基因重组肠三叶因子对肠上皮细胞修复和保护作用的研究

Protective and reparative effects of recombinant intestinal trefoil factor on intestinal epithelial cells

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 explore the protective and reparative effects of recombinant intestinal trefoil factor on intestinal epithelial cells. Methods ： Firstly, different concentrations of ITF were added to IEC-6 cells, and MTT assay was performed. Secondly, wounds were established in confluent monolayers of IEC-6 cells, and cells migrating across the wound border were counted in a blind manner. Lastly, burn serum of wistar rats was isolated at 24h after thermal injury. The changes of ALT, AST and LDH were detected after the application of the stimulation, Results ： MTT assay showed that ITF did not obviously stimulate proliferation of IEC-6 cells. However, ITF enhanced migration activity 2-3 fold. In addition, ITF could protect IEC-6 cells from injury induced by burn serum. Conclusion： ITF is demon-strated to promote the healing of gastric mucosal lesions by enhancing migration activity and protect the epithelial layer of the gastrointestinal tract from damage. And the can enhance the protective effects.