Department of Applied Biology, Kyoto Institute of Technology, Sakyo-ku, Kyoto, 606 8585, Japan.
Department of Chemistry and Materials Engineering, Kansai University, Suita, Osaka, 564 8680, Japan.
Biochem Biophys Res Commun. 2020 Sep 17;530(2):362-366. doi: 10.1016/j.bbrc.2020.07.141. Epub 2020 Aug 12.
Activated hepatic stellate cells (HSCs) play a central role in fibrillary collagen production, the primary cause of liver fibrosis. Although it is known that primary cultured HSCs are activated by plastic culture dish stiffness, HSC activation and quiescent-state-reversion mechanisms are still unclear. In this study, we used cultured normal rat HSCs on 3.2 kPa collagen normal liver stiffness equivalent gel, to determine whether high glucose or high succinate conditions induce HSC activation, and examined whether activated HSCs reverted to a quiescent state when suppressed by GPR91 antagonists or TGF-β1 receptor inhibitors. We measured the gene expression levels of α-SMA and type I collagen HSC activation markers using real-time PCR. Our data indicated that high glucose conditions induced HSC activation, and showed that under continuous high glucose exposure HSC activation progressed. A GPR91 antagonist, 2 d, and a TGF-β1 receptor inhibitor, SB525334, suppressed the Col1α mRNA expression level of these activated HSCs. Similarly, under extended high succinate exposure, 2 d and SB525334 reduced Col1α mRNA expression levels of activated HSCs. From the above, we determined that even though HSCs had already been activated by high glucose or succinate conditions which persisted after activation, the GPR91 antagonist and the TGF-β1 receptor inhibitor were able to reduce the production of type I collagen from activated HSCs.
活化的肝星状细胞（HSCs）在纤维胶原产生中起核心作用，这是肝纤维化的主要原因。尽管已知原代培养的 HSCs 被塑料培养皿的刚性激活，但 HSC 的激活和静止状态恢复机制仍不清楚。在这项研究中，我们使用在 3.2kPa 胶原正常肝硬度等效凝胶上培养的正常大鼠 HSCs，以确定高葡萄糖或高琥珀酸盐条件是否诱导 HSC 激活，并检查当被 GPR91 拮抗剂或 TGF-β1 受体抑制剂抑制时，激活的 HSCs 是否恢复到静止状态。我们使用实时 PCR 测量 HSC 激活标志物α-SMA 和 I 型胶原的基因表达水平。我们的数据表明高葡萄糖条件诱导 HSC 激活，并表明在持续的高葡萄糖暴露下，HSC 激活会进展。GPR91 拮抗剂 2d 和 TGF-β1 受体抑制剂 SB525334 抑制这些活化 HSCs 的 Col1α mRNA 表达水平。同样，在延长的高琥珀酸盐暴露下，2d 和 SB525334 降低了活化 HSCs 的 Col1α mRNA 表达水平。由此我们确定，即使 HSCs 已经被高葡萄糖或琥珀酸盐条件激活，并且在激活后持续存在，GPR91 拮抗剂和 TGF-β1 受体抑制剂仍能够减少活化 HSCs 产生的 I 型胶原。
