目的利用大鼠小肠上皮细胞系(IEC-6),研究单味中药提取物对小肠上皮细胞增殖和葡萄糖吸收转运功能的调控作用。方法藿香、苍术、黄柏和石膏等4种中药被定向提取,MTT细胞增殖试验筛选提取物的适宜作用质量浓度,测定各处理组细胞的葡萄糖吸收率和Na^+,K^+-SATP酶活性,同时采用荧光定量PCR技术分析细胞中钠-葡萄糖共转运载体(SGLT1)、葡萄糖转运载体2(GLUT2)和Na^+,K^+-ATP酶的mRNA表达量。结果各中药提取物对大鼠小肠上皮细胞增殖的影响与其作用质量浓度有关。50μg/mL苍术挥发油和10μg/mL黄柏生物碱可提高IEC-6细胞葡萄糖的吸收功能,并显著上调了SGLTl和Na^+,K^+ -ATP酶mRNA表达量;50μg/mL藿香挥发油促进了细胞的葡萄糖吸收,但仅对GLUT2的基因表达有上调效应(P〈0.05);而50μg/mL石膏水提物对细胞葡萄糖的吸收无促进作用(P〉0.05)。结论苍术挥发油、藿香挥发油和黄柏生物碱对细胞增殖和葡萄糖吸收均有促进作用,但其影响机制并不完全一样。
Objective To study the effects of Chinese herb extracts on cell proliferation and glucose absorption in intestinal epithelial cell line (IEC-6 cells) of rats. Methods The extracts of four Chinese herbs, including Herba Agastaches (HA), Rhizoma Atractylodis (RA), Cortex Phellodendri (CP), and Gypsum Fibrosum (GF), were made. Their suitable concentration on the cell proliferation in IEC-6 cells was determined by MTT method, and glucose absorption and the activity of Na^+ , K^+-ATPase in IEC-6 cells were assayed. A method of real time PCR was applied to the determination of SGLT1 and GLUT2 mRNA expression in the cells. Results Chinese herb extracts treatment altered the cell proliferation in a dose-dependent manner. Moreover, RA volatile oil (50μg/mL) and CP alkaloid (10μg/mL) treatment increased glucose absorption and the activity of Na^+ ,K^+-ATPase genes (P〈0.05). HA Volatile oil (50μg/ mL)treatment improved glucose absorption and up-regulated the gene expression of GLUT2. However, GF water extract (50 μg/mL) treatment did not affect the glucose absorption and transport in IEC-6 cells (P〉0.05). Conclusion The three extracts treatment, including RA volatile oil, CP aklaloid, and HA volatile oil, could increase glucose absorption and the expression of glucose transport carrier genes, but their regulative mechanism are not totally the same.