Department of Molecular Gastroenterology and Hepatology, Graduate School of Medical Science, Kyoto Prefectural University of Medicine, Kawaramachi-Hirokoji, Kamigyo-ku, Kyoto, Japan.
Biol Pharm Bull. 2011;34(11):1659-65. doi: 10.1248/bpb.34.1659.
Heat shock protein (HSP) 47 may play an important role in the pathogenesis of intestinal fibrosis. Daikenchuto (DKT), a traditional Japanese herbal (Kampo) medicine, has been reported to ameliorate intestinal inflammation. The aims of this study were to determine time-course profiles of several parameters of fibrosis in a rat model, to confirm the HSP47-expressing cells in the colon, and finally to evaluate DKT's effects on intestinal fibrosis. Colitis was induced in male Wistar rats weighing 200 g using an enema of trinitrobenzene sulfonic acid (TNBS). HSP47 localization was determined by immunohistochemistry. Colonic inflammation and fibrosis were assessed by macroscopic, histological, morphometric, and immunohistochemical analyses. Colonic mRNA expression of transforming growth factor β1 (TGF-β1), HSP47, and collagen type I were assessed by real time-polymerase chain reaction (PCR). DKT was administered orally once a day from 8 to 14 d after TNBS administration. The colon was removed on the 15th day. HSP47 immunoreactivity was coexpressed with α-smooth muscle actin-positive cells located in the subepithelial space. Intracolonic administration of TNBS resulted in grossly visible ulcers. Colonic inflammation persisted for 6 weeks, and fibrosis persisted for 4 weeks after cessation of TNBS treatment. The expression levels of mRNA and proteins for TGF-β1, HSP47, and collagen I were elevated in colonic mucosa treated with TNBS. These fibrosis markers indicated that DKT treatment significantly inhibited TNBS-induced fibrosis. These findings suggest that DKT reduces intestinal fibrosis associated with decreasing expression of HSP47 and collagen content in the intestine.
热休克蛋白 47（HSP47）可能在肠道纤维化的发病机制中发挥重要作用。大建中汤（DKT）是一种传统的日本草药（汉方药），已被报道可改善肠道炎症。本研究的目的是确定纤维化的几个参数在大鼠模型中的时程谱，确认结肠中 HSP47 表达细胞，并最终评估 DKT 对肠道纤维化的作用。使用三硝基苯磺酸（TNBS）灌肠法诱导雄性 Wistar 大鼠结肠炎。通过免疫组织化学法确定 HSP47 的定位。通过宏观、组织学、形态计量学和免疫组织化学分析评估结肠炎症和纤维化。通过实时聚合酶链反应（PCR）评估转化生长因子β1（TGF-β1）、HSP47 和胶原 I 的结肠 mRNA 表达。在 TNBS 给药后第 8 至 14 天每天口服给予 DKT。第 15 天切除结肠。HSP47 免疫反应性与位于上皮下空间的α-平滑肌肌动蛋白阳性细胞共表达。TNBS 腔内给药导致肉眼可见的溃疡。结肠炎症持续 6 周，TNBS 治疗停止后纤维化持续 4 周。用 TNBS 处理的结肠黏膜中 TGF-β1、HSP47 和胶原 I 的 mRNA 和蛋白表达水平升高。这些纤维化标志物表明 DKT 治疗可显著抑制 TNBS 诱导的纤维化。这些发现表明 DKT 通过降低 HSP47 和胶原含量来减少与肠道相关的纤维化。
