食品科学 ›› 2020, Vol. 41 ›› Issue (13): 148-159.doi: 10.7506/spkx1002-6630-20190528-339

• 营养卫生 • 上一篇    下一篇

纳豆枯草芽孢杆菌LNUB236所产纳豆激酶的溶栓活性

朱俊丰,刘帅,赵鹏燕,赵健,李佳增,王天琦,王丽,刘宏生   

  1. (1.辽宁大学生命科学院,辽宁 沈阳 110036;2.辽宁大学药学院,辽宁 沈阳 110036)
  • 出版日期:2020-07-15 发布日期:2020-07-29
  • 基金资助:
    沈阳市科技创新“双百工程”百项重大科技成果转化项目(Z17-5-078); 辽宁省科技厅大型仪器设备共享服务平台能力建设基金项目(kjhx2017028); 辽宁省高等学校国(境)外培养项目(2018LNGXGJWPY-YB006);沈阳市科技局科技成果转化推进计划项目(17-65-7-00); 辽宁省教育厅青年项目(LQN201711);辽宁省重点研发计划项目(2019JH2/10300041)

Thrombolytic Activity of Nattokinase Produced by Bacillus subtilis natto LNUB236

ZHU Junfeng, LIU Shuai, ZHAO Pengyan, ZHAO Jian, LI Jiazeng, WANG Tianqi, WANG Li, LIU Hongsheng   

  1. (1. School of Life Sciences, Liaoning University, Shenyang 110036, China; 2. School of Pharmacy, Liaoning University, Shenyang 110036, China)
  • Online:2020-07-15 Published:2020-07-29

摘要: 采用碱液浸提法从仙草中提取粗多糖JMBP-C,通过超滤、离子交换和凝胶柱层析纯化获得一种中性多糖JMBP-N和两种酸性多糖JMBP-A1和JMBP-A2。采用H2O2对人体肝细胞LO2进行氧化损伤,构建LO2细胞的氧化损伤模型。通过测定细胞存活率、乳酸脱氢酶(lactate dehydrogenase,LDH)、谷胱甘肽过氧化物酶(glutathione peroxidase,GSH-Px)、超氧化物歧化酶(superoxide dismutase,SOD)、过氧化氢酶(catalase,CAT)活力和丙二醛(malondialdehyde,MDA)含量探讨JMBP-C、JMBP-A1和JMBP-A2对氧化损伤细胞的保护作用。结果表明,3 种多糖能显著增加氧化损伤细胞的存活率,有效降低LDH的释放量,2 mg/mL时,仙草多糖均能极显著降低损伤细胞LDH活力(P<0.01),JMBP-A1质量浓度为0.5 mg/mL时,细胞存活率比模型组增加49.47%。仙草多糖能提高GSH-Px、SOD和CAT活力,且呈现一定的量效关系。JMBP-A2质量浓度为0.125 mg/mL时能将GSH-Px活力提高到正常细胞的69.20%,高于阳性对照,2 mg/mL的JMBP-A1将SOD和CAT活力分别恢复到正常对照组的80.32%和88.14%,极显著高于模型组(P<0.01),效果接近阳性对照。同时,3 种多糖样品均能有效减少MDA生成,0.5 mg/mL的JMBP-A1使受损细胞的MDA含量降低了31.50%,与阳性对照效果相当。综上,仙草多糖可增加细胞内抗氧化酶活性,降低脂质过氧化物生成,有明显的氧化损伤保护作用。

关键词: 仙草多糖, 细胞, 氧化损伤, 抗氧化酶

Abstract: Objective: The aim of this study is to evaluate the thrombolytic effects of nattokinase (NK) produced by Bacillus subtilis natto LNUB236 in vitro and in vivo. Methods: The thrombolytic effect of NK in vitro was determined by the dissolution rate of blood clots. Meanwhile, the in vivo effect was assessed by using a carrageenan-induced rat tail thrombosis model and a ferric chloride-induced carotid arterial thrombosis model as well as measuring mouse tail bleeding time. The toxicity of NK was measured by acute toxicity test (14 days) and chronic toxicity test (28 and 90 days) in rats. Results: NK significantly increased the dissolution rate of blood clots, while it significantly reduced the length of carrageenan-induced rat tail thrombus and the mass of ferric chloride-induced-carotid thrombus (P < 0.05). In addition, NK significantly prolonged the tail bleeding time of mice. In the acute and chronic toxicity tests, the tissues of the mice in the NK group had no pathological damage, and blood biochemical indexes were in the normal range. Conclusion: The NK produced by Bacillus subtilis natto LNUB236 exerts a thrombolytic activity both in vitro and in vivo, without any toxicity for the body. This study can lay the foundation for the development of functional foods containing NK produced by Bacillus subtilis natto LNUB236.

Key words: nattokinase, thrombolytic activity, toxicity

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