白术糖复合物对IEC-6细胞分化及绒毛蛋白表达的影响

Effects of Atractylodes macrocephala Monosaccharide Composition on Cytodifferentiation and Villin Expression of IEC-6 Cells in vitro

目的:观察白术糖复合物对小肠隐窝细胞(IEC-6)细胞分化及分化标志物之一绒毛蛋白(villin)表达的影响。方法:IEC-6细胞分为空白对照组、阳性对照药胃泌素组(250μg/L)、白术糖复合物组(62.5、125、250、500、1000 mg/L),经相应处理后。光镜下观察细胞大体形态;透射电镜下观察细胞超微结构;RT-PCR法检测villin mRNA表达;免疫荧光法检测villin蛋白表达。结果:①给药7 d,光镜下空白对照组细胞处于幼稚状态;胃泌素和白术糖复合物组细胞处于分化状态;②给药20 d,电镜下空白对照组胞核较大,常染色质比例较高,细胞边缘仅见少量微绒毛;胃泌素和白术糖复合物组胞核较小,常/异染色质比例协调,细胞边缘生成大量微绒毛;③给药6 h,空白对照组细胞villin mRNA表达非常少,胃泌素组和白术糖复合物组villin mRNA表达均明显高于空白对照组;④给药7 d,免疫荧光检测发现空白对照组细胞仅有很少villin蛋白表达;胃泌素组表达非常强烈;白术糖复合物组表达稍弱于胃泌素组,但有特异性绒毛蛋白聚集点。结论:白术糖复合物有通过上调IEC-6细胞绒毛蛋白表达及分布而促其分化的作用。

Objective：To study the influence of Atractylodes macrocephala monosaccharide composition（AMMC） on IEC-6 cells by observing cytodifferentiation and expression of villin.Method：Observed cell morphous under light microscope,observed cell ultramicrostructure under transmission electron microscope,evaluated villin mRNA by way of RT-PCR,and detected protein of villin by immunofluorescence.Cells were divided into the groups including blank control,positive control（given with gastrin 250 μg/L） and AMMC groups（given with AMMC 62.5,125,250,500,1 000 mg/L respectively）.Results：1.Cells in blank control group were immature under light microscope,which were in state of differentiation in groups with gastrin and different concentration of AMMC.2.Cells in blank control group had bigger nucleuses with higher ratio of heterochromatin and much less villin at the edge of cells,which in groups with gastrin and different concentration of AMMC had smaller nucleuses with normal ratio of heterochromatin /heterochromatin and abundance of villin at the edge of cells.3.Villin mRNA of IEC-6 cells in groups with gastrin and different concentration of AMMC were obviously much more than that of cells in blank control group 6 h afeer treatment.4.Little protein of villin was detected by immunofluorescence in blank control group,which was much brighter in group with gastrin,and less bright in group with different concentration of AMMC,in which specific villin assembled.Conclusion：AMMC can promote cytodifferentiation by up-regulating the expression and distribution of villin in IEC-6 cells.