Department of General Surgery, Southeast University Medical School, Nanjing, Jiangsu 210009, P.R. China.
Int J Mol Med. 2017 Aug;40(2):337-346. doi: 10.3892/ijmm.2017.3016. Epub 2017 Jun 9.
Surgeries, particularly ileocecal resection (ICR), are often required in the treatment of Crohn's disease (CD). However, recurrences are common for patients who undergo ICR, and anastomotic fibrosis is the main cause of re-operation. The present study aimed to investigate the therapeutic effects of triptolide (TPL) in ameliorating fibrosis following ileocolonic anastomosis. A model of IL‑10‑/‑ mice undergoing ICR was used to study postsurgical inflammation and fibrosis of anastomosis. For this purpsose, interleukin (IL)‑10‑/‑ mice were randomly divided into 3 groups as follows: the control group, the saline‑treated group subjected to ICR (ST‑ICR) and the TPL‑treated group subjected to ICR (TT‑ICR). Wild‑type (WT) mice of matching ages were assigned to the WT group. The effects of TPL treatment on ileocolonic anastomosis were determined by histopathological evaluation, western blot analysis and ELISA. The analysis of the effects of TPL treatment on microRNA‑16‑1 (miR‑16‑1) and heat shock protein 70 (HSP70) expression was carried out by RT‑qPCR and western blot analysis. Compared with the control group, significantly higher inflammation scores following anastomosis were observed in the ST‑ICR group (P<0.05), although reversion was observed in the TT‑ICR group, which was consistent with changes in the area of CD4+ cell infiltration. The elevated fibrosis scores and the overexpression of procollagen I and III in the ST‑ICR group were all inhibited by TPL. With an increase in the severity of inflammation and fibrosis, the levels of IL‑6, tumor necrosis factor‑α (TNF‑α) and transforming growth factor‑β1 (TGF‑β1) increased; however, a significant decrease in these levels was observed following treatment with TPL (P<0.05). The results of RT‑qPCR revealed that the upregulated miR‑16‑1 levels in the ST‑ICR group were significantly reduced by TPL. HSP70, which can be inhibited by miR-16-1, ameliorates anastomotic inflammation and fibrosis. Thus, the present study demonstrates that TPL exerts a protective effect against fibrosis following anastomosis in CD. The miR‑16‑1/HSP70 signaling pathway, which can be regulated by TPL, may thus represent a novel therapeutic option in CD that deserves further investigation.
手术，特别是回肠结肠切除术（ICR），通常是克罗恩病（CD）治疗的需要。然而，接受 ICR 的患者常复发，吻合口纤维化是再次手术的主要原因。本研究旨在探讨雷公藤内酯醇（TPL）在改善回肠结肠吻合术后纤维化中的治疗效果。使用 IL-10-/-小鼠进行 ICR 模型来研究手术后吻合口的炎症和纤维化。为此，IL-10-/-小鼠被随机分为 3 组：对照组、接受 ICR 的生理盐水处理组（ST-ICR）和接受 ICR 的 TPL 处理组（TT-ICR）。匹配年龄的野生型（WT）小鼠被分配到 WT 组。通过组织病理学评估、Western blot 分析和 ELISA 确定 TPL 治疗对回肠结肠吻合术的影响。通过 RT-qPCR 和 Western blot 分析分析 TPL 治疗对 microRNA-16-1（miR-16-1）和热休克蛋白 70（HSP70）表达的影响。与对照组相比，ST-ICR 组吻合后炎症评分明显升高（P<0.05），而 TT-ICR 组则有所逆转，与 CD4+细胞浸润面积的变化一致。ST-ICR 组胶原 I 和胶原 III 的纤维化评分升高和过度表达均被 TPL 抑制。随着炎症和纤维化程度的加重，IL-6、肿瘤坏死因子-α（TNF-α）和转化生长因子-β1（TGF-β1）水平升高；然而，TPL 治疗后这些水平显著下降（P<0.05）。RT-qPCR 结果显示，ST-ICR 组上调的 miR-16-1 水平被 TPL 显著降低。HSP70 可被 miR-16-1 抑制，可改善吻合口炎症和纤维化。因此，本研究表明 TPL 对 CD 吻合口后纤维化具有保护作用。TPL 可调节的 miR-16-1/HSP70 信号通路可能是 CD 的一种新的治疗选择，值得进一步研究。
