唐古特大黄多糖促进肠上皮细胞的增殖、移行和分化

Effects of Rheum tanguticum polysaccharide in promoting HIE Ccell proliferation, differentiation and migration

目的 观察唐古特大黄多糖对正常人肠上皮细胞(HIEC)增殖、移行和分化的影响,探讨其肠粘膜可能的修复机制。方法 HIEC细胞接种后24h,加入不同浓度的大黄多糖,加药后24h,MTT法观察细胞增殖情况,HE染色法和碱性磷酸酶活性的测定观察细胞分化情况,细胞接种后待长成单细胞层后,在细胞层上作一圆形损伤区,并加入大黄多糖,分别于加药后0、12、24、36、48h倒置显微镜下观察细胞移行情况并拍照,采用图像分析定量测定缺损区随时间变化的情况。结果 大黄多糖可剂量依赖性地促进HIEC细胞的增殖、移行和分化,与正常对照组比较,细胞增殖、移行以及碱性磷酸酶活性差异均具有显著性(P< 0 .05或P<0. 01)。结论 大黄多糖可能通过促进肠上皮细胞的增殖移行和分化来促进肠粘膜的损伤修复。

Aim To explore the possible mechanism of Rheum tanguticum polysaccharide (RTP) in repairing mucosal membrane by observing its effects on the proliferation, differentiation and migration in normal human intestinal epithelial cell line (HIEC) in vitro.Methods 24 h after cell inoculation, RTP were added into the cultured media with different concentrations. MTT assay was used to observe the effect of RTP on cell proliferation, while H&E staining and alkaline phosphatase (ALP) activity measurement were introduced to assess cell differentiation. After the cultured HIEC cells formed a complete monolayer,an artificial wound with a cell free area of constant size was made in the center of the mucosal cell sheet. RTP were added just after artificial wounding and its effects on cell migration were monitored under phase-contrast microscopy and photographs were taken every 12 h for 48 h after wounding. And changes in the cell-free area over time were quantified by using an image analyzer.Results RTP dose-dependently enhanced the proliferation, migration and differentiation of HIEC cells. Compared with Normal group, there existed significant differences on cell proliferation, migration and ALP activity(P<0.05 or P<0.01).Conclusion RTP might improve intestinal mucosal repair by enhancing intestinal epithelial cell proliferation, migration and differentiation.