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目的观察三清降糖方对α-葡萄糖苷酶活性的抑制作用,为临床应用三清降糖方治疗糖尿病提供客观依据,并为进一步开发新药奠定基础。方法 (1)利用正交实验建立离体小肠肠囊外翻法观察α-葡萄糖苷酶抑制作用的反应体系,选用L934正交实验表筛选反应体系中底物类型、底物浓度、反应时间的最佳条件。(2)分组及实验方法:取Wistar雄性大鼠15只,处死后取空肠及回肠段制作78段肠囊,分为13组:阴性对照组、拜糖平不同浓度组(0.0032mg/ml、0.016mg/ml、0.08mg/ml、0.4mg/ml、2mg/ml、10mg/ml)6组,三清降糖方不同浓度组(0.016mg/ml、0.08mg/ml、0.4mg/ml、2mg/ml、10mg/ml、50mg/ml)6组,每组6段。拜糖平组和三清降糖方组分别在反应体系中加入不同浓度的相应药物,阴性对照组中不加任何药物。37℃震荡水浴120min,取出肠囊,测肠囊内液体的葡萄糖含量,计算药物对葡萄糖吸收的抑制率,比较各组间抑制率的差异。结果 (1)正交试验结果显示:最佳反应体系的条件为底物选用麦芽糖,浓度为0.25mol/L,反应时间为120min。(2)拜糖平不同浓度组的葡萄糖生成量经单因素方差分析,组间差异有统计学意义(P<0.05),其中浓度为10mg/ml的拜糖平6组对葡萄糖生成量的抑制作用最强,抑制率达到84.4%。(3)三清降糖方不同浓度组的葡萄糖生成量经单因素方差分析,组间比较差异有统计学意义(P<0.05)。其中浓度为50mg/ml的三清降糖方6组对葡萄糖生成量的抑制作用最强,抑制率达到74.00%。与浓度为10mg/ml拜糖平6组相当(P>0.05)。结论三清降糖方能够明显抑制α-葡萄糖苷酶活性,其抑制作用随着浓度的增大而增强,其抑制效应呈现剂量依赖性。抑制α-葡萄糖苷酶活性是三清降糖方降糖作用的机制之一。
Objective To observe the inhibitory effect of Sanqing Jiangtang Recipe on α-glucosidase activity and provide an objective basis for the clinical application of Sanqiang Jiangtang Recipe in the treatment of diabetes and lay a foundation for the further development of new drugs. Methods (1) Orthogonal experiment was used to establish the reaction system of in vitro small intestine enteral eversion to observe the inhibitory effect of α-glucosidase. L934 orthogonal test was used to screen the substrate type, substrate concentration, reaction time Optimal conditions. (2) Grouping and experimental methods: Fifteen Wistar male rats were sacrificed and 78 jejunal sacs were made from jejunum and ileum after sacrifice. They were divided into 13 groups: negative control group, different concentrations of cyproterone (0.0032mg / ml, (0.016mg / ml, 0.08mg / ml, 0.4mg / ml, 2mg / ml and 10mg / ml) 2mg / ml, 10mg / ml, 50mg / ml) 6 groups, 6 in each group. Yipangping group and Sanqing Hypoglycemic group were added to the corresponding concentration of the corresponding drugs in the reaction system, the negative control group without any drug. 37 ℃ shock water bath 120min, remove the gut, measuring the content of liquid in the gut of the intestinal fluid to calculate the inhibition rate of drug absorption of glucose, inhibit the difference between the groups. Results (1) The results of orthogonal test showed that the optimal conditions for the reaction system were as follows: maltose was used as the substrate, the concentration was 0.25mol / L and the reaction time was 120min. (2) The levels of glucose in different concentrations of albuterol group were analyzed by one-way ANOVA, the difference was statistically significant (P <0.05), among which Glucose level was inhibited The strongest effect, the inhibition rate reached 84.4%. (3) The glucose production of different concentration groups of Sanqing Hypoglycemic group were analyzed by one-way ANOVA, the difference between the two groups was statistically significant (P <0.05). Among them, the concentration of 50mg / ml Sanqing Jiangtang Fang 6 group had the strongest inhibitory effect on the amount of glucose production, the inhibition rate reached 74.00%. It was comparable to the 6 groups at the concentration of 10 mg / ml (P> 0.05). Conclusion Sanqiang Jiangtang Prescription can significantly inhibit the activity of α-glucosidase, and its inhibitory effect increases with increasing concentration, and its inhibitory effect appears in a dose-dependent manner. Inhibition of α-glucosidase activity is Sanqing hypoglycemic side hypoglycemic one of the mechanisms.