Neural stem cell transplantation in the stomach rescues gastric function in neuronal nitric oxide synthase-deficient mice
Neural stem cell transplantation in the stomach rescues gastric function in neuronal nitric oxide synthase-deficient mice
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DOI：
10.1053/j.gastro.2005.08.055
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发表时间：
2005-12-01
期刊：
影响因子：
29.4
通讯作者：
Pasricha, PJ
中科院分区：
文献类型：
--
作者：
Micci, MA;Kahrig, KM;Pasricha, PJ
背景与目的：一氧化氮是肠神经系统中主要的抑制性神经递质。一氧化氮能神经元的丧失或功能障碍与运动的严重破坏、顽固性症状和长期痛苦有关。本研究的目的是评价幽门内移植神经干细胞（NSCs）对胃轻瘫的遗传模型nNOS（-/-）小鼠胃排空和幽门功能的影响。研究方法：从经转基因工程改造以表达绿色荧光蛋白的胚胎小鼠中分离NSC，并将其移植到nNOS（-/-）小鼠的幽门中。移植的细胞在幽门切片中可见，并通过免疫荧光染色进一步表征。移植后一周，使用酚红法测量胃排空到非营养餐，并通过测量幽门条在无肾上腺素能、非胆碱能条件下对电场刺激（EFS）作出反应的器官浴中的松弛来评估幽门功能。结果：移植后1周，移植的神经干细胞分化为神经元，并表达神经元型一氧化氮合酶。与注射载体的对照相比，接受NSC的小鼠的胃排空显著增加（49.67%对35.09%;通过Student t检验，P <0.01）。EFS诱导的幽门条松弛也显著增加（双向方差分析P <0.01）。一氧化氮合酶抑制剂N-G-硝基-L-精氨酸甲酯和神经元阻滞剂河豚毒素阻断了EFS诱导的舒张，表明所观察到的效果是NO介导的和神经源性的。结论：我们的研究结果支持神经干细胞移植作为一种可行的治疗选择神经肠道疾病的潜力。
Background & Aims: Nitric oxide is a major inhibitory neurotransmitter in the enteric nervous system. Loss or dysfunction of nitrinergic neurons is associated with serious disruptions of motility, intractable symptoms, and long-term suffering. The aim of this study was to evaluate the effect of intrapyloric transplantation of neural stem cells (NSCs) on gastric emptying and pyloric function in nNOS(-/-) mice, a well-established genetic model of gastroparesis. Methods: NSCs were isolated from embryonic mice transgenically engineered to express green fluorescent protein and transplanted into the pylorus of nNOS(-/-) mice. Grafted cells were visualized in pyloric sections and further characterized by immunofluorescence staining. One week posttransplantation, gastric emptying to a non-nutrient meal was measured using the phenol red method and pyloric function was assessed by measuring the relaxation of pyloric strips in an organ bath in response to electrical field stimulation (EFS) under no adrenergic, noncholinergic conditions. Results: One week following implantation, grafted NSCs differentiated into neurons and expressed neuronal nitric oxide synthase. Gastric emptying was significantly increased in mice that received NSCs as compared with vehicle-injected controls (49.67% vs 35.09%; P < .01 by Student t test). EFS-induced relaxation of pyloric strips was also significantly increased (P < .01 by 2-way analysis of variance). The nitric oxide synthase inhibitor N-G-nitro-L-arginine methyl ester and the neuronal blocker tetrodotoxin blocked the EFS-induced relaxation, indicating that the observed effect is NO mediated and neuronally derived. Conclusions: Our results support the potential of NSC transplantation as a viable therapeutic option for neuroenteric disorders.