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目的:探讨基因重组肠三叶因子(ITF)对肠上皮细胞(IEC-6)是否具有促进修复和保护的作用。方法:制备不同浓度的重组ITF。首先,采用MTT法观察ITF对IEC-6细胞增殖活性的影响;其次,制作IEC-6细胞机械损伤模型,观察ITF对IEC-6细胞移行速率的影响;最后,采用烧伤血清致IEC-6细胞损伤模型,观察ITF对IEC-6细胞的保护作用。结果:各种浓度的ITF对IEC-6细胞具有一定的促增殖作用,但差异无统计学意义(P=0.19)。ITF可加快IEC-6细胞移行速度,是对照组的2~3倍(P<0.01)。ITF可明显降低烧伤血清致IEC-6细胞损伤程度,反映细胞损伤的谷草转氨酶(AST)、乳酸脱氢酶(LDH)活性显著下降,且作用持续12 h。联合使用黏蛋白(Mucin)可显著加强ITF的细胞保护作用,谷丙转氨酶(ALT)、AST、LDH活性显著降低,且保护作用延长至48 h(P<0.01)。结论:ITF可加快细胞移行速度,促进受损肠黏膜屏障的重建,对肠上皮细胞具有明显的保护作用。单独使用ITF对细胞增殖影响不明显,与Mucin联合使用,其保护作用明显增强。
Objective: To investigate whether recombinant human intestinal trefoil factor (ITF) can promote the repair and protection of intestinal epithelial cells (IEC-6). Methods: Different concentrations of recombinant ITF were prepared. First, MTT assay was used to observe the effect of ITF on the proliferation of IEC-6 cells. Secondly, IEC-6 cells were prepared and the effects of ITF on the migration rate of IEC-6 cells were observed. Finally, IEC-6 cells The model of injury was observed and the protective effect of ITF on IEC-6 cells was observed. Results: Various concentrations of ITF had a certain effect of promoting proliferation of IEC-6 cells, but the difference was not statistically significant (P = 0.19). ITF accelerated IEC-6 cell migration 2 to 3 times that of the control group (P <0.01). ITF could significantly reduce the degree of injury of IEC-6 cells induced by burn serum, and significantly decrease the activities of aspartate aminotransferase (AST) and lactate dehydrogenase (LDH) in cell injury, and the effect lasted 12 h. Combination of Mucin can significantly enhance the cytoprotective effect of ITF. The activities of ALT, AST and LDH were significantly reduced, and the protective effect was prolonged to 48 h (P <0.01). Conclusion: ITF can speed up cell migration and promote the reconstruction of impaired intestinal mucosal barrier, which has a significant protective effect on intestinal epithelial cells. The use of ITF alone had no significant effect on cell proliferation. In combination with Mucin, its protective effect was significantly enhanced.