樟树不同器官中多糖抗氧化、免疫调节活性的研究
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摘要
采用先喷雾干燥后醇沉的方法制备樟树根多糖(CCRP)、樟树茎多糖(CCSP)、樟树叶多糖(CCLP)以及樟树果实多糖(CCFP),并用苯酚硫酸法检测各多糖的含量,通过测定多糖清除DPPH自由基、ABTS自由基的能力以及还原能力评价抗氧化活性。然后通过体外实验评估其对RAW 264.7巨噬细胞免疫调节活性。研究发现:CCRP、CCSP、CCLP和CCFP多糖的含量依次为52.35%、58.56%、56.62%和62.36%,果实多糖含量最高,抗氧化能力评价中,相同器官中粗多糖比精制多糖抗氧化活性能力强。同一浓度下,果实多糖对DPPH自由基清除能力最强,当多糖提取物浓度为2 mg/mL,清除率达到87.65%,其次是根多糖,清除率为79.63%;多糖提取物还原能力较强的部位是果实和根,在多糖浓度为2 mg/mL时还原能力分别达到最大0.98和0.85;但是各器官多糖对ABTS自由基的清除能力较弱。另一方面,CCRP、CCLP、CCFP和CCSP均可显著提高RAW 264.7细胞内一氧化氮(NO)的浓度和细胞因子如前列腺素E2(PGE2)和肿瘤坏死因子(TNF-α)的分泌。免疫调节活性大小顺序为樟树果实多糖>樟树叶多糖>樟树根多糖>樟树茎多糖。CCRP、CCSP、CCLP和CCFP均具有抗氧化活性并可以提高免疫力,在功能食品的开发中,可以作为一个潜在的抗氧化、免疫调节剂。
Spray of drying-alcohol precipitation was used to prepare polysaccharides from root,stem,leaf and fruit of Cinamomum camphora,and the contents of polysaccharide were detected by phenol-sulfuric acid method.The in vitro antioxidant activity of polysaccharide was investigated by measuring the total reducing power and scavenging activities against DPPH and ABTS.Their in vitro immunological activity were evaluated with macrophage RAW264.7.The results showed that the contents of polysaccharides in CCRP,CCSP,CCLP and CCFP were 52.35%,58.56%,56.62% and 62.36% respectively,and the fruit had the highest polysaccharide content. In the evaluation of antioxidant capacity,crude polysaccharides had stronger antioxidant activity than purified polysaccharides in the same organs.Under the same concentration,the fruit polysaccharide had the strongest scavenging ability to DPPH free radical. When the concentration of polysaccharide extract was 2 mg/mL,the clearance rate was 87.65%,the second was root polysaccharide,the clearance rate was 79.63%,the organs with high reduction capacity of the polysaccharide extract were fruit and root,and the reduction ability of the polysaccharide was 2 mg/mL to the maximum of 0.98 and 0.85,respectively.but the scavenging capacity of Polysaccharides from various organs to ABTS free radicals was weak.The results indicated that CCRP,CCLP,CCFP and CCSP could significantly promote the concentrations of intracellular nitric oxide(NO)and cytokines such as prostaglandin E2(PGE2)and tumor necrosis factor-α(TNF-α) in RAW 264.7 cells,and the order of immunological activity was:CCFP > CCLP > CCRP >CCSP.Taken together,these results suggest that CCFP can improve immunity,and could be explored as a potential immunomodulatory agent in functional food.
Spray of drying-alcohol precipitation was used to prepare polysaccharides from root,stem,leaf and fruit of Cinamomum camphora,and the contents of polysaccharide were detected by phenol-sulfuric acid method.The in vitro antioxidant activity of polysaccharide was investigated by measuring the total reducing power and scavenging activities against DPPH and ABTS.Their in vitro immunological activity were evaluated with macrophage RAW264.7.The results showed that the contents of polysaccharides in CCRP,CCSP,CCLP and CCFP were 52.35%,58.56%,56.62% and 62.36% respectively,and the fruit had the highest polysaccharide content. In the evaluation of antioxidant capacity,crude polysaccharides had stronger antioxidant activity than purified polysaccharides in the same organs.Under the same concentration,the fruit polysaccharide had the strongest scavenging ability to DPPH free radical. When the concentration of polysaccharide extract was 2 mg/mL,the clearance rate was 87.65%,the second was root polysaccharide,the clearance rate was 79.63%,the organs with high reduction capacity of the polysaccharide extract were fruit and root,and the reduction ability of the polysaccharide was 2 mg/mL to the maximum of 0.98 and 0.85,respectively.but the scavenging capacity of Polysaccharides from various organs to ABTS free radicals was weak.The results indicated that CCRP,CCLP,CCFP and CCSP could significantly promote the concentrations of intracellular nitric oxide(NO)and cytokines such as prostaglandin E2(PGE2)and tumor necrosis factor-α(TNF-α) in RAW 264.7 cells,and the order of immunological activity was:CCFP > CCLP > CCRP >CCSP.Taken together,these results suggest that CCFP can improve immunity,and could be explored as a potential immunomodulatory agent in functional food.
引文
[1]胡文杰，高捍东，江香梅.响应面法优化樟树叶精油水蒸汽蒸馏提取工艺[J].江西农业大学学报，2013,35(1):144-151.Hu W J,Gao H D,Jiang X M.Optimization of extraction techniques for the essential oil in leaves in Cinnamomum camphora by response surface methodology[J].Acta Agriculturae Universitatis Jiangxiensis,2013,35(1):144-151.
[2]张峰，毕良武，赵振东.樟树植物资源分布及化学成分研究进展[J].天然产物研究与开发，2017,29(3):517-531.Zhang F,Bi L W,Zhao Z D.Review on plant resources and chemical composition of Camphor tree[J].Natural Product Research Development,2017,29(3):517-531.
[3]杨芳，郑青，赵曼丽，等.响应面法优化樟树籽油脂质体的制备工艺[J].食品科学，2013,34(2):125-130.Yang F,Zheng Q,Zhao M L,et al.Optimization of preraration process for Cinnamomum camphora seed oil liposome[J].Food Science,2013,34(2):125-130.
[4]郭丹，曾解放，范国荣，等.樟树精油的化学成分及生物活性研究进展[J].生物质化学工程，2015,49(1):53-57.Guo D,Zeng J F,Fan G R,et al.Research progress in chemical compositions and bioactivity of essential oil from Cinnamomum camphora[J].Biomass Chemical Engineering,2015,49(1):53-57.
[5]尹爱武，田润，李探芳，等.樟树木化学成分研究[J].天然产物研究与开发，2014,26(6):876-878.Yin A W,Tian R,Li T F,et al.Studies on the chemical constituents of the stem of Cinamomum camphora[J].Natural Product Research Development,2014,26(6):876-878.
[6]周海旭.樟树叶中木脂素提取分离及其生物活性研究[D].长沙：中南林业科技大学，2017.Zhou H X.Study on the extraction,purificatiom and biological activities of lignans from Cinnamomum camphora leaf[D].Changsha:Central South University of Forestry and Technology,2017.
[7]Roszaini K,Azah M A N,Mailina J,et al.Toxicity and antitermite activity of the essential oils from Cinnamomum camphora,Cymbopogon nardus,Melaleuca cajuputi and Dipterocarpus sp.against Coptotermes curvignathus[J].Wood Science and Technology,2013,47(6):1273-1284.
[8]Zhou H,Ren J,Li Z.Antibacterial activity and mechanism of pinoresinol from Cinnamomum camphora leaves against foodrelated bacteria[J].Food Control,2017,79:192-199.
[9]C?r D,Knez?，Knez Hrn?i?M.Antitumour,antimicrobial,antioxidant and antiacetylcholinesterase effect of ganoderma lucidum terpenoids and polysaccharides:a review[J].Molecules,2018,23(3):649-658.
[10]Zong A,Cao H,Wang F.Anticancer polysaccharides from natural resources:a review of recent research[J].Carbohydrate Polymers,2012,90(4):1395-1410.
[11]Liu J,Willf?r S,Xu C.A review of bioactive plant polysaccharides:biological activities,functionalization,and biomedical applications[J].Bioactive Carbohydrates and Dietary Fibre,2015,5(1):31-61.
[12]Galluzzi L,BuquéA,Kepp O,et al.Immunogenic cell death in cancer and infectious disease[J].Nature Reviews Immunology,2017,17(2):97-111.
[13]Tartey S,Takeuchi O.Pathogen recognition and Toll-like receptor targeted therapeutics in innate immune cells[J].International Reviews of Immunology,2017,36(2):57-73.
[14]Berraondo P,Minute L,Ajona D,et al.Innate immune mediators in cancer:between defense and resistance[J].Immunological Reviews,2016,274(1):290-306.
[15]Zhang L,Wang C C.Inflammatory response of macrophages in infection[J].Hepatobiliary&Pancreatic Diseases International,2014,13(2):138-152.
[16]Wang J,Hu S,Nie S,et al.Reviews on mechanisms of in vitro antioxidant activity of polysaccharides[J].Oxidative Medicine and Cellular Longevity,2016(64):11-13.
[17]张晓艳，王波，黄攀，等.马齿苋不同器官多糖提取物的抗氧化性研究[J].食品工业科技，2017,38(5):130-133.Zhang X Y,Wang B,Huang P,et al.Study on the antioxidant activities of polysaccharide extracts from different organs of Portulace oleracea L.[J].Science and Technology of Food Industry,2017,38(5):130-133.
[18]任颖朗，季德胜，张云林，等.龙须菜多糖的提取及其免疫调节活性研究[J].现代食品科技，2017,33(10):45-51.Ren Y L,Ji D S,Zhang Y L,et al.Extraction of polysaccharides from Gracilaria lemaneiformis and their immunoregulatory activity[J].Modern Food Science and Technology,2017,33(10):45-51.
[19]张丹，李丹，许启泰，等.槟榔提取物不同部位的抗氧化性比较及成分研究[J].食品工业科技，2015,36(2):102-104.Zhang D,Li D,Xu Q T,et al.Comparative study of antioxidant activity and ingredient in different parts of Areca catechu L.extract[J].Science and Technology of Food Industry,2015,36(2):102-104.
[20]徐丛玥，林款，梁征，等.不同提取方法对米邦塔仙人掌粗多糖体外抗氧化性的影响[J].食品工业科技，2018(3):56-60.Xu C Y,Lin K,Liang Z,et al.Effect of different extraction methods on the antioxidant activity of crude polysaccharide from Opuntia Milpa Alta Haw in vitro[J].Science and Technology of Food Industry,2018(3):56-60.
[21]陈铭，薛琪，沈婷，等.超微粉碎对莲子心活性成分提取率及抗氧化能力的影响[J].现代食品科技，2016,32(11):228-234.Chen M,Xue Q,Shen T,et al.Effects of ultrafine grinding on the extraction rate and antioxidant properties of the active ingredients of Lotus plumule[J].Modern Food Science and Technology,2016,32(11):228-234.
[22]Hu W C,Wu L,Qiang Q,et al.The dichloromethane fraction from Mahonia bealei(Fort.)Carr.leaves exerts an anti-inflammatory effect both in vitro and in vivo[J].Journal of Ethnopharmacology,2016,188:134-143.
[23]Wu L,Li X,Hu W,et al.5-Methoxyl aesculetin abrogates lipopolysaccharide-induced inflammation by suppressing MAPK and AP-1 pathways in RAW 264.7 cells[J].International Journal of Molecular Sciences,2016,17(3):315-342.
[24]蔡英卿，赖钟雄，方煜，等.枇杷不同器官多糖含量的比较[J].河北工程大学学报（自然科学版），2007,24(3):60-63.Cai Y Q,Lai Z X,Fang Y,et al.Analyses of polysaccharide contents in different organs of Eriobotrya japonica L.[J].Journal of Hebei University of Engineering(Natural Science Edition),2007,24(3):60-63.
[25]王娜，王凯旋，李静，等.樟树叶片水溶性粗多糖提取及抗氧化活性研究[J].北方园艺，2010(24):70-73.Wang N,Wang K X,Li J,et al.Study on extraction and antioxidant activity of water-soluble coarse polysaccharides from Camphor Tree[J].Northern Horticulture,2010(24):70-73.
[26]付湘晋，王挥，冉晓敏，等.樟树叶多酚提取物体外清除自由基及抗油脂氧化活性研究[J].食品工业科技，2013,34(19):108-110.Fu X J,Wang H,Ran X M,et al.Antioxidant activity of polyphenolic compounds in Camphor tree leaves[J].Science and Technology of Food Industry,2013,34(19):108-110.
[27]许小向，洪艳平，胡捷敏，等.松针多糖微波提取工艺及抗氧化性研究[J].食品工业科技，2015,36(23):222-227.Xu X Q,Hong Y P,Hu J M,et al.Study on microwave extraction technology and antioxidant activity of pine needle polysaccharose[J].Science and Technology of Food Industry,2015,36(23):222-227.
[28]Yu J,Hu M,Wang Y,et al.Extraction,partial characterization and bioactivity of polysaccharides from Senecio scandens Buch.-Ham[J].International Journal of Biological Macromolecules,2018,109:535-543.
[29]Wen Z S,Xiang X W,Jin H X,et al.Composition and anti-inflammatory effect of polysaccharides from Sargassum horneri in RAW264.7 macrophages[J].International Journal of Biological Macromolecules,2016,88:403-413.
[30]杨瑾，陈怡，刘影，等.樟树成熟果皮中总多糖的提取工艺研究[J].山东化工，2018,47(11):46-54.Yang J,Chen Y,Liu Y,et al.Study on the extraction technology of total polysaccharides from mature pericarp of Cinnamomum camphora[J].Shandong Chemical Industry,2018,47(11):46-54.
[31]王先.芳樟叶黄酮和多糖的提取分离、结构鉴定及抗氧化活性研究[D].厦门：厦门大学，2008.Wang X.Extraction,separation,identification and antioxidant activities of flavonoid and polysaccharide from Cinnamomum camphora leaves[D].Xiamen:Xiamen University,2008.
[2]张峰，毕良武，赵振东.樟树植物资源分布及化学成分研究进展[J].天然产物研究与开发，2017,29(3):517-531.Zhang F,Bi L W,Zhao Z D.Review on plant resources and chemical composition of Camphor tree[J].Natural Product Research Development,2017,29(3):517-531.
[3]杨芳，郑青，赵曼丽，等.响应面法优化樟树籽油脂质体的制备工艺[J].食品科学，2013,34(2):125-130.Yang F,Zheng Q,Zhao M L,et al.Optimization of preraration process for Cinnamomum camphora seed oil liposome[J].Food Science,2013,34(2):125-130.
[4]郭丹，曾解放，范国荣，等.樟树精油的化学成分及生物活性研究进展[J].生物质化学工程，2015,49(1):53-57.Guo D,Zeng J F,Fan G R,et al.Research progress in chemical compositions and bioactivity of essential oil from Cinnamomum camphora[J].Biomass Chemical Engineering,2015,49(1):53-57.
[5]尹爱武，田润，李探芳，等.樟树木化学成分研究[J].天然产物研究与开发，2014,26(6):876-878.Yin A W,Tian R,Li T F,et al.Studies on the chemical constituents of the stem of Cinamomum camphora[J].Natural Product Research Development,2014,26(6):876-878.
[6]周海旭.樟树叶中木脂素提取分离及其生物活性研究[D].长沙：中南林业科技大学，2017.Zhou H X.Study on the extraction,purificatiom and biological activities of lignans from Cinnamomum camphora leaf[D].Changsha:Central South University of Forestry and Technology,2017.
[7]Roszaini K,Azah M A N,Mailina J,et al.Toxicity and antitermite activity of the essential oils from Cinnamomum camphora,Cymbopogon nardus,Melaleuca cajuputi and Dipterocarpus sp.against Coptotermes curvignathus[J].Wood Science and Technology,2013,47(6):1273-1284.
[8]Zhou H,Ren J,Li Z.Antibacterial activity and mechanism of pinoresinol from Cinnamomum camphora leaves against foodrelated bacteria[J].Food Control,2017,79:192-199.
[9]C?r D,Knez?，Knez Hrn?i?M.Antitumour,antimicrobial,antioxidant and antiacetylcholinesterase effect of ganoderma lucidum terpenoids and polysaccharides:a review[J].Molecules,2018,23(3):649-658.
[10]Zong A,Cao H,Wang F.Anticancer polysaccharides from natural resources:a review of recent research[J].Carbohydrate Polymers,2012,90(4):1395-1410.
[11]Liu J,Willf?r S,Xu C.A review of bioactive plant polysaccharides:biological activities,functionalization,and biomedical applications[J].Bioactive Carbohydrates and Dietary Fibre,2015,5(1):31-61.
[12]Galluzzi L,BuquéA,Kepp O,et al.Immunogenic cell death in cancer and infectious disease[J].Nature Reviews Immunology,2017,17(2):97-111.
[13]Tartey S,Takeuchi O.Pathogen recognition and Toll-like receptor targeted therapeutics in innate immune cells[J].International Reviews of Immunology,2017,36(2):57-73.
[14]Berraondo P,Minute L,Ajona D,et al.Innate immune mediators in cancer:between defense and resistance[J].Immunological Reviews,2016,274(1):290-306.
[15]Zhang L,Wang C C.Inflammatory response of macrophages in infection[J].Hepatobiliary&Pancreatic Diseases International,2014,13(2):138-152.
[16]Wang J,Hu S,Nie S,et al.Reviews on mechanisms of in vitro antioxidant activity of polysaccharides[J].Oxidative Medicine and Cellular Longevity,2016(64):11-13.
[17]张晓艳，王波，黄攀，等.马齿苋不同器官多糖提取物的抗氧化性研究[J].食品工业科技，2017,38(5):130-133.Zhang X Y,Wang B,Huang P,et al.Study on the antioxidant activities of polysaccharide extracts from different organs of Portulace oleracea L.[J].Science and Technology of Food Industry,2017,38(5):130-133.
[18]任颖朗，季德胜，张云林，等.龙须菜多糖的提取及其免疫调节活性研究[J].现代食品科技，2017,33(10):45-51.Ren Y L,Ji D S,Zhang Y L,et al.Extraction of polysaccharides from Gracilaria lemaneiformis and their immunoregulatory activity[J].Modern Food Science and Technology,2017,33(10):45-51.
[19]张丹，李丹，许启泰，等.槟榔提取物不同部位的抗氧化性比较及成分研究[J].食品工业科技，2015,36(2):102-104.Zhang D,Li D,Xu Q T,et al.Comparative study of antioxidant activity and ingredient in different parts of Areca catechu L.extract[J].Science and Technology of Food Industry,2015,36(2):102-104.
[20]徐丛玥，林款，梁征，等.不同提取方法对米邦塔仙人掌粗多糖体外抗氧化性的影响[J].食品工业科技，2018(3):56-60.Xu C Y,Lin K,Liang Z,et al.Effect of different extraction methods on the antioxidant activity of crude polysaccharide from Opuntia Milpa Alta Haw in vitro[J].Science and Technology of Food Industry,2018(3):56-60.
[21]陈铭，薛琪，沈婷，等.超微粉碎对莲子心活性成分提取率及抗氧化能力的影响[J].现代食品科技，2016,32(11):228-234.Chen M,Xue Q,Shen T,et al.Effects of ultrafine grinding on the extraction rate and antioxidant properties of the active ingredients of Lotus plumule[J].Modern Food Science and Technology,2016,32(11):228-234.
[22]Hu W C,Wu L,Qiang Q,et al.The dichloromethane fraction from Mahonia bealei(Fort.)Carr.leaves exerts an anti-inflammatory effect both in vitro and in vivo[J].Journal of Ethnopharmacology,2016,188:134-143.
[23]Wu L,Li X,Hu W,et al.5-Methoxyl aesculetin abrogates lipopolysaccharide-induced inflammation by suppressing MAPK and AP-1 pathways in RAW 264.7 cells[J].International Journal of Molecular Sciences,2016,17(3):315-342.
[24]蔡英卿，赖钟雄，方煜，等.枇杷不同器官多糖含量的比较[J].河北工程大学学报（自然科学版），2007,24(3):60-63.Cai Y Q,Lai Z X,Fang Y,et al.Analyses of polysaccharide contents in different organs of Eriobotrya japonica L.[J].Journal of Hebei University of Engineering(Natural Science Edition),2007,24(3):60-63.
[25]王娜，王凯旋，李静，等.樟树叶片水溶性粗多糖提取及抗氧化活性研究[J].北方园艺，2010(24):70-73.Wang N,Wang K X,Li J,et al.Study on extraction and antioxidant activity of water-soluble coarse polysaccharides from Camphor Tree[J].Northern Horticulture,2010(24):70-73.
[26]付湘晋，王挥，冉晓敏，等.樟树叶多酚提取物体外清除自由基及抗油脂氧化活性研究[J].食品工业科技，2013,34(19):108-110.Fu X J,Wang H,Ran X M,et al.Antioxidant activity of polyphenolic compounds in Camphor tree leaves[J].Science and Technology of Food Industry,2013,34(19):108-110.
[27]许小向，洪艳平，胡捷敏，等.松针多糖微波提取工艺及抗氧化性研究[J].食品工业科技，2015,36(23):222-227.Xu X Q,Hong Y P,Hu J M,et al.Study on microwave extraction technology and antioxidant activity of pine needle polysaccharose[J].Science and Technology of Food Industry,2015,36(23):222-227.
[28]Yu J,Hu M,Wang Y,et al.Extraction,partial characterization and bioactivity of polysaccharides from Senecio scandens Buch.-Ham[J].International Journal of Biological Macromolecules,2018,109:535-543.
[29]Wen Z S,Xiang X W,Jin H X,et al.Composition and anti-inflammatory effect of polysaccharides from Sargassum horneri in RAW264.7 macrophages[J].International Journal of Biological Macromolecules,2016,88:403-413.
[30]杨瑾，陈怡，刘影，等.樟树成熟果皮中总多糖的提取工艺研究[J].山东化工，2018,47(11):46-54.Yang J,Chen Y,Liu Y,et al.Study on the extraction technology of total polysaccharides from mature pericarp of Cinnamomum camphora[J].Shandong Chemical Industry,2018,47(11):46-54.
[31]王先.芳樟叶黄酮和多糖的提取分离、结构鉴定及抗氧化活性研究[D].厦门：厦门大学，2008.Wang X.Extraction,separation,identification and antioxidant activities of flavonoid and polysaccharide from Cinnamomum camphora leaves[D].Xiamen:Xiamen University,2008.
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