Song Hou-Pan, Li Ru-Liu, Wang Yi-Yu, Tu Xiao-Hua, Deng Jiao, Chen Wei-Wen
Zhongguo Zhong Xi Yi Jie He Za Zhi. 2016 Jul;36(7):861-866.
To observe the effects of methanol extracts from Atractylodes macro- cephalae Rhizoma (AMR) on the proliferation and migration of IEC-6 cell (small intestinal epithelial cells) and the expression of phospholipase C-γ1 (PLC-γ1) , and to explore the mechanism of AMR (a Chinese herb capable of invigorating Pi replenishing qi) for promoting repair of gastrointestinal mucosal injury.
IEC-6 cells were divided into the blank group, the positive control (spermidine, SPD; 5 μmol/L) group, AMR extracts groups (50, 100, and 200 mg/L). The alpha-difluoromethylornithine (DFMO, polyamines synthesis inhibitor) group, the SPD +DFMO group, AMR extracts (50, 100, and 200 mg/L) +DF- MO groups were set up in stress test. IEC-6 cells were cultured by adherence for 24 h,and then treated with AMR extracts for appropriate periods of time. Effects of IEC-6 cell proliferation after action of AMR extracts were detected by Real-time Cell Analyzer (RTCA). The effect of AMR extracts on IEC-6 cell migration number was detected using scratch method. mRNA and protein expressions of PLC-γ1 levels were detected by fluorescent quantitative polymerase chain reaction ( RT-qPCR) and Western blot respectively.
Compared with the blank group, AMR extracts showed no obvious effect on IEC-6 cell proliferation (P >0. 05). But SPD and AMR extracts (100 and 200 mg/L) not only promoted IEC-6 cell migration (P <0. 01), but also improved mRNA and protein expressions of PLC-γl in the process of cell migration (P <0. 01). Compared with the DFMO group, SPD and AMR extracts (100 and 200 mg/L) could reverse inhibitory effects of DFMO on cell migration, and mRNA and protein expressions of PLC-γl (all P <0. 01).
AMR extracts played roles in repairing gastrointestinal mucosal injury possibly by promoting polyamine mediated intestinal epithelial cell migration, and its effect on intestinal epithelial cell proliferation was not main potentcy.
观察白术甲醇提取物对IEC-6细胞（小肠上皮细胞）增殖、迁移及磷脂酶C-γ1（PLC-γ1）表达的影响，探讨白术这一健脾益气中药促进胃肠黏膜损伤修复的机制。
将IEC-6细胞分为空白组、阳性对照组（亚精胺，SPD；5 μmol/L）、白术提取物组（50、100和200 mg/L）。应激试验中设置α-二氟甲基鸟氨酸（DFMO，多胺合成抑制剂）组、SPD +DFMO组、白术提取物（50、100和200 mg/L）+DFMO组。IEC-6细胞贴壁培养24 h，然后用白术提取物处理适当时间。采用实时细胞分析仪（RTCA）检测白术提取物作用后IEC-6细胞的增殖情况。采用划痕法检测白术提取物对IEC-6细胞迁移数量的影响。分别通过荧光定量聚合酶链反应（RT-qPCR）和蛋白质免疫印迹法检测PLC-γ1水平的mRNA和蛋白表达。
与空白组相比，白术提取物对IEC-6细胞增殖无明显影响（P>0.05）。但SPD及白术提取物（100和200 mg/L）不仅促进IEC-6细胞迁移（P<0.01），还能提高细胞迁移过程中PLC-γ1的mRNA和蛋白表达（P<0.01）。与DFMO组相比，SPD及白术提取物（100和200 mg/L）可逆转DFMO对细胞迁移及PLC-γ1的mRNA和蛋白表达的抑制作用（均P<0.01）。
白术提取物可能通过促进多胺介导的肠上皮细胞迁移发挥修复胃肠黏膜损伤的作用，其对肠上皮细胞增殖的作用并非主要效能。
