滋肾清肝方及其有效成分对二肽基肽酶-4抑制作用的实验研究

Vitro Screening Study of Inhibitory Effect of Zishen Qinggan Formula and its Effective Ingredients on DPP-4

目的:研究滋肾清肝方及其有效成分(2,3,5,4'-四羟基二苯乙烯-2-O-β-D-葡萄糖苷,1-脱氧野尻霉素和决明子苷、红镰霉素龙胆二糖苷、橙黄决明素、甲基钝叶决明素、白藜芦醇)对二肽基肽酶4(DPP-4)活性的影响,研究其发挥降糖作用的可能机制。方法:建立DPP-4抑制剂的体外筛选模型,对酶量、底物浓度、最适p H和反应时间进行优化,选择磷酸西格列汀片作为阳性对照药,通过建立的筛选模型对以上成分进行筛选,计算抑制率及IC50。结果:红镰霉素龙胆二糖苷、决明子苷、橙黄决明素、甲基钝叶决明素对DPP-4的抑制率<10%,白藜芦醇、2,3,5,4'-四羟基二苯乙烯-2-O-β-D-葡萄糖苷、1-脱氧野尻霉素、滋肾清肝方提取物、磷酸西格列汀对DPP-4的IC50值分别为(48.805±12.874)、(2.845±0.428)、(0.305±0.002)、(1.628±0.324)、(0.003±0.0005)mmol/L。结论:滋肾清肝方及其有效成分具有不同程度的DPP-4抑制作用,抑制DPP-4可能为其调节血糖的作用机制。

Objective：To study the inhibitory effect of Zishen Qinggan Formula and its effective ingredients（2,3,5,4＇-tetrahydroxystilbene-2-O-β-D-glucopyranoside,l-deoxynojirimycin,cassiaside,rubrofusarin gentiobioside,aurantio-obtusin,chryso-obtusin,resveratrol）on DPP-4,and study the possible mechanism of its hypoglycemic effect. Method：Establish vitro screening method of DPP-4 inhibitor,and the amount of enzyme,substrate concentration,the optimum p H and reaction time were optimized. And select the Sitagliptin Phosphate tablets as positive control drug. Through the screening model has been established,calculate the inhibition rate,calculate IC50. Result：The inhibition rate of DPP-4 of cassiaside,rubrofusarin gentiobioside,aurantio-obtusin,chryso-obtusin are less than 10%. IC50 value for the DPP-4 of resveratrol,2,3,5,4＇-tetrahydroxystilbene-2-O-β-D-glucopyranoside,l-deoxynojirimycin,extract of Zishen Qinggan formula and Sitagliptin Phosphate are（48.805±12.874） mmol/L,（2.845±0.428） mmol/L,（0.305±0.002） mmol/L,（1.628±0.324） mmol/L,（0.003±0.0005） mmol/L. Conclusions：Zishen Qinggan Formula and its effective ingredients have certain inhibitory effect on DPP-4,and Jasminoidin has the obvious inhibitory action to the DPP-4,and DPP-4 inhibitory effect may be the mechanism for regulating blood sugar.