白术多糖调控钙离子以促进细胞迁移及E-钙黏蛋白表达的研究

Research of Atractylodes macrocephala Polysaccharides in Promoting Cell Migration and E-cadherin Expression Through Regulating Calcium Ions

目的观察白术多糖对大鼠小肠上皮细胞(IEC-6)钙离子水平、细胞迁移和E-钙黏蛋白表达的影响,探讨益气健脾中药白术促进胃肠黏膜损伤修复的作用机制。方法:流式细胞仪检测白术多糖对细胞游离钙离子水平([Ca^(2+)]cyt)的影响;划痕法复制细胞迁移模型,观察白术多糖对细胞迁移的影响;Western blot法和免疫荧光法检测白术多糖对细胞E-钙黏蛋白表达的影响。结果白术多糖(25,50,100 mg·L^(-1))可增加细胞[Ca^(2+)]cyt、促进细胞迁移、提高E-钙黏蛋白表达,与正常对照组比较,差异有统计学意义(P<0.05,P<0.01);可逆转多胺合成抑制剂α-二氟甲基鸟氨酸(DFMO)所致的[Ca^(2+)]cyt降低、细胞迁移抑制和E-钙黏蛋白表达减少,与DFMO组比较,差异有统计学意义(P<0.05,P<0.01)。结论白术多糖可通过提高细胞钙离子水平以促进细胞迁移和E-钙黏蛋白表达,为探讨益气健脾中药白术修复胃肠黏膜损伤的作用机制提供参考。

Objective To investigate the effect of Atractylodes macrocephala polysaccharides（AMP） on calcium ion level, cell migration and E-cadherin expression in mice intestinal epithelial cells（IEC-6）, and to explore the mechanism of Rhizoma Atractylodis Macrocephalae,a Chinese herbal medicine with the actions of strengthening qi and invigorating spleen in promoting the recovery of gastrointestinal mucosa from wounding. Methods The effect of AMP on the free calcium ion concentration（[Ca（2＋）]cyt）in IEC-6 was detected by flow cytometry. Cell migration model established by scratch damage was used for the observation of the effect of AMKP on IEC-6 cell migration. Immunofluorescence method and Western blot method were used to detect the influence of AMP on the expression of E-cadherin. Results The results showed that AMP（25,50 or 100 mg·L（-1）） could increase [Ca（2＋）]cyt,promote cell migration and enhance E-cadherin expression（P 0.05 or P 0.01 compared with the normal control group）. AMP could reverse the reduction of [Ca（2＋）]cytand E-cadherin expression as well as the inhibition of cell migration induced by polyamines synthesis inhibitor DFMO（P 0.05 or P 0.01 compared with DFMO model group）. Conclusion The results indicate that AMP can increase [Ca（2＋）]cytto promote cell migration and E-cadherin expression,which will provide reference for studying Rhizoma Atractylodis Macrocephalae in promoting the recovery of gastrointestinal mucosa from wounding.