红豆越橘总三萜的纯化及体外抗炎活性

doi:10.3969/j.issn.1000-2006.201809010

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南京林业大学学报（自然科学版） ›› 2019, Vol. 62 ›› Issue (04): 132-138.doi: 10.3969/j.issn.1000-2006.201809010

红豆越橘总三萜的纯化及体外抗炎活性

樊梓鸾^1,2,赵梓煣¹,赵翔¹,陈凯莉¹,付玉杰^3*

(1. 东北林业大学林学院,黑龙江哈尔滨 150040; 2. 黑龙江省森林食品资源利用重点实验室, 黑龙江哈尔滨 150040; 3. 东北林业大学森林植物生态学教育部重点实验室黑龙江哈尔滨 150040)

出版日期:2019-07-22 发布日期:2019-07-22
基金资助:
收稿日期:2018-09-11 修回日期:2019-03-13 基金项目:黑龙江省科学自然基金项目(QC2016021); 中国博士后科学基金项目(2016M600239); 中央高校基本科研业务费专项资金项目(2572019BA09); 国家自然科学基金项目(31170510)。第一作者:樊梓鸾(fzl_1122@163.com ),ORCID(0000-0003-3031-2876)。*通信作者:付玉杰(985461289@qq.com ),教授,博士,ORCID(0000-0002-0445-0

Purification of total triterpenoids from lingonberry and anti-inflammatory activity in vitro

FAN Ziluan^1,2, ZHAO Zirou^l, ZHAO Xiang¹, CHEN Kaili¹, FU Yujie^3*

(1. College of Forestry,Northeast Forestry University, Harbin 150040, China; 2. Key Laboratory of Forest Food Resources Utilization of Heilongjiang Province, Harbin 150040, China; 3. Key Laboratory of Forest Plant Ecology, Ministry of Education, Northeast Forestry University, Harbin 150040, China)

Online:2019-07-22 Published:2019-07-22

摘要/Abstract

摘要： 【目的】为了综合开发利用红豆越橘果实,通过大孔吸附树脂-Sephadex LH-20纯化工艺获得纯度较高的红豆越橘总三萜化合物,并分析此三萜化合物的体外抗炎活性。【方法】以野生矮丛红豆越橘为原料,首先采用静态吸附-解析实验和动态吸附-解析实验筛选大孔吸附树脂,优化最佳工艺,确定最佳上样质量浓度、pH、上样体积、上样流速以及洗脱液浓度; 然后采用Sephadex LH-20羟丙基葡聚糖凝胶二次纯化,获得高纯度的红豆越橘总三萜; 采用对透明质酸酶和牛血清白蛋白的变性抑制率为抗炎评价指标,分析红豆越橘总三萜的抗炎活性。【结果】比较7种大孔树脂对红豆越橘总三萜的纯化效果,结果显示,X-5树脂最适合初级纯化,最佳纯化工艺优化结果为上样质量浓度1.5 mg/mL、样液pH 6、上样量为4/3 BV、上样流速1 BV/h、80%(体积分数)的乙醇进行洗脱,红豆越橘总三萜的纯度由原来的5.13%提高到29.46%。进一步采用Sephadex LH-20二次纯化获得纯度为(43.25±0.31)%的红豆越橘总三萜,抗炎活性结果显示,对透明质酸酶和牛血清白蛋白变性抑制率分别可达(81.5±1.37)%、(72.59 ±1.84)%。【结论】红豆越橘是一种营养丰富的浆果,通过二次纯化技术获得纯度较高的三萜类化合物,并初步证实红豆越橘总三萜具有一定的抗炎活性。

Abstract: 【Objective】 In order to further develop and utilize lingonberry(Vaccinium vitis-idaea)fruits, the total triterpenoids of lingonberry were obtained, and the purification process was optimized through using macroporous adsorption resin and secondary purification of with Sephadex LH-20. The in vitro anti-inflammatory activity of this triterpene compound was analyzed. 【Method】 The experiment was based on the wild buckwheat-hung lingonberry as the raw material. The a static adsorption-analysis experiment, and the dynamic adsorption -analysis experiment were used to select the macroporous adsorption resin, optimization of the best process, and determination of the best sample concentration, pH 6, sample volume, sample flow rate, and ethanol concentration of the eluent. The secondary purification used Sephadex LH-20 hydroxypropyl dextran gel to obtain high-purity lingonberry total triterpene. Using hyaluronidase and the inhibition of serum albumin denaturation as an anti-inflammatory evaluation index to evaluate the anti-inflammatory activity of the total triterpenes of from lingonberry. 【Result】 The purification effect of seven macroporous resins on the total triterpenes of lingonberry was compared. The results showed that the X-5 resin was most suitable for the primary purification. The optimum purification process was the sample concentration of 1.5 mg/mL, elution at pH 6, loading volume 4/3 BV, loading flow rate of 1 BV/h, and 80% ethanol as eluant. The purity of the total triterpenes of lingonberry increased from 5.13% to 29.46%. Further purification using Sephadex LH-20 obtained a purity of(43.25±0.31)%. The total anti-inflammatory activity of the lingonberry was(81.5±1.37)% for the hyaluronidase and(72.59±1.84)% for the denaturation of bovine serum albumin. 【Conclusion】 Lingonberry is a nutrient-rich berry. High-purity triterpenoids were obtained through the secondary purification technology, and it was preliminarily showed that the total triterpenes of lingonberry had significant anti-inflammatory activity. Our results provided a certain theoretical basis for the further development and utilization of lingonberry.

中图分类号:

TS255.4

樊梓鸾,赵梓煣,赵翔,等. 红豆越橘总三萜的纯化及体外抗炎活性[J]. 南京林业大学学报（自然科学版）, 2019, 62(04): 132-138.

FAN Ziluan, ZHAO Zirou^l, ZHAO Xiang, CHEN Kaili, FU Yujie. Purification of total triterpenoids from lingonberry and anti-inflammatory activity in vitro[J].Journal of Nanjing Forestry University (Natural Science Edition), 2019, 62(04): 132-138.DOI: 10.3969/j.issn.1000-2006.201809010.

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红豆越橘总三萜的纯化及体外抗炎活性

Purification of total triterpenoids from lingonberry and anti-inflammatory activity in vitro

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