酶法辅助热水浸提刺梨多糖工艺优化及其抗肿瘤活性研究

唐健波 吕都 彭梅 范建新 潘牧 陈朝军 王辉 杨娟

唐健波,吕都,彭梅,等. 酶法辅助热水浸提刺梨多糖工艺优化及其抗肿瘤活性研究[J]. 食品工业科技,2021,42(18):98−105. doi:  10.13386/j.issn1002-0306.2021030147
引用本文: 唐健波,吕都,彭梅,等. 酶法辅助热水浸提刺梨多糖工艺优化及其抗肿瘤活性研究[J]. 食品工业科技,2021,42(18):98−105. doi:  10.13386/j.issn1002-0306.2021030147
TANG Jianbo, LÜ Du, PENG Mei, et al. Optimization of Enzymatic-assisted Thermal Water Extraction of Rosa roxburghii Polysaccharide and Its Antitumor Activity[J]. Science and Technology of Food Industry, 2021, 42(18): 98−105. (in Chinese with English abstract). doi:  10.13386/j.issn1002-0306.2021030147
Citation: TANG Jianbo, LÜ Du, PENG Mei, et al. Optimization of Enzymatic-assisted Thermal Water Extraction of Rosa roxburghii Polysaccharide and Its Antitumor Activity[J]. Science and Technology of Food Industry, 2021, 42(18): 98−105. (in Chinese with English abstract). doi:  10.13386/j.issn1002-0306.2021030147

酶法辅助热水浸提刺梨多糖工艺优化及其抗肿瘤活性研究

doi: 10.13386/j.issn1002-0306.2021030147
基金项目: 贵州省优质特色粮食产业发展专项(黔农财[2019]21号);贵州省工信厅刺梨专项(黔财农[2019]261号)
详细信息
    作者简介:

    唐健波(1988−),男,硕士研究生,助理研究员,研究方向:特色食品研究与开发,E-mail:startar2020@163.com

    通讯作者:

    杨娟(1971−),女,博士,研究员,研究方向:天然产物研究及开发,E-mail:yangxz2002@126.com

  • 中图分类号: TS201.1

Optimization of Enzymatic-assisted Thermal Water Extraction of Rosa roxburghii Polysaccharide and Its Antitumor Activity

  • 摘要: 以刺梨干果为原材料,研究酶法辅助热水浸提刺梨多糖的提取工艺及其抗肿瘤活性。通过单因素实验结合响应面法优化刺梨多糖的提取工艺,并通过建立S180实体瘤模型对最优条件下提取的刺梨多糖进行抗肿瘤活性研究。结果表明,酶法辅助热水浸提刺梨多糖的最优工艺条件为:酶解pH6.2,酶解时间4 h,酶解温度41 ℃,酶添加浓度1.62%,液料比30:1 mL/g,酶解反应完成后90 ℃热水浸提3 h。在此工艺条件下,刺梨多糖得率为6.32%±0.12%。抗肿瘤活性研究表明,在灌胃剂量为200 mg/kg时,酶法辅助热水浸提刺梨多糖对S180肿瘤小鼠的抑瘤率为52.13%±1.84%,具有明显的抗肿瘤活性,并能显著(P<0.05)提高肿瘤小鼠的白细胞数量、胸腺指数和脾脏指数,说明在此工艺条件下提取的刺梨多糖,具有一定的提升肿瘤小鼠免疫能力和抗肿瘤的作用,可作为潜在的功能性食品添加或天然抗肿瘤药物来源。
  • 图  1  酶解pH对刺梨多糖得率的影响

    Figure  1.  Effect of enzymatic hydrolysis pH on the yield of RRTP

    图  2  酶解时间对刺梨多糖得率的影响

    Figure  2.  Effect of enzymatic hydrolysis time on the yield of RRTP

    图  3  酶解温度对刺梨多糖得率的影响

    Figure  3.  Effect of enzymatic hydrolysis temperature on the yield of RRTP

    图  4  酶添加浓度对刺梨多糖得率的影响

    Figure  4.  Effect of enzyme dosage on the yield of RRTP

    图  5  不同因素交互作用对刺梨多糖得率影响的三维响应面图

    Figure  5.  3D-response surface plots of the interaction of different variables on the yield of RRTP

    表  1  响应面试验因素及水平设计

    Table  1.   Factors and levels of response surface methodology

    水平因素
    A(酶解pH)B(酶解时间,h)C(酶解温度,℃)D(酶添加浓度,%)
    -15.43.0301.0
    05.84.0401.5
    16.25.0502.0
    下载: 导出CSV

    表  2  响应面试验设计及结果

    Table  2.   Design and results of Box-Benhnken experiment

    实验号ABCD刺梨多糖得率(%)
    100006.15
    200−115.52
    30−1−105.23
    4100−15.85
    501105.11
    60−1015.54
    7−10−105.41
    800006.09
    900−1−15.37
    10−11005.26
    11−10105.24
    1200006.11
    1301−105.45
    1410−105.91
    1500005.99
    16010−15.45
    1710016.19
    18−100−15.55
    1911006.05
    201−1006.09
    210−1105.55
    22001−15.49
    2300115.45
    24−10015.50
    250−10−15.67
    2610106.06
    2701015.50
    28−1−1005.47
    2900006.14
    下载: 导出CSV

    表  3  响应面模型方差分析

    Table  3.   ANOVA for response surface quadratic model

    方差来源平方和自由度均方FP显著性
    模型3.05140.2245.32<0.0001***
    A1.1511.15239.79<0.0001***
    B0.04410.0449.230.0088**
    C8.333E-00618.333E-0061.733E-0030.9674
    D8.533E-00318.533E-0031.770.2041
    AB7.225E-00317.225E-0031.500.2405
    AC0.02610.0265.320.0368*
    AD0.03810.0387.910.0139*
    BC0.1110.1122.640.0003**
    BD8.100E-00318.100E-0031.680.2153
    CD9.025E-00319.025E-0031.880.1923
    A20.02010.0204.090.0626
    B20.7110.71148.07<0.0001***
    C21.0611.06219.96<0.0001***
    D20.3810.3879.37<0.0001***
    残差0.067144.809E-003
    失拟项0.051105.101E-0031.250.4479
    纯误差0.01644.050E-003
    总离差3.1228
    注:*:表示差异显著(P<0.05);**:表示差异高度显著(P<0.01);***:表示差异极其显著(P<0.001)。
    下载: 导出CSV

    表  4  刺梨多糖对荷S180小鼠抑瘤作用、外周血白细胞及其脏器的影响

    Table  4.   Effect of PPRT on tumor inhibition, peripheral blood leucocyte and viscera index on S180 tumor-bearing mice

    分组剂量(mg/kg)抑瘤率(%)白细胞数(×109/L)胸腺指数脾脏指数
    CT组016.89±1.702.30±0.357.11±0.42
    CTX组3562.32±1.09a10.40±1.34**1.97±0.30*5.54±0.32**
    H-PS组40027.36±1.38d19.31±1.64**##2.72±0.33**##7.79±0.36**##
    M-PS组20052.13±1.84b20.23±1.13**##2.41±0.24##7.46±0.49##
    L-PS组10044.20±2.03c24.53±1.74**##2.29±0.327.67±0.69*##
    注:*:表示与CT组比较,P<0.05;**:表示与CT组比较,P<0.01;#:表示与CTX组比较,P<0.05;##:表示与CTX组比较,P<0.01。
    下载: 导出CSV
  • [1] Xu J W, Vidyarthi S K, Bai W B, et al. Nutritional constituents, health benefits and processing of Rosa roxburghii: A review[J]. Journal of Functional Foods,2019,60:103456. doi:  10.1016/j.jff.2019.103456
    [2] Chen G J, Kan J Q. Characterization of a novel polysaccharide isolated from Rosa roxburghii Tratt fruit and assessment of its antioxidant in vitro and in vivo[J]. International Journal of Biological Macromolecules,2018,107:166−174. doi:  10.1016/j.ijbiomac.2017.08.160
    [3] 周宏炫, 黄颖, 谭书明, 等. 刺梨多酚对急性酒精中毒大鼠的解酒护肝作用[J/OL]. 食品科学, 2020, doi: 10.7506/spkx1002-6630-20200 828-384.

    Zhou H X, Huang Y, Tan S M, et al. Anti-alcoholic and hepatoprotective effects of polyphenols from Rosa roxburghii Tratt in acute alcoholism rats[J/OL]. Food Science, 2020, doi: 10.7506/spkx1002-6630-20200 828-384.
    [4] 夏仕青, 张爱华. 刺梨的营养保健功能及其开发利用研究进展[J]. 贵州医科大学学报,2018,43(10):1129−1132, 1153. [Xia S Q, Zhang A H. Research progress on nutrition and health function and exploitation of Rosa roxburghii Tratt[J]. Journal of Guizhou Medical University,2018,43(10):1129−1132, 1153.
    [5] 杨宗玲, 李晗, 范方宇, 等. 超声辅助酶法提取无籽刺梨果渣中黄酮的工艺优化及其抗氧化活性[J/OL]. 食品工业科技, 2021, doi:  10.13386/j.issn1002-0306.2020090273.

    Yang Z L, Li H, Fan F Y, et al. Ultrasound-assisted enzymatic extraction of flavonoids from Rosa sterilis’pomace and its antioxidant[J/OL]. Science and Technology of Food Industry, 2021, doi:  10.13386/j.issn1002-0306.2020090273.
    [6] 李跃红, 冉茂乾, 徐孟怀, 等. 不同产地刺梨果实品质分析与模糊综合评判[J]. 安徽农业科学,2020,48(17):202−205. [Li Y H, Ran M Q, Xu M H, et al. Analysis of fruit quality and fuzzy comprehensive evaluation ofRosa roxburghii Tratt from different habitats[J]. Journal of Anhui Agricultural Sciences,2020,48(17):202−205. doi:  10.3969/j.issn.0517-6611.2020.17.052
    [7] He J Y, Zhang Y H, Ma N, et al. Comparative analysis of multiple ingredients in Rosa roxburghii and R. sterilis fruits and their antioxidant activities[J]. Journal of Functional Foods,2016,27:29−41. doi:  10.1016/j.jff.2016.08.058
    [8] Chen Y, Liu Z J, Liu J, et al. Inhibition of metastasis and invasion of ovarian cancer cells by crude polysaccharides fromRosa roxburghii tratt in vitro[J]. Asian Pacific Journal of Cancer Prevention,2014,15(23):10351−10354.
    [9] Xu S J, Zhang F, Wang L J, et al. Flavonoids of Rosa roxburghii Tratt offers protection against radiation induced apoptosis and inflammation in mouse thymus[J]. Apoptosisi,2018,23:470−483. doi:  10.1007/s10495-018-1466-7
    [10] Xu P, Cai X H, Zhang W B, et al. Flavonoids ofRosa roxburghii Tratt exhibit radioprotection and anti-apoptosis properties via the Bcl-2(Ca2+)/Caspase-3/PARP-1 pathway[J]. Apoptpsis,2016,21:1125−1143. doi:  10.1007/s10495-016-1270-1
    [11] 简崇东, 唐雄林, 黄晓华, 等. 脑梗死患者口服刺梨汁抗动脉粥样硬化临床研究[J]. 亚太传统医药,2017,13(3):136−137. [Jian C D, Tang X L, Huang X H, et al. Clinical research of anti-atherosclerosis by oral Rosa roxburghii Tratt juice in patients with acute cerebral infarction[J]. Asia-Pacific Traditional Medicine,2017,13(3):136−137.
    [12] 涂永丽, 谭书明, 周宏炫, 等. 刺梨口服液对消化不良小鼠胃肠动力的促进作用[J]. 现代食品科技,2021,37(1):17−23. [Tu Y L, Tan S M, Zhou H X, et al. Effect of Rosa roxburghii Tratt oral liquid on gastrointestinal motility in dyspepsia mice[J]. Modern Food Science and Technology,2021,37(1):17−23.
    [13] 邹胜, 徐溢, 张庆. 天然植物多糖分离纯化技术研究现状和进展[J]. 天然产物研究与开发,2015,27(8):1501−1509. [Zou S, Xu Y, Zhang Q. Review on extraction and purification technology of polysaccharides from natural plants[J]. Natural Product Research and Development,2015,27(8):1501−1509.
    [14] 李秀秀, 尚静, 杨曦, 等. 多糖的增稠、胶凝及乳化特性研究进展[J/OL]. 食品科学, 2021, doi:  10.7506/spkx1002-6630-20200617-239.

    Li X X, Shang J, Yang X, et al. Rheology of food polysaccharides-thickening, gelling and emulsifying properties[J/OL]. Food Science, 2021, doi: 10.7506/spkx1002-6630-20200617-239.
    [15] 唐健波, 肖雄, 杨娟, 等. 响应面优化超声辅助提取刺梨多糖工艺研究[J]. 天然产物研究与开发,2015,27(2):314−320. [Tang J B, Xiao X, Yang J, et al. Optimization of ultrasonic-assisted extraction of polysaccharides from Rosa roxburghii Tratt using response surface methodology[J]. Natural Product Research and Development,2015,27(2):314−320.
    [16] Zong S, Li J L, Ye Z Y, et al. Lachnum polysaccharide suppresses S180 sarcoma by boosting anti-tumor immune responses and skewing tumor-associated macrophages toward M1 phenotype[J]. International Journal of Biological Macromolecules,2020,144:1022−1033. doi:  10.1016/j.ijbiomac.2019.09.179
    [17] Cai B C, Pan J Y, Chen H, et al. Oyster polysaccharides ameliorate intestinal mucositis and improve metabolism in 5-fluorouracil- treated S180 tumour-bearing mice[J]. Carbohydrate Polymers,2021,256:117545. doi:  10.1016/j.carbpol.2020.117545
    [18] 贾福怀, 涂宏建, 王俊, 等. 超声-闪式协同提取白及须根多糖工艺优化及其抗肿瘤活性[J]. 食品工业科技,2019,40(20):188−195, 208. [Jia F H, Tu H J, Wang J, et al. Optimization of ultrasonic-flash synergistic extraction and antitumor activity of polysaccharide from fibrous root of Bletilla striata[J]. Science and Technology of Food Industry,2019,40(20):188−195, 208.
    [19] 姚佳, 彭梅, 肖雄, 等. 仙茅多糖对小鼠S180实体瘤的顺铂增敏作用及其机制[J]. 华西药学杂志,2014,29(2):132−134. [Yao J, Peng M, Xiao X, et al. Efficacy enhancing effect and mechanisms of Curculigo orchioides polysaccharide on S180 tumor-bearing mice treated by Cisplatin[J]. West China Journal of Pharmaceutical Sciences,2014,29(2):132−134.
    [20] 高涛, 罗振宇, 罗黄洋, 等. 川明参多糖的超声辅助酶法提取及其体外抗氧化活性[J]. 食品研究与开发,2020,41(23):86−92. [Gao T, Luo Z Y, Luo H Y, et al. Ultrasound-assisted enzymatic extraction of polysaccharides from chuanminshen violaceum and its antioxidant acticity in virto[J]. Food Research and Development,2020,41(23):86−92.
    [21] Guo Y, Shang H M, Zhao J C, et al. Enzyme-assisted extraction of a cup plant (Silphium perfoliatum L.) polysaccharide and its antioxidant and hypoglycemic activities[J]. Process Biochemistry,2020,92:17−28. doi:  10.1016/j.procbio.2020.03.005
    [22] 吕长鑫, 李萌萌, 徐晓明, 等. 响应面分析法优化纤维素酶提取紫苏多糖工艺[J]. 食品科学,2013,34(2):6−10. [Lü C X, Li M M, Xu X M, et al. Optimization of enzymatic extraction conditions for crude polysaccharides from perilla leaves by response surface methodology[J]. Food Science,2013,34(2):6−10.
    [23] Hu H G, Zhao Q L, Xie H J, et al. Polysaccharides from pineapple pomace: new insight into ultrasonic-cellulase synergistic extraction and hypoglycemic activities[J]. International Journal of Biological Macromolecules,2019,121:1213−1226. doi:  10.1016/j.ijbiomac.2018.10.054
    [24] Madondo M T, Quinn M, Plebanski M. Low dose cyclophosphamide: Mechanisms of T cell modulation[J]. Cancer Treatment Reviews,2016,42:3−9. doi:  10.1016/j.ctrv.2015.11.005
    [25] Strati P, Wierda W, Burger J, et al. Myelosuppression after frontline fludarabine, cyclophosphamide, and rituximab in patients with chronic lymphocytic leukemia: analysis of persistent and new-onset cytopenia[J]. Cancer,2013,119(21):3805−3811. doi:  10.1002/cncr.28318
    [26] Molodykh O P, Sorokina I V, Vinogradova E V, et al. Ultrastructure of the liver in response to cyclophosphamide and triterpenoids[J]. Bulletin of Experimental Biology and Medicine,2020,160:400−405.
    [27] Nevine R T, Amin H A, Sultan A A. The protective effect of Moringa oleifera leaves against cyclophosphamide-induced urinary bladder toxicity in rats[J]. Tissue and Cell,2015,47(1):94−104. doi:  10.1016/j.tice.2014.12.002
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  • 收稿日期:  2021-03-12
  • 网络出版日期:  2021-08-04
  • 刊出日期:  2021-09-14

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