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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 observe the effect of Atractylodes macrocephala complex on the differentiation and differentiation of IEC-6 cells and villin, a marker of differentiation. Methods: IEC-6 cells were divided into blank control group, positive control gastrin group (250μg / L), atractylodes macrocephala complex group (62.5,125,250,500,1000 mg / L), after corresponding treatment. The morphology of cells was observed under light microscope. The ultrastructure of cells was observed by transmission electron microscopy. The expression of villin mRNA was detected by RT-PCR and the expression of villin by immunofluorescence. Results: ①The cells in the blank control group were in a naive state under the light microscope for 7 days. The cells in the group of gastrin and atractylodes macrocephala were in a differentiated state. ② After 20 days of administration, the nuclei of the blank control group were larger under electron microscope. The proportion of chromatin was high, only a few microvilli were seen at the edge of the cell. The nuclei of gastrin and atractylodes macrocephala were smaller, the ratio of common / heterochromatin was coordinated and a large number of microvilli were formed on the edge of cells. Villin mRNA expression of group cells was very little, villin mRNA expression of gastrin group and Atractylodes macrocephala complex were significantly higher than that of blank control group. ④After administration for 7 days, there was only a few villin protein expression in the blank control group detected by immunofluorescence ; Gastrin expression was very strong; Atractylodes glycosides group slightly weaker expression of gastrin group, but there is a specific villin aggregation point. CONCLUSION: Atractylodes macrocephala complex can promote differentiation through up-regulating the expression and distribution of villi in IEC-6 cells.