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目的探讨紫花芸豆胰蛋白酶抑制剂的分离纯化及降糖作用。方法通过分离提纯得到紫花芸豆胰蛋白酶抑制剂,后建立四氧嘧啶糖尿病动物模型,取雄性昆明种小鼠15例,随机分为正常组、四氧嘧啶糖尿病模型组与胰蛋白酶抑制剂组,各5例。四氧嘧啶糖尿病模型组小鼠腹腔注射四氧嘧啶后予以紫花芸豆胰蛋白酶抑制,胰蛋白酶抑制剂组小鼠予以紫花芸豆胰蛋白酶抑制,正常组小鼠注射0.9%氯化钠溶液。观察3组小鼠的血糖变化情况。结果当p H值为4.5和7.5时,紫花芸豆胰蛋白酶抑制剂的提取率高,75℃下紫花芸豆胰蛋白酶抑制剂的活性回收率和纯化倍数最高,p H=8,馏分比例30%~60%时的活性强,又因收集的样品应为酸性蛋白,故后续纯化过程中考虑加硫酸铵饱和30%后,继续加至60%,收集此时沉淀进行溶解透析为下一步离子交换所用;紫花芸豆胰蛋白酶抑制剂可降低四氧嘧啶诱导的血糖升高,同时降低总胆固醇(TC),但对高密度脂蛋白胆固醇(HDL-C)和三酰甘油(TG)的效果不明显。结论紫花芸豆胰蛋白酶抑制剂治疗糖尿病有一定的临床效果。
OBJECTIVE To study the isolation, purification and hypoglycemic effect of purple kidney bean trypsin inhibitor. Methods The purple kidney beans trypsin inhibitor was isolated and purified, and alloxan diabetic model was established. Fifteen male Kunming mice were randomly divided into normal group, alloxan-induced diabetic model group and trypsin inhibitor group 5 cases. Alloxan diabetic mice were injected with alloxan after intraperitoneal injection of purple kidney beans trypsin inhibition, trypsin inhibitor mice were purple kidney beans trypsin inhibition, normal mice were injected with 0.9% sodium chloride solution. The changes of blood glucose in 3 groups of mice were observed. Results When the p H value was 4.5 and 7.5, the extraction rate of purple kidney beans trypsin inhibitor was high, and the activity recovery and purification fold of purple kidney beans trypsin inhibitor was the highest at 75 ℃ with p H = 8 and the fraction ratio of 30% 60% when the activity is strong, and because the collected samples should be acidic protein, so follow-up purification process to consider adding 30% ammonium sulfate saturation, continue to add to 60%, the precipitation was collected at this time dissolved dialysis used for the next ion exchange ; Purple kidney beans trypsin inhibitor can reduce alloxan-induced hyperglycemia, while reducing total cholesterol (TC), but the effect of high-density lipoprotein cholesterol (HDL-C) and triglyceride (TG) is not obvious. Conclusion Purple kidney beans trypsin inhibitor for the treatment of diabetes have a certain clinical effect.