發(fā)布時間：2018-01-19 02:40

  本文關(guān)鍵詞： 固醇調(diào)節(jié)元件結(jié)合蛋白-1c 五層龍 果糖 脂肪肝　出處：《重慶醫(yī)科大學(xué)》2014年碩士論文　論文類型：學(xué)位論文

【摘要】：目的：五層龍?zhí)崛∥铮⊿OR）為治療肥胖和糖尿病的傳統(tǒng)中草藥，這兩種疾病都與脂肪肝密切相關(guān)。已有研究證實SOR具有改善脂肪肝的作用，但是其機(jī)制尚不清楚。本文旨在研究SOR改善高果糖所致大鼠脂肪肝的分子機(jī)制。 方法：選取24只健康SD大鼠隨機(jī)分為4組（n=6），飼養(yǎng)10周：(1)空白對照組:自由飲水；(2)果糖對照組:給予10%果糖水(w/v,每天配制)；(3)SOR低濃度藥物組:果糖溶液+SOR提取物5mg/kg；(4)SOR高濃度藥物組:果糖溶液+SOR提取物20mg/kg。每三天記一次體重，并根據(jù)體重變化調(diào)整用藥組大鼠給藥量。用藥組起始果糖水濃度均為10%，每天記錄攝食量與飲用果糖水量，并計算果糖攝入量，根據(jù)果糖對照組近三天果糖攝入量，調(diào)整果糖水濃度使用藥組大鼠果糖攝入量與果糖對照組一致。于第10周末，夜間禁食14h后，稱重，乙醚麻醉下，摘除眼球取血，分離血漿檢測TC、TG、血糖、NEFA及胰島素含量；處死大鼠，取肝臟稱重，行肝臟H.E染色和油紅O染色，，同時檢測肝臟TG、TC含量；用TRIzol提取肝組織總RNA，逆轉(zhuǎn)錄獲得cDNA，用Real-Time PCR方法檢測肝臟脂質(zhì)合成轉(zhuǎn)錄因子SREBP-1c、ChREBP及其下游相關(guān)基因FAS、ACC-1、SCD-1、LPK和脂質(zhì)氧化相關(guān)基因PPAR-α、CPT-1α、ACO及PPAR-γ、CD36的mRNA水平；提取肝細(xì)胞核蛋白，用Western blot方法檢測調(diào)節(jié)脂質(zhì)合成的兩個關(guān)鍵轉(zhuǎn)錄因子SREBP-1c和ChREBP的蛋白表達(dá)量。實驗數(shù)據(jù)應(yīng)用StatView統(tǒng)計軟件進(jìn)行分析，以均數(shù)±標(biāo)準(zhǔn)誤（x±SEM）表示，t檢驗進(jìn)行兩組數(shù)據(jù)的差異比較。 結(jié)果： 1.生化指標(biāo)：與空白對照組相比，果糖對照組的血漿TC、TG、血糖及胰島素含量明顯增加，而NEFA顯著下降（P＜0.05）；SOR用藥組相對于果糖對照組，血漿TC、TG、血糖、胰島素及NEFA均無明顯改變（P＞0.05）。 2．肝相關(guān)參數(shù)：各組大鼠肝重與肝TC含量無差異（P＞0.05）；但果糖對照組與空白對照組相比，肝重與體重比、肝TG含量明顯升高（P＜0.05）；SOR（20mg/kg）顯著下調(diào)果糖引起的肝TG含量的升高（P＜0.05）。 3.形態(tài)學(xué)顯示：（1）H.E染色顯示，果糖對照組肝細(xì)胞空泡化的程度較空白對照組明顯增強(qiáng)，SOR（20mg/kg）明顯改善肝細(xì)胞空泡化的程度。（2）ORO染色顯示，與空白對照組比較，果糖對照組ORO染色陽性面積區(qū)域及陽性面積率明顯增加（P＜0.05），SOR（20mg/kg）具有明顯的改善作用（P＜0.05）。 4. Real-Time PCR結(jié)果：果糖對照組相對于空白對照組，顯著上調(diào)肝臟SREBP-1c、FAS、ACC-1、SCD-1、ChREBP及LPK mRNA的表達(dá)并下調(diào)ACO與CPT-1α mRNA的表達(dá)（P＜0.05），對PPAR-γ、PPAR-α、及CD36mRNA無顯著性影響（P＞0.05）；SOR（20mg/kg）明顯抑制肝臟SREBP-1c、FAS、ACC-1、SCD-1及ChREBP mRNA的過表達(dá)（P＜0.05），但對LPK、PPAR-γ、PPAR-α、ACO、CPT-1α及CD36mRNA無明顯改變（P＞0.05）。 5．Western blot分析顯示：與空白對照組比較，果糖對照組細(xì)胞核SREBP-1c和ChREBP的蛋白含量增加（P＜0.05），SOR（20mg/kg）明顯抑制SREBP-1c核蛋白的表達(dá)（P＜0.05），但沒有顯著改變ChREBP核蛋白含量（P＞0.05）。 結(jié)論 1. SOR（20mg/kg）能夠改善高果糖所致大鼠脂肪肝； 2.肝臟SREBP-1c是SOR改善高果糖誘導(dǎo)所致大鼠脂肪肝的重要分子靶點。
[Abstract]:Objective: five layer (SOR) to extract the Dragon herbs for treatment of obesity and diabetes in the tradition, these two diseases are closely related to fatty liver. It has been proved that SOR can improve the fatty liver function, but the mechanism is not clear. This paper aims to study the molecular mechanism of SOR to improve the fatty liver of rats induced by high fructose.
Methods: 24 healthy SD rats were randomly divided into 4 groups (n=6), fed for 10 weeks: (1) control group: free drinking water; (2) the control group were given 10% fructose fructose water (w/v, daily preparation); (3) SOR of low concentration drug group: fructose solution extract of +SOR 5mg/kg; (4) SOR of high dose group: fructose solution extract of +SOR 20mg/kg. every three days on a weight, and the dosage is adjusted according to the weight changes of the rats in treatment group. Treatment group initial fructose water concentration was 10%, daily food intake and drinking water and fructose, fructose intake is calculated, according to the fructose in the control group three days of fructose intake, adjust the concentration of water use fructose and fructose fructose intake group rats in control group. In the tenth week, the night after fasting 14h, weighing, ether anesthesia, removal of the eye blood plasma separation detection of TC, TG, blood glucose, NEFA and insulin content; the rats were sacrificed and the liver weight Liver, H.E staining and oil red O staining, and detection of liver TG, TC content; the total RNA was extracted from liver tissue by TRIzol, reverse transcription cDNA, using Real-Time PCR method to detect the liver lipogenic transcription factor SREBP-1c, ChREBP and its downstream genes FAS, ACC-1, SCD-1, LPK and lipid oxidation related genes of PPAR- alpha, CPT-1 alpha, ACO and PPAR- CD36 gamma, mRNA levels; from the liver nuclear protein expression detection, regulation of lipid synthesis of two key transcription factor SREBP-1c and ChREBP protein by Western blot method. The experimental data using statistical software StatView analysis, mean standard error (x + SEM) said, t test for the comparison of data between the two groups.
Result錛

本文編號：1442302