橙皮苷与陈皮酶转化工艺研究及其对正常小鼠小肠推进作用的比较

Enzymatic transformation of hesperidin and Citri Reticulatae Pericarpium and comparison of their promoting effects on small intestine of normal mice

目的:建立橙皮苷酶转化最佳工艺并应用于陈皮酶转化;比较橙皮苷和陈皮酶转化前后产物对正常小鼠小肠推进作用,为陈皮酶转化研究提供依据。方法:以橙皮苷酶转化产物橙皮素单葡萄糖苷含量为指标,采用单因素方法考察酶用量、pH、反应时间和温度,建立其最佳工艺,并用于陈皮的酶转化实验;采用正常小鼠促小肠动力实验比较陈皮酶转化前后橙皮苷及其酶解产物橙皮素单葡萄糖苷的药效。结果:橙皮苷酶转化最佳工艺为:酶使用量0.1 g, pH=4,反应时间60 min,温度65℃;在小肠推进实验中,与空白组对比,陈皮酶转化后与橙皮素单葡萄糖苷各剂量组及橙皮苷高、中剂量组有显著性差异(P<0.05),其中陈皮酶转化后高、中剂量组及橙皮苷高剂量组与橙皮素单葡萄糖苷各剂量组均有极显著差异(P<0.01);组间对比,陈皮酶转化前后中剂量组存在显著差异(P<0.05),陈皮酶转化前后高剂量组存在极显著差异(P<0.01);橙皮素单葡萄糖苷高剂量组与橙皮苷高剂量组对比有显著性差异(P<0.05)。结论:同摩尔数的橙皮素单葡萄糖苷对正常小鼠小肠推进作用优于橙皮苷;陈皮酶转化后小肠推进作用优于陈皮。该酶解方法操作简便、反应可控,预测性良好,可进一步用于橙皮苷和陈皮的酶转化研究。

Objective: To establish the best process of enzymatic transformation of hesperidin and apply it to Citri Reticulatae Pericarpium, and further compare the small intestine propulsive effect of hesperidin and Pericarpium Citri Reticulatae before and after enzymatic transformation in normal mice, so as to provide a basis for the study on the enzymatic transformation of Citri Reticulatae Pericarpium. Methods: Using the content of hesperetin-7-O-glucoside as the index, the optimal process parameters of enzymatic transformation were investigated by single factor methods, including enzyme dosage, pH, reaction time and temperature. The best process was established and applied in enzymatic transformation test of Citri Reticulatae Pericarpium. The effects of Citri Reticulatae Pericarpium before and after enzymatic transformation, hesperidin and its hydrolysate hesperetin-7-O-glucoside were compared using promoting small intestine motility test in normal mice. Results: The optimized condition under which the transformation rate of hesperetin-7-O-glucoside was the highest is: enzyme dosage of 0.1 g, pH value of 4, reaction time of 60 min, temperature of 65 ℃. In small intestine propulsion experiment, compared with the blank group, there were significant differences in each dose groups of Citri Reticulatae Pericarpium after enzymatic transformation and hesperidin-7-O-glucoside, high-and medium-dose groups of hesperidin(P<0.05), among which there were extremely significant differences in high-dose, medium-dose groups of Citri Reticulatae Pericarpium after enzymatic transformation, high-dose group of hesperidin and each dose groups of hesperidin-7-O-glucoside(P<0.01). Compared between groups, there was extremely significant difference between middle-dose group of Citri Reticulatae Pericarpium before and after enzymatic transformation(P<0.05);there was extremely significant difference between high-dose group of Citri Reticulatae Pericarpium before and after enzymatic transformation(P<0.01);there was significant difference between high-dose group of hesperidin-7-O-glucoside and high-dose group of hesperidin(P<0.05). Conclusion: Hesperidin-7-O-glucoside with the same mole number has better intestinal propulsion than hesperidin in normal mice;the intestinal propulsive effect of Citri Reticulatae Pericarpium after enzymatic transformation was better than Citri Reticulatae Pericarpium. The method is simple, controllable and predictable, it can be further used in the study of the enzymatic transformation of Citri Reticulatae Pericarpium.