肠道菌群对抗精神病药源性代谢紊乱的影响研究

发布时间：2018-03-14 03:17

  本文选题：精神分裂症　切入点：肠道菌群　出处：《郑州大学》2017年硕士论文　论文类型：学位论文

【摘要】：目的1.动态观察首发未用药精神分裂症患者用药前后肠道微生物、炎性细胞因子及代谢指标的变化,进一步探讨抗精神病药物对代谢指标、炎症细胞因子、肠道微生物的影响。2.通过对首发未用药精神分裂症患者用药前后不同时期肠道微生物与代谢指标、炎性细胞因子等进行关联性研究,以探讨肠道微生物与抗精神病药物引起的代谢异常的关系。方法1.选取2015年1月至2016年8月于郑州大学第一附属医院精神医学科住院的40例首发精神分裂症患者(患者组)和35名健康志愿者(健康对照组),患者组在入组当日详细询问病史,收集一般临床资料,并进行体格检查及精神检查,同时进行PANSS量表评定,于住院第二天清晨采血、收集粪便标本、测量身高、体重、计算体重指数(BMI),出院后对入组患者进行6周、12周、24周的随访,收集资料及粪便、血标本同上。2.使用葡萄糖氧化酶法(GOD)检测空腹血糖(FBG)水平,使用酶色度法测定血清空腹甘油三酯(TG)、胆固醇(CHOL)、高密度脂蛋白(HDL)和低密度脂蛋白(LDL)的水平;采用电化学发光法检测血清空腹胰岛素(INS)、使用酶联免疫法(ELISA)检测血清空腹细胞因子IL-6、IL-10的水平。3.肠道微生物总DNA的提取:按照德国QIAGEN公司提供的QIAamp粪便DNA提取试剂盒说明书操作,粪便总DNA浓度和纯度使用紫外分光光度计检测以保存备用。4.粪便肠道微生物(拟杆菌属、乳酸杆菌属、双歧杆菌属)的绝对拷贝数采用荧光定量PCR(QPCR)法进行测定:引物设计以16SrRNA序列为依据,常规PCR扩增目的片段,并回收、纯化常规PCR产物,构建载体。使用SYBR Green I荧光染料,根据扩增的阈值及阈值循环数,绘制标准曲线从而计算出肠道微生物(拟杆菌属、乳酸杆菌属、双歧杆菌属)的拷贝数。5.采用SPSS 20.0软件统计分析数据。使用K-S单样本检验计量资料是否为正态分布,若服从正态分布,结果以均数±标准差(X±S)来表示,两组间参数的比较采用两独立样本t检验;若不服从正态分布,采用非参数Mann-Whitney U检验,结果以中位数来表示。采用卡方检验或连续校正的卡方检验方法分析计数资料,结果以百分比[例(%)]来表示。肠道各细菌菌属与糖脂代谢指标及炎症因子等的相关性经检验符合正态分布的采用Pearson相关分析,不符合正态分布的采用Spearman相关分析。患者组内用药前后生物学指标的变化(连续性计量资料)使用方差分析,差异有统计学意义以双侧P0.05来表示。结果1.共入组患者组40例(其中5例脱落,35例完成随访),健康对照组35例,两组性别、年龄、文化程度的差异均无统计学意义(P均0.05)。2.两组基线期体重、BMI、FBG、INS、CHOL、TG、HDL、LDL水平无明显差异(P均0.05)3.两组间基线血清炎性细胞因子及肠道菌群水平比较:(1)炎症因子水平比较:血清IL-6水平患者组[(32.69±10.83)pg/ml]明显高于健康对照组[(24.65±5.51)pg/ml],IL-10水平患者组[(27.40±9.89)pg/ml]明显低于对照组[(32.93±9.18)pg/ml],差异有统计学意义(p均0.05)。(2)两组间粪便双歧杆菌属、拟杆菌属、乳酸杆菌属及乳酸杆菌属/拟杆菌属、双歧杆菌属/拟杆菌属水平比较:患者组双歧杆菌属水平[(6.86±0.76)lg copies/g wet fecals]低于健康对照组[(8.05±0.68)lg copies/g wet fecals],差异有统计学意义(p0.001),患者组拟杆菌属[(8.85±1.31)lg copies/g wet fecals]高于健康对照组[(7.36±1.62)lg copies/g wet fecals],差异有统计学意义(p0.001),患者组乳酸杆菌属(8.65±0.79)低于健康对照组(8.84±0.70),差异无统计学意义(p=0.168),患者组乳酸杆菌属/拟杆菌属(1.12±0.05)、双歧杆菌属/拟杆菌属(0.79±0.16)均较健康对照组水平低(1.27±0.35)、(1.16±0.32),统计学差异显著(p均0.001)。4.患者组治疗后不同时期糖脂代谢、炎症因子、肠道微生物水平的比较:(1)糖脂代谢指标变化:患者组治疗后FBG升高,6周、12周、24周[(4.56±0.58)、(4.62±0.50)、(4.52±0.58)mmol/L]均高于基线水平[(4.3±0.65)mmol/L],差异有统计学意义(p均0.05);CHOL水平升高,6周、12周、24周[(4.73±1.15)、(4.54±0.90)、(4.41±0.84)mmol/L]均高于基线水平[(3.83±0.85)mmol/L],差异有统计学意义(p均0.05);TG水平升高,6周、12周、24周[(1.35±0.75)、(1.58±0.76)、(1.41±0.93)mmol/L]均高于基线水平[(0.97±0.50)mmol/L],差异有统计学意义(p均0.05);HDL水平改变无明显差异(p0.05),LDL水平升高,6周、12周、24周[(2.81±0.90)、(2.85±0.78)、(2.48±0.70)mmol/L]均高于基线水平[(2.21±0.73)mmol/L],差异有统计学意义(p均0.05)。患者组不同时期体重指标的比较:体重增加,6周、12周、24周[(59.56±10.19)、(63.07±10.59)、(64.91±10.04)kg/m2]均高于基线水平[(55.20±9.56)kg],有统计学差异(p0.05)。BMI升高,6周、12周、24周[(23.76±4.60)、(24.44±4.59)、(25.29±4.65)kg/m2]均高于基线水平[(21.76±4.07)kg/m2],有统计学差异(p0.05)。(2)炎性细胞因子水平的改变:患者组服药后IL-6水平先下降后升高,基线、6周、12周、24周[(32.69±10.83)、(31.36±7.46)、(36.00±10.16)、(37.73±11.31)pg/ml],差异有统计学意义(p均0.05);IL-10水平缓慢升高6周、12周、24周[(28.64±9.06)、(31.01±7.97)、(35.06±13.97)pg/ml]均高于基线[(27.40±9.89)pg/ml],统计学差异性显著(p均0.05)。(3)肠道菌群水平的比较:患者组服用抗精神病药物治疗后拟杆菌属整体水平降低6周、12周、24周[(8.59±0.84)、(7.43±0.74)、(7.62±0.62)lg copies/g wet fecals]均低于基线水平[(8.85±1.31)lg copies/g wet fecals],差异有统计学意义(F=22.368,p0.001);乳酸杆菌属缓慢升高,6周、12周、24周[(8.91±1.03)、(8.91±0.84)、(8.97±0.69)lg copies/g wet fecals],均高于基线水平[(8.65±0.79)lg copies/g wet fecals]差异有统计学意义(F=55.279,p0.001);双歧杆菌属水平先降低,6周、12周[(6.56±0.58)、(6.56±0.69)lg copies/g wet fecals],后缓慢恢复,24周[(6.84±0.62)lg copies/g wet fecals],均低于基线[(6.86±0.76)lg copies/g wet fecals],差异有统计学意义(F=99.948,p0.001);乳酸杆菌属/拟杆菌属的比例缓慢升高,6周(1.05±0.15)改变不明显、12周、24周(1.21±0.16)、(1.19±0.14)均高于基线(1.12±0.05),差异有统计学意义(F=9.467,p0.001);双歧杆菌属/拟杆菌属比例先降低后升高6周(0.77±0.12)、12周(0.89±0.13)、24周(0.90±0.01),差异有统计学意义(F=1274.264,p0.001)。5.相关分析:(1)基线期肠道微生物与糖脂代谢指标、炎症因子、体重、BMI无明显相关性(p均0.05)。(2)治疗6周患者组拟杆菌属拷贝数与体重、BMI、FBG、TCHO负相关(r=-0.370,-0.340,-0.481,-0.462,p均0.05),乳酸杆菌属的拷贝数与BMI、TG正相关(r=-0.348,0.373,p均0.05),双歧杆菌属的拷贝数与体重、BMI、TCHO、TG、LDL负相关(r=-0.420,-0.415,-0.394,-0.404,-0.443,p均0.05);(3)治疗12周患者拟杆菌属拷贝数与体重、BMI负相关(r=-0.373,-0.429,p均0.05),乳酸杆菌属拷贝数与体重、BMI正相关(r=0.358,0.407,p0.05),双歧杆菌属拷贝数与体重、BMI、TCHO、TG均负相关(r=-0.379,-0.503,-0.447,-0.411,p均0.05);(4)治疗24周患者组拟杆菌属拷贝数与体重、BMI、FBG、TCHO、TG、LDL均负相关(r=-0.472,-0.470,-0.407,-0.499,-0.395,-0.441,p均0.05),乳酸杆菌属的拷贝数与BMI正相关(r=0.402,p0.05),双歧杆菌属的拷贝数与体重、BMI、FBG、TCHO、TG、LDL负相关(r=-0.446,-0.466,-0.438,-0.475,-0.404,-0.436,p均0.05);(5)药物治疗后肠道微生物与IL-6、IL-10相关性不明显(p均0.05)。结论1.首发精神分裂症患者存在肠道菌群失调及细胞因子介导的免疫紊乱,二者间相互作用,可能在精神分裂症的发生发展中起着一定的作用。2.抗精神病药物治疗后引起体重增加、糖脂代谢异常、炎症因子水平异常及肠道微生物组成改变。3.抗精神病药物引起的体重增加,糖脂代谢异常可能与肠道微生物组成及免疫状态的改变有关。
[Abstract]:Objective To observe the dynamic 1. first-episode medication before and after treatment in patients with intestinal microflora of mental division, changes of inflammatory cytokines and metabolic indicators, further explore the effect of antipsychotic drugs on metabolism, inflammatory cytokines,.2. intestinal microorganisms based on first-episode schizophrenic patients before and after treatment in different periods of gut microbiota and metabolic parameters. Correlation of inflammatory cytokines, to explore the relationship between intestinal microflora and antipsychotic induced metabolic abnormalities. Methods from January 2015 to August 2016 1. in the spirit of the medical department of the First Affiliated Hospital of Zhengzhou University 40 schizophrenic patients (patient group) and 35 healthy volunteers (healthy control group). Patients in the group the detailed history, collected the clinical data and physical examination and mental examinations. At the same time scale PANSS Evaluation on hospital day second morning blood samples collected fecal samples, measurement of height, weight, body mass index (BMI), the patients were discharged after 6 weeks, 12 weeks, 24 weeks of follow-up, data collection and feces, blood samples using.2.. The method of glucose oxidase (GOD) fasting blood glucose (FBG the level of serum triglyceride), using enzymatic colorimetric method (TG), cholesterol (CHOL), high density lipoprotein (HDL) and low density lipoprotein (LDL) level; electrochemiluminescence detection of serum fasting insulin (INS), using enzyme-linked immunosorbent assay (ELISA) detection of serum fasting cytokines IL-6, the extraction of IL-10.3. level intestinal microbial total DNA extraction kit according to QIAamp fecal DNA supplied by the German company QIAGEN, fecal DNA concentration and purity using UV spectrophotometer detection to save standby.4. fecal intestinal microorganisms (bacteroides, Lactobacillus, Bifidobacterium) the absolute copy number by fluorescence quantitative PCR (QPCR) method: primers were designed by 16SrRNA sequences based on conventional PCR amplification, purification and recovery, the product of conventional PCR, using SYBR Green vector. I fluorescent dye, according to the amplification threshold and the threshold cycle number. To calculate the standard curve of intestinal microflora (Bacteroides, Lactobacillus and Bifidobacterium) copy number.5. SPSS 20 software was used for statistical analysis of data. The use of K-S single sample test measurement data for normal distribution, if the normal distribution, results mean standard deviation (X + S) to said, compared with two groups of parameters between two independent samples t test; if you do not obey the normal distribution, using non parametric Mann-Whitney U test results in the median said. Analysis using chi square test or chi square test method of continuous correction Count data, results in the percentage of cases (%)] [to express. The intestinal bacteria from correlation with glucose and lipid metabolism and inflammatory factors such as the test with normal distribution using the Pearson correlation analysis, does not meet the normal distribution using Spearman correlation analysis. The changes of biological indexes before and after treatment in the patients with continuous (measurement data using analysis of variance), the difference was statistically significant in terms of bilateral P0.05. Results a total of 1. patients in group 40 cases (including 5 cases of loss, 35 patients completed the follow-up), healthy control group of 35 cases, two groups of gender, age, culture degree difference were not statistically significant (P 0.05).2. two group baseline weight, BMI, FBG, INS, CHOL, TG, HDL, no significant differences in the level of LDL (P 0.05) compared with baseline serum inflammatory cytokines and intestinal flora of 3. levels between the two groups: (1) compare the level of inflammatory factors: the level of serum IL-6 Group [(32.69 + 10.83 PG) /ml] was significantly higher than the healthy control group [(24.65 + 5.51) pg/ml], IL-10 levels of the patients with [(27.40 + 9.89) pg/ml] was significantly lower than the control group [(32.93 + 9.18) pg/ml], the difference was statistically significant (P < 0.05). (2) between the two groups of fecal Bifidobacterium, Bacteroides, Lactobacillu bacteria and Lactobacillus / Bacteroides, Bifidobacterium / Bacteroides level: Patients with Bifidobacterium level [(6.86 + 0.76) LG copies/g wet fecals] lower than that of the control group [(8.05 + 0.68) LG copies/g wet fecals], the difference was statistically significant (p0.001), Bacteroides group is [(8.85 + 1.31) LG copies/g wet fecals] higher than that of the control group [(7.36 + 1.62) LG copies/g wet fecals], the difference was statistically significant (p0.001), Lactobacillus group (8.65 + 0.79) is lower than the control group (8.84 + 0.70), the difference was not statistically significant (p=0.168), group Lactobacillus / Bacteroides 灞，

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