Amelioration of murine dextran sulfate sodium-induced colitis by ex vivo extracellular superoxide dismutase gene transfer
Amelioration of murine dextran sulfate sodium-induced colitis by ex vivo extracellular superoxide dismutase gene transfer
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DOI：
10.1097/01.mib.0000225335.68614.73
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发表时间：
2006-07-01
影响因子：
4.9
通讯作者：
Niitsu, Yoshiro
中科院分区：
文献类型：
--
作者：
Oku, Takatomi;Iyama, Satoshi;Niitsu, Yoshiro
背景：虽然炎症性肠病的病因尚未完全阐明，但推测活性氧参与其中。细胞外超氧化物歧化酶（EC-SOD）是SOD的同工酶，主要在体液中发挥作用。我们研究了一个离体EC-SOD基因转移到葡聚糖硫酸钠（DSS）诱导的colitis mice.Materials and Methods的疗效：实验性结肠炎诱导Balb/c小鼠提供DSS在无菌蒸馏水提供所需的。从Balb/c小鼠胚胎中获得同基因成纤维细胞，并逆转录病毒转导hEC-SOD基因。经酶联免疫吸附试验证实，这些工程细胞在培养基中分泌EC-SOD，并接种于DSS处理的小鼠背部皮下。通过疾病活动指数（DAI：体重、直肠出血和粪便粘稠度）、组织学疾病严重程度分级和细胞因子（肿瘤坏死因子-α、白细胞介素-1 β）产生水平评价结肠粘膜损伤。8-通过免疫组织化学染色评估粘膜组织中的羟基脱氧鸟苷（8-OHdG）水平。丙二醛（MDA）测定使用比色assay.Results：一个显着的改善，观察到DAI评分和组织学的严重程度，以及粘膜组织水平的炎性细胞因子，8-OHdG，和MDA的小鼠与那些没有基因治疗相比，不仅在轻度结肠炎模型，但也在一个严重的结肠炎模型。在这些models.Conclusions治疗小鼠的生存显着延长：体外转移的EC-SOD基因是可行的DSS诱导的结肠炎的治疗。
Background: Although the etiology of inflammatory bowel disease has not been fully clarified, reactive oxygen species is speculated to be involved. Extracellular superoxide dismutase (EC-SOD), an isozyme of SODs, is known to function mainly in body fluids. We investigated the efficacy of an ex vivo EC-SOD gene transfer into dextran sulfate sodium (DSS)-induced colitis mice.Materials and Methods: Experimental colitis was induced by providing Balb/c mice with DSS in sterile distilled water provided as desired. The syngenic fibroblasts were obtained from Balb/c mice embryos and retrovirally transduced with the hEC-SOD gene. These engineered cells were confirmed to secrete EC-SOD in culture medium by enzyme-linked immunosorbent assay and were inoculated subcutaneously in the backs of DSS-treated mice. Mucosal injury of the colon was evaluated by the disease activity index (DAI: body weight, rectal bleeding, and stool consistency), grading of histologic disease severity, and levels of cytokine (tumor necrosis factor-alpha, interieukin-1 beta) production. 8-Hydroxydeoxyguanosine (8-OHdG) levels in the mucosal tissue were assessed by immunohistochemical staining. Malondialdehyde (MDA) was measured using a colorimetric assay.Results: A significant improvement was observed in DAI score and histologic severity as well as in mucosal tissue levels of inflammatory cyrokines, 8-OHdG, and MDA of mice treated with the EC-SOD gene as compared with those without gene therapy, not only in a mild colitis model but also in a severe colitis model. Survival of treated mice in these models was significantly prolonged.Conclusions: Ex vivo transfer of the EC-SOD gene was feasible for treatment of DSS-induced colitis.