【作者】 宋厚盼； 李茹柳； 王一寓； 曾丹； 张磊； 邓娇； 陈炜璇； 陈蔚文；

【Author】 SONG Hou-pan;LI Ru-liu;WANG Yi-yu;ZENG Dan;ZHANG Lei;DENG Jiao;CHEN Wei-xuan;CHEN Wei-wen;Spleen-Stomach Institute,Guangzhou University of Chinese Medicine;

【机构】 广州中医药大学脾胃研究所；

【摘要】 目的:考察白术提取物（AMK）对小肠上皮IEC-6细胞迁移过程多胺介导信号通路中钙离子调控的影响,探讨白术促进胃肠黏膜损伤修复的作用机制。方法:在刀片划痕法细胞迁移模型上观察白术提取物对细胞迁移的影响;流式细胞仪检测细胞质游离钙离子水平([Ca²⁺]cyt);荧光定量PCR法检测钙通道蛋白瞬时受体电位阳离子通道1（TRPC1）mRNA表达;Western Blot法检测TRPC1蛋白表达。结果:①白术提取物（100mg/L和200mg/L,下同）能促进IEC-6细胞迁移、增加[Ca²⁺]cyt、提高TRPC1 mRNA和蛋白表达（P<0.05,P<0.01）。②白术提取物可逆转多胺合成抑制剂（DFMO）所致的细胞迁移抑制、[Ca²⁺]cyt降低、TRPC1 mRNA和蛋白表达降低（P<0.01）。③若去除了细胞外的Ca²⁺(使用无Ca²⁺培养液),则不但空白对照组出现显著的细胞迁移抑制（P<0.01）,且白术提取物促进细胞迁移的作用程度也明显弱于使用含Ca²⁺培养液时（P<0.01）。结论:白术提取物促进小肠上皮细胞迁移的作用与影响多胺介导信号通路中钙离子调控有关,主要作用途径可能是通过促进细胞外Ca²⁺内流而增加[Ca²⁺]cyt,另一较弱的作用途径可能是增加细胞内钙库释放Ca²⁺。更多还原

【Abstract】 Objective:To investigate the effects of Atractylodes macrocephala koidz extract（AMK） on polyamine mediated calcium signaling pathway during intestinal epithelial cell migration,and explore the mechanism of AMK on promoting gastrointestinal mucosal repair after wounding.Methods:Cell migration was observed and calculated on a wounding model of IEC-6 cells induced by a single-edge razor blade;The cytosolic free Ca²⁺ concentration([Ca²⁺]cyt) was detected by flow cytometry;The expression of TRPClmRNA and protein levels were assessed by RT-qPCR and Western blot analysis,respectively.Results:①Treatment with AMK（100mg/L and 200mg/L） caused an acceleration of cell migration,an elevation of[Ca²⁺]cyt,and significant increases in TRPC1 mRNA and protein levels（P<0.05,P<0.01）.②AMK reversed the inhibition of cell migration,reduction of[Ca²⁺]cyt,and decreased TRPC1 mRNA and protein expression induced by polyamine inhibitor DFMO（P<0.01）.③When removal of extracellular Ca²⁺ by using of Ca²⁺-free media,the cell migration was significantly inhibited both in the control and AMK groups（P<0.01）.Conclusion:AMK promoting intestinal epithelial cell migration is related to its role of influencing polyamine mediated Ca²⁺ signaling pathway,which mainly promotes extracellular Ca²⁺ influx and increases[Ca²⁺]cyt,another weaker approach may increase[Ca²⁺]cyt by elevating intracellular Ca²⁺ store release.更多还原