Polymorphism of Gut Microbiota in High Altitude Tibetan Patients with Coronary Artery Heart Disease
-
摘要:
目的 分析青藏高原藏族冠心病(coronary artery heart disease, CHD)患者肠道菌群的分布和组成特征。 方法 2018年9月至2020年9月, 连续招募世居青藏高原(海拔3600~4500 m)的藏族CHD患者[高海拔藏族CHD患者(high altitude Tibetan CHD, HTC)]和藏族健康人群[高海拔藏族健康人群(high altitude Tibetan normal, HTN)], 长期居住于西宁(海拔2260 m)的汉族CHD患者[中海拔汉族CHD患者(middle altitude Han CHD, MHC)]和武汉(海拔13 m)汉族CHD患者[低海拔汉族CHD患者(low altitude Han CHD, LHC)]。其中HTC与MHC均来自青海大学附属医院心内科住院患者, HTN均来自青海大学附属医院体检中心, LHC均来自华中科技大学附属协和医院心内科住院患者。收集研究对象粪便组织标本, 对肠道菌群16S rRNA V3~V4区进行DNA测序并进行生物信息学分析。 结果 共入选符合纳入和排除标准的CHD患者36例(HTC 8例、MHC 14例、LHC 14例), HTN 34例。α多样性分析显示, HTC与HTN的肠道菌群Shannon指数无显著性差异(P=0.091);HTC的肠道菌群Shannon指数最高, MHC次之, LHC最低(P=0.025)。β多样性分析显示, HTC的肠道菌群分布与HTN、MHC、LHC均存在显著差异。在对肠道菌群组成成分的分析中, HTC在门水平、属水平亦显示出不同于MHC、LHC的特征, 其致病菌如链球菌(Streptococcus)、埃希氏菌_志贺氏菌(Escherichia_Shigella)和克雷伯氏菌(Klebsiella)的相对丰度下降, 有益菌如粪杆菌(Faecalibacterium)、普氏菌(Prevotella)、链型杆菌(Catenibacterium)和乳酸杆菌(Lactobacillus)的相对丰度升高。 结论 高海拔藏族CHD患者肠道菌群呈现出不同于同海拔藏族健康人群以及中、低海拔CHD患者的多态性。 Abstract:Objective To analyze the distribution and composition of intestinal flora in Tibetan patients with coronary artery heart disease (CHD) on the Qinghai-Tibet Plateau. Methods From September 2018 to September 2020, following patients were recruited: Tibetan CHD patients living in the area of Qinghai-TibetPlateau [altitude 3600-4500 m, Tibetan patients at high altitudes with CHD (HTC)], healthy people [(normal Tibetans at high altitudes(HTN)], Han patients with CHD living in Xining [altitude 2260 m, Han CHD at a middle altitude (MHC)] and Wuhan [altitude 13 m, Han CHD at a low altitude(LHC)], for a long time. Among them, HTC and MHC were all from inpatients of the Department of Cardiology, Affiliated Hospital of Qinghai University. HTN were all from the Physical Examination Center of the Affiliated Hospital of Qinghai University, and LHC were all from inpatients of the Department of Cardiology, the Union Hospital Affiliated to Huazhong University of Science and Technology. The fecal samples were collected, and the 16S rRNA V3-V4 regions of the intestinal flora were DNA sequenced and bioinformatic analysis was performed. Results A total of 36 CHD patients (8 HTC, 14 MHC, 14 LHC) and 34 HTN patients that met the inclusion and exclusion criteria were enrolled. α-diversity analysis showed that there was no significant difference in the Shannon index of intestinal flora between HTC and HTN (P=0.091), the Shannon index of intestinal flora in HTC was the highest, followed by MHC, and the lowest in LHC(P=0.025). β-diversity analysis showed that the intestinal flora distribution of HTC was significantly different from that of HTN, MHC and LHC. In the analysis of the composition of intestinal flora, HTC also showed different characteristics from MHC and LHC at the phylum level and genus water. The relative abundance of its pathogenic bacteria, i.e. Streptococcus, Escherichia_Shigella and Klebsiella decreased; the beneficial bacteria, i.e. Faecalibacterium, Prevotella, Catenibacterium and Lactobacillus, were increased in relative abundance. Conclusions The intestinal flora of high-altitude Tibetan patients with CHD showed polymorphisms that were different from those of healthy Tibetans at the same altitude and CHD patients at medium and low altitudes. -
Key words:
- coronary artery heart disease /
- gut microbiota /
- high altitude /
- Tibetan
作者贡献:高中山、任明、马玉兰进行研究构思与设计、可行性分析以及论文撰写与修订;高中山、朱露露进行标本、文献资料收集;马玉兰负责论文的质量控制及审校。利益声明:所有作者均声明不存在利益冲突 -
图 1 高海拔藏族冠心病患者和高海拔藏族健康人群肠道菌群多样性比较
A.稀疏曲线(Shannon指数);B. α多样性(Shannon指数);C. β多样性(主坐标分析);HTN、HTC: 同表 1;PCoA: 主坐标分析
图 3 不同海拔、民族冠心病患者肠道菌群在门水平上的组成差异
A.肠道菌群优势菌门分布;B.Firmicutes和Bacteroidetes相对丰度比值;C~E.门水平肠道菌群组成差异;HTC、MHC、LHC: 同表 1;F/B: Firmicutes/Bacteroidetes
图 4 不同海拔、民族冠心病患者肠道菌群属水平分布和组成差异
A. 肠道菌群分布的LEfSe图;B.物种丰度热图示肠道菌群在属水平上的组成;HTC、MHC、LHC:同表 1
表 1 患者一般临床资料比较
指标 HTN(n=34) HTC(n=8) MHC(n=14) LHC(n=14) P值 年龄(x±s,岁) 52.4±6.5 57.6±9.7 58.4±12.1 54.9±8.3 0.130 男性[n(%)] 23(67.6) 5(62.5) 10(62.5) 10(71.4) 0.968 吸烟[n(%)] 8(23.5) 1(12.5) 8(57.1) 5(35.7) 0.084 体质量指数(x±s,kg/m2) 23.7±4.5 24.4±1.9 23.9±1.2 25.1±2.1 0.606 收缩压(x±s,mm Hg) 119.2±4.4 121.0±12.1 121.2±6.3 124.9±14.2 0.219 舒张压(x±s,mm Hg) 76.5±6.3 75.1±5.8 78.6±5.4 75.6±7.8 0.540 总胆固醇(x±s,mmol/L) 4.23±1.04 3.89±0.88 3.34±0.49* 3.30±0.89* 0.003 甘油三酯(x±s,mmol/L) 1.52±0.73 1.92±0.66 1.58±0.57 1.54±0.68 0.510 高密度脂蛋白胆固醇(x±s,mmol/L) 1.03±0.22 1.04±0.34 0.85±0.18 1.05±0.27 0.093 低密度脂蛋白胆固醇(x±s,mmol/L) 2.82±0.94 2.10±0.94* 1.97±0.56* 1.84±0.79* 0.001 HTC:高海拔藏族冠心病患者;HTN:高海拔藏族健康人群;MHC:中海拔汉族冠心病患者;LHC:低海拔汉族冠心病患者;*与HTN比较,差异具有统计学意义 -
[1] Mingji C, Onakpoya IJ, Perera R, et al. Relationship between altitude and the prevalence of hypertension in Tibet: a systematic review[J]. Heart, 2015, 101: 1054-1060. doi: 10.1136/heartjnl-2014-307158 [2] Faeh D, Gutzwiller F, Bopp M, et al. Lower mortality from coronary heart disease and stroke at higher altitudes in Switzerland[J]. Circulation, 2009, 120: 495-501. doi: 10.1161/CIRCULATIONAHA.108.819250 [3] Fujimoto N, Matsubayashi K, Miyahara T, et al. The risk factors for ischemic heart disease in Tibetan highlanders[J]. Jpn Heart J, 1989, 30: 27-34. doi: 10.1536/ihj.30.27 [4] 次仁罗布, 姜铁民, 金峰, 等. 西藏高海拔地区冠心病患者冠状动脉病变特点及其介入治疗疗效观察[J]. 世界最新医学信息文摘, 2018, 18: 13-18. https://www.cnki.com.cn/Article/CJFDTOTAL-WMIA201879006.htm [5] Tang WH, Kitai T, Kitai T, et al. Gut Microbiota in Cardiovascular Health and Disease[J]. Circ Res, 2017, 120: 1183-1196. doi: 10.1161/CIRCRESAHA.117.309715 [6] Jia Z, Zhao X, Liu X, et al. Impacts of the Plateau Environment on the Gut Microbiota and Blood Clinical Indexes in Han and Tibetan Individuals[J]. mSystems, 2020, 5: 1-16. [7] Caporaso JG, Kuczynski J, Stombaugh J, et al. QⅡME allows analysis of high-throughput community sequencing data[J]. Nat Methods, 2010, 7: 335-336. doi: 10.1038/nmeth.f.303 [8] Magoč T, Salzberg SL. FLASH: fast length adjustment of short reads to improve genome assemblies[J]. Bioinformatics, 2011, 27: 2957-2963. doi: 10.1093/bioinformatics/btr507 [9] Edgar RC, Haas BJ, Clemente JC, et al. UCHIME improves sensitivity and speed of chimera detection[J]. Bioinformatics, 2011, 27: 2194-2200. doi: 10.1093/bioinformatics/btr381 [10] Edgar RC. Search and clustering orders of magnitude faster than BLAST. Bioinformatics[J]. Bioinformatics, 2010, 26: 2460-2461. doi: 10.1093/bioinformatics/btq461 [11] 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: 590-596. [12] Gasmi A, Mujawdiya PK, Pivina L, et al. Relationship between gut microbiota, gut hyperpermeability, and obesity[J]. Curr Med Chem, 2020, 27: 1-13. doi: 10.2174/092986732701200218105010 [13] 胡海兵, 崔立, 郭靓骅, 等. 基于高通量测序技术的冠心病患者肠道菌群多样性研究[J]. 上海交通大学学报(农业科学版), 2016, 34: 1-19. https://www.cnki.com.cn/Article/CJFDTOTAL-SHNX201602001.htm [14] Arseneault-Bréard J, Rondeau I, Gilbert K, et al. Combination of Lactobacillus helveticus R0052 and Bifidobacterium longum R0175 reduces post-myocardial infarction depression symptoms and restores intestinal permeability in a rat model[J]. Br J Nutr, 2012, 107: 1793-1799. doi: 10.1017/S0007114511005137 [15] Lam V, Su J, Koprowski S, et al. Intestinal microbiota determine severity of myocardial infarction in rats[J]. FASEB J, 2012, 26: 1727-1735. doi: 10.1096/fj.11-197921 [16] Ott SJ, El Mokhtari NE, Musfeldt M, et al. Detection of diverse bacterial signatures in atherosclerotic lesions of patients with coronary heart disease[J]. Circulation, 2006, 113: 929-937. doi: 10.1161/CIRCULATIONAHA.105.579979 [17] Koren O, Spor A, Felin J, et al. Human oral, gut, and plaque microbiota in patients with atherosclerosis[J]. Proc Natl ACHD Sci USA, 2011, 108: 4592-4598. doi: 10.1073/pnas.1011383107 [18] Wu P, Chen JN, Chen JJ, et al. Trimethylamine N-oxide promotes apoE-/- mice atherosclerosis by inducing vascular endothelial cell pyroptosis via the SDHB/ROS pathway[J]. J Cell Physiol, 2020, 235: 6582-6591. doi: 10.1002/jcp.29518 [19] 朱华, 李卓, 苏磊, 等. 冠心病人源肠道菌群小鼠模型的建立及评价[J]. 中国实验动物学报, 2019, 27: 716-724. https://www.cnki.com.cn/Article/CJFDTOTAL-ZGSD201906004.htm [20] Zhu LL, Ma ZJ, Ren M, et al. Distinct Features of Gut Microbiota in High-Altitude Tibetan and Middle-Altitude Han Hypertensive Patients[J]. Cardiol Res Pract, 2020, 2020: 1957843. [21] Schneeberger M, Everard A, Gómez-Valadés AG, et al. Akkermansia muciniphila inversely correlates with the onset of inflammation, altered adipose tissue metabolism and metabolic disorders during obesity in mice[J]. Sci Rep, 2015, 5: 16643. doi: 10.1038/srep16643 [22] Ma Y, Zhu L, Ma Z, et al. Distinguishing feature of gut microbiota in Tibetan highland coronary artery disease patients and its link with diet[J]. Sci Rep, 2021, 11: 18486. doi: 10.1038/s41598-021-98075-9 [23] Courtney HS, Pownall HJ. The structure and function of serum opacity factor: a unique streptococcal virulence determinant that targets high-density lipoproteins[J]. J Biomed Biotechnol, 2010, 2010: 956071. [24] Zhang B, Wang X, Xia R, et al. Gut microbiota in coronary artery disease: a friend or foe?[J]. Biosci Rep, 2020, 40: 1-11. [25] Li J, Zhao F, Wang Y, et al. Gut microbiota dysbiosis contributes to the development of hypertension[J]. Microbiome, 2017, 5: 14. doi: 10.1186/s40168-016-0222-x [26] Zhang M, Zhou L, Wang Y, et al. Faecalibacterium prausnitzii produces butyrate to decrease c-Myc-related metabolism and Th17 differentiation by inhibiting histone deacetylase 3[J]. Int Immunol, 2019, 31: 499-514. doi: 10.1093/intimm/dxz022 [27] De Vadder F, Kovatcheva-Datchary P, Zitoun C, et al. Microbiota-Produced Succinate Improves Glucose Homeostasis via Intestinal Gluconeogenesis[J]. Cell Metab, 2016, 24: 151-157. doi: 10.1016/j.cmet.2016.06.013 [28] Kelly TN, Bazzano LA, Ajami NJ, et al. Gut Microbiome Associates With Lifetime Cardiovascular Disease Risk Profile Among Bogalusa Heart Study Participants[J]. Circ Res, 2016, 1198: 956-964. [29] Park YE, Kim MS, Shim KW, et al. Effects of Lactobacillus plantarum Q180 on Postprandial Lipid Levels and Intestinal Environment: A Double-Blind, Randomized, Placebo-Controlled, Parallel Trial[J]. Nutrients, 2020, 12: 255. doi: 10.3390/nu12010255 [30] Malik M, Suboc TM, Tyagi S, et al. Lactobacillus plantarum 299v Supplementation Improves Vascular Endothelial Function and Reduces Inflammatory Biomarkers in Men With Stable Coronary Artery Disease[J]. Circ Res, 2018, 123: 1091-1102. doi: 10.1161/CIRCRESAHA.118.313565 [31] Gan XT, Ettinger G, Huang CX, et al. Probiotic administration attenuates myocardial hypertrophy and heart failure after myocardial infarction in the rat[J]. Circ Heart Fail, 2014, 7: 491-499. doi: 10.1161/CIRCHEARTFAILURE.113.000978 [32] Liu H, Chen X, Hu X, et al. Alterations in the gut microbiome and metabolism with coronary artery disease severity[J]. Microbiome, 2019, 7: 68. doi: 10.1186/s40168-019-0683-9