【作者】 刘冰冰； 郑棋月； 和磊； 黄漫青； 张楠； 孔玲； 高珊； 鲜波； 李国君； 任建武；

【Author】 Liu Bingbing;Zheng Qiyue;He Lei;Huang manqing;Zhang nan;Kong ling;Gao shan;Bo Xian;Li Guojun;Ren Jianwu;CAS-MPG Partner institute for Computational Biology;College of Biological Sciences and Biotechnology,Beijing Forestry University;Beijing University of Agriculture;Beijing Centers for Disease Control,Beijing Research Center of Preventive Medicine,Beijing Key Laboratory of Diagnostic and Traceability Technologies for Food Poisoning;The capital medical university school of public health;

【机构】 中科院上海生命科学院计算生物所； 北京林业大学生物科学与技术学院； 北京农学院； 北京市疾病预防控制中心北京市预防医学研究中心北京市食物中毒诊断溯源技术重点实验室； 首都医科大学公共卫生学院；

【摘要】 目的本文对黑果枸杞原花青素(Lyctum ruthenicum Murray,简称原花青素)提取工艺进行了酶法优化,并以秀丽隐杆线虫(Caenorhabditis elegans,简称线虫)作为模式生物,研究黑果枸杞原花青素对线虫的急性毒性效应,以及延长寿命的功效作用及其初步机制,为原花青素用于功能性保健品开发利用奠定实验基础。方法采用纤维素酶法结合正交分析优化黑果枸杞原花青素的提取工艺:采用线虫急性毒性试验、寿命实验及氧化应激和热应激实验研究其毒性、寿命影响及其可能机制。结果正交分析表明,黑果枸杞原花青素最高得率为9.47mg/g时的最佳提取工艺条件为酶解液pH值3.4,酶解温度30C,酶解时间30min,酶浓度4%。急性毒性实验结果表明,浓度为50μg/mL及300μg/mL的原花青素作用24h对线虫均无急性毒性效应。线虫寿命实验表明,持续饲喂50μg/mL原花青素的实验组,线虫平均寿命与对照组相比延长了22.88%(P<0.01)。氧化应激与热应激条件下,持续饲喂50.μg/mL原花青素的实验组与对照组相比,平均寿命分别延长了44.28%(P<0.05)与24.46%(P<0.01)。结论酶法提取工艺优化后,黑果枸杞原花青素的得率为9.47mg/g。毒性和功效实验结果表明,黑果枸杞原花青素(浓度<300μg/mL)对线虫没有急性毒性作用,原花青素(50μg/mL)可延长线虫寿命,其功效可能是通过提高线虫的氧化应激及热应激能力而实现的。更多还原

【Abstract】 Objective:In this paper,we discussed the optimum extraction condition of Lycium ruthenicum Murray procyanidins,researched the effect and preliminary mechanism of procyanidins on model organism Caenorhabditis elegans in terms of toxicity and aging,aiming at laying a experimental foundation for health products.Method:The cellulose enzymatic extraction combined with orthogonal analysis were used for optimum extraction of procyanidins.We studied the mechanism of procyanidins’ effect on C.elegans by the method of acute toxicity,lifespan,oxidative stress and heat stress.Result:The orthogonal analysis indicated that when we got the highest procyanidins proportion of 9.47 mg/g,the optimum extraction conditions were as follows:the hydrolyzate pH is 3.4,the reaction temperature is 30℃,the hydrolyzate time is 30 min,and the enzyme concentration is 4%.Acute toxicity assay showed that 50μg/mL and 300μg/mL of procyanidins had no acute toxicity on C.elegans in 24 h.In C.elegans assay,compared with the control group,the 50μg/mL group prolonged the mean lifespan by 22.88%(P<0.01).In the condition of oxidative and heat stress,the50μg/mL group prolonged the mean lifespan by 44.28%(P<005) and 24.46%(P<0.01) respectively.Conclusion:We got the procyanidins of 9.47 mg/g after procyanidins’ enzymatic optimum extraction.The toxicity and efficacy assay showed that procyanidins(concentration≤300μg/mL) had no toxicity on C.elegans,procyanidins(50μg/mL) could extend the mean lifespan of C.elegans by improving the ability of tolerance stress.更多还原