目的 利用Caco-2細(xì)胞模型比較知母鹽炙前后的降血糖作用，并考察其作用機(jī)制，為鹽知母炮制原理的闡明提供依據(jù)。方法 建立Caco-2單層細(xì)胞模型，采用CCK-8細(xì)胞活力試劑盒檢測5 000、2 500、500、250、50、25、5、2.5、0.5、0.25 μg/mL生、鹽知母水煎液細(xì)胞毒性，來確定藥物最大毒性劑量。以蔗糖酶和麥芽糖酶底物，葡萄糖氧化酶法測定葡萄糖質(zhì)量濃度，計(jì)算抑制率，比較其對Caco-2單層細(xì)胞上蔗糖酶和麥芽糖酶活力的影響。葡萄糖吸收實(shí)驗(yàn)測定剩余葡萄糖的質(zhì)量濃度，比較生、鹽知母對Caco-2細(xì)胞吸收和轉(zhuǎn)運(yùn)葡萄糖的影響。結(jié)果 當(dāng)生、鹽知母質(zhì)量濃度≤250 μg/mL時，Caco-2細(xì)胞可進(jìn)行細(xì)胞實(shí)驗(yàn)。實(shí)驗(yàn)選取了50、250 μg/mL兩個質(zhì)量濃度。生、鹽知母水煎液50、250 μg/mL對蔗糖酶、麥芽糖酶活性均有抑制作用，且鹽知母水煎液對蔗糖酶、麥芽糖酶的抑制作用均優(yōu)于生知母。生、鹽知母水煎液均有降低細(xì)胞對葡萄糖吸收的趨勢，且呈現(xiàn)劑量相關(guān)性，但效果不顯著。鹽知母水煎液對葡萄糖吸收的抑制作用略好于生知母（P<0.05）。結(jié)論 鹽知母對Caco-2細(xì)胞的降糖作用優(yōu)于生知母，其降糖機(jī)制可能主要體現(xiàn)在α-葡萄糖苷酶上，而對葡萄糖吸收作用影響較小。

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.

國家自然科學(xué)基金資助項(xiàng)目（81102810）；遼寧省自然科學(xué)基金資助項(xiàng)目（2013020165）