Wang Hua, Sun Rui-Ting, Li Yang, Yang Yue-Feng, Xiao Feng-Jun, Zhang Yi-Kun, Wang Shao-Xia, Sun Hui-Yan, Zhang Qun-Wei, Wu Chu-Tse, Wang Li-Sheng
Department of Experimental Hematology, Beijing Institute of Radiation Medicine, Beijing, 100850, PR China.
Department of Experimental Hematology, Beijing Institute of Radiation Medicine, Beijing, 100850, PR China; College of Life Science and Bioengineering, Beijing University of Technology, Beijing, 100022, PR China.
PLoS One. 2015 May 1;10(5):e0124420. doi: 10.1371/journal.pone.0124420. eCollection 2015.
Effective therapeutic strategies to address intestinal complications after radiation exposure are currently lacking. Mesenchymal stem cells (MSCs), which display the ability to repair the injured intestine, have been considered as delivery vehicles for repair genes. In this study, we evaluated the therapeutic effect of hepatocyte growth factor (HGF)-gene-modified MSCs on radiation-induced intestinal injury (RIII).
Female 6- to 8-week-old mice were radiated locally at the abdomen with a single 13-Gy dose of radiation and then treated with saline control, Ad-HGF or Ad-Null-modified MSCs therapy. The transient engraftment of human MSCs was detected via real-time PCR and immunostaining. The therapeutic effects of non- and HGF-modified MSCs were evaluated via FACS to determine the lymphocyte immunophenotypes; via ELISA to measure cytokine expression; via immunostaining to determine tight junction protein expression; via PCNA staining to examine intestinal epithelial cell proliferation; and via TUNEL staining to detect intestinal epithelial cell apoptosis.
The histopathological recovery of the radiation-injured intestine was significantly enhanced following non- or HGF-modified MSCs treatment. Importantly, the radiation-induced immunophenotypic disorders of the mesenteric lymph nodes and Peyer's patches were attenuated in both MSCs-treated groups. Treatment with HGF-modified MSCs reduced the expression and secretion of inflammatory cytokines, including tumor necrosis factor alpha (TNF-α) and interferon-gamma (IFN-γ), increased the expression of the anti-inflammatory cytokine IL-10 and the tight junction protein ZO-1, and promoted the proliferation and reduced the apoptosis of intestinal epithelial cells.
Treatment of RIII with HGF-gene-modified MSCs reduces local inflammation and promotes the recovery of small intestinal histopathology in a mouse model. These findings might provide an effective therapeutic strategy for RIII.
目前缺乏有效的治疗策略来应对辐射暴露后的肠道并发症。间充质干细胞（MSCs）具有修复受损肠道的能力，已被视为修复基因的递送载体。在本研究中，我们评估了肝细胞生长因子（HGF）基因修饰的MSCs对辐射诱导的肠道损伤（RIII）的治疗效果。
6至8周龄雌性小鼠腹部接受单次13 Gy剂量的局部辐射，然后分别接受生理盐水对照、Ad-HGF或Ad-Null修饰的MSCs治疗。通过实时PCR和免疫染色检测人MSCs的短暂植入。通过流式细胞术（FACS）评估未修饰和HGF修饰的MSCs的治疗效果，以确定淋巴细胞免疫表型；通过酶联免疫吸附测定（ELISA）测量细胞因子表达；通过免疫染色确定紧密连接蛋白表达；通过增殖细胞核抗原（PCNA）染色检查肠上皮细胞增殖；通过末端脱氧核苷酸转移酶介导的缺口末端标记（TUNEL）染色检测肠上皮细胞凋亡。
未修饰或HGF修饰的MSCs治疗后，辐射损伤肠道的组织病理学恢复显著增强。重要的是，两个MSCs治疗组中辐射诱导的肠系膜淋巴结和派尔集合淋巴结的免疫表型紊乱均得到缓解。HGF修饰的MSCs治疗降低了包括肿瘤坏死因子α（TNF-α）和干扰素-γ（IFN-γ）在内的炎性细胞因子的表达和分泌，增加了抗炎细胞因子白细胞介素-10（IL-10）和紧密连接蛋白闭合蛋白1（ZO-1）的表达，并促进了肠上皮细胞的增殖，减少了其凋亡。
在小鼠模型中，用HGF基因修饰的MSCs治疗RIII可减轻局部炎症，促进小肠组织病理学恢复。这些发现可能为RIII提供一种有效的治疗策略。
