目的 利用Caco-2细胞模型比较知母盐炙前后的降血糖作用，并考察其作用机制，为盐知母炮制原理的阐明提供依据。方法 建立Caco-2单层细胞模型，采用CCK-8细胞活力试剂盒检测5 000、2 500、500、250、50、25、5、2.5、0.5、0.25 μg/mL生、盐知母水煎液细胞毒性，来确定药物最大毒性剂量。以蔗糖酶和麦芽糖酶底物，葡萄糖氧化酶法测定葡萄糖质量浓度，计算抑制率，比较其对Caco-2单层细胞上蔗糖酶和麦芽糖酶活力的影响。葡萄糖吸收实验测定剩余葡萄糖的质量浓度，比较生、盐知母对Caco-2细胞吸收和转运葡萄糖的影响。结果 当生、盐知母质量浓度≤250 μg/mL时，Caco-2细胞可进行细胞实验。实验选取了50、250 μg/mL两个质量浓度。生、盐知母水煎液50、250 μg/mL对蔗糖酶、麦芽糖酶活性均有抑制作用，且盐知母水煎液对蔗糖酶、麦芽糖酶的抑制作用均优于生知母。生、盐知母水煎液均有降低细胞对葡萄糖吸收的趋势，且呈现剂量相关性，但效果不显著。盐知母水煎液对葡萄糖吸收的抑制作用略好于生知母（P<0.05）。结论 盐知母对Caco-2细胞的降糖作用优于生知母，其降糖机制可能主要体现在α-葡萄糖苷酶上，而对葡萄糖吸收作用影响较小。

Objective To compare the hypoglycemic effects of Anemarrhena Rhizoma before and after processed with salt-water by Caco-2 cell model, and explore its mechanism, so as to provide reference for explain the processing principle of Anemarrhena Rhizoma. Methods Caco-2 cell models were established. Cytotoxicitits of Anemarrhena Rhizoma and salt Anemarrhena Rhizoma with concentrations of 5 000, 2 500, 500, 250, 50, 25, 5, 2.5, 0.5, and 0.25 μg/mL were determined with CCK-8 cell viability kit, and obtained the maximum toxic dose. The GOP-POD method was used to determine the concentration of sucrose and maltase activity, and to calculate the inhibition rate, and the effects of Caco-2 on the activity of invertase and enzyme in the monolayer cells were compared. Residual glucose were measured by glucose test method, and effects of Anemarrhena Rhizoma and salt Anemarrhena Rhizoma on glucose transport and uptake in Caco-2 cell were compared. Results Caco-2 cell models were selected to used in the flowing tests when they were treated by Anemarrhena Rhizoma and salt Anemarrhena Rhizoma with concentrations below 250 μg/mL. The optimized concentrations of 50 and 250 μg/mL were finally selected. Anemarrhena Rhizoma and salt Anemarrhena Rhizoma (50 and 250 μg/mL) had inhibitory effects on sucrase and maltase activity, and the inhibition of salt Anemarrhena Rhizoma were better than those of Anemarrhena Rhizoma. Anemarrhena Rhizoma and salt Anemarrhena Rhizoma had the trend to reduce absorb of glucose, and showed a dose related, but the effects were not significant. Glucose absorptions of salt Anemarrhena Rhizoma were slightly stronger than those of Anemarrhena Rhizoma (P<0.05). Conclusion Hypoglycemic effects of salt Anemarrhena Rhizoma on Caco-2 cell are stronger than those of Anemarrhena Rhizoma, and mechanism may be mainly reflected according to α-glucosidase, but effect on glucose uptake is small.

国家自然科学基金资助项目（81102810）；辽宁省自然科学基金资助项目（2013020165）