本文選題：精神分裂癥　切入點：腸道菌群　出處：《鄭州大學(xué)》2017年碩士論文　論文類型：學(xué)位論文

【摘要】：目的1.動態(tài)觀察首發(fā)未用藥精神分裂癥患者用藥前后腸道微生物、炎性細(xì)胞因子及代謝指標(biāo)的變化,進(jìn)一步探討抗精神病藥物對代謝指標(biāo)、炎癥細(xì)胞因子、腸道微生物的影響。2.通過對首發(fā)未用藥精神分裂癥患者用藥前后不同時期腸道微生物與代謝指標(biāo)、炎性細(xì)胞因子等進(jìn)行關(guān)聯(lián)性研究,以探討腸道微生物與抗精神病藥物引起的代謝異常的關(guān)系。方法1.選取2015年1月至2016年8月于鄭州大學(xué)第一附屬醫(yī)院精神醫(yī)學(xué)科住院的40例首發(fā)精神分裂癥患者(患者組)和35名健康志愿者(健康對照組),患者組在入組當(dāng)日詳細(xì)詢問病史,收集一般臨床資料,并進(jìn)行體格檢查及精神檢查,同時進(jìn)行PANSS量表評定,于住院第二天清晨采血、收集糞便標(biāo)本、測量身高、體重、計算體重指數(shù)(BMI),出院后對入組患者進(jìn)行6周、12周、24周的隨訪,收集資料及糞便、血標(biāo)本同上。2.使用葡萄糖氧化酶法(GOD)檢測空腹血糖(FBG)水平,使用酶色度法測定血清空腹甘油三酯(TG)、膽固醇(CHOL)、高密度脂蛋白(HDL)和低密度脂蛋白(LDL)的水平;采用電化學(xué)發(fā)光法檢測血清空腹胰島素(INS)、使用酶聯(lián)免疫法(ELISA)檢測血清空腹細(xì)胞因子IL-6、IL-10的水平。3.腸道微生物總DNA的提取:按照德國QIAGEN公司提供的QIAamp糞便DNA提取試劑盒說明書操作,糞便總DNA濃度和純度使用紫外分光光度計檢測以保存?zhèn)溆谩?.糞便腸道微生物(擬桿菌屬、乳酸桿菌屬、雙歧桿菌屬)的絕對拷貝數(shù)采用熒光定量PCR(QPCR)法進(jìn)行測定:引物設(shè)計以16SrRNA序列為依據(jù),常規(guī)PCR擴(kuò)增目的片段,并回收、純化常規(guī)PCR產(chǎn)物,構(gòu)建載體。使用SYBR Green I熒光染料,根據(jù)擴(kuò)增的閾值及閾值循環(huán)數(shù),繪制標(biāo)準(zhǔn)曲線從而計算出腸道微生物(擬桿菌屬、乳酸桿菌屬、雙歧桿菌屬)的拷貝數(shù)。5.采用SPSS 20.0軟件統(tǒng)計分析數(shù)據(jù)。使用K-S單樣本檢驗計量資料是否為正態(tài)分布,若服從正態(tài)分布,結(jié)果以均數(shù)±標(biāo)準(zhǔn)差(X±S)來表示,兩組間參數(shù)的比較采用兩獨立樣本t檢驗;若不服從正態(tài)分布,采用非參數(shù)Mann-Whitney U檢驗,結(jié)果以中位數(shù)來表示。采用卡方檢驗或連續(xù)校正的卡方檢驗方法分析計數(shù)資料,結(jié)果以百分比[例(%)]來表示。腸道各細(xì)菌菌屬與糖脂代謝指標(biāo)及炎癥因子等的相關(guān)性經(jīng)檢驗符合正態(tài)分布的采用Pearson相關(guān)分析,不符合正態(tài)分布的采用Spearman相關(guān)分析�；颊呓M內(nèi)用藥前后生物學(xué)指標(biāo)的變化(連續(xù)性計量資料)使用方差分析,差異有統(tǒng)計學(xué)意義以雙側(cè)P0.05來表示。結(jié)果1.共入組患者組40例(其中5例脫落,35例完成隨訪),健康對照組35例,兩組性別、年齡、文化程度的差異均無統(tǒng)計學(xué)意義(P均0.05)。2.兩組基線期體重、BMI、FBG、INS、CHOL、TG、HDL、LDL水平無明顯差異(P均0.05)3.兩組間基線血清炎性細(xì)胞因子及腸道菌群水平比較:(1)炎癥因子水平比較:血清IL-6水平患者組[(32.69±10.83)pg/ml]明顯高于健康對照組[(24.65±5.51)pg/ml],IL-10水平患者組[(27.40±9.89)pg/ml]明顯低于對照組[(32.93±9.18)pg/ml],差異有統(tǒng)計學(xué)意義(p均0.05)。(2)兩組間糞便雙歧桿菌屬、擬桿菌屬、乳酸桿菌屬及乳酸桿菌屬/擬桿菌屬、雙歧桿菌屬/擬桿菌屬水平比較:患者組雙歧桿菌屬水平[(6.86±0.76)lg copies/g wet fecals]低于健康對照組[(8.05±0.68)lg copies/g wet fecals],差異有統(tǒng)計學(xué)意義(p0.001),患者組擬桿菌屬[(8.85±1.31)lg copies/g wet fecals]高于健康對照組[(7.36±1.62)lg copies/g wet fecals],差異有統(tǒng)計學(xué)意義(p0.001),患者組乳酸桿菌屬(8.65±0.79)低于健康對照組(8.84±0.70),差異無統(tǒng)計學(xué)意義(p=0.168),患者組乳酸桿菌屬/擬桿菌屬(1.12±0.05)、雙歧桿菌屬/擬桿菌屬(0.79±0.16)均較健康對照組水平低(1.27±0.35)、(1.16±0.32),統(tǒng)計學(xué)差異顯著(p均0.001)。4.患者組治療后不同時期糖脂代謝、炎癥因子、腸道微生物水平的比較:(1)糖脂代謝指標(biāo)變化:患者組治療后FBG升高,6周、12周、24周[(4.56±0.58)、(4.62±0.50)、(4.52±0.58)mmol/L]均高于基線水平[(4.3±0.65)mmol/L],差異有統(tǒng)計學(xué)意義(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]均高于基線水平[(3.83±0.85)mmol/L],差異有統(tǒng)計學(xué)意義(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]均高于基線水平[(0.97±0.50)mmol/L],差異有統(tǒng)計學(xué)意義(p均0.05);HDL水平改變無明顯差異(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]均高于基線水平[(2.21±0.73)mmol/L],差異有統(tǒng)計學(xué)意義(p均0.05)�；颊呓M不同時期體重指標(biāo)的比較:體重增加,6周、12周、24周[(59.56±10.19)、(63.07±10.59)、(64.91±10.04)kg/m2]均高于基線水平[(55.20±9.56)kg],有統(tǒng)計學(xué)差異(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]均高于基線水平[(21.76±4.07)kg/m2],有統(tǒng)計學(xué)差異(p0.05)。(2)炎性細(xì)胞因子水平的改變:患者組服藥后IL-6水平先下降后升高,基線、6周、12周、24周[(32.69±10.83)、(31.36±7.46)、(36.00±10.16)、(37.73±11.31)pg/ml],差異有統(tǒng)計學(xué)意義(p均0.05);IL-10水平緩慢升高6周、12周、24周[(28.64±9.06)、(31.01±7.97)、(35.06±13.97)pg/ml]均高于基線[(27.40±9.89)pg/ml],統(tǒng)計學(xué)差異性顯著(p均0.05)。(3)腸道菌群水平的比較:患者組服用抗精神病藥物治療后擬桿菌屬整體水平降低6周、12周、24周[(8.59±0.84)、(7.43±0.74)、(7.62±0.62)lg copies/g wet fecals]均低于基線水平[(8.85±1.31)lg copies/g wet fecals],差異有統(tǒng)計學(xué)意義(F=22.368,p0.001);乳酸桿菌屬緩慢升高,6周、12周、24周[(8.91±1.03)、(8.91±0.84)、(8.97±0.69)lg copies/g wet fecals],均高于基線水平[(8.65±0.79)lg copies/g wet fecals]差異有統(tǒng)計學(xué)意義(F=55.279,p0.001);雙歧桿菌屬水平先降低,6周、12周[(6.56±0.58)、(6.56±0.69)lg copies/g wet fecals],后緩慢恢復(fù),24周[(6.84±0.62)lg copies/g wet fecals],均低于基線[(6.86±0.76)lg copies/g wet fecals],差異有統(tǒng)計學(xué)意義(F=99.948,p0.001);乳酸桿菌屬/擬桿菌屬的比例緩慢升高,6周(1.05±0.15)改變不明顯、12周、24周(1.21±0.16)、(1.19±0.14)均高于基線(1.12±0.05),差異有統(tǒng)計學(xué)意義(F=9.467,p0.001);雙歧桿菌屬/擬桿菌屬比例先降低后升高6周(0.77±0.12)、12周(0.89±0.13)、24周(0.90±0.01),差異有統(tǒng)計學(xué)意義(F=1274.264,p0.001)。5.相關(guān)分析:(1)基線期腸道微生物與糖脂代謝指標(biāo)、炎癥因子、體重、BMI無明顯相關(guān)性(p均0.05)。(2)治療6周患者組擬桿菌屬拷貝數(shù)與體重、BMI、FBG、TCHO負(fù)相關(guān)(r=-0.370,-0.340,-0.481,-0.462,p均0.05),乳酸桿菌屬的拷貝數(shù)與BMI、TG正相關(guān)(r=-0.348,0.373,p均0.05),雙歧桿菌屬的拷貝數(shù)與體重、BMI、TCHO、TG、LDL負(fù)相關(guān)(r=-0.420,-0.415,-0.394,-0.404,-0.443,p均0.05);(3)治療12周患者擬桿菌屬拷貝數(shù)與體重、BMI負(fù)相關(guān)(r=-0.373,-0.429,p均0.05),乳酸桿菌屬拷貝數(shù)與體重、BMI正相關(guān)(r=0.358,0.407,p0.05),雙歧桿菌屬拷貝數(shù)與體重、BMI、TCHO、TG均負(fù)相關(guān)(r=-0.379,-0.503,-0.447,-0.411,p均0.05);(4)治療24周患者組擬桿菌屬拷貝數(shù)與體重、BMI、FBG、TCHO、TG、LDL均負(fù)相關(guān)(r=-0.472,-0.470,-0.407,-0.499,-0.395,-0.441,p均0.05),乳酸桿菌屬的拷貝數(shù)與BMI正相關(guān)(r=0.402,p0.05),雙歧桿菌屬的拷貝數(shù)與體重、BMI、FBG、TCHO、TG、LDL負(fù)相關(guān)(r=-0.446,-0.466,-0.438,-0.475,-0.404,-0.436,p均0.05);(5)藥物治療后腸道微生物與IL-6、IL-10相關(guān)性不明顯(p均0.05)。結(jié)論1.首發(fā)精神分裂癥患者存在腸道菌群失調(diào)及細(xì)胞因子介導(dǎo)的免疫紊亂,二者間相互作用,可能在精神分裂癥的發(fā)生發(fā)展中起著一定的作用。2.抗精神病藥物治療后引起體重增加、糖脂代謝異常、炎癥因子水平異常及腸道微生物組成改變。3.抗精神病藥物引起的體重增加,糖脂代謝異�？赡芘c腸道微生物組成及免疫狀態(tài)的改變有關(guān)。
[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，

本文編號：1609363