原花青素B_2对D-半乳糖模型小鼠肠道菌群的影响
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摘要
以D-半乳糖皮下注射诱导小鼠衰老模型,探究原花青素B_2对衰老小鼠肠道菌群结构的影响。将C57/BL小鼠随机分为对照组、衰老组和原花青素B_2组,7周后测定其十二指肠及结肠组织中总抗氧化能力、超氧化物歧化酶活力、过氧化物酶活力和丙二醛含量,利用Illumina MiSeq高通量测序技术检测其肠道菌群16S rDNA基因,探索原花青素B_2与衰老小鼠肠道菌群之间的关系,利用气相色谱-质谱联用仪检测小鼠粪便短链脂肪酸(shortchainfattyacids,SCFAs)含量。结果表明:D-半乳糖模型小鼠的十二指肠及结肠的抗氧化酶活力显著下降,MDA含量显著上升(P<0.05);在原花青素B_2干预后,小鼠体内的多种抗氧化酶活力上升,MDA含量显著降低(P<0.05),与对照组没有显著差异(P>0.05);利用高通量测序技术发现3组小鼠肠道菌群中主要发生变化的是norank-f-Bacteroidales-s24-7、Bacteroides、Blautia、unclassified-Lachnospiraeae、Ruminiclostridium、Lachnospiraeae-NK4A136、Roseburia 7种菌属;相比于对照组,衰老组norank-f-Bacteroidales-s24-7的相对丰度显著降低,其他菌属的相对丰度显著上升(P<0.05);与衰老组相比,原花青素B_2组norank-f-Bacteroidales-s24-7、Roseburia、Lachnospiraeae-NK4A136和Bacteroides的相对丰度显著上升,而Blautia、unclassified-Lachnospiraeae和Ruminiclostridium的相对丰度显著降低(P<0.05)。原花青素B_2可提高衰老小鼠体内SCFAs含量,改善肠道环境,进而延缓衰老进程。
The objective of the study is to investigate the effect of procyanidins B_2 on the intestinal microflora in D-galactose-induced aging mouse model. Thirty C57/BL mice were randomly and equally divided into control, aging model and procyanidins B_2 treatment groups. After an experimental period of seven weeks, total antioxidant capacity, superoxide dismutase activity and catalase activity, and malondialdehyde(MDA) content as a lipid peroxidation marker were detected in the duodenum and colon. High-throughput sequencing of the 16 S rDNA gene of the intestinal flora was performed using an Illumina MiSeq platform to explore the relationship between procyanidin B_2 and the intestinal flora in aging mice. The contents of short-chain fatty acids(SCFAs) in mouse feces were determined by gas chromatography-mass spectrometry. The results showed that the activity of antioxidant enzymes was significantly decreased in the duodenum and colon of aging mice,and the content of MDA was significantly increased(P < 0.05). After intervention with procyanidins B_2, antioxidant enzyme and MDA levels were restored to values not significantly different from normal(P > 0.05). The major differential intestinal microorganisms were norank-f-Bacteroidales-s24-7, Bacteroides, Blautia, unclassified-Lachnospiraeae, Ruminiclostridium,Lachnospiraeae-NK4A136, and Roseburia among all groups of mice. Compared with the control group, the proportion of norank-f-Bacteroidales-s24-7 in the aging group was decreased significantly, and the proportions of the other strains were increased significantly(P < 0.05). Compared with the aging group, the proportions of norank-f-Bacteroidales-s24-7,Roseburia, Lachnospiraeae-NK4 A136 and Bacteroides were significantly increased in the mice treated with procyanidins B_2,while the proportion of Blautia, unclassified-Lachnospiraeae, and Ruminiclostridium was significantly decreased(P < 0.05).Accordingly, procyanidins B_2 could regulate the contents of SCFAs in aging mice to improve the intestinal environment,thereby delaying the aging process.
The objective of the study is to investigate the effect of procyanidins B_2 on the intestinal microflora in D-galactose-induced aging mouse model. Thirty C57/BL mice were randomly and equally divided into control, aging model and procyanidins B_2 treatment groups. After an experimental period of seven weeks, total antioxidant capacity, superoxide dismutase activity and catalase activity, and malondialdehyde(MDA) content as a lipid peroxidation marker were detected in the duodenum and colon. High-throughput sequencing of the 16 S rDNA gene of the intestinal flora was performed using an Illumina MiSeq platform to explore the relationship between procyanidin B_2 and the intestinal flora in aging mice. The contents of short-chain fatty acids(SCFAs) in mouse feces were determined by gas chromatography-mass spectrometry. The results showed that the activity of antioxidant enzymes was significantly decreased in the duodenum and colon of aging mice,and the content of MDA was significantly increased(P < 0.05). After intervention with procyanidins B_2, antioxidant enzyme and MDA levels were restored to values not significantly different from normal(P > 0.05). The major differential intestinal microorganisms were norank-f-Bacteroidales-s24-7, Bacteroides, Blautia, unclassified-Lachnospiraeae, Ruminiclostridium,Lachnospiraeae-NK4A136, and Roseburia among all groups of mice. Compared with the control group, the proportion of norank-f-Bacteroidales-s24-7 in the aging group was decreased significantly, and the proportions of the other strains were increased significantly(P < 0.05). Compared with the aging group, the proportions of norank-f-Bacteroidales-s24-7,Roseburia, Lachnospiraeae-NK4 A136 and Bacteroides were significantly increased in the mice treated with procyanidins B_2,while the proportion of Blautia, unclassified-Lachnospiraeae, and Ruminiclostridium was significantly decreased(P < 0.05).Accordingly, procyanidins B_2 could regulate the contents of SCFAs in aging mice to improve the intestinal environment,thereby delaying the aging process.
引文
[1]CEVENINI E,CARUSO C,CCANDORE G.Age-related inflammation:the contribution of different organs,tissues and systems.how to face it for therapeutic approaches[J].Current Pharmaceutical Design,2010,16(6):609-618.DOI:10.2174/138161210790883840.
[2]BIAGI E,CANDELA M,FAIRWEATHER-TAIT S,et al.Ageing of the human metaorganism:the microbial counterpart[J].Age,2012,34(1):247-267.DOI:10.1007/s11357-011-9217.
[3]DAVINELLI S,WILLCOX D C,SCAPAGNINI G.Extending healthy ageing:nutrient sensitive pathway and centenarian population[J].Immunity&Ageing,2012,9:1-9.DOI:10.1186/1742-4933-9-9.
[4]BIAGI E,NYLUND L,CANDELA M,et al.Correction:through ageing,and beyond:gut microbiota and inflammatory status in seniors and centenarians[J].PLoS ONE,2010,5(5):e10667.DOI:10.1371/journal.pone.0010667.
[5]CHASSARD C,LACROIX C.Carbohydrates and the human gut microbiota[J].Current Opinion Clinical Nutrtion and Metabolic Care,2013,16(4):453-460.DOI:10.1097/MCO.0b013e3283619e63.
[6]SCHIFFRIN E J,MORLEY J E,DONNET-HUGHES A,et al.The inflammatory status of the elderly:the intestinal contribution[J].Mutation Research-Fundamental and Molecular Mechanisms of Mutagenesis,2010,690(1/2):50-56.DOI:10.1016/j.mrfmmm.2009.07.011.
[7]ACOSTAESTRAD A B A,GUTIéRREZURIBE J A,SERNASALDíVAR S O.Bound phenolics in foods,a review[J].Food Chemistry,2014,152:46-55.DOI:10.1016/j.foodchem.2013.11.093.
[8]SIRK T W,FRIEDMAN M,BROWN E F.Molecular binding of black tea theaflavins to biological membranes:relationship to bioactivities[J].Journal of Agricultural Food Chemistry,2011,59(8):3780-3787.DOI:10.1021/jf2006547.
[9]CARDONA F,ANDRéS-LACUEVA C,TULIPANI S,et al.Benefits of polyphenols on gut microbiota and implications in human health[J].Journal of Nutritional Biochemistry,2013,24(8):1415-1422.DOI:10.1016/j.jnutbio.2013.05.001.
[10]崔福顺,张华,李官浩,等.美味牛肝菌黄酮类提取物体内抗氧化作用研究[J].食品科技,2014,39(8):201-205.DOI:10.13684/j.cnki.spkj.2014.08.044.
[11]吴美媛,余甜女,王喜周.猴头菇多糖对D-半乳糖小鼠体内抗氧化作用的研究[J].食品研究与开发,2016,37(10):55-57.DOI:10.3969/j.issn.1005-6521.2016.10.014.
[12]FORMAN H J.Redox signaling:an evolution from free radicals to aging[J].Free Radical Biology and Medicine,2016,97:398-407.DOI:10.1016/j.freeradbiomed.2016.07.003.
[13]MATSUMOTO M,BENNO Y.Anti-inflammatory metabolite production in the gut from the consumption of probiotic yogurt containing Bifidobacterium animalis subsp.lactis LKM512[J].Bioscience Biotechnology and Biochemistry,2006,70(6):1287-1292.DOI:10.1271/bbb.50464.
[14]HIDENORI H,MITSUO S,MAKI K,et al.Molecular analysis of fecal microbiota in elderly individuals using 16S rDNA library and T-RFLP[J].Microbiology and Immunology,2003,47(8):557-570.DOI:10.1111/j.1348-0421.
[15]M?KIVUOKKO H,TIIHONEN K,TYNKKYNEN S,et al.The effect of age and non-steroidal anti-inflammatory drugs on human intestinal microbiota composition[J].British Journal of Nutrition,2010,103(2):227-234.DOI:10.1017/S0007114509991553.
[16]CLAESSON M J,CUSACK S,O’SULLIVAN O,et al.Composition,variability,and temporal stability of the intestinal microbiota of the elderly[J].Proceedings of the National Academy of Sciences of the United States of America,2011,108(Suppl 1):4586-4591.DOI:10.1073/pnas.1000097107.
[17]BEZIRTZOGLOU E,STAVROPOULOU E.Immunology and probiotic impact of the newborn and young children intestinal microflora[J].Anaerobe,2011,17(6):369-374.DOI:10.1016/j.anaerobe.2011.03.010.
[18]CAPORASO J G,LAUBER C L,COSTELLO E K,et al.Moving pictures of the human microbiome[J].Genome Biology,2011,12(5):1-8.DOI:10.1186/gb-2011-12-5-r50.
[19]GOULET O.Potential role of the intestinal microbiota in programming health and disease[J].Nutrition Reviews,2015,73(Suppl 1):32-40.DOI:10.1093/nutrit/nuv039.
[20]LEY R E.Obesity and the human microbiome[J].Current Opinion in Gastroenterology,2010,26(1):5-11.DOI:10.1097/MOG.0b013e328333d751.
[21]LEY R E,TURNBAUGH P J,KLEIN S,et al.Microbial ecology:human gut microbes associated with obesity[J].Nature,2006,444:1022-1023.DOI:10.1038/4441022a.
[22]GREENBLUM S,TURNBAUGH P J,BORENSTEIN E.Metagenomic systems biology of the human gut microbiome reveals topological shifts associated with obesity and inflammatory bowel disease[J].Proceedings of the National Academy of Sciences,2011,109(2):594-599.DOI:10.1073/pnas.1116053109.
[23]MARIAT D,FIRMESSE O,LEVENEZ F,et al.The Firmicutes/Bacteroidetes ratio of the human microbiota changes with age[J].BMC Microbiology,2009,9:1-11.DOI:10.1186/1471-2180-9-123.
[24]YUE S Y,CHINNAPANDI B,GE H,et al.A Lactobacillus cocktail changes gut flora and reduces cholesterolemia and weight gain in hyperlipidemia mice[J].Symbiosis,2014,2(2):1-14.
[25]王芳.广西巴马长寿老人肠道菌群及其与膳食纤维多糖饮食关联性研究[D].南宁:广西大学,2015:53-57.
[26]ZHAO Liang,XU Wentao,IBRAHIM S A,et al.Effects of age and region on fecal microflora in elderly subjects living in Bama,Guangxi,China[J].Current Microbiology,2011,62(1):64-70.DOI:10.1007/s00284-010-9676-4.
[27]WANG T T,CAI G X,QIU Y P,et al.Structural segregation of gut microbiota between colorectal cancer patients and healthy volunteers[J].ISME Journal,2012,6(2):320-329.DOI:10.1038/ismej.2011.109.
[28]ZWIELEHNER J,LISZT K,HANDSCHUR M,et al.Combined PCR-DGGE fingerprinting and quantitative-PCR indicates shifts in fecal population sizes and diversity of Bacteroides,Bifidobacteria and Clostridium cluster IV in institutionalized elderly[J].Experimental Gerontology,2009,44(6/7):440-446.DOI:10.1016/j.exger.2009.04.002.
[29]DHARMARAJAN T S,SOHAGIA A,PITCHUMONI C S,et al.The gastrointestinal system and aging[M].New York:Springer,2012:33-47.DOI:10.1007/978-1-4419-1623-5_5.
[30]SALONEN A,LAHTI L,SALOJ?RVI J,et al.Impact of diet and individual variation on intestinal microbiota composition and fermentation products in obese men[J].ISME Journal,2014,8(11):2218-2230.DOI:10.1038/ismej.2014.63.
[31]BEEK C M V D,LENAERTS K,AVESAAT M V,et al.670Expression of the short-chain fatty acid receptors GPR41 and GPR43throughout the human ileum and colon[J].Gastroenterology,2015,148(4):S128-S129.DOI:10.1016/S0016-5085(15)30445-5.
[32]UNGER M M,SPIEGEL J,DILLMANN K U,et al.Short chain fatty acids and gut microbiota differ between patients with Parkinson’s disease and age-matched controls[J].Parkinsonism&Related Disorders,2016,32:66-72.DOI:10.1016/j.parkreldis.2016.08.019.
[33]FULOP T,LARBI A,DUPUIS G,et al.Immunosenescence and inflamm-aging as two sides of the same coin:friends or foes?[J].Frontiers in Immunology,2017,8:1960-1090.DOI:10.3389/fimmu.2017.01960.
[2]BIAGI E,CANDELA M,FAIRWEATHER-TAIT S,et al.Ageing of the human metaorganism:the microbial counterpart[J].Age,2012,34(1):247-267.DOI:10.1007/s11357-011-9217.
[3]DAVINELLI S,WILLCOX D C,SCAPAGNINI G.Extending healthy ageing:nutrient sensitive pathway and centenarian population[J].Immunity&Ageing,2012,9:1-9.DOI:10.1186/1742-4933-9-9.
[4]BIAGI E,NYLUND L,CANDELA M,et al.Correction:through ageing,and beyond:gut microbiota and inflammatory status in seniors and centenarians[J].PLoS ONE,2010,5(5):e10667.DOI:10.1371/journal.pone.0010667.
[5]CHASSARD C,LACROIX C.Carbohydrates and the human gut microbiota[J].Current Opinion Clinical Nutrtion and Metabolic Care,2013,16(4):453-460.DOI:10.1097/MCO.0b013e3283619e63.
[6]SCHIFFRIN E J,MORLEY J E,DONNET-HUGHES A,et al.The inflammatory status of the elderly:the intestinal contribution[J].Mutation Research-Fundamental and Molecular Mechanisms of Mutagenesis,2010,690(1/2):50-56.DOI:10.1016/j.mrfmmm.2009.07.011.
[7]ACOSTAESTRAD A B A,GUTIéRREZURIBE J A,SERNASALDíVAR S O.Bound phenolics in foods,a review[J].Food Chemistry,2014,152:46-55.DOI:10.1016/j.foodchem.2013.11.093.
[8]SIRK T W,FRIEDMAN M,BROWN E F.Molecular binding of black tea theaflavins to biological membranes:relationship to bioactivities[J].Journal of Agricultural Food Chemistry,2011,59(8):3780-3787.DOI:10.1021/jf2006547.
[9]CARDONA F,ANDRéS-LACUEVA C,TULIPANI S,et al.Benefits of polyphenols on gut microbiota and implications in human health[J].Journal of Nutritional Biochemistry,2013,24(8):1415-1422.DOI:10.1016/j.jnutbio.2013.05.001.
[10]崔福顺,张华,李官浩,等.美味牛肝菌黄酮类提取物体内抗氧化作用研究[J].食品科技,2014,39(8):201-205.DOI:10.13684/j.cnki.spkj.2014.08.044.
[11]吴美媛,余甜女,王喜周.猴头菇多糖对D-半乳糖小鼠体内抗氧化作用的研究[J].食品研究与开发,2016,37(10):55-57.DOI:10.3969/j.issn.1005-6521.2016.10.014.
[12]FORMAN H J.Redox signaling:an evolution from free radicals to aging[J].Free Radical Biology and Medicine,2016,97:398-407.DOI:10.1016/j.freeradbiomed.2016.07.003.
[13]MATSUMOTO M,BENNO Y.Anti-inflammatory metabolite production in the gut from the consumption of probiotic yogurt containing Bifidobacterium animalis subsp.lactis LKM512[J].Bioscience Biotechnology and Biochemistry,2006,70(6):1287-1292.DOI:10.1271/bbb.50464.
[14]HIDENORI H,MITSUO S,MAKI K,et al.Molecular analysis of fecal microbiota in elderly individuals using 16S rDNA library and T-RFLP[J].Microbiology and Immunology,2003,47(8):557-570.DOI:10.1111/j.1348-0421.
[15]M?KIVUOKKO H,TIIHONEN K,TYNKKYNEN S,et al.The effect of age and non-steroidal anti-inflammatory drugs on human intestinal microbiota composition[J].British Journal of Nutrition,2010,103(2):227-234.DOI:10.1017/S0007114509991553.
[16]CLAESSON M J,CUSACK S,O’SULLIVAN O,et al.Composition,variability,and temporal stability of the intestinal microbiota of the elderly[J].Proceedings of the National Academy of Sciences of the United States of America,2011,108(Suppl 1):4586-4591.DOI:10.1073/pnas.1000097107.
[17]BEZIRTZOGLOU E,STAVROPOULOU E.Immunology and probiotic impact of the newborn and young children intestinal microflora[J].Anaerobe,2011,17(6):369-374.DOI:10.1016/j.anaerobe.2011.03.010.
[18]CAPORASO J G,LAUBER C L,COSTELLO E K,et al.Moving pictures of the human microbiome[J].Genome Biology,2011,12(5):1-8.DOI:10.1186/gb-2011-12-5-r50.
[19]GOULET O.Potential role of the intestinal microbiota in programming health and disease[J].Nutrition Reviews,2015,73(Suppl 1):32-40.DOI:10.1093/nutrit/nuv039.
[20]LEY R E.Obesity and the human microbiome[J].Current Opinion in Gastroenterology,2010,26(1):5-11.DOI:10.1097/MOG.0b013e328333d751.
[21]LEY R E,TURNBAUGH P J,KLEIN S,et al.Microbial ecology:human gut microbes associated with obesity[J].Nature,2006,444:1022-1023.DOI:10.1038/4441022a.
[22]GREENBLUM S,TURNBAUGH P J,BORENSTEIN E.Metagenomic systems biology of the human gut microbiome reveals topological shifts associated with obesity and inflammatory bowel disease[J].Proceedings of the National Academy of Sciences,2011,109(2):594-599.DOI:10.1073/pnas.1116053109.
[23]MARIAT D,FIRMESSE O,LEVENEZ F,et al.The Firmicutes/Bacteroidetes ratio of the human microbiota changes with age[J].BMC Microbiology,2009,9:1-11.DOI:10.1186/1471-2180-9-123.
[24]YUE S Y,CHINNAPANDI B,GE H,et al.A Lactobacillus cocktail changes gut flora and reduces cholesterolemia and weight gain in hyperlipidemia mice[J].Symbiosis,2014,2(2):1-14.
[25]王芳.广西巴马长寿老人肠道菌群及其与膳食纤维多糖饮食关联性研究[D].南宁:广西大学,2015:53-57.
[26]ZHAO Liang,XU Wentao,IBRAHIM S A,et al.Effects of age and region on fecal microflora in elderly subjects living in Bama,Guangxi,China[J].Current Microbiology,2011,62(1):64-70.DOI:10.1007/s00284-010-9676-4.
[27]WANG T T,CAI G X,QIU Y P,et al.Structural segregation of gut microbiota between colorectal cancer patients and healthy volunteers[J].ISME Journal,2012,6(2):320-329.DOI:10.1038/ismej.2011.109.
[28]ZWIELEHNER J,LISZT K,HANDSCHUR M,et al.Combined PCR-DGGE fingerprinting and quantitative-PCR indicates shifts in fecal population sizes and diversity of Bacteroides,Bifidobacteria and Clostridium cluster IV in institutionalized elderly[J].Experimental Gerontology,2009,44(6/7):440-446.DOI:10.1016/j.exger.2009.04.002.
[29]DHARMARAJAN T S,SOHAGIA A,PITCHUMONI C S,et al.The gastrointestinal system and aging[M].New York:Springer,2012:33-47.DOI:10.1007/978-1-4419-1623-5_5.
[30]SALONEN A,LAHTI L,SALOJ?RVI J,et al.Impact of diet and individual variation on intestinal microbiota composition and fermentation products in obese men[J].ISME Journal,2014,8(11):2218-2230.DOI:10.1038/ismej.2014.63.
[31]BEEK C M V D,LENAERTS K,AVESAAT M V,et al.670Expression of the short-chain fatty acid receptors GPR41 and GPR43throughout the human ileum and colon[J].Gastroenterology,2015,148(4):S128-S129.DOI:10.1016/S0016-5085(15)30445-5.
[32]UNGER M M,SPIEGEL J,DILLMANN K U,et al.Short chain fatty acids and gut microbiota differ between patients with Parkinson’s disease and age-matched controls[J].Parkinsonism&Related Disorders,2016,32:66-72.DOI:10.1016/j.parkreldis.2016.08.019.
[33]FULOP T,LARBI A,DUPUIS G,et al.Immunosenescence and inflamm-aging as two sides of the same coin:friends or foes?[J].Frontiers in Immunology,2017,8:1960-1090.DOI:10.3389/fimmu.2017.01960.