發(fā)布時間：2018-03-11 09:22

  本文選題：高脂血癥　切入點：胰島素抵抗　出處：《承德醫(yī)學院》2017年碩士論文　論文類型：學位論文

【摘要】：目的:糖尿病腎病(Diabetic Nephropathy,DN)是糖尿病(DM)常見的慢性并發(fā)癥,亦是引起終末期腎病(end-stage renal disease,ESRD)的重要原因。其發(fā)病機制復雜,近年來發(fā)現(xiàn)炎癥反應在DN中發(fā)揮重要作用。TNF-α及IL-6是主要的炎性標記物,而調控和轉錄很多炎癥因子表達的核轉錄因子-κB(Nuclear factor-kappa B,NF-κB)是聯(lián)系肥胖、炎癥的重要機制。DN還存在不同程度的糖脂代謝紊亂,常協(xié)同致病,其中也與炎癥密不可分。脂質代謝紊亂能誘導細胞產(chǎn)生炎癥反應,繼而發(fā)生胰島素抵抗(insulin resistance,IR),IR可刺激多種炎性細胞因子釋放,加重腎小球肥大,此外,IR帶來的高胰島素血癥還可使腎小球血流量增加,腎小球毛細血管靜水壓升高,處于高灌注狀態(tài),進而引起蛋白尿。過多的脂質在腎小球沉積,還可導致腎小球硬化。GLP-1受體激動劑(Glucagon like peptide-1 receptor agonist)利拉魯肽是目前治療糖尿病的新藥,研究發(fā)現(xiàn)該藥可改善IR、減輕體重,并可降低糖尿病大鼠尿蛋白、減輕腎小球高濾過和腎臟過度肥大從而改善腎功能,但具體機制不詳。本研究通過觀察高脂喂養(yǎng)IR大鼠腎功能、尿蛋白、血脂,血清及腎組織中NF-κB、TNF-α及IL-6的表達,探討糖尿病前期IR階段腎臟炎癥因子的表達變化,及與血脂等的相關性,并用利拉魯肽干預治療探討利拉魯肽對腎臟炎癥進程的影響及機制,為其在糖尿病腎病及其他腎臟疾病中的應用提供理論依據(jù)。方法:成年雄性Wistar大鼠54只(230-260g),隨機分為對照組(C)普通飼料喂養(yǎng)16只與模型組高脂飼料喂養(yǎng)38只,8周后,各組取5只大鼠行高胰島素-正常葡萄糖鉗夾試驗計算葡萄糖輸注率(glucose infusion rate,GIR),評價胰島素抵抗,判定造模成功后,將模型組隨機分為高脂組(H)、利拉魯肽低劑量組(Li-1)(100μg/kg/d)和利拉魯肽高劑量組(Li-2)(200μg/kg/d),每組11只。前兩組用生理鹽水皮下注射;后兩組分別用利拉魯肽100μg/kg/d及200μg/kg/d皮下注射。干預2周后,每組再取5只大鼠行鉗夾試驗計算葡萄糖輸注率。每組剩余的6只大鼠用代謝籠收集24 h尿液,后腹主動脈放血處死,取腎臟標本行透射電鏡觀察腎臟病理結構以及應用Real time-PCR技術測定腎組織NF-κB、TNF-α及IL-6m RNA表達,自動生化分析儀檢測血肌酐(Scr)、尿素氮(BUN)、總膽固醇(TCH)、甘油三酯(TG)、低密度脂蛋白膽固醇(LDL-C),免疫比濁法測定24h尿微量白蛋白(MAU),Elisa法測定血清NF-κB、TNF-α及IL-6蛋白濃度。結果:1.四組大鼠一般情況比較與對照組相比,高脂組大鼠Scr、BUN、MAU、TCH、TG、LDL-C均明顯升高(P0.05);與高脂組相比,利拉魯肽低劑量組大鼠Scr、BUN、MAU、TG、LDL-C均降低(P0.05),利拉魯肽高劑量組大鼠Scr、BUN、MAU、TCH、TG、LDL-C明顯降低(P0.05);與利拉魯肽低劑量組相比,利拉魯肽高劑量組Scr、BUN、MAU、TG、LDL-C進一步降低(P0.05)。2.四組大鼠葡萄糖輸注率比較藥物干預前:與對照組GIR(30.21±2.51mg/kg.min)相比,高脂組GIR(20.41±3.64mg/kg.min)下降,差異有統(tǒng)計學意義(P0.05)。利拉魯肽干預2周后:與對照組GIR(30.77±2.37mg/kg.min)相比,高脂組GIR(18.83±1.95 mg/kg.min)下降,差異有統(tǒng)計學意義(P0.05)。與高脂組相比,利拉魯肽低劑量組GIR(21.55±2.46mg/kg.min)升高(P0.05),與低劑量組相比,利拉魯肽高劑量組GIR(25.80±1.40mg/kg.min)進一步升高,差異均有統(tǒng)計學意義(P0.05)3.四組大鼠血清NF-κB、TNF-α和IL-6蛋白濃度的比較與對照組相比,高脂組血清NF-κB、TNF-α和IL-6蛋白濃度均升高(P0.05);與高脂組相比,利拉魯肽干預組NF-κB、TNF-α和IL-6蛋白濃度均降低(P0.05);且利拉魯肽高劑量組比利拉魯肽低劑量組,NF-κB、TNF-α和IL-6蛋白濃度進一步降低(P0.05)。4.四組大鼠腎臟組織NF-κB、TNF-α和IL-6 m RNA的比較與對照組相比,高脂組腎臟組織NF-κB、TNF-α和IL-6 m RNA的表達均升高(P0.05);與高脂組相比,利拉魯肽干預后腎臟組織NF-κB、TNF-α和IL-6 m RNA的表達均降低(P0.05);利拉魯肽高劑量組與利拉魯肽低劑量組相比,腎臟組織NF-κB、TNF-α和IL-6 m RNA的表達進一步降低(P0.05)。5.四組大鼠腎臟組織電鏡超微結構比較透射電鏡結果顯示(放大10000倍):對照組大鼠腎小球結構完整,基底膜(GBM)清晰,足突分布整齊、間隙明顯;高脂組大鼠腎小球足突腫脹,部分融合、脫落,基底膜增厚;利拉魯肽干預可改善上述改變,尤以利拉魯肽高劑量組明顯。6.相關性分析研究結果血清TNF-α和IL-6蛋白濃度與MAU呈正相關(r值=0.802、0.748,P0.05);腎組織TNF-αm RNA表達與TCH、TG呈明顯的正相關(r值分別為0.878、0.817,P0.05);腎組織IL-6 m RNA表達與TCH、TG呈明顯正相關(r值分別為0.806、0.854,P0.05);腎組織NF-κB m RNA表達與TNF-α和IL-6 m RNA表達呈正相關(r值分別為0.810、0.867,P0.05)。結論:1高脂飲食可使大鼠發(fā)生脂代謝紊亂及胰島素抵抗,并可通過增加腎臟NF-κB和炎癥因子TNF-α和IL-6表達激活炎癥反應,增加微量白蛋白尿,加重腎損傷。2利拉魯肽可呈劑量依賴性減輕脂代謝紊亂,改善胰島素抵抗,減少微量白蛋白尿,保護腎功能。3利拉魯肽可呈劑量依賴性抑制腎臟NF-κB表達,下調腎組織炎癥因子TNF-α和IL-6表達,改善腎功能。
[Abstract]:Objective: diabetic nephropathy (Diabetic Nephropathy DN) is a common chronic complication of diabetes mellitus (DM), is also the cause of end-stage renal disease (end-stage renal, disease, ESRD) is an important cause. Its pathogenesis is complex, in recent years found that inflammation play an important role of.TNF- alpha and IL-6 is the main inflammatory markers in DN in the regulation of transcription and expression of many inflammatory factors of nuclear factor kappa B (Nuclear factor-kappa B, NF- K B) is linked to obesity, inflammation is an important mechanism of.DN glucose and lipid metabolic disorder in different degree, which is often synergistic, and inflammation are inseparable. Lipid metabolism can induce cells to produce inflammatory reaction. Then the occurrence of insulin resistance (insulin, resistance, IR, IR) can stimulate a variety of inflammatory cytokines release, glomerular hypertrophy, in addition, hyperinsulinemia caused by IR can make the glomerular blood flow increased, glomerular Capillary hydrostatic pressure, in a high perfusion state, thereby causing proteinuria. Excessive lipid in glomerular deposition, can also lead to glomerular sclerosis.GLP-1 receptor agonist (Glucagon like peptide-1 receptor agonist) is currently the liraglutide treatment of diabetes drugs, the study found that the drug can improve IR, reduce weight, and can reduce the urinary protein in diabetic rats, reduce glomerular filtration and renal hypertrophy and improve renal function, but the specific mechanism is unknown. This study by observing the renal function, IR rats were fed high fat urine protein, blood lipid, blood and kidney tissue of NF- kappa B, expression of TNF- and IL-6, to investigate the expression of inflammatory factors in Kidney Early IR stage of diabetes mellitus and its correlation with serum lipids, and liraglutide treatment to investigate the effects of liraglutide on renal inflammatory processes and mechanisms, as in diabetic nephropathy and other kidney 鐤劇梾涓殑搴旂敤鎻愪緵鐞嗚渚濇嵁.鏂規(guī)硶:鎴愬勾闆勬，

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