蛋氨酸限制对高脂饮食小鼠肠道氧化还原状态、炎症和菌群的影响
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摘要
目的:研究饮食蛋氨酸限制对高脂饮食小鼠肠道氧化还原状态、炎症和菌群的影响。方法:将27只SPF级雄性C57BL/6小鼠随机分为3组,分别为正常饮食组(C:0.86%(质量分数,下同)蛋氨酸、4%猪油)、高脂饮食组(HM:0.86%蛋氨酸、20%猪油)、高脂蛋氨酸限制组(LM:0.17%蛋氨酸、20%猪油),每周测定小鼠体质量,12周实验结束后处死小鼠,并取血液、回肠、盲肠和结肠样品,测定血浆胆固醇(total cholesterol,TC)、甘油三酯(triglyceride,TG)、高密度脂蛋白胆固醇(high-density lipoprotein cholesterol,HDL-C)、低密度脂蛋白胆固醇(low-density lipoprotein cholesterol,LDL-C)、脂多糖(lipopolysaccharide,LPS)和LPS结合蛋白(LPS-binding protein,LBP)的含量;测定回肠和结肠组织中氧化应激相关指标;用实时荧光定量聚合酶链式反应测定回肠炎症基因肿瘤坏死因子-α(tumour necrosis factor-α,TNF-α)、白介素-6(interleukin-6,IL-6)mRNA的表达水平;提取盲肠内容物DNA,用限制性末端酶切的方法分析小鼠盲肠内容物中菌群的变化;提取结肠内容物DNA,用高通量测序分析小鼠结肠内容物中菌群的变化。结果:与HM组相比,LM组小鼠体质量、血浆TC、TG和LDL-C水平显著降低(P<0.05,P<0.01),HDL-C水平极显著增加(P<0.01);回肠总抗氧化能力和还原型谷胱甘肽/氧化型谷胱甘肽(glutathione/oxidizided glutathione,GSH/GSSG)的比值显著增加(P<0.05);结肠GSH/GSSG比值显著增加(P<0.05),丙二醛含量显著降低(P<0.05);血浆LPS和LBP水平显著降低(P<0.05,P<0.01);回肠TNF-α、IL-6 mRNA的表达水平显著下调(P<0.05);盲肠菌群Shannon-Weiner指数与均匀度指数显著上升(P<0.05),结肠菌群中双歧杆菌和颤螺杆菌丰度显著增加(P<0.05)。结论:饮食蛋氨酸限制具有显著改善高脂饮食小鼠肠道组织氧化还原状态、炎症反应和菌群结构的作用。
Purpose: To investigate the effect of dietary methionine restriction(MR) on gut redox status, inflammation and microbiota in high-fat diet-fed mice. Methods: Twenty-seven male C57BL/6 mice were randomly and equally divided into control group(C: 0.86% methionine and 4% lard oil), high-fat diet group(HM: 0.86% methionine and 20% lard oil),high-fat diet with MR group(LM: 0.17% methionine and 20% lard oil). Body mass was recorded weekly. After 12 weeks,the mice were killed to collect blood plasma, cecum, ileum and colon. Plasma total cholesterol(TC), triglyceride(TG),low-density lipoprotein cholesterol(LDL-C), high-density lipoprotein cholesterol(HDL-C), lipopolysaccharide(LPS),and LPS-binding protein(LBP) contents were examined. Some indicators of oxidative stress in ileum and colon were determined. The expression of tumour necrosis factor-α(TNF-α) and interleukin-6(IL-6) mRNA in ileum were determined by polymerase chain reaction(PCR). DNA was extracted from cecal contents for analysis of microbiota changes by terminal restriction fragment length polymorphism. DNA was extracted from colonic contents for analysis of microbiota changes by high throughput sequencing. Results: Compared with HM, LM significantly decreased body mass, plasma TC, TG and LDL-C, and colonic malondialdehyde(MDA) level(P < 0.05, P < 0.01). In contrast, LM significantly increased plasma HDL-C(P < 0.01), total antioxidant capacity(T-AOC) and glutathione/oxidized glutathione(GSH/GSSG) ratio in ileum(P < 0.05), and GSH/GSSG ratio and MDA level in colon(P < 0.05). Similarly, LM significantly decreased the plasma levels of LPS and LBP, the expression of TNF-α and IL-6 mRNA in ileum(P < 0.05), but significantly increased the Shannon-Weiner index and even index of cecal microbiota(P < 0.05) and the relative abundance of Bifidobacterium and Oscillospira in colonic contents(P < 0.05). Conclusion: MR can improve the intestinal redox status, inflammatory response and gut microbiota in high-fat diet-fed mice.
Purpose: To investigate the effect of dietary methionine restriction(MR) on gut redox status, inflammation and microbiota in high-fat diet-fed mice. Methods: Twenty-seven male C57BL/6 mice were randomly and equally divided into control group(C: 0.86% methionine and 4% lard oil), high-fat diet group(HM: 0.86% methionine and 20% lard oil),high-fat diet with MR group(LM: 0.17% methionine and 20% lard oil). Body mass was recorded weekly. After 12 weeks,the mice were killed to collect blood plasma, cecum, ileum and colon. Plasma total cholesterol(TC), triglyceride(TG),low-density lipoprotein cholesterol(LDL-C), high-density lipoprotein cholesterol(HDL-C), lipopolysaccharide(LPS),and LPS-binding protein(LBP) contents were examined. Some indicators of oxidative stress in ileum and colon were determined. The expression of tumour necrosis factor-α(TNF-α) and interleukin-6(IL-6) mRNA in ileum were determined by polymerase chain reaction(PCR). DNA was extracted from cecal contents for analysis of microbiota changes by terminal restriction fragment length polymorphism. DNA was extracted from colonic contents for analysis of microbiota changes by high throughput sequencing. Results: Compared with HM, LM significantly decreased body mass, plasma TC, TG and LDL-C, and colonic malondialdehyde(MDA) level(P < 0.05, P < 0.01). In contrast, LM significantly increased plasma HDL-C(P < 0.01), total antioxidant capacity(T-AOC) and glutathione/oxidized glutathione(GSH/GSSG) ratio in ileum(P < 0.05), and GSH/GSSG ratio and MDA level in colon(P < 0.05). Similarly, LM significantly decreased the plasma levels of LPS and LBP, the expression of TNF-α and IL-6 mRNA in ileum(P < 0.05), but significantly increased the Shannon-Weiner index and even index of cecal microbiota(P < 0.05) and the relative abundance of Bifidobacterium and Oscillospira in colonic contents(P < 0.05). Conclusion: MR can improve the intestinal redox status, inflammatory response and gut microbiota in high-fat diet-fed mice.
引文
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[29]KOZáKOVáH,REHáKOVáZ,KOLáNSKáJ.Bifidobacterium bifidum monoassociation of gnotobiotic mice:effect on enterocyte brush-border enzymes[J].Folia Microbiologica,2001,46(6):573-576.
[30]OKADA Y,TSUZUKI Y,R,KOMOTO S,et al.Anti-inflammatory effects of the genus Bifidobacterium on macrophages by modification of phospho-I kappaB and SOCS gene expression[J].International Journal of Experimental Pathology,2010,90(2):131-140.DOI:10.1111/j.1365-2613.2008.00632.x.
[31]KONIKOFF T,GOPHNA U.Oscillospira:a central,enigmatic component of the human gut microbiota[J].Trends in Microbiology,2016,24(7):523-524.DOI:10.1016/j.tim.2016.02.015.
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[33]DERRIEN M,COLLADO M C,BENAMOR K,et al.The Mucin degrader Akkermansia muciniphila is an abundant resident of the human intestinal tract[J].Applied&Environmental Microbiology,2008,74(5):1646-1648.DOI:10.1128/AEM.01226-07.
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[2]LEIBEL R L,ROSENBAUM M,HIRSCH J.Changes in energy expenditure resulting from altered body weight[J].New England Journal of Medicine,1995,332(10):621-628.DOI:10.1056/NEJM199503093321001.
[3]AKIYAMA T,TACHIBANA I,SHIROHARA H,et al.Highfat hypercaloric diet induces obesity,glucose intolerance and hyperlipidemia in normal adult male Wistar rat[J].Diabetes Research and Clinical Practice,1996,31(1/2/3):27-35.DOI:10.1016/0168-8227(96)01205-3.
[4]SASATOMI Y,TADA M,UESUGI N,et al.Obesity associated with hypertension or hyperlipidemia accelerates renal damage[J].Pathobiology,2001,69(2):113-118.DOI:10.1159/000048764.
[5]冉兴无,李晓松,童南伟,等.中国肥胖人群体脂分布特点及其与心血管危险因素的关系[J].四川大学学报(医学版),2004,35(5):699-703.DOI:10.3760/j:issn:0253-9624.2005.05.006.
[6]BASTARD J P,MAACHI M,LAGATHU C,et al.Recent advances in the relationship between obesity,inflammation,and insulin resistance[J].European Cytokine Network,2006,1(17):4-12.
[7]QIAO Yi,SUN Jin,DING Yinyi,et al.Alterations of the gut microbiota in high-fat diet mice is strongly linked to oxidative stress[J].Applied Microbiology&Biotechnology,2013,97(4):1689-1697.DOI:10.1007/s00253-012-4323-6.
[8]LUTGENDORFF F,AKKERMANS L M,SODERHOLM J D.The role of microbiota and probiotics in stress-induced gastro-intestinal damage[J].Current Molecular Medicine,2008,8(4):282-298.DOI:10.2174/156652408784533779.
[9]CUCCHIARA S,STRONATI L,ALOI M.Interactions between intestinal microbiota and innate immune system in pediatric inflammatory bowel disease[J].Journal of Clinical Gastroenterology,2012,46(9):64-66.DOI:10.1097/MCG.0b013e31826a857f.
[10]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.
[11]B?CKHED F,MANCHESTER J K,SEMENKOVICH C F,et al.Mechanisms underlying the resistance to diet-induced obesity in germfree mice[J].Proceedings of the National Academy of Sciences of the United States of America,2007,104(3):979-984.DOI:10.1073/pnas.0605374104.
[12]TURNBAUGH P J,LEY R E,MAHOWALD M A,et al.An obesityassociated gut microbiome with increased capacity for energy harvest[J].Nature,2006,444:1027-1031.DOI:10.1038/nature05414.
[13]武阳丰,马冠生,胡永华,等.中国居民的超重和肥胖流行现状[J].中华预防医学杂志,2005,39(5):316-320.DOI:10.3760/j:issn:0253-9624.2005.05.006.
[14]ORENTREICH N,MATIAS J R,DEFELICE A,et al.Low methionine ingestion by rats extends life span[J].Journal of Nutrition,1993,123(2):269-274.
[15]ABLES G P,PERRONE C E,ORENTREICH D,et al.Methioninerestricted C57BL/6J mice are resistant to diet-induced obesity and insulin resistance but have low bone density[J].PLoS ONE,2012,7(12):e51357.DOI:10.1371/journal.pone.0051357.
[16]SUN L,SADIGHI AKHA A A,MILLER R A,et al.Life-span extension in mice by preweaning food restriction and by methionine restriction in middle age[J].The Journals of Gerontology,Series A:Biological Sciences and Medical Sciences,2009,64(7):711-722.DOI:10.1093/gerona/glp051.
[17]CAVUOTO P,FENECH M F.A review of methionine dependency and the role of methionine restriction in cancer growth control and lifespan extension[J].Cancer Treatment Reviews,2012,38(6):726-736.DOI:10.1016/j.ctrv.2012.01.004.
[18]SANZ A,CARO P,AYALA V,et al.Methionine restriction decreases mitochondrial oxygen radical generation and leak as well as oxidative damage to mitochondrial DNA and proteins[J].FASEB Journal,2006,20(8):1064-1073.DOI:10.1096/fj.05-5568com.
[19]王雅楠,张佳红,郭海涛,等.蛋氨酸限制和胶原蛋白肽对高脂饮食小鼠脂代谢和氧化应激的联合作用[J].食品科学,2018,39(9):108-115.DOI:10.7506/spkx1002-6630-201809017.
[20]WANG Panpan,LI Ya,XIAO Hang,et al.Isolation of lactobacillus reuteri from Peyer’s patches and their effects on sIgA production and gut microbiota diversity[J].Molecular Nutrition&Food Research,2016,60(9):2020-2030.DOI:10.1002/mnfr.201501065.
[21]LEES E K,KROL E,GRANT L,et al.Methionine restriction restores a younger metabolic phenotype in adult mice with alterations in fibroblast growth factor 21[J].Aging Cell,2014,13(5):817-827.DOI:10.1111/acel.12238.
[22]ELSHORBAGY A K,VALDIVIA-GARCIA M,MATTOCKS D A,et al.Cysteine supplementation reverses methionine restriction effects on rat adiposity:significance of stearoyl-coenzyme A desaturase[J].Journal of Lipid Research,2011,52(1):104-112.DOI:10.1194/jlr.M010215.
[23]BALAMURUGAN G,SHANTHA A.Effect of Erythrina variegata seed extract on hyperlipidemia elicited by high-fat diet in Wistar rats[J].Journal of Pharmacy and Bioallied Sciences,2010,2(4):350-355.DOI:10.4103/0975-7406.72139.
[24]CABREIRO F,AU C,LEUNG K Y,et al.Metformin retards aging in C.elegans by altering microbial folate and methionine metabolism[J].Cell,2013,153(1):228-239.DOI:10.1016/j.cell.2013.02.035.
[25]ZHANG Y,FISCHER K E,SOTO V,et al.Obesity-induced oxidative stress,accelerated functional decline with age and increased mortality in mice[J].Archives of Biochemistry&Biophysics,2015,576:39-48.DOI:10.1016/j.abb.2014.12.018.
[26]DE LA SERRE,ELLIS C L,LEE J,et al.Propensity to high-fat diet-induced obesity in rats is associated with changes in the gut microbiota and gut inflammation[J].American Journal of Physiology Gastrointestinal&Liver Physiology,2010,299(2):G440-G448.DOI:10.1152/ajpgi.00098.2010.
[27]B?CKHED F,DING H,WANG T,et al.The gut microbiota as an environmental factor that regulates fat storage[J].Proceedings of the National Academy of Sciences of the United States of America,2004,101(44):15718-15723.DOI:10.1073/pnas.0407076101.
[28]WU G D,CHEN J,HOFFMANN C,et al.Linking long-term dietary patterns with gut microbial enterotypes[J].Science,2011,334:105-158.DOI:10.1080/19490976.2016.1270809.
[29]KOZáKOVáH,REHáKOVáZ,KOLáNSKáJ.Bifidobacterium bifidum monoassociation of gnotobiotic mice:effect on enterocyte brush-border enzymes[J].Folia Microbiologica,2001,46(6):573-576.
[30]OKADA Y,TSUZUKI Y,R,KOMOTO S,et al.Anti-inflammatory effects of the genus Bifidobacterium on macrophages by modification of phospho-I kappaB and SOCS gene expression[J].International Journal of Experimental Pathology,2010,90(2):131-140.DOI:10.1111/j.1365-2613.2008.00632.x.
[31]KONIKOFF T,GOPHNA U.Oscillospira:a central,enigmatic component of the human gut microbiota[J].Trends in Microbiology,2016,24(7):523-524.DOI:10.1016/j.tim.2016.02.015.
[32]TIMS S,DEROM C,JONKERS D M,et al.Microbiota conservation and BMI signatures in adult monozygotic twins[J].ISME Journal,2013,7(4):707-717.DOI:10.1038/ismej.2012.146.
[33]DERRIEN M,COLLADO M C,BENAMOR K,et al.The Mucin degrader Akkermansia muciniphila is an abundant resident of the human intestinal tract[J].Applied&Environmental Microbiology,2008,74(5):1646-1648.DOI:10.1128/AEM.01226-07.
[34]GANESH B P,KLOPFLEISCH R,LOH G,et al.Commensal Akkermansia muciniphila exacerbates gut inflammation in Salmonella Typhimurium-infected gnotobiotic mice[J].PLoS ONE,2013,8(9):e74963.DOI:10.1371/journal.pone.0074963.
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