Inhibitory effects of interferons on pancreatic stellate cell activation

瞄准：分析并且比较干扰素(IFN ) 的效果胰腺的星形的房间(PSC ) 上的 -alpha,IFN 贝它，和 IFN-gamma 激活在试管内并且阐明 IFN 的分子的基础行动。方法：PSC 与 recombinant 老鼠 IFN 从老鼠的胰腺的织物被孤立、有教养、刺激。房间增长和骨胶原合成被测量 5-bromo-2'-deoxyuridine (BrdU ) 的加入进 DNA 估计并且(3H ) 脯氨酸进醋性的酸可溶的蛋白质分别地。Apoptotic 房间被 FACS 分析(sub-G1 山峰方法) 决定。myofibroblastic PSC 显型的展览被 alpha 光滑的肌肉肌动朊(alpha- SMA ) 的免疫污点分析监视表示。估计信号变换器的激活和抄写(STAT ) 的使活跃之物，用 phospho-STAT-specific 抗体的西方的污点被执行。在 STAT1 功能上的研究，蛋白质的表示被 siRNA 禁止。结果：IFN 贝它和 IFN-gamma 然而并非 IFN-alpha 显著地减少了 PSC 增长和骨胶原合成。IFN-gamma 是清楚地禁止了 alpha-SMA 表示的唯一的 IFN。在使用的试验性的条件下面， apoptotic 细胞死亡的提高的率都没响应任何 IFN 处理被观察。IFN 贝它和 IFN-gamma 导致了 STAT1 和 STAT3 酷氨酸磷酸化的猛烈增加，当 IFN-alpha 的效果弱得多时。有 siRNA 的 STAT1 表示的抑制与 IFN-gamma 的显著地减少的生长禁止的效果被联系。结论：IFN 贝它并且特别地 IFN-gamma 在 PSC 激活在试管内上显示禁止的效果并且应该关于他们的在试管内效率被测试。由 IFN-gamma 行动的生长抑制要求 STAT1。

AIM： To analyze and to compare the effects of interferon （IFN）-α, IFN-β, and IFN-γ on pancreatic stellate cell （PSC） activation/n vitro and to elucidate the molecular basis of IFN action. METHODS： PSCs were isolated from rat＇s pancreatic tissue, cultured and stimulated with recombinant rat IFNs. Cell proliferation and collagen synthesis were assessed by measuring the incorporation of 5-bromo-2＇ -deoxyuridine （BrdU） into DNA and [^3H]-proline into acetic acid-soluble proteins, respectively. Apoptotic ceils were determined by FACS analysis （sub-G1 peak method）. Exhibition of the myofibroblastic PSC phenotype was monitored by immunoblot analysis of （α-smooth muscle actin （α-SMA） expression. To assess the activation of signal transducer and activator of transcription （STAT）, Western blots using phospho- STAT-specific antibodies were performed. In studies on STAT1 function, expression of the protein was inhibited by siRNA. RESULTS： IFN-β and IFN-γ, but not IFN-α significantly diminished PSC proliferation and collagen synthesis. IFN-γ, was the only IFN that clearly inhibited α-SMA expression. Under the experimental conditions used, no enhanced rate of apoptotic cell death was observed in response to any IFN treatment. IFN-β and IFN-γ, induced a strong increase of STAT1 and STAT3 tyrosine phosphorylation, while the effect of IFN-α was much weaker. Inhibition of STAT1 expression with siRNA was associated with a significantly reduced growth-inhibitory effect of IFN-γ. CONCLUSION： IFN-β and particularly IFN-γ, display inhibitory effects on PSC activation in vitro and should be tested regarding their in vitro efficiency. Growth inhibition by IFN-γ action requires STAT1.