发酵桑叶糖蛋白提取工艺优化及活性研究

doi:10.20047/j.issn1673-7210.2023.19.02

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摘要目的筛选发酵桑叶糖蛋白（FMLGP）的最佳提取工艺并评价其活性。方法以蛋白浓度为指标，采用单因素结合响应面法优化FMLGP提取工艺；通过琼脂糖纤维蛋白平板法检测纤溶活性；利用ox-LDL诱导巨噬细胞RAW264.7泡沫化，实验分为空白组、模型组和FMLGP高、中、低剂量组（终浓度分别为15、10、5 mg/ml），细胞培养24 h，采用油红染色法考察各组细胞内脂质蓄积程度。结果 FMLGP最佳提取条件为PBS缓冲液、料液比1∶6 g/ml、于25℃提取4 h，蛋白浓度为（57.92±1.14）mg/ml，纤溶活性为（1 780.66±76.49）IU/ml，15 mg/ml FMLGP对巨噬泡沫细胞有明显促进脂质排出作用。结论本研究确定了FMLGP的最佳提取工艺，通过纤溶活性及巨噬细胞泡沫化实验提示FMLGP具有促进脂质排出作用。

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	孙丹丹　　孙银玲　　郑宏宇　　丁纯洁　　赵娢　　宋吉鹏　　陈丽艳

关键词 ：发酵桑叶, 糖蛋白, 响应面法, 活性

Abstract：Objective To determine the optimal extraction process of fermented mulberry leaf glycoprotein (FMLGP), and to evaluate the activity of glycoprotein. Methods Taking protein concentration as index, the extraction process of FMLGP was optimized by single factor combined with response surface method. Fibrinolytic activity was detected by agarose fibrin plate method. Macrophage RAW264.7 foam was induced by ox-LDL. The experiment was divided into blank group, model group, and FMLGP high-dose, medium-dose, and low-dose groups (the final concentration was 15, 10, and 5 mg/ml, respectively). The cells were cultured for 24 h and the degree of lipid accumulation in each group was investigated by Oil red staining. Results The optimal extraction conditions of FMLGP were PBS buffer solution, solid-liquid ratio of 1∶6 g/ml, extraction at 25℃ for 4 h, protein concentration of (57.92±1.14) mg/ml, fibrinolytic activity of (1 780.66±76.49) IU/ml, and 15 mg/ml FMLGP could significantly promote lipid excretion in macrophage foam cells. Conclusion In this study, the optimal extraction process of FMLGP is determined, and the fibrinolytic activity and macrophage foaming experiments shows that FMLGP had potential anti-atherosclerosis effects.

Key words： Fermented mulberry leave Glycoprotein Response surface methodology Activity

基金资助:国家自然科学基金资助项目（U20A20400）；
国家中药材产业技术体系岗位任务书（CARS-21）；
黑龙江省省属科研院所科研业务费项目（CZKYF2021A002）；
黑龙江省卫生健康委科研课题（20211313050162）。

通讯作者: 陈丽艳（1971-），女，硕士，主任药师，硕士生导师；研究方向：中药发酵炮制工艺及健康产品的研究开发。

作者简介: 孙丹丹（1995-），女，黑龙江省中医药科学院中药研究所2020级中药学专业在读硕士研究生；研究方向：中药发酵。

引用本文:

孙丹丹　　孙银玲　　郑宏宇　　丁纯洁　　赵娢　　宋吉鹏　　陈丽艳. 发酵桑叶糖蛋白提取工艺优化及活性研究[J]. 中国医药导报, 2023, 20(19): 10-13,34.
SUN Dandan SUN Yinling ZHENG Hongyu DING Chunjie ZHAO Han SONG Jipeng CHEN Liyan. Study of optimization and activity of extraction process of fermented mulberry leaf glycoprotein. 中国医药导报, 2023, 20(19): 10-13,34.

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