藤茶中二氢杨梅素和杨梅素对体外胃蛋白酶模型建立及活性评价

Establishment of in Vitro Pepsin Model of Dihydromyricetin and Myricetin in Rattan Tea and Activity Evaluation Effect Analysis

为了解藤茶中二氢杨梅素、杨梅素对胃蛋白酶活性的影响,通过探索对体外胃蛋白酶模型建立的影响因素确定建立条件,选用胃蛋白酶—血红蛋白为反应体系,在一定条件下分析胃蛋白酶作用时间、胃环境模拟成分(NaCl溶液、盐酸溶液)、终止剂三氯乙酸、血红蛋白、胃蛋白酶活性的影响因素以及二氢杨梅素和杨梅素对胃蛋白酶活性的影响。结果表明,胃蛋白酶作用时间为10 min、胃环境模拟成分(0.1 mol/L NaCl溶液和0.065 mol/L盐酸溶液)、5 m L血红蛋白溶液(1%)、1 mL胃蛋白酶(0.10 mg/mL)。另外,以不同浓度的二氢杨梅素、杨梅素为实验组,以酶活性变化指标为考察组,考察二氢杨梅素和杨梅素对胃蛋白酶活性的影响。结果为当二氢杨梅素用量为0.05mg时,对0.1 mg胃蛋白酶有最大抑制作用,空白组酶活力(u)为10632.3229,实验组酶活力(u)为695.6809,抑制率为93.46%;当杨梅素用量为0.15 mg时,对0.1 mg蛋白酶有最大抑制作用,空白组酶活力(u)为10632.3229,实验组酶活力(u)为2260.9629,抑制率为78.74%。由此可知,二氢杨梅素和杨梅素对胃蛋白酶均有明显的抑制作用,且在同剂量的黄酮参与反应中,二氢杨梅素的抑制率比杨梅素的大,这为后期的具体应用奠定了一定的理论基础。

In order to understand the rattan tea dihydrogen arbutus pigment,myricetin influence on pepsin activity,through the exploration of the influence factors of pepsin in vitro model to establish conditions were determined,the selection of pepsin,hemoglobin as reaction system,under certain conditions to explore stomach protease action time,environmental simulation component(NaCl solution,hydrochloric acid solution),termination of trichloroacetic acid,hemoglobin and the factors influencing the activity of pepsin and dihydrogen arbutus pigment and myricetin on pepsin activity,the results show that:The action time of pepsin was 10 min,the simulated components of gastric environment(0.1 mol/L NaCl solution and 0.065 mol/L hydrochloric acid solution),5 mL hemoglobin solution(1%),and 1 mL pepsin(0.10 mg/mL).In addition,the effects of dihydromyricetin and myricetin at different concentrations on pepsin activity were investigated by taking dihydromyricetin and myricetin as the experimental group and the change of enzyme activity as the study group.The results showed that dihydromyricetin had the largest inhibitory effect on 0.1 mg pepsin when the dosage was 0.05 mg,the enzyme activity(u)of the blank group was 10632.3229,and the enzyme activity(u)of the experimental group was 695.6809,with an inhibition rate of 93.46%.When the dosage of myricetin was 0.15 mg,it had the maximum inhibition effect on the protease of 0.1 mg,the enzyme activity(u)of the blank group was 10632.3229,and the enzyme activity(u)of the experimental group was 2260.9629,and the inhibition rate was 78.74%.It can be seen that both dihydromyricetin and myricetin have obvious inhibitory effects on pepsin,and the inhibition rate of dihydromyricetin is higher than that of myricetin in the reaction with the same dose of flavonoids,which provides certain theoretical basis for the specific application in the later stage.