基于高通量测序的精神分裂症患者肠道菌群多样性
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摘要
目的探讨精神分裂症患者肠道菌群多样性变化。方法收集16例精神分裂症患者(schizophrenia,Sch)与18例健康者(healthy donor,HD)的粪便样本,提取肠道菌群基因组,扩增16SrRNA基因,运用Illumina平台进行测序。对测序结果进行多样性和物种组成差异分析。结果精神分裂症患者组肠道菌群在门、纲、目、科、属、种、分类操作单元(operational taxonomic units,OTUs)水平的群落种类数目少于健康对照组。Alpha多样性指标中患者组的Ace指数(226.58±31.67)、Chao1指数(222.29±34.45)和Shannon指数(2.66±0.69)明显低于健康对照组(293.63±56.07、302.2±57.99、3.59±0.36),差异均有统计学意义(Ps<0.001),而Simpson指数(0.20±0.17)明显高于健康对照组(0.07±0.03),差异有统计学意义(P<0.01);Beta多样性分析显示两组研究对象肠道菌群样本可被鉴别区分。精神分裂症患者组与健康对照组样本在门和属水平上肠道菌群组成和含量有差异,其中患者组拟杆菌门和软壁菌门占比明显低于健康组,差异有统计学意义(21.76%vs.27.05%,P=0.008;0.00%vs.0.20%,P=0.033),而放线菌门明显高于健康组(13.52%vs.2.88%,P=0.020),差异有统计学意义;患者组拟杆菌属和柔嫩梭菌属占比明显低于健康组(9.15%vs.20.60%,P=0.031;3.29%vs.9.58%,P=0.005),差异有统计学意义,而双歧杆菌属、普雷沃菌属和巨单胞菌属明显高于健康组(10.89%vs.1.78%,P=0.025;10.88%vs.1.98%,P=0.046;10.78%vs.2.69%,P=0.026),差异有统计学意义。结论基于16SrRNA的高通量测序有助于分析精神分裂症患者肠道菌群多样性变化,为研究肠道菌群与精神分裂症的关系提供新的思路和理论依据。
Objective To explore the changes of gut microbiota in patients with schizophrenia.Methods Fresh fecal samples were collected from schizophrenia patients(n=16)and healthy controls(n=18)for DNA extraction,16 SrRNA amplification and Illumina sequencing.Analyses of diversity and species differences between two groups were based on the sequencing data.Results Compared to the healthy controls,the gut microbiota species in schizophrenia patients decreased at the levels of phylum,class,order,family,genus,species and operational taxonomic units(OTUs).The Ace estimator,Chao1 estimator and Shannon index of alpha diversity were significantly lower in schizophrenia patients(226.58±31.67,222.29±34.45,2.66±0.69)compared with the healthy controls(293.63±56.07,302.2±57.99,3.59±0.36,Ps<0.001),while the Simpson index was significantly higher(0.20±0.17 vs 0.07±0.03,P<0.01).Analyses ofβ-diversity was able to differentiate the schizophrenia patients from the healthy controls based on their gut microbiota.The compositions and proportions of gut microbiota in the schizophrenia group were different from those in controls at the levels of phylum and genus.At phylum level,Bacteroidetes and Tenericutes significantly reduced in schizophrenia patients compared to healthy controls(21.76%vs.27.05%,P=0.008;0.00%vs.0.20%,P=0.033),while Actinobacteria increased in schizophrenia patients compared to healthy controls(13.52%vs.2.88%,P=0.020).The genus Bacteroides and genus Faecalibacteriumsignificantly decreased in schizophrenia patients compared with the healthy controls(9.15% vs.20.60%,P=0.031;3.29%vs.9.58%,P=0.005),whereas genus Bifidobacterium,genus Prevotella-9and genus Megamonas significantly increased in schizophrenia patients compared with the healthy controls(10.89% vs.1.78%,P=0.025;10.88% vs.1.98%,P=0.046;10.78% vs.2.69%,P=0.026).Conclusion High-throughput sequencing of 16 SRNA is conducive to analysis of gut microbiota diversity,which can provide new ideas and theoretical foundations for studies on the relationship between gut microbiota and schizophrenia.
Objective To explore the changes of gut microbiota in patients with schizophrenia.Methods Fresh fecal samples were collected from schizophrenia patients(n=16)and healthy controls(n=18)for DNA extraction,16 SrRNA amplification and Illumina sequencing.Analyses of diversity and species differences between two groups were based on the sequencing data.Results Compared to the healthy controls,the gut microbiota species in schizophrenia patients decreased at the levels of phylum,class,order,family,genus,species and operational taxonomic units(OTUs).The Ace estimator,Chao1 estimator and Shannon index of alpha diversity were significantly lower in schizophrenia patients(226.58±31.67,222.29±34.45,2.66±0.69)compared with the healthy controls(293.63±56.07,302.2±57.99,3.59±0.36,Ps<0.001),while the Simpson index was significantly higher(0.20±0.17 vs 0.07±0.03,P<0.01).Analyses ofβ-diversity was able to differentiate the schizophrenia patients from the healthy controls based on their gut microbiota.The compositions and proportions of gut microbiota in the schizophrenia group were different from those in controls at the levels of phylum and genus.At phylum level,Bacteroidetes and Tenericutes significantly reduced in schizophrenia patients compared to healthy controls(21.76%vs.27.05%,P=0.008;0.00%vs.0.20%,P=0.033),while Actinobacteria increased in schizophrenia patients compared to healthy controls(13.52%vs.2.88%,P=0.020).The genus Bacteroides and genus Faecalibacteriumsignificantly decreased in schizophrenia patients compared with the healthy controls(9.15% vs.20.60%,P=0.031;3.29%vs.9.58%,P=0.005),whereas genus Bifidobacterium,genus Prevotella-9and genus Megamonas significantly increased in schizophrenia patients compared with the healthy controls(10.89% vs.1.78%,P=0.025;10.88% vs.1.98%,P=0.046;10.78% vs.2.69%,P=0.026).Conclusion High-throughput sequencing of 16 SRNA is conducive to analysis of gut microbiota diversity,which can provide new ideas and theoretical foundations for studies on the relationship between gut microbiota and schizophrenia.
引文
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[13] Song M,Chan AT.Diet,gut microbiota,and colorectal cancer prevention:A review of potential mechanisms and promising targets for future research[J].Curr Colorectal Cancer Rep,2017,13(6):429-439.
[14] Jiang H,Ling Z,Zhang Y,et al.Altered fecal microbiota composition in patients with major depressive disorder[J].Brain Behav Immun,2015,48:186-194.
[15] Miquel S,Leclerc M,Martin R,et al.Identification of metabolic signatures linked to anti-inflammatory effects of Faecalibacterium prausnitzii[J].MBio,2015,6(2):pii:e00300-15.
[16] Castro-Nallar E,Bendall ML,Pérez-Losada M,et al.Composition,taxonomy and functional diversity of the oropharynx microbiome in individuals with schizophrenia and controls[J].Peer J,2015,3:e1140.
[17] Kawahara T,Takahashi T,Oishi K,et al.Consecutive oral administration of Bifidobacterium longum MM-2improves the defense system against influenza virus infection by enhancing natural killer cell activity in a murine model[J].Microbiol Immunol,2015,59(1):1-12.
[18] Mokrozub VV,Lazarenko LM,Sichel LM,et al.The role of beneficial bacteria wall elasticity in regulating innate immune response[J].EPMA J,2015,6(1):13.
[19] PAN Junxi,XIE Peng.Status and prospects of neuropsychological microbiome[J].Chin Bull Life Sci,2017,29(7):669-681.(in Chinese)潘俊希,谢鹏.神经精神疾病微生物组研究现状和展望[J].生命科学,2017,29(7):669-681.
[20]刘萍,罗本燕.肠道微生态与中枢神经系统疾病的相关性[J].中国神经精神疾病杂志,2016,42(4):251-254.
[21] Berding K,Donovan SM.Microbiome and nutrition in autism spectrum disorder:Current knowledge and research needs[J].Nutr Rev,2016,74(12):723-736.
[22] Evans SJ,Bassis CM,Hein R,et al.The gut microbiome composition associates with bipolar disorder and illness severity[J].J Psychiatr Res,2017,87:23-29.
[23] Zheng P,Zeng B,Zhou C,et al.Gut microbiome remodeling induces depressive-like behaviors through a pathway mediated by the host′s metabolism[J].Mol Psychiatry,2016,21(6):786-796.
[24] Jiang H,Ling Z,Zhang Y,et al.Altered fecal microbiota composition in patients with major depressive disorder[J].Brain Behav Immun,2015,48:186-194.
[25] Schnorr SL,Bachner HA.Integrative therapies in anxiety treatment with special emphasis on the gut microbiome[J].Yale J Biol Med,2016,89(3):397-422.
[26] LI Yongsheng,LU Yanjin,CHEN Jiaqiang,et al.Changes of intestinal floras in elderly patients with first-episode untreated schizophrenia[J].Int J Lab Med,2017,38(6):808-809.(in Chinese)李永生,卢燕金,陈家强,等.老年首发未经治疗精神分裂症患者肠道菌群的变化[J].国际检验医学杂志,2017,38(6):808-809.
[27] SHEN Bianhong,TAO Yunhai,ZHU Chunyan.The role of intestinal flora in the pathogenesis of schizophrenia and its relationship with inflammatory factors[J].Chin J General Pract,2018,16(2):276-278.(in Chinese)申变红,陶云海,朱春燕.肠道菌群比例在精神分裂症发病中的作用及其与炎症因子的关系[J].中华全科医学,2018,16(2):276-278.
[28] Yuan XX,Zhang PF,Wang YP,et al.Changes in metabolism and microbiota after 24-week risperidone treatment in drug na6ve,normal weight patients with first episode schizophrenia[J].Schizophr Res,2018,Pii:S0920-9964(18)30274-3.
[29] ZHU Yanyan,WEN Jianxun,REN Xiaomeng,et al.The distribution characteristics of intestinal flora in patients with gastric cancer[J].Chin J Microecol,2017,29(6):655-658.(in Chinese)朱燕燕,温建勋,任晓萌,等.胃癌患者肠道菌群的分布特点分析[J].中国微生态学杂志,2017,29(6):655-658.
[30] Shen Y,Xu J,Li Z,et al.Analysis of gut microbiota diversity and auxiliary diagnosis as a biomarker in patients with schizophrenia:A cross-sectional study[J].Schizophr Res,2018,pii:S0920-9964(18)30002-1.
[2] SUN Zhenxiao,YU Xiangfen.Research progress of mortality and risk factors in first-episode schizophrenia[J].Chin J Clin(Electronic),2015,9(2):309-312.(in Chinese)孙振晓,于相芬.首发精神分裂症患者的死亡率及其危险因素研究进展[J].中华临床医师杂志(电子版),2015,9(2):309-312.
[3]张馨月,姚晶晶,吕一丁,等.精神分裂症的发病机制及治疗靶点的研究进展[J].国际精神病学杂志,2018,45(2):201-204.
[4] Romijn JA,Corssmit EP,Havekes LM,et al.Gut-brain axis[J].Curr Opin Clin Nutr Metab Care,2008,11(4):518-521.
[5] LIU Dandan,LIU Yinhui,TANG Li.Gut microbiota and neuropsychiatric disorders[J].Chin J Microecol,2017,29(7):850-854.(in Chinese)刘丹丹,刘银辉,唐立.肠道菌群与神经精神疾病[J].中国微生态学杂志,2017,29(7):850-854.
[6] Caso JR,Balanzamartinez V,Palomo T,et al.The microbiota and gut-brain axis:Contributions to the immunopathogenesis of schizophrenia[J].Curr Pharm Des,2016,22(40):6122-6133.
[7] WANG Biao,ZHAO Heping,GAO Wenjie,et al.Diversity analysis of intestinal microbial flora in osteoporosis patients[J].Chin J Osteoporosis,2017,23(6):715-718.(in Chinese)王飙,赵和平,高文杰,等.骨质疏松症患者肠道菌群多样性分析[J].中国骨质疏松杂志,2017,23(6):715-718.
[8] Magoˇc T,Salzberg SL.Flash:Fast length adjustment of short reads to improve genome assemblies[J].Bioinformation,2011,27(21):2957-2963.
[9] Edgar RC.UPARSE:Highly accurate OTU sequences from microbial amplicon reads[J].Nat Methods,2013,10(10):996-998.
[10] Quast C,Pruesse E,Yilmaz P,et al.The SILVA ribosomal RNA gene database project:Improved data processing and web-based tools[J].Nucleic Acids Res,2013,41(D1):D590-596.
[11] Jung SP,Kang H.Assessment of microbial diversity bias associated with soil heterogeneity and sequencing resolution in pyrosequencing analyses[J].J Microbiol,2014,52(7):574-580.
[12] Lozupone CA,Hamady M,Kelley ST,et al.Quantitative and qualitative beta diversity measures lead to different insights into factors that structure microbial communities[J].Appl Environ Microbiol,2007,73(5):1576-1585.
[13] Song M,Chan AT.Diet,gut microbiota,and colorectal cancer prevention:A review of potential mechanisms and promising targets for future research[J].Curr Colorectal Cancer Rep,2017,13(6):429-439.
[14] Jiang H,Ling Z,Zhang Y,et al.Altered fecal microbiota composition in patients with major depressive disorder[J].Brain Behav Immun,2015,48:186-194.
[15] Miquel S,Leclerc M,Martin R,et al.Identification of metabolic signatures linked to anti-inflammatory effects of Faecalibacterium prausnitzii[J].MBio,2015,6(2):pii:e00300-15.
[16] Castro-Nallar E,Bendall ML,Pérez-Losada M,et al.Composition,taxonomy and functional diversity of the oropharynx microbiome in individuals with schizophrenia and controls[J].Peer J,2015,3:e1140.
[17] Kawahara T,Takahashi T,Oishi K,et al.Consecutive oral administration of Bifidobacterium longum MM-2improves the defense system against influenza virus infection by enhancing natural killer cell activity in a murine model[J].Microbiol Immunol,2015,59(1):1-12.
[18] Mokrozub VV,Lazarenko LM,Sichel LM,et al.The role of beneficial bacteria wall elasticity in regulating innate immune response[J].EPMA J,2015,6(1):13.
[19] PAN Junxi,XIE Peng.Status and prospects of neuropsychological microbiome[J].Chin Bull Life Sci,2017,29(7):669-681.(in Chinese)潘俊希,谢鹏.神经精神疾病微生物组研究现状和展望[J].生命科学,2017,29(7):669-681.
[20]刘萍,罗本燕.肠道微生态与中枢神经系统疾病的相关性[J].中国神经精神疾病杂志,2016,42(4):251-254.
[21] Berding K,Donovan SM.Microbiome and nutrition in autism spectrum disorder:Current knowledge and research needs[J].Nutr Rev,2016,74(12):723-736.
[22] Evans SJ,Bassis CM,Hein R,et al.The gut microbiome composition associates with bipolar disorder and illness severity[J].J Psychiatr Res,2017,87:23-29.
[23] Zheng P,Zeng B,Zhou C,et al.Gut microbiome remodeling induces depressive-like behaviors through a pathway mediated by the host′s metabolism[J].Mol Psychiatry,2016,21(6):786-796.
[24] Jiang H,Ling Z,Zhang Y,et al.Altered fecal microbiota composition in patients with major depressive disorder[J].Brain Behav Immun,2015,48:186-194.
[25] Schnorr SL,Bachner HA.Integrative therapies in anxiety treatment with special emphasis on the gut microbiome[J].Yale J Biol Med,2016,89(3):397-422.
[26] LI Yongsheng,LU Yanjin,CHEN Jiaqiang,et al.Changes of intestinal floras in elderly patients with first-episode untreated schizophrenia[J].Int J Lab Med,2017,38(6):808-809.(in Chinese)李永生,卢燕金,陈家强,等.老年首发未经治疗精神分裂症患者肠道菌群的变化[J].国际检验医学杂志,2017,38(6):808-809.
[27] SHEN Bianhong,TAO Yunhai,ZHU Chunyan.The role of intestinal flora in the pathogenesis of schizophrenia and its relationship with inflammatory factors[J].Chin J General Pract,2018,16(2):276-278.(in Chinese)申变红,陶云海,朱春燕.肠道菌群比例在精神分裂症发病中的作用及其与炎症因子的关系[J].中华全科医学,2018,16(2):276-278.
[28] Yuan XX,Zhang PF,Wang YP,et al.Changes in metabolism and microbiota after 24-week risperidone treatment in drug na6ve,normal weight patients with first episode schizophrenia[J].Schizophr Res,2018,Pii:S0920-9964(18)30274-3.
[29] ZHU Yanyan,WEN Jianxun,REN Xiaomeng,et al.The distribution characteristics of intestinal flora in patients with gastric cancer[J].Chin J Microecol,2017,29(6):655-658.(in Chinese)朱燕燕,温建勋,任晓萌,等.胃癌患者肠道菌群的分布特点分析[J].中国微生态学杂志,2017,29(6):655-658.
[30] Shen Y,Xu J,Li Z,et al.Analysis of gut microbiota diversity and auxiliary diagnosis as a biomarker in patients with schizophrenia:A cross-sectional study[J].Schizophr Res,2018,pii:S0920-9964(18)30002-1.