党参和甘草糖复合物对IEC-6细胞迁移和膜电位的影响

Effect of Codonopsis and Glycyrrhiza glycoconjugates on migration and membrane potential of IEC-6 cells

目的观察党参糖复合物和甘草糖复合物对小肠上皮细胞IEC-6迁移和细胞膜电位的影响,探讨益气健脾中药党参和甘草促进胃肠黏膜损伤修复的作用机制。方法在正常或钾通道抑制剂4-氨基吡啶(4-AP)负荷下,分别加入党参和甘草糖复合物(25~200 mg·L^(-1))与IEC-6细胞培养24 h,相差显微镜下观察细胞迁移数,流式细胞仪检测细胞膜电位。结果与细胞正常对照组比较,党参和甘草糖复合物(50和100 mg·L^(-1))可提高细胞迁移数(P<0.01,P<0.05)。与正常对照组比较,4-AP可减少细胞迁移数(P<0.01);与4-AP模型组比较,党参和甘草糖复合物(50~200 mg·L^(-1))可逆转4-AP所致的细胞迁移抑制(P<0.01)。流式细胞仪检测结果表明,与正常对照组比较,党参和甘草糖复合物(50 mg·L^(-1))可提高细胞膜电位(P<0.01),增加细胞膜超极化水平;与正常对照组比较,4-AP模型组细胞膜电位降低(P<0.01),增加细胞膜去极化水平;与4-AP模型组比较,党参和甘草糖复合物(100和200 mg·L^(-1))可逆转4-AP所致的细胞膜去极化(P<0.01)。结论党参和甘草促进胃肠黏膜损伤修复的作用机制,可能与其糖复合物影响小肠上皮细胞迁移的多胺介导钾通道激活信号通路有关。

OBJECTIVE! To observe the effect of Codonopsis and Glycyrrhiza glycoconjugates on migration and membrane potential of small intestinal epithelial cells （IEC-6）, and to explore the promoting effects of Yiqi jianpi herb Codonopsis and Glycyrrhiza on gastrointestinal mucosal injury repair and the underlying mechanisms. METHODS Under normal conditions or loaded with the inhibitor of potassium channel 4-aminopyridine （4-AP）, IEC-6 cells were treated with Coclonopsis and Glycyrrhiza glycoconjugates （25-200 mg ·L-1） for 24 h, respectively. IEC-6 cell migration was observed by the phase contrast microscope and cell membrane potential was detected by flow cytometry. RESULTS Codonopsis and Glycyrrhiza glycoconjugates （50 and 100 mg·L-1） increased the number of migrated IEC-6 cell compared with normal control group（P〈0.01,P〈0.05）. Compared with normal control group, 4-AP reduced the number of migrated IEC-6 cell （P〈0.01）. Codonopsis and Glycyrrhiza glycoconjugates （50-200 mg·L-1） reversed cell migration inhibited by 4-AP significantly when compared with 4-AP model group（P〈0.01）. The results of flow cyometry analysis showed that the cell membrane poten- tial was increased after treatment with Codonopsis and Glycyrrhiza glycoconjugates （50 rag· L-1 ） compared with normal control group ~nd resulted in an increase in cell membrane hyperpolarization （P〈0.01）. Compared with normal control group, 4-AP decreased the cell membrane potential （P〈0.01）and resulted in cell membrane depolarization. Also, compared with 4-AP model group, cell membrane depolarization induced by 4-AP was rever,ed by treatment with Codonopsis and Glycyrrhiza glycoconjugates （ 100 and 200 rag· L-1）. CONCLUSION Coclonopsis and Glycyrrhiza glycoconjugates may promote gastrointestinal mucosal injury repair and the mechanisms may involve the activation of signaling pathways by affecting polyamine-dependent intestinal epithelial cell migration voltage-gated K＋ channels.