Culture on a Fragmin/protamine-Coated Plate Suppresses the Collagen Type IαI and TGF-β1 mRNA Expression of Rat Hepatic Stellate RI-T Cells
Culture on a Fragmin/protamine-Coated Plate Suppresses the Collagen Type IαI and TGF-β1 mRNA Expression of Rat Hepatic Stellate RI-T Cells
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DOI：
10.1292/jvms.12-0396
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发表时间：
2013-05-01
影响因子：
1.2
通讯作者：
Sakaue, Motoharu
中科院分区：
文献类型：
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作者：
Sekiguchi, Hitomi;Hemmi, Natsuko;Sakaue, Motoharu
肝星状细胞(HSCs)在正常肝脏细胞内保存维生素A。当肝脏受损时，HSCs转化为肌成纤维细胞样细胞，然后增殖并增加其胶原的表达。在塑料平板上培养，造血干细胞会自发激活。为了保持HSC处于静止状态，胶原蛋白的低表达，通常采用细胞外基质(ECM)包被的方法，如Matrigel包衣或富含层粘连蛋白的包衣。Kishimoto等人。[14]据报道，Fragmin(R)/鱼精蛋白微粒(F/P-MPS)具有吸收细胞外基质等与肝素结合的细胞因子的能力。因此，我们研究了在F/P-MPS涂层平板上的培养是否维持了RI-T细胞(来源于大鼠HSCs)的静止状态，包括对胶原表达的抑制。我们发现，在F/P-MPS涂层平板上培养的RI-T细胞中，与未涂层和Matrigel涂层平板上的培养相比，RI-T细胞中I型胶原αI和转化生长因子-β1的mRNA水平显著受到抑制。结论：F/P-MPS包埋法有利于维持HSC中Ⅰ型胶原和转化生长因子-β1mRNA的低表达。
Hepatic stellate cells (HSCs) intracellularly preserve vitamin A in the normal liver. When the liver is damaged, HSCs transform into myofibroblast-like cells, and then proliferate and increase their expression of collagen. Cultured on a plastic plate, HSCs spontaneously activate. To maintain HSCs in a quiescent state with low expression of collagen, coating methods with extracellular matrixes (ECMs) such as Matrigel-coating or laminin-rich coating are commonly used for HSC cultivation. Kishimoto et al. [14] reported that Fragmin (R)/protamine microparticles (F/P-MPs) have the ability to absorb heparin-binding cytokines like ECMs. Therefore, we examined whether the cultivation on an F/P-MPs,coated plate maintains the quiescent state of RI-T cells (derived from rat HSCs) including the suppression of collagen expression. We found that the mRNA levels of collagen type I alpha I and TGF-beta 1 in RI-T cells were significantly suppressed in the cultivation on F/P-MPs-coated plates compared to cultures on noncoated and Matrigel-coated plates. We conclude that the F/P-MPs coating method is useful for maintaining with low expressions of collagen I alpha I and TGF-beta 1 mRNA levels in HSCs.