Third Department of Internal Medicine, Nara Medical University, Kashihara, Nara, Japan.
World J Gastroenterol. 2009 Nov 28;15(44):5533-40. doi: 10.3748/wjg.15.5533.
To investigate the possible use of the multiple cytokine production modulator, Y-40138, as a novel immunotherapy in the rat nonalcoholic steatohepatitis (NASH) model.
We allocated 6-wk-old male F344 rats to choline-supplemented, L-amino acid-defined (CSAA) diet (control group), CSAA diet + Y-40138 (control + Y-40138 group), choline-deficient, L-amino acid-defined (CDAA) diet (NASH group), or CDAA diet + Y-40138 (NASH + Y-40138 group). In each group, we measured the plasma alanine aminotransferase (ALT) levels, and the plasma and liver levels of tumor necrosis factor-alpha (TNF-alpha), interferon-gamma (IFN-gamma), and interleukin-10 (IL-10). Tissue specimens of phosphate buffered saline-perfused liver were subjected to hematoxylin and eosin staining, Azan staining, Sirius red staining, and immunohistochemical staining (for Kupffer cells and TNF-alpha). We then extracted Kupffer cells from the collagenase-perfused livers using the Percoll gradient centrifugation method, and measured the TNF-alpha levels in the supernatant (in vitro TNF-alpha production by Kupffer cells) using an enzyme-linked immunosorbent assay kit.
In comparison to the NASH group, serum ALT elevation was mild, production of serum and liver TNF-alpha and IFN-gamma was inhibited, and IL-10 production was increased in the NASH + Y-40138 group. Amelioration of liver histology was also noted in the NASH + Y-40138 group. Kupffer cell immunohistochemical staining revealed no differences between groups, whereas TNF-alpha immunohistochemical staining showed fewer stained cells in the NASH + Y-40138 group than in the NASH group. The TNF-alpha levels in the in-vitro Kupffer cell culture supernatant were lower in the NASH + Y-40138 group than in the NASH group.
Administration of Y-40138 to NASH model rats reduced hepatic inflammation and suppressed fibrosis. These results indicate that the multiple cytokine production modulator, Y-40138, is promising as a novel treatment for NASH.
研究新型免疫疗法 Y-40138 在大鼠非酒精性脂肪性肝炎（NASH）模型中的应用潜力。
将 6 周龄雄性 F344 大鼠分为胆碱补充型、必需氨基酸限定型（CSAA）饮食（对照组）、CSAA 饮食+Y-40138（对照+Y-40138 组）、胆碱缺乏型、必需氨基酸限定型（CDAA）饮食（NASH 组）或 CDAA 饮食+Y-40138（NASH+Y-40138 组）。在每组中，我们测量了血浆丙氨酸氨基转移酶（ALT）水平，以及血浆和肝脏中肿瘤坏死因子-α（TNF-α）、干扰素-γ（IFN-γ）和白细胞介素-10（IL-10）水平。用磷酸盐缓冲盐水灌注肝脏组织标本，进行苏木精和伊红染色、锇酸染色、天狼星红染色和免疫组织化学染色（用于库普弗细胞和 TNF-α）。然后，我们使用聚蔗糖梯度离心法从胶原酶灌注的肝脏中提取库普弗细胞，并使用酶联免疫吸附测定试剂盒测量上清液中的 TNF-α 水平（库普弗细胞的 TNF-α 体外产生）。
与 NASH 组相比，血清 ALT 升高较轻，血清和肝脏 TNF-α 和 IFN-γ 的产生受到抑制，IL-10 的产生增加，NASH+Y-40138 组的肝组织学也得到改善。库普弗细胞免疫组织化学染色显示各组之间无差异，而 TNF-α 免疫组织化学染色显示 NASH+Y-40138 组染色细胞少于 NASH 组。NASH+Y-40138 组库普弗细胞体外培养上清液中的 TNF-α 水平低于 NASH 组。
Y-40138 给药可减少 NASH 模型大鼠的肝炎症和抑制纤维化。这些结果表明，多细胞因子产生调节剂 Y-40138 有望成为 NASH 的一种新治疗方法。
