Abstract：
OBJECTIVE: Hirschsprung disease-associated enterocolitis (HAEC) is a common life-threatening complication of Hirschsprung disease (HSCR). We aimed to investigate the effectiveness, long-term safety and the underlying mechanisms of Mesenchymal stem cells (MSCs) based therapy for HAEC.
METHODS: Specimens from HSCR and HAEC patients were used to assess the inflammatory condition. Ednrb knock-out mice was used as HAEC model. MSCs was intraperitoneally transplanted into HAEC mice. The therapy effects, long-term outcome, safety and toxicity and the mechanism of MSCs on the treatment of HAEC were explored in vivo and in vitro.
RESULTS: Intestinal M1 macrophages infiltration and severe inflammation condition were observed in HAEC. After the injection of MSCs, HAEC mice showed significant amelioration of the inflammatory injury and inhibition of M1 macrophages infiltration. The expression levels of pro-inflammatory cytokines (TNF-α and IFN-γ) were decreased and anti-inflammatory cytokines (IL-10 and TGF-β) were increased. In addition, we found that effective MSCs homing to the inflamed colon tissue occurred without long-term toxicity response. However, COX-2 inhibitor could diminish the therapeutic effects of MSCs. Using MSCs and macrophages co-culture system, we identified that MSCs could alleviate HAEC by inhibiting M1 macrophages activation through COX-2-dependent MAPK/ERK signaling pathway.
CONCLUSIONS: MSCs ameliorate HAEC by reducing M1 macrophages polarization via COX-2 mediated MAPK/ERK signaling pathway, thus providing novel insights and potentially promising strategy for the treatment or prevention of HAEC.
METHODS: Specimens from HSCR and HAEC patients were used to assess the inflammatory condition. Ednrb knock-out mice was used as HAEC model. MSCs was intraperitoneally transplanted into HAEC mice. The therapy effects, long-term outcome, safety and toxicity and the mechanism of MSCs on the treatment of HAEC were explored in vivo and in vitro.
RESULTS: Intestinal M1 macrophages infiltration and severe inflammation condition were observed in HAEC. After the injection of MSCs, HAEC mice showed significant amelioration of the inflammatory injury and inhibition of M1 macrophages infiltration. The expression levels of pro-inflammatory cytokines (TNF-α and IFN-γ) were decreased and anti-inflammatory cytokines (IL-10 and TGF-β) were increased. In addition, we found that effective MSCs homing to the inflamed colon tissue occurred without long-term toxicity response. However, COX-2 inhibitor could diminish the therapeutic effects of MSCs. Using MSCs and macrophages co-culture system, we identified that MSCs could alleviate HAEC by inhibiting M1 macrophages activation through COX-2-dependent MAPK/ERK signaling pathway.
CONCLUSIONS: MSCs ameliorate HAEC by reducing M1 macrophages polarization via COX-2 mediated MAPK/ERK signaling pathway, thus providing novel insights and potentially promising strategy for the treatment or prevention of HAEC.
摘要：
目的：先天性巨结肠病相关小肠结肠炎（HAEC）是先天性巨结肠病（HSCR）常见的危及生命的并发症。我们的目的是调查有效性，基于间充质干细胞（MSCs）的HAEC治疗的长期安全性和潜在机制。
方法：使用来自HSCR和HAEC患者的样本来评估炎症状况。使用Ednrb敲除小鼠作为HAEC模型。将MSCs腹膜内移植到HAEC小鼠中。治疗效果，长期结果，对MSCs治疗HAEC的作用机制和安全性进行了研究。
结果：在HAEC中观察到肠M1巨噬细胞浸润和严重的炎症状况。注射MSCs后,HAEC小鼠显示出炎性损伤的显著改善和M1巨噬细胞浸润的抑制。促炎细胞因子（TNF-α和IFN-γ）的表达水平降低，抗炎细胞因子（IL-10和TGF-β）增加。此外,我们发现,有效的MSCs归巢至发炎的结肠组织,没有发生长期毒性反应.然而,COX-2抑制剂可降低MSCs的治疗效果。使用MSCs和巨噬细胞共培养系统，我们发现MSCs可以通过COX-2依赖性MAPK/ERK信号通路抑制M1巨噬细胞的活化,从而缓解HAEC.
结论：MSCs通过COX-2介导的MAPK/ERK信号通路减少M1巨噬细胞的极化，从而改善HAEC，从而为HAEC的治疗或预防提供了新的见解和潜在的有希望的策略。
方法：使用来自HSCR和HAEC患者的样本来评估炎症状况。使用Ednrb敲除小鼠作为HAEC模型。将MSCs腹膜内移植到HAEC小鼠中。治疗效果，长期结果，对MSCs治疗HAEC的作用机制和安全性进行了研究。
结果：在HAEC中观察到肠M1巨噬细胞浸润和严重的炎症状况。注射MSCs后,HAEC小鼠显示出炎性损伤的显著改善和M1巨噬细胞浸润的抑制。促炎细胞因子（TNF-α和IFN-γ）的表达水平降低，抗炎细胞因子（IL-10和TGF-β）增加。此外,我们发现,有效的MSCs归巢至发炎的结肠组织,没有发生长期毒性反应.然而,COX-2抑制剂可降低MSCs的治疗效果。使用MSCs和巨噬细胞共培养系统，我们发现MSCs可以通过COX-2依赖性MAPK/ERK信号通路抑制M1巨噬细胞的活化,从而缓解HAEC.
结论：MSCs通过COX-2介导的MAPK/ERK信号通路减少M1巨噬细胞的极化，从而改善HAEC，从而为HAEC的治疗或预防提供了新的见解和潜在的有希望的策略。
