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目的:探讨骨髓间充质干细胞(MSC)对大鼠小肠缺血-再灌注(I/R)损伤时肠屏障的保护作用。方法:Ficoll法分离培养大鼠的MSC。36只大鼠随机分为对照组和治疗组,每组18只。治疗组在建模后60 min,通过尾静脉将1×106/ml的MSC注射至大鼠体内,对照组注射等量的等渗盐水。于输注后6、24和72 h分批处死大鼠,获取血液、小肠组织等标本。检测血液IL-6、IL-10、肿瘤坏死因子-α(TNF-α)、丙二醛(MDA)、超氧化物歧化酶(SOD)的浓度,同时观察小肠组织病理改变。结果:治疗组大鼠的血清IL-6、TNF-α水平在治疗后6 h和24 h显著低于对照组(P<0.05);IL-10则显著高于对照组(P<0.05);MDA水平在各时间点组间均有显著性差异(P<0.05),SOD在治疗后24 h组间有显著性差异(P<0.01)。治疗组小肠组织荧光显微镜下观察可见DAPI阳性细胞。病理和免疫组化显示,治疗组小肠黏膜损伤在6和24 h较对照组轻,24和72 h肠黏膜组织细胞增殖较对照组明显(P<0.05)。结论:移植的MSC能迁移并定植于受损肠道,对肠I/R损伤有一定的保护作用。MSC能调节大鼠肠I/R损伤后引起的全身性炎症反应。
Objective: To investigate the protective effect of bone marrow mesenchymal stem cells (MSCs) on the intestinal barrier in rats with intestinal ischemia reperfusion (I / R) injury. Methods: The isolated and cultured rat MSCs were isolated by Ficoll method. 36 rats were randomly divided into control group and treatment group, 18 rats in each group. At 60 min after the model was established in the treatment group, 1 × 10 6 / ml MSCs were injected into the rats through the tail vein, and the control group was injected with the same amount of isotonic saline. After 6, 24 and 72 h infusion rats were killed in batches, access to blood, small intestine and other specimens. The concentrations of IL-6, IL-10, TNF-α, MDA and SOD in the blood were detected and the histopathological changes in small intestine were observed. Results: The serum levels of IL-6 and TNF-α in the treatment group were significantly lower than those in the control group at 6 and 24 h after treatment (P <0.05), while the levels of IL-10 in the treatment group were significantly higher than those in the control group (P <0.05). There were significant differences in the levels of MDA at all time points (P <0.05), and there was a significant difference in SOD between the 24 h groups after treatment (P <0.01). In the treatment group, DAPI positive cells were observed under the fluorescence microscope. The pathological and immunohistochemical results showed that the injury of intestinal mucosa in treatment group was lighter than that in control group at 6 and 24 h, and the proliferation of intestinal mucosa in 24 and 72 h group was significantly higher than that in control group (P <0.05). CONCLUSION: The transplanted MSC can migrate and colonize the damaged gut, and have some protective effect on intestinal I / R injury. MSC modulates the systemic inflammatory response induced by intestinal I / R injury in rats.