Physiological ranges of matrix rigidity modulate primary mouse hepatocyte function in part through hepatocyte nuclear factor 4 alpha.
Physiological ranges of matrix rigidity modulate primary mouse hepatocyte function in part through hepatocyte nuclear factor 4 alpha.
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DOI：
10.1002/hep.28450
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发表时间：
2016-07
期刊：
影响因子：
--
通讯作者：
Chang TT
中科院分区：
文献类型：
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作者：
Desai SS;Tung JC;Zhou VX;Grenert JP;Malato Y;Rezvani M;Español-Suñer R;Willenbring H;Weaver VM;Chang TT
基质刚性对细胞行为有重要影响，在肝纤维化过程中增加;然而，其对原代肝细胞功能的影响尚不清楚。我们假设纤维化肝脏中基质刚性增加会激活肝细胞中的机械转导，并导致肝脏特异性功能的抑制。为了在细胞水平上确定基质硬度的生理相关范围，我们对正常和纤维化肝脏的肝小叶进行了详细的原子力显微镜分析。我们确定正常的肝基质硬度约为150 Pa，在纤维化肝脏中纤维胶原沉积附近的区域增加到1- 6 kPa。原代肝细胞在可调刚度的胶原基质上的体外培养表明，基质刚度的纤维化水平对细胞骨架张力有深远的影响，并显着抑制肝细胞特异性功能。正常肝硬度通过肝细胞核因子4 α（HNF 4 α）维持功能性基因调节，而纤维化基质硬度抑制HNF 4 α转录网络。纤维化水平的基质硬度通过粘着斑激酶（FAK）激活原代肝细胞中的机械转导。此外，阻断Rho/Rho相关蛋白激酶（ROCK）通路可从在硬基质上培养的肝细胞中拯救HNF 4 α表达。纤维化水平的基质硬度显著抑制肝细胞特异性功能，部分是通过抑制Rho/ROCK途径介导的HNF 4 α转录网络。基质刚性在调节肝细胞功能中的作用的增加将促进我们对肝硬化中肝细胞功能障碍机制的理解，并刺激慢性肝病新治疗方法的开发。
Matrix rigidity has important effects on cell behavior and is increased during liver fibrosis; however, its effect on primary hepatocyte function is unknown. We hypothesized that increased matrix rigidity in fibrotic livers would activate mechanotransduction in hepatocytes and lead to inhibition of hepatic-specific functions. To determine the physiologically relevant ranges of matrix stiffness at the cellular level, we performed detailed atomic force microscopy analysis across liver lobules from normal and fibrotic livers. We determined that normal liver matrix stiffness was around 150Pa and increased to 1–6kPa in areas near fibrillar collagen deposition in fibrotic livers. In vitro culture of primary hepatocytes on collagen matrix of tunable rigidity demonstrated that fibrotic levels of matrix stiffness had profound effects on cytoskeletal tension and significantly inhibited hepatocyte-specific functions. Normal liver stiffness maintained functional gene regulation by hepatocyte nuclear factor 4 alpha (HNF4α) whereas fibrotic matrix stiffness inhibited the HNF4α transcriptional network. Fibrotic levels of matrix stiffness activated mechanotransduction in primary hepatocytes through focal adhesion kinase (FAK). In addition, blockade of the Rho/Rho-associated protein kinase (ROCK) pathway rescued HNF4α expression from hepatocytes cultured on stiff matrix. Fibrotic levels of matrix stiffness significantly inhibit hepatocyte-specific functions in part by inhibiting the HNF4α transcriptional network mediated through the Rho/ROCK pathway. Increased appreciation of the role of matrix rigidity in modulating hepatocyte function will advance our understanding of the mechanisms of hepatocyte dysfunction in liver cirrhosis and spur development of novel treatments for chronic liver disease.