，中间刚度采用稳定的中间表型。软支持物上的静止细胞能够在暴露于硬支持物的情况下进行成肌纤维细胞分化。刚度依赖性分化需要粘附到基质蛋白和机械张力的产生。转化生长因子-β处理增强了在刚性支持物上的分化，但不是必需的。HSC在体外分化为肌成纤维细胞主要是作为基质的物理性质而不是化学性质的函数。HSC需要具有机械刚性的基质，与基质蛋白粘附并产生机械张力，以分化。这些发现表明，肝硬度的改变是推动纤维化进展的关键因素。
Olsen AL, Bloomer SA, Chan EP, Gaca MDA, Georges PC, Sackey B, Uemura M, Janmey PA, Wells RG. Hepatic stellate cells require a stiff environment for myofibroblastic differentiation. Am J Physiol Gastrointest Liver Physiol 301: G110-G118, 2011. First published April 28, 2011; doi:10.1152/ajpgi.00412.2010.-The myofibroblastic differentiation of hepatic stellate cells (HSC) is a critical event in liver fibrosis and is part of the final common pathway to cirrhosis in chronic liver disease from all causes. The molecular mechanisms driving HSC differentiation are not fully understood. Because macroscopic tissue stiffening is a feature of fibrotic disease, we hypothesized that mechanical properties of the underlying matrix are a principal determinant of HSC activation. Primary rat HSC were cultured on inert polyacrylamide supports of variable but precisely defined shear modulus (stiffness) coated with different extracellular matrix proteins or poly-L-lysine. HSC differentiation was determined by cell morphology, immunofluorescence staining, and gene expression. HSC became progressively myofibroblastic as substrate stiffness increased on all coating matrices, including Matrigel. The degree rather than speed of HSC activation correlated with substrate stiffness, with cells cultured on supports of intermediate stiffness adopting stable intermediate phenotypes. Quiescent cells on soft supports were able to undergo myofibroblastic differentiation with exposure to stiff supports. Stiffness-dependent differentiation required adhesion to matrix proteins and the generation of mechanical tension. Transforming growth factor-beta treatment enhanced differentiation on stiff supports, but was not required. HSC differentiate to myofibroblasts in vitro primarily as a function of the physical rather than the chemical properties of the substrate. HSC require a mechanically stiff substrate, with adhesion to matrix proteins and the generation of mechanical tension, to differentiate. These findings suggest that alterations in liver stiffness are a key factor driving the progression of fibrosis.