【摘要】：背景：非酒精性脂肪性肝病(non-alcoholic fatty liver disease, NAFLD)是指除乙醇和其他明確的肝損害因素所致的，以彌漫性肝細(xì)胞大泡性脂肪變?yōu)橹饕卣鞯呐R床病理綜合征。根據(jù)肝內(nèi)脂肪變、炎癥和纖維化的程度，將NAFLD分為單純性脂肪肝、脂肪性肝炎、脂肪性肝硬化。 NAFLD的發(fā)病機(jī)制目前尚未完全明確，目前研究認(rèn)為NAFLD的發(fā)病與胰島素抵抗、脂代謝異常、脂質(zhì)過氧化等密切相關(guān)。而隨著研究的不斷深入，有學(xué)者認(rèn)為腸道菌群的改變可以通過直接調(diào)控機(jī)體脂肪合成與儲存相關(guān)基因的表達(dá)，影響能量吸收和儲存，促進(jìn)肥胖形成和胰島素抵抗等機(jī)制影響能量代謝[1]，參與NAFLD的發(fā)生和發(fā)展。 禁食誘導(dǎo)脂肪因子（fasting-induced adipose factor，F(xiàn)iaf）基因主要負(fù)責(zé)編碼脂蛋白脂肪酶（lipoprteinlipase，LPL）抑制因子，LPL可以水解在脂蛋白顆粒的甘油三酯，促進(jìn)脂質(zhì)沉積。Fiaf正是通過抑制LPL，從而抑制甘油三酯循環(huán)。有研究表明在動物體內(nèi)，F(xiàn)iaf可被腸道菌群調(diào)控，腸桿菌的增加可以抑制Fiaf的表達(dá)，使得LPL表達(dá)增多，導(dǎo)致脂質(zhì)沉積，但其具體機(jī)制需進(jìn)一步的研究[2,23]。也有研究表明2型糖尿病患者外周血中Fiaf減少，且與胰島素抵抗相關(guān)[24]。因此，我們就NAFLD患者腸道菌群的變化，及其是否與Fiaf因子、胰島素抵抗有關(guān)進(jìn)行初步研究。 目的：檢測NAFLD患者腸道菌群的變化，測定血漿Fiaf因子的水平，分析腸道菌群與Fiaf因子、胰島素抵抗的相關(guān)性，探討腸道菌群在非NAFLD發(fā)生發(fā)展中的作用。 方法：本研究共收集在我院門診和體檢部經(jīng)臨床、實(shí)驗(yàn)室、B超確診為非酒精性脂肪型肝病患者30例（病例組），在我院門診體檢結(jié)果為健康并且愿意接受實(shí)驗(yàn)的成年人30例（對照組）。NAFLD診斷標(biāo)準(zhǔn)參照2010年中華醫(yī)學(xué)會肝臟病學(xué)分會脂肪肝和酒精性肝病學(xué)組制定的非酒精性脂肪性肝病診療指南[3]。所有研究對象均除外近期使用腸道微生態(tài)制劑、酸奶、使用抗生素等可能影響腸道菌群的制劑。常規(guī)測量兩組的身高、體重、腹圍，計(jì)算BMI值。選擇腸道菌群中5種具有代表性的細(xì)菌進(jìn)行培養(yǎng)，分別計(jì)數(shù)病例組和對照組腸道菌群中這5種細(xì)菌的數(shù)量，用以評價腸道菌群的變化狀況。收集所有研究對象清晨空腹抽取的靜脈血血清，采用全自動生化分析儀測定血清谷丙轉(zhuǎn)氨酶（ALT）、谷草轉(zhuǎn)氨酶(AST)、總膽固醇（CHOL）、甘油三酯（TG）、低密度脂蛋白（LDL）、空腹血糖（FPG）以及空腹胰島素（FINS）水平。使用平衡穩(wěn)態(tài)模型的評價法，計(jì)算胰島素抵抗指數(shù)（homeostatic model assessment-insulin resistance,HOMA-IR），評估胰島素抵抗程度。通過RT-PCR方法檢測兩組血漿中Fiaf的mRNA的表達(dá)。根據(jù)檢測結(jié)果，分析腸道菌群數(shù)量、Fiaf的含量、HOMA-IR的相關(guān)性。 結(jié)果:1兩組腸道菌群數(shù)量比較：與對照組相比，NAFLD組變形桿菌和腸球菌數(shù)量明顯增加（9.20±0.20vs8.73±0.41，7.70±0.55vs7.18±0.36，P＜0.05），雙歧桿菌、擬桿菌、乳桿菌數(shù)量明顯減少（8.89±0.28vs9.16±0.22，7.29±0.36vs7.94±0.15，9.21±0.36vs9.76±0.22，P＜0.05）。2兩組年齡、性別、BMI、腹圍比較：兩組在年齡、性別上差異無意義（P0.05），相比正常對照組，NAFLD組研究對象的腹圍、BMI均明顯升高（90.5±8.86vs81.67±8.89，27.42±3.14vs24.22±2.37，P＜0.05）。3兩組血清學(xué)指標(biāo)比較：與對照組相比，病例組ALT（37.75±29.93vs11.17±1.72）、AST（27.96±18.99vs10.83±1.47）、CHOL（5.17±0.80vs4.44±0.54）、TG（2.16±1.04vs0.94±0.41）水平明顯升高，，均P＜0.05；病例組Fiaf水平明顯降低（1.21±0.05vs1.26±0.03，P＜0.05），病例組HOMA-IR明顯升高（4.91±1.21vs2.86±0.51，P＜0.05）。兩組LDL差異無統(tǒng)計(jì)學(xué)意義。4相關(guān)性分析：5種腸道菌群中只有變形桿菌數(shù)量與血清FiafmRNA表達(dá)水平呈顯著性負(fù)相關(guān)（r=-0.734，P0.01），血漿FiafmRNA表達(dá)水平和HOMA-IR呈顯著性負(fù)相關(guān)（r=-0.819，P0.01），變形桿菌數(shù)量和HOMA-IR呈正相關(guān)（r=0.614，P0.01）。 結(jié)論：1NAFLD患者存在腸道菌群的失調(diào)，表現(xiàn)為雙歧桿菌、擬桿菌、乳桿菌減少，變形桿菌、腸球菌增加。2變形桿菌數(shù)量與Fiaf因子的表達(dá)水平呈顯著性負(fù)相關(guān)，與HOMA-IR呈正相關(guān)，而Fiaf因子的表達(dá)水平與HOMA-IR呈顯著性負(fù)相關(guān)。3表明在NAFLD的發(fā)病過程中，腸道菌群失調(diào)可能通過抑制Fiaf因子的表達(dá)引起IR，參與NAFLD的發(fā)生發(fā)展。
[Abstract]:BACKGROUND: Non-alcoholic fatty liver disease (NAFLD) is a clinicopathological syndrome characterized by diffuse hepatocyte bullous steatosis, which is caused by the removal of ethanol and other definite liver damage factors. Steatohepatitis, fatty liver cirrhosis.
Nowadays, the pathogenesis of NAFLD has not been fully clarified. Current studies suggest that the pathogenesis of NAFLD is closely related to insulin resistance, abnormal lipid metabolism and lipid peroxidation. Receiving and storing, promoting the formation of obesity and insulin resistance and other mechanisms affect energy metabolism, participate in the occurrence and development of NAFLD.
Fasting-induced adipose factor (Fiaf) gene is mainly responsible for encoding lipoprotein lipase (LPL) inhibitors, LPL can hydrolyze triglyceride in lipoprotein particles, promote lipid deposition. Fiaf is by inhibiting LPL, thereby inhibiting the triglyceride cycle. Studies have shown that in animals, Fiaf Enterobacteriaceae can inhibit the expression of Fiaf, increase the expression of LPL and lead to lipid deposition, but the specific mechanism needs further study [2,23]. Some studies have shown that the decrease of Fiaf in peripheral blood of patients with type 2 diabetes mellitus is related to insulin resistance [24]. Whether it is related to Fiaf factor and insulin resistance is preliminarily studied.
Objective: To detect the changes of intestinal flora in patients with NAFLD, determine the level of plasma Fiaf factor, analyze the correlation between intestinal flora and Fiaf factor, insulin resistance, and explore the role of intestinal flora in the development of non-NAFLD.
Methods: A total of 30 cases of non-alcoholic fatty liver disease (case group) diagnosed by clinical, laboratory and B-mode ultrasound were collected in the outpatient and physical examination department of our hospital. 30 healthy adults (control group) were willing to accept the experiment. The diagnostic criteria of NAFLD were referred to fatty liver disease of Chinese Medical Association in 2010. All subjects except those who had recently used intestinal microbial agents, yoghurt, antibiotics and other agents that may affect intestinal flora. The height, weight, abdominal circumference of the two groups were routinely measured and BMI values were calculated. Five representative bacteria were selected from the intestinal flora. The bacteria were cultured and counted to evaluate the changes of intestinal flora in the case group and the control group. The venous blood serum was collected from all the subjects on an empty stomach in the morning. The serum ALT, AST, CHOL, glycerol were measured by automatic biochemical analyzer. Levels of triglyceride (TG), low density lipoprotein (LDL), fasting blood glucose (FPG) and fasting insulin (FINS) were measured. Home ostatic model assessment-insulin resistance (HOMA-IR) was used to evaluate insulin resistance. The expression of Fiaf mRNA in plasma was detected by RT-PCR. According to the test results, the number of intestinal flora, the content of Fiaf and the correlation of HOMA-IR were analyzed.
Results: 1. The number of intestinal flora in NAFLD group was significantly higher than that in control group (9.20.20 vs 8.73.41, 7.70.55 vs 7.18.36, P < 0.05). The number of bifidobacteria, Bacteroides and Lactobacillus decreased significantly (8.89.28 vs 9.16.22, 7.29.36 vs 7.94.15, 9.21.36 vs 9.76.22, P < 0.05). Age, sex, BMI, abdominal circumference: There was no significant difference in age and sex between the two groups (P 0.05). Compared with the normal control group, the abdominal circumference and BMI of the NAFLD group were significantly higher (90.5 The levels of AST, AST, AST, AST, CHOL, CHOL, CHOL, CHOL, CHOL, CHOL, CHOL (5.17 (5.17 (0.80 vs 4.44 (0.44 (0.54), TG (2.16 (1.16 (1.04vs 0.94 (.94 (0.41) 41) were significantly increased (P < 0.05), Fiaflevels in case group were significantly decreased (1.21 ((1.21 (0.0.05vs 1.26 (.03), P < 0.03) and HOMA-IR was significantly increased (4.91 (4.91 (1.91 (1.21 1.21 1.21 1.21 vs 2.21 21.21.21 vs 2.21.86 In the meantime, it is necessary to study the relationship between the two. Analysis: Among the five intestinal flora, only Proteus was negatively correlated with serum Fiaf mRNA expression (r = - 0.734, P 0.01), plasma Fiaf mRNA expression was negatively correlated with HOMA-IR (r = - 0.819, P 0.01), and the number of Proteus was positively correlated with HOMA-IR (r = 0.614, P 0.01).
CONCLUSION: 1. The intestinal flora of NAFLD patients is imbalance, showing bifidobacterium, bacteroides, Lactobacillus decrease, Proteus and Enterococcus increase. 2. The number of Proteus is negatively correlated with the expression level of Fiaf factor, and positively correlated with HOMA-IR, while the expression level of Fiaf factor is negatively correlated with HOMA-IR. During the course of disease, intestinal flora disorder may induce IR by inhibiting the expression of Fiaf factor and participate in the occurrence and development of NAFLD.
【學(xué)位授予單位】：河北醫(yī)科大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2014
【分類號】：R575.5

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