摘要
目的以Ednrb基因敲除小鼠为动物模型,探讨应用牙髓干细胞(dental pulp stem cells,DPSCs)治疗先天性巨结肠相关性小肠结肠炎(Hirschsprung disease associated enterocolitis,HAEC)的疗效,为临床治疗HAEC寻找新途径。方法将同龄内皮素受体B基因突变鼠(花斑鼠)按随机数字法平均分为实验组及对照组,每组8只。实验组采用DPSCs腹腔注射的方式,细胞计数为0.5×10^6/kg,对照组注射同等量PBS缓冲液。正常组采用同龄野生型小鼠(8只),不做任何处理。对各组小鼠的体重变化、4周时存活情况进行观察记录,对各组小鼠肠炎发生情况进行病理评分。同时观察TNF-α、IFN-γ、TGF-β以及IL-10等炎症因子表达情况,采用免疫荧光技术检测肠道CD4、CD20、CD56及CD68等相关免疫细胞表面标记物表达情况。结果实验组小鼠寿命明显长于对照组[(26.2±2.5) d比(22.0±3.0) d],4周生存率明显高于对照组(62.50%比16.70%),组间比较,上述指标比较,差异均有统计学意义(P<0.05)。2周龄时,对照组小鼠血浆中促炎因子TNF-α、IFN-γ含量分别为35.21±2.08和32.31±7.41,与正常组29.52±0.19和18.94±0.49比较,明显升高,且差异有统计学意义(P<0.05)。对照组抑炎因子TGF-β、IL-10含量分别为41.38±2.39和53.89±4.90,较正常组14.84±0.02和43.56±0.97明显降低,且差异有统计学意义(P<0.05)。而DPSCs移植后,实验组TNF-α及IFN-γ含量分别为31.31±0.63和24.75±2.76,与对照组35.21±2.08和32.31±7.41比较,含量明显下降,且差异有统计学意义(P<0.05)。实验组TGF-β、IL-10含量分别为122.97±0.90和69.00±0.28,较对照组14.84±0.20和41.38±2.39明显升高,组间比较,差异有统计学意义(P<0.05)。同时,实验组与对照组比较,CD4、CD20、CD56及CD68等免疫细胞表面标记物表达均有降低。结论牙髓干细胞具有免疫调节、抑制组织炎症介质释放,减轻组织炎症损伤等作用,可明显减轻缓解HAEC的症状,可为临床治疗HAEC提�
Objective To employ an animal model of Ednrb knockout mice for exploring the efficacy of the treatment using dental pulp stem cells (DPSCs) in Hirschsprung's disease associated enterocolitis (HAEC). Methods The experiment group received an intraperitoneal injection of DPSCs with a cell count of 0.5×10^6/kg.The control group received an injection of the same amount of PBS buffer and the normal group included wild-type mice with the same age but without any treatment.Changes in body weight of each group of mice and 4-week survival were recorded.For each group, gross macroscopic intestinal changes were observed and the pathogenesis of enteritis was evaluated.The expressions of such proinflammatory factors as TNF-α, TGF-β, IFN-γ, TGF-β and IL-10 and such immune cell surface markers as CD4, CD20, CD56 and CD68 in intestinal tract were detected by immunofluorescence. Results The lifespan of experiment group was significantly longer than that of control group [(26.2±2.5) vs.(22.0±3.0) days] and 4-week survival rate was significantly higher than that of control group (62.50% vs.16.70%). When the above parameters were compared, the between-group differences were statistically significant (P<0.05). At an age of 2 weeks, the plasma levels of TNF-α and IFN-γ were 35.21±2.08 and 32.31±7.41 in control group and both were significantly higher than those in normal group 29.52±0.19 and 18.94±0.49 (P<0.05). The levels of TGF-β and IL-10 were 41.38±2.39 and 53.89±4.90 in control group and both were significantly lower than 14.84±0.02 and 43.56±0.97 in normal group respectively.And the differences were statistically significant (P<0.05). After transplanting DPSCs, the contents of TNF-α and IFN-γ were 31.31±0.63 and 24.75±2.76 in experiment group versus 35.21±2.08 and 32.31±7.41 in control group.The contents decreased significantly and the difference was statistically significant (P<0.05). The contents of TGF-β and IL-10 were 122.97±0.90 and 69.00±0.28 in experiment group and both were significantly higher
作者
蒙信尧
陈绪勇
肖俊
朱天琦
冯杰雄
Meng Xinyao;Chen Xuyong;Xiao Jun;Zhu Tianqi;Feng Jiexiong(Department of Pediatric Surgery, Affiliated Tongji Hospital, Tongji Medical College, Huazhong University of Science & Technology, Wuhan 430030, China)
出处
《中华小儿外科杂志》
CSCD
北大核心
2019年第3期263-268,共6页
Chinese Journal of Pediatric Surgery
基金
国家自然科学基金(81700448)。
关键词
小肠结肠炎
先天性巨结肠
牙髓
干细胞
免疫抑制
Hirschsprung disease associated enterocolitis
Hirschsprung’s disease
Dental Pulp
Stem Cells
Immunodepression
