Therapeutic strategy for intractable inflammatory bowel diseases by mesenchymal stem cells

包括少量间充质干细胞(MSCs)在内的骨髓源性细胞对临床人类和实验动物结肠炎具有治疗作用。其详细机制(S)可能部分是由粘膜再生介导的，因为骨髓间充质干细胞具有向多个细胞部分分化的潜力。但MSCs被认为还有其他功能，如抗炎和粘膜再生，因为抗炎系统参与了结肠炎的修复。我们观察了骨髓间充质干细胞对葡聚糖硫酸钠(DSS)诱导的大鼠急性结肠炎的治疗作用和抗炎作用。近交系雄性Lewis大鼠饮水中分别灌服0、1、2、4%的DSS，连续7d诱发实验性结肠炎。骨髓从胫骨和股骨中挤出。然后分离其单个核细胞，在含10%胎牛血清的低糖DMEM中培养，用于MSCs的生长。在给药…后的0、2和4天更多的DSS、MSCs(5×10^6细胞)经尾静脉注射。我们每天检查食物和水的摄入量、大便情况和体重。7d后，取大鼠全结肠，用实时荧光定量RT-α法检测大鼠炎性细胞因子肿瘤坏死因子-β、IL-1、IL-10和COX2mRNA的表达。我们通过波形蛋白和α-平滑肌肌动蛋白的免疫组织化学染色和细胞表面标志物，如骨髓前体细胞标志物CD90，而不是CD45、HLADR、CD11b和CD31，用流式细胞仪技术证实了MSC的特征。通过对大鼠体重、食欲下降、便血和结肠长度缩短的评估，确定DSS的最佳剂量为4%。MSC治疗改善了小鼠的便血和体重下降，并显著抑制了结肠长度的缩短。在骨髓间充质干细胞处理的大鼠直肠局部炎性细胞因子α和IL-1β的表达明显减少，分别为40%和15%。局部COX2的表达也被抑制到15%。抗炎细胞因子IL-10的表达也下降至25%。在远端结肠(直肠轻度口侧)，MSC处理的结肠细胞因子的表达也有类似的趋势。提示MSC可能通过抗炎作用对实验性结肠炎有治疗作用。较少

Bone marrow-derived cells including a small amount of mesenchymal stem cells (MSCs) had therapeutic effects for clinical human and experimental animal colitis. Its detailed mechanism(s) may be partly mediated by mucosal regeneration, since MSCs have potential for differentiation to several parts of cells. But MSCs was thought to have other functions such as anti-inflammation as well as mucosal regeneration, because anti-inflammatory system is involved in the repair of colitis. We examined the therapeutic efficacy and anti-inflammatory effects of bone marrow-derived MSCs for dextran sulfate sodium (DSS)-induced acute colitis in rats. Experimental colitis was induced by orally administration of 0, 1, 2, or 4% DSS in drinking water for 7 days in inbred male Lewis rats. Bone marrow was extruded from tibias and femurs. Then, its mononuclear cells were isolated and cultured in low-glucose DMEM containing 10% fetal bovine serum for MSCs outgrowth. On 0, 2, and 4 days after the administration … More of DSS, MSCs (5 X 10^6 cells) were injected via tail vein. We checked the volumes of food and water intake, stool condition, and body weight everyday. On day 7, total colon was excised and each colonic mRNA expression of inflammatory cytokines such as TNF-α, IL-1β, IL-10, and COX2 was measured by real time RT-PCR method. We confirmed the MSC's characterization by both the immunostaining for vimentin and α-smooth muscle actin and the cell surface markers such as CD90, the bone marrow progenitor cell marker, but not CD45, HLA-DR, CD11b, nor CD31 using flow cytometric technique. Optimal dose of DSS for the rats used was confirmed at 4% by the assessing for loss of body weight and appetite, bloody fluid stool, and the shortening of colon length. MSC treatment improved the bloody stool and body weight loss, and significantly inhibited the shortening of colon length. At the rectum of MSC-treated rats, expressions of local inflammatory cytokines such as TNF-α and IL-1β were markedly decreased to about 40 and 15%. Local COX2 expression was also suppressed to 15%. IL-10, an anti-inflammatory cytokine, expression was also, decreased to 25%. At the distal colon cite (slightly oral side of rectum), similar tendency was observed about the expressions of cytokines in the MSC-treated colons. These findings suggested that MSC could have the therapeutic efficacy for the experimental colitis via anti-inflammatory functions. Less