膳食干預腸道菌群預防代謝性疾病

1、膳食干預腸道菌群預防代謝性疾病膳食干預腸道菌群預防代謝性疾病Dietary Modulation of Gut Microbiota for Preventive management of Metabolic Diseases趙立平趙立平 Liping Zhao上海交通大學生命科學技術(shù)學院上海交通大學生命科學技術(shù)學院上海系統(tǒng)生物醫(yī)學研究中心上海系統(tǒng)生物醫(yī)學研究中心Shanghai Jiao Tong UniversityHuman Superorganisms正確認識自己的身體正確認識自己的身體“超級生物體超級生物體”諾貝爾獎得主-里德伯格, Science 2000Joshua Leder

2、bergNobel Laureate 1958人體內(nèi)生活著1-2公斤細菌 90%在腸道里人體的細胞組成 10%人細胞+90%細菌細胞人體的基因組成 2.5萬個人的基因 3百多萬個微生物基因 豐富而活躍的代謝活動腸道菌群腸道菌群人體后天獲得的人體后天獲得的“第二個基因組第二個基因組”Gut Microbiota-Second genome acquired after birth食品食品藥品藥品人基因組人基因組腸道微生物組腸道微生物組健康健康疾病疾病體質(zhì)體質(zhì)亞健康亞健康先天遺傳后天養(yǎng)成Shanghai Jiao Tong University“慢性病的腸道起源慢性病的腸道起源”“人體系統(tǒng)生物學方

3、法人體系統(tǒng)生物學方法”Shanghai Jiao Tong University慢性病的腸道起源Gut Origin of Chronic DiseasesIIya MechnikovNobel Laureate 1908“Later he took up the study of the flora of the human intestine and developed atheory that senility is due to poisoning of the body by the products of certain of these bacteria. To prevent

4、the multiplication of these organisms he proposed a diet containing milk fermented by bacilli which produce large amounts of lactic acid and for a time this diet became widely popular.”-/Symbiosis vs. Dysbiosis“糞毒入血， 百病蜂起”中醫(yī)觀念“Fecal toxins into bloodstream will lead to all kinds

5、of diseases” Concept from Traditional Chinese Medicine (TCM)細胞毒素、遺傳毒素、免疫毒素-肝腸循環(huán)-腸屏障受損Shanghai Jiao Tong University腸道菌群結(jié)構(gòu)失調(diào)與代謝綜合征腸道菌群結(jié)構(gòu)失調(diào)與代謝綜合征Cani, 2009Shanghai Jiao Tong University腸道菌群控制能量代謝基因表達腸道菌群控制能量代謝基因表達Backhed, F., et al., The gut microbiota as an environmental factor that regulates fat stora

6、ge. Proc Natl Acad Sci U S A, 2004. 101:15718-15723. Backhed, F., et al., Mechanisms underlying the resistance to diet-induced obesity in germ-free mice. Proc Natl Acad Sci U S A, 2007. 104: 979-984.Turnbaugh, P.J., et al., An obesity-associated gut microbiome with increased capacity for energy harv

7、est. Nature, 2006. 444(7122): p. 1027-1031.Turnbaugh, P.J., et al., Diet-induced obesity is linked to marked but reversible alterations in the mouse distal gut microbiome. Cell Host Microbe, 2008. 3: 213-223.Jeffery Gordon Shanghai Jiao Tong University肥胖肥胖糖尿病糖尿病炎性衰老炎性衰老 低度、系統(tǒng)性的慢性炎癥低度、系統(tǒng)性的慢性炎癥 慢性炎癥與衰

8、老和老年病的關(guān)系慢性炎癥與衰老和老年病的關(guān)系Inflammation and Age-related Diseases代謝性疾病代謝性疾病Endotoxin-the most common immunotoxin內(nèi)毒素-最常見的免疫毒素內(nèi)毒素Molecular Inflammation induced by endotoxin內(nèi)毒素誘導的分子炎癥Shanghai Jiao Tong UniversityInflammation induced by diet-disrupted gut microbiota菌群結(jié)構(gòu)失調(diào)誘發(fā)慢性炎癥菌群結(jié)構(gòu)失調(diào)誘發(fā)慢性炎癥Cani, P.D., Amar, J.

9、, Iglesias, M.A., Poggi, M., Knauf, C., Bastelica, D. et al. (2007a) Metabolic endotoxemia initiates obesity and insulin resistance. Diabetes 56: 1761-1772.Cani, P.D., Neyrinck, A.M., Fava, F., Knauf, C., Burcelin, R.G., Tuohy, K.M. et al. (2007b) Selective increases of bifidobacteria in gut microfl

10、ora improve high-fat-diet-induced diabetes in mice through a mechanism associated with endotoxaemia. Diabetologia 50: 2374-2383 Cani, P.D, et al.,(2008) Changes in gut microbiota control metabolic endotoxemia-induced inflamation in high-fat diet-induced obesity and diabetes in mice. Diabetes 57: 147

11、0-1482Cani, P.D, et al.,(2009) Changes in gut microbiota control inflammation in obese mice through a mechanism involving GLP-2-driven improvement of gut permeability. Gut 58:1091-1103Shanghai Jiao Tong UniversityApoa-1 knock-out mouseApoa-1 基因敲除鼠基因敲除鼠C57BL/6 miceApoa-1 基因基因敲除敲除Apoa-1-/- mice敲除小鼠敲除小

12、鼠 Apoa-l gene的策略的策略 (Williamson, 1992, PNAS)Apolipoprotein A-高密度脂蛋白水平低易感動脈粥樣硬化先天性胰島素抵抗Shanghai Jiao Tong University動物實驗高脂飼料高脂飼料植物飼料植物飼料敲除鼠敲除鼠壞基因植物飼料植物飼料 對照鼠對照鼠好基因高脂飼料高脂飼料 兩種基因兩種基因 X 兩種飼料兩種飼料 飼喂飼喂6個月個月壞食物好食物壞食物好食物Shanghai Jiao Tong UniversityFood intakeGlucose Tolerance Test Growth CurvePhenotype Dat

13、a好基因+壞食物病得最重！Shanghai Jiao Tong UniversityDiet is shaping gut microbiota飲食決定腸道菌群結(jié)構(gòu)，基因作用很小Wt/NC野生型野生型/植物飼料植物飼料Apoa-/- /NC敲除鼠敲除鼠/植物飼料植物飼料Wt/HFD野生型野生型/高脂飼料高脂飼料Apoa-/- /HFD敲除鼠敲除鼠/高脂飼料高脂飼料Shanghai Jiao Tong University新一代革命性測序技術(shù)新一代革命性測序技術(shù)15100 臺老一代測序儀1個技術(shù)員，1臺新設備NATURE METHODS ，16 | VOL.5 NO.1 | JANUARY 20

14、08 Shanghai Jiao Tong University% of Total Sequences ApoA- -/- high fat dietApoA- -/- normal chow Wild-type high fat dietWild-type normal chow*P0.01 *P0.91LDL- cholesterol （mmol/L）3.42 3.153.420.00-4.16blood pressure（mm Hg）100/140 80/12075/120 80/120Aspartate aminotransferase（U/L）122.0 513110-47LBP

15、(g/ml)(I)8.132.445.76_IL-1beta (pg/ml)(I)0.090.110.07_IL-6 (pg/ml)(I)6.564.392.72_Adiponectin (ng/ml)(A)1839.411925.634145.38_Shanghai Jiao Tong University16S rRNA gene V3-DGGE Pattern M 0d 4w 9w 13w18w 23w PCAShanghai Jiao Tong University0d 4w9w13w18w23wAbundance distribution of predominant phyloty

16、pes A pathogenShanghai Jiao Tong UniversityMetagenomic Sequencing以腸道菌群為靶標的慢性病早期營養(yǎng)干預 -30天 0天 4周 9周 13周 18周 23周干預期 維持期 基線 （n=163）（n=123） （n=101） （n=93）123位合格志愿者BMI30，中心型II度肥胖癥患者 （F:M=69:54） WHO中國臨床試驗注冊中心通過倫理審查并注冊，中國臨床試驗注冊中心通過倫理審查并注冊，注冊號為注冊號為ChiCTR-TRC-00000353 Shanghai Jiao Tong University菌群參與代謝性疾病發(fā)展的

17、分子過程保護腸屏障的細菌種群水平下降，條件致病菌的種群水平上升；腸道的通透性上升；血液中的細菌抗原載荷量升高；炎性水平升高；胰島素抵抗出現(xiàn)；血糖、血脂、血壓調(diào)控機制受損。Shanghai Jiao Tong University相關(guān)分子標示物的使用菌群結(jié)構(gòu)的454高通量測序分析腸屏障功能檢查 乳果糖（lactulose）尿中排泄率甘露醇（mannitol）尿中排泄率比值法 血液中的細菌抗原載荷量 脂多糖結(jié)合蛋白LBP的濃度測定炎性水平測定 C反應蛋白CRP IL-1、IL-6、TNF-和脂聯(lián)素 胰島素抵抗測定 空腹胰島素 穩(wěn)態(tài)模型的胰島素抵抗指數(shù)（HOMA-IR）=空腹胰島素（IU/L）空腹血

18、糖（mmol/L）/22.5。 Shanghai Jiao Tong University空腹血糖糖化血紅蛋白甘油三酯總膽固醇高密度脂蛋白低密度脂蛋白血壓谷草轉(zhuǎn)氨酶 丙氨酸氨基轉(zhuǎn)移酶 -谷氨酰轉(zhuǎn)移酶 相關(guān)臨床標示物的使用Shanghai Jiao Tong University保護腸屏障的細菌增加保護腸屏障的細菌增加條件致病菌下降條件致病菌下降* * Proteobacteria Bacteroidetes Fusobacteria ActinobacteriaFirmicutes010204060% (percentage)Phylum -30d 9w 23wShanghai Jiao To

19、ng University腸屏障功能 -30d9w23w0.000.010.020.030.040.060.080.100.12L/M ratioP-values 0.05*P-values 0.01* *Shanghai Jiao Tong University脂多糖結(jié)合蛋白（LBP）-30d9w23w-100102030405060100120LBP(g/ml)P-values 0.05*P-values 0.01* *Shanghai Jiao Tong UniversityC反應蛋白（CRP）-30d9w23w-202468101216202410CRP(mg/L)0P-values

20、0.05*P-values 0.01* *Shanghai Jiao Tong University炎性因子-30d9w23w-10123420406080TNF-(pg/ml)-30d9w23w-101234510152025IL-6(pg/ml)-30d9w23w-0.10.01.5IL-1(pg/ml)-30d9w23w024681015202530Adiponectin(g/ml)P-values 0.05*P-values 0.01* *Shanghai Jiao Tong University 糖穩(wěn)態(tài)-30d9w23w3.03.54.04.55.

21、05.56.06.57.07.56.1FPG(mmol/L)3.9-30d9w23w-50510152025303510015020027FINS(IU/ml)6-30d9w23w2.53.03.54.04.55.05.56.06.57.05.8HbA1c(%)3.8-30d9w23w-2024681020304050HOMA-IRP-values 0.05*P-values 0.01* *Shanghai Jiao Tong University血脂-30d9w23w-101234102030401.7TG(mmol/L)0-30d9w23w123456785.17TC(mmol/L)3-3

22、0d9w23w0.81.01.82.02.2HDL-C(mmol/L)0.91-30d9w23w01234564.16LDL-C(mmol/L)0P-values 0.05*P-values 0.01* *Shanghai Jiao Tong University肝功能-30d9w23w010203040506047AST(U/L)10-30d9w23w-10010203040506010012041ALT(U/L)0-30d9w23w-100102030405060708012015018056GGT(U/L)0P-values 0.05*P-values

23、 0.01* *Shanghai Jiao Tong University血壓-30d9w23w80100120140160180200SBP(mm Hg)-30d9w23w5060708090100110120130140DBP(mm Hg)P-values 0.05*P-values 0.01* *Shanghai Jiao Tong University志愿者體重變化情況 Weight BMI -30d 9w 23w Time Point -30d 9w 23w Time Point * * * * P-values 0.05All vs. -30d 9w vs. 23w * *kgkg

24、/m84.980.478.031.830.129.09周周: 減重中位值減重中位值4.5 kg, 最高最高 15.5 kg23周周: 減重中位值減重中位值 6.9 kg, 最高最高 20.3 kgShanghai Jiao Tong University以菌群為靶點的健康監(jiān)測新技術(shù)以菌群為靶點的健康監(jiān)測新技術(shù)腸道菌群結(jié)構(gòu)檢測 腸屏障保護菌 毒素、抗原產(chǎn)生菌腸道菌毒素檢測 免疫毒性 細胞毒性 遺傳毒性腸屏障功能檢測血液抗原載荷量測定炎性反應檢測內(nèi)毒素結(jié)合蛋白內(nèi)毒素結(jié)合蛋白C反應蛋白反應蛋白空腹胰島素空腹胰島素Shanghai Jiao Tong University樣品采集情況 StoolUri

25、neBloodUrine for Gut permeability testDun hua fangfirst trial1611616968Dun hua fangsecond trial70703030Coal central hospital287287123123He ping hospital1961968484Total714714305305Shanghai Jiao Tong University基于功能元基因組學的人體系統(tǒng)生物學分析基于功能元基因組學的人體系統(tǒng)生物學分析ConcentrationPrimary MoleculesSecondary MoleculesResorptionDilutionFiltration1234567ppmhippurateureaallantoincreatininehippurate2-oxoglutaratecitrateTMAOsuccinatefumaratewatercreatininetaurineMetabolism-MetabonomeMicrobiome-MetagenomeProteome-ImmunityVibration, noise, exhaust gas