苦参素联合骨髓间充质干细胞对溃疡性结肠炎大鼠结肠组织的修复作用

发布时间：2018-10-16 14:18

【摘要】：目的 探讨骨髓间充质干细胞(Mesenchymal stem cells,MSCs)以及苦参素联合MSCs对溃疡性结肠炎(Ulcerative colitis, UC)大鼠结肠组织的修复作用。 方法 从雄性大鼠骨髓中提取MSCs。将40只雌性大鼠随机分为正常组（A）、模型组（B）、MSCs组（C）及MSCs加苦参素组（D），每组10只。B、C、D组用三硝基苯磺酸/乙醇复合法建立UC模型，24h后，A组和B组分别经尾静脉输入生理盐水1mL；C组、D组分别经尾静脉输入1×106MSC s悬液1mL；D组给予苦参素肌肉注射2周。留取结肠组织标本，，进行病理学观察，用PCR技术和原位荧光杂交法检测C、D组是否存在MSCs Y染色体性别决定区域(sex determining region Y,SRY)；免疫组织化学、Western blotting检测各组结肠组织bFGF的蛋白水平；RT-PCR技术分析结肠组织TNFα、IL-1β及bFGF mRNA的表达。 结果 C、D组结肠组织中可见SRY阳性；A组bFGF mRNA和蛋白的表达及TNFα、IL-1βmRNA的表达均较低；较A组，B、C、D组结肠组织中bFGF及TNFα、IL-1β的表达均增高(P 0.05)；较B组，C、D组bFGF表达增高，TNFα、IL-1β表达下降(P 0.05)；较C组，D组bFGF表达增高，TNFα、IL-1β表达下降(P 0.05)。 结论 MSCs能在UC大鼠结肠组织中定植，能够抑制UC大鼠结肠组织中的炎症反应、修复损伤的结肠黏膜,其作用机制可能与其下调结肠组织中TNFα、IL-1β mRNA的表达、上调bFGF mRNA及蛋白的表达有关；苦参素联合MSCs治疗大鼠UC有协同增强作用。
[Abstract]:Objective to investigate the repair effect of bone marrow mesenchymal stem cells (Mesenchymal stem cells,MSCs) and matrine combined with MSCs on colonic tissue of (Ulcerative colitis, UC) rats with ulcerative colitis. Methods MSCs. was extracted from male rat bone marrow. Forty female rats were randomly divided into normal group (A), model group), (B), MSCs group (C) group) and MSCs + matrine group (D), group, 10 rats in each group). The UC model was established by trinitrobenzenesulfonic acid / ethanol compound method in group B and C group. 24 hours later, 1 mL of normal saline was injected into group A and group B respectively through tail vein. Group C and group D were injected with 1 脳 106MSC s suspension through tail vein for 2 weeks. The colonic tissue samples were collected for pathological observation, and PCR technique and in situ fluorescence hybridization were used to detect the presence of (sex determining region Y sex determining region of MSCs Y chromosome in group C ~ (2 +), and immunohistochemical, Western blotting was used to detect the protein level of bFGF in colon tissue of each group. The expression of TNF 伪, IL-1 尾 and bFGF mRNA in colon tissue was analyzed by RT-PCR technique. Results the expression of SRY, TNF 伪, IL-1 尾 mRNA, bFGF, TNF 伪 and IL-1 尾 in colon tissue of group A were lower than those in group A, and the expression of bFGF and TNF 伪, IL-1 尾 in group A were higher than those in group A (P 0.05). Compared with group B, the expression of bFGF and TNF 伪 and IL-1 尾 in group C were higher than those in group B, and the expression of bFGF and TNF 伪 and IL-1 尾 in group D were higher than those in group C (P 0.05). Conclusion MSCs can colonize the colon tissue of UC rats, inhibit the inflammatory reaction in the colon tissue of UC rats, repair the damaged colon mucosa, and its mechanism may be related to the down-regulation of the expression of TNF 伪 and IL-1 尾 mRNA in the colon tissue. The up-regulation of bFGF mRNA and protein expression was related to the synergistic effect of oxymatrine combined with MSCs on UC in rats.
【学位授予单位】：辽宁医学院
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R574.62

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