典型甜菊糖苷的抗氧化和抗炎活性
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摘要
本文以6种代表性甜菊糖苷及其肠道代谢物甜菊醇以及甜菊醇的重排异构体异甜菊醇为对象,研究了上述甜菊化合物的抗氧化性及其对鼠巨噬细胞Raw264.7的细胞毒性和抗炎活性。结果表明,所试甜菊糖苷在100μg/mL内对鼠巨噬细胞的Raw264.7均无细胞毒性,也不会诱导炎症反应;其中异甜菊醇抗炎效果最优,100μg/mL时相对于脂多糖刺激后产生的NO含量降低了约30.7%。所试甜菊糖苷对DPPH·和·OH两种自由基的清除率最高分别为24.8%和13.8%,其对DPPH·的清除能力具有结构相关性,但对·OH清除率均低,无结构相关性;而甜菊酚对DPPH·和ABTS~+·清除率分别为92.1%和29.6%,即甜菊提取物的自由基清除率主要由甜菊酚产生。
In this experiment,the cytotoxicity and anti-inflammatory activity of stevia compounds on mouse macrophage Raw264.7 cells was investigated with 6 representative steviol glycosides along with their intestinal metabolite steviol as well as steviol's rearrangement product isosteviol. Meanwhile,the antioxidant activity of the above stevia compounds and their cytotoxicity and anti-inflammatory activity against Raw264.7 of murine macrophages were investigated. The results showed that the stevia compounds had no cytotoxicity,nor induced inflammatory reaction in the range of 100 μg/mL. Isosteviol showed the best anti-inflammatory effect,when the isosteviol concentration was 100 μg/mL,the NO contents produced by stimulation with lipopolysaccharide was reduced about 30.7%. The highest free radical scavenging abilities of the assayed steviol glycosides were 24.8%(DPPH·)and 13.8%(·OH),respectively. There had a structural correlation with the scavenging ability on DPPH free radicals,while there was no structural correlation on hydroxyl free radical clearance. When the experiment was carried out with stevia phenols,the DPPH· and ABTS~+· scavenging rates were 92.1% and 29.6%,respectively. Therefore,the antioxidant activity of stevia extracts could be mainly caused by the stevia phenols.
In this experiment,the cytotoxicity and anti-inflammatory activity of stevia compounds on mouse macrophage Raw264.7 cells was investigated with 6 representative steviol glycosides along with their intestinal metabolite steviol as well as steviol's rearrangement product isosteviol. Meanwhile,the antioxidant activity of the above stevia compounds and their cytotoxicity and anti-inflammatory activity against Raw264.7 of murine macrophages were investigated. The results showed that the stevia compounds had no cytotoxicity,nor induced inflammatory reaction in the range of 100 μg/mL. Isosteviol showed the best anti-inflammatory effect,when the isosteviol concentration was 100 μg/mL,the NO contents produced by stimulation with lipopolysaccharide was reduced about 30.7%. The highest free radical scavenging abilities of the assayed steviol glycosides were 24.8%(DPPH·)and 13.8%(·OH),respectively. There had a structural correlation with the scavenging ability on DPPH free radicals,while there was no structural correlation on hydroxyl free radical clearance. When the experiment was carried out with stevia phenols,the DPPH· and ABTS~+· scavenging rates were 92.1% and 29.6%,respectively. Therefore,the antioxidant activity of stevia extracts could be mainly caused by the stevia phenols.
引文
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[19]Cai C E,Yang Y Y,Zhao M L,et al. Extraction and antioxidation of polysaccharide from Porphyra Haitanensis using response surface method[J]. Pakistan Journal of Botany,2017,49(3):1137-1141.
[20]Zou Y,Hou X Y. Extraction optimization,composition analysis,and antioxidation evaluation of polysaccharides from white jelly mushroom,Tremella fuciformis(Tremellomycetes)[J]. International Journal of Medicinal Mushrooms,2017,19(12):1113-1121.
[21]Can Z,Balta? N. Bioactivity and enzyme inhibition properties of Stevia rebaudiana[J]. Gurrent Enzyme Inhibition,2016,12:188-194.
[22]Boonkaewwan C,Ao M,Toskulkao C,et al. Specific immunomodulatory and secretory activities of stevioside(SVS)and steviol(SVO)in intestinal cells[J]. Faseb Journal,2007,21(6):A1319-A1319.
[23]Boonkaewwan C,Ao M,Toskulkao C,et al. Specific immunomodulatory and secretory activities of stevioside and steviol in intestinal cells[J]. Journal of Agricultural and Food Chemistry,2008,56(10):3777-3784.
[24]Li F Y,Fu Y H,Liu B,et al. Stevioside suppressed inflammatory cytokine secretion by downregulation of NF-kappa B and MAPK signaling pathways in LPS-stimulated RAW264.7 cells[J]. Inflammation,2012,35(5):1669-1675.
[2]López V,Perez S,Vinuesa A,et al. Stevia rebaudiana ethanolic extract exerts better antioxidant properties and antiproliferative effects in tumour cells than its diterpene glycoside stevioside[J]. Food and Function,2016,7(4):2107-2113.
[3]Yu H,Yang G Q,Sato M,et al. Antioxidant activities of aqueous extract from Stevia rebaudiana stem waste to inhibit fish oil oxidation and identification of its phenolic compounds[J]. Food Chemistry,2017,232:379-386.
[4]Ortiz-Viedma J,Romero N,Puente L,et al. Antioxidant and antimicrobial effects of stevia(Stevia rebaudiana Bert.)extracts during preservation of refrigerated salmon paste[J]. European Journal of Lipid Science and Technology,2017,119(10):1600467.
[5]Prata C,Zambonin L,Rizzo B,et al. Glycosides from Stevia rebaudiana Bertoni possess insulin-mimetic and antioxidant activities in rat cardiac fibroblasts[J]. Oxidative Medicine and Cellular Longevity,2017,2017:1-13.
[6]Mikhailov A L,Nevmerzhitskaya J Y,Timofeeva O A. The antioxidant properties of stevioside under the influence of heavy metals[J]. Research Journal of Pharmaceutical Biological and Chemical Sciences,2016,7(5):1721-1727.
[7]Boonkaewwan C,Toskulkao C,Vongsakul M. Anti-inflammatory and immunomodulatory activities of stevioside and its metabolite steviol on THP-1 cells[J]. Journal of Agricultural and Food Chemistry,2006,54(3):785-789.
[8]Boonkaewwan C,Burodom A. Anti-inflammatory and immunomodulatory activities of stevioside and steviol on colonic epithelial cells[J]. Journal of the Science of Food and Agriculture,2013,93(15):3820-3825.
[9]Choi D H,Cho U M,Hwang H S. Anti-inflammation effect of rebaudioside a by inhibition of the mapk and NF-κB signal pathway in raw264.7 macrophage[J]. Journal of Applied Biological Chemistry,2018,61(2):205-211.
[10]Zhang Q,Yang H,Li Y,et al. Toxicological evaluation of ethanolic extract from Stevia rebaudiana Bertoni leaves:Genotoxicity and subchronic oral toxicity[J]. Regulatory Toxicology and Pharmacology,2017,86253-259.
[11]Wan H D,Tao G J,Kim D,et al. Enzymatic preparation of a natural sweetener rubusoside from specific hydrolysis of stevioside with beta-galactosidase from Aspergillus sp.[J]. Journal of Molecular Catalysis B-Enzymatic,2012,8212-17.
[12]Wood H B,Allerton R,Diehl H W,et al. Stevioside. i. the structure of the glucose moieties[J]. Journal of Organic Chemistry,1955,20(7):875-883.
[13]Chen J M,Xia Y M,Wan H D,et al. A complete specific cleavage of glucosyl and ester linkages of stevioside for preparingsteviol with a beta-galactosidase from Sulfolobus solfataricus[J]. Journal of Molecular Catalysis B-Enzymatic,2014,105:126-131.
[14]张宗英,胡学一,夏咏梅. 盐酸或氨基磺酸催化水解斯替夫苷制备甜菊醇[J]. 精细化工,2014,31(4):540-544.
[15]Brand-Williams W,Cuvelier M E,Berset C. Use of a free-radical method to evaluate antioxidant activity[J]. Food Science and Technology-Lebensmittel-Wissenschaft & Technologie,1995,28(1):25-30.
[16]Rowley D A,Halliwell B. Superoxide-dependent formation of hydroxyl radicals from NADH and NADPH in the presence of iron salts[J]. Febs Letters,1982,142(1):39-41.
[17]Re R,Pellegrini N,Proteggente A,et al. Antioxidant activity applying an improved ABTS radical cation decolorization assay[J]. Free Radical Biology and Medicine,1999,26(9):1231-1237.
[18]Pan F,Su T J,Liu Y,et al. Extraction,purification and antioxidation of a polysaccharide from Fritillaria unibracteata var. wabuensis[J]. International Journal of Biological Macromolecules,2018,112:1073-1083.
[19]Cai C E,Yang Y Y,Zhao M L,et al. Extraction and antioxidation of polysaccharide from Porphyra Haitanensis using response surface method[J]. Pakistan Journal of Botany,2017,49(3):1137-1141.
[20]Zou Y,Hou X Y. Extraction optimization,composition analysis,and antioxidation evaluation of polysaccharides from white jelly mushroom,Tremella fuciformis(Tremellomycetes)[J]. International Journal of Medicinal Mushrooms,2017,19(12):1113-1121.
[21]Can Z,Balta? N. Bioactivity and enzyme inhibition properties of Stevia rebaudiana[J]. Gurrent Enzyme Inhibition,2016,12:188-194.
[22]Boonkaewwan C,Ao M,Toskulkao C,et al. Specific immunomodulatory and secretory activities of stevioside(SVS)and steviol(SVO)in intestinal cells[J]. Faseb Journal,2007,21(6):A1319-A1319.
[23]Boonkaewwan C,Ao M,Toskulkao C,et al. Specific immunomodulatory and secretory activities of stevioside and steviol in intestinal cells[J]. Journal of Agricultural and Food Chemistry,2008,56(10):3777-3784.
[24]Li F Y,Fu Y H,Liu B,et al. Stevioside suppressed inflammatory cytokine secretion by downregulation of NF-kappa B and MAPK signaling pathways in LPS-stimulated RAW264.7 cells[J]. Inflammation,2012,35(5):1669-1675.