本文關(guān)鍵詞： 糖尿病腎病 雷公藤多苷 氧化應激　出處：《新鄉(xiāng)醫(yī)學院》2014年碩士論文　論文類型：學位論文

【摘要】：背景糖尿病腎病(diabetic nephropathy, DN)為糖尿病最常見的慢性并發(fā)癥之一,是引起發(fā)達國家終末期腎衰竭的主要原因,腎小球細胞外基質(zhì)(extracelluar matrix,ECM)增生、基底膜增厚,’腎小管間質(zhì)纖維化是其基本病理特征,最終可導致腎衰竭。近年來臨床和研究證實,DN不僅在成人中的發(fā)病率增高,而且在兒童中的總體發(fā)病趨勢也明顯增加,因此兒童糖尿病腎病已成為嚴重威脅著兒童身心健康的代謝性疾病。但其具體機制尚不明確,近年來研究發(fā)現(xiàn)氧化應激(oxitative stress,OS)在DN發(fā)生、發(fā)展中發(fā)揮著重要作用。雷公藤多苷(tripterygium wilfordii polyglycoside, TWP)是從雷公藤植物中提取的總甙,其主要成分為環(huán)氧二萜內(nèi)酯類化合物。研究表明TWP可通過抑制細胞和體液免疫,從多個環(huán)節(jié)抑制免疫應答過程。李軍華等[[24]研究報道,TWP對腸黏膜組織具有抗氧化作用,那么TWP對腎組織是否具有抗氧化作用經(jīng)查新國內(nèi)尚未見報道。本研究以此為切入點,在建立DN大鼠模型的基礎(chǔ)上,分析TWP對DN大鼠腎臟氧化應激反應的影響,研究TWP對DN大鼠腎臟的保護作用,為TWPDN提供實驗數(shù)據(jù)和理論基礎(chǔ)。 目的本研究擬通過建立DN大鼠模型,通過觀察TWP對DN大鼠飲食、飲水、毛發(fā)、體重、精神狀態(tài)等一般狀況及血糖、肌酐清除率(creatinine clearance,Ccr)、尿素氮(blood urea nitrogen,BUN)、24h尿微量白蛋白排泄率(urinary albumin excretion rate,UAER)等生化指標的影響,分析TWP對DN大鼠腎組織丙二醛(Malondialdehyde, MDA)含量和谷胱甘肽過氧化物酶(glutathione peroxidase, GSH-Px)活性及血清過氧化氫酶(catalase, CAT)活性和血清超氧陰離子(superoxide oxygen anion, O2-)水平的影響,觀察TWP對DN大鼠腎臟病理改變的影響,探討TWP對DN大鼠腎臟保護作用,為TWP治療DN提供實驗數(shù)據(jù)和理論依據(jù)。 方法85只健康雄性(Sprague-Dawley,SD)大鼠,隨機分為正常組(13只)和DN大鼠模型制作組(72只),模型制作組大鼠按52mg-kg-1左下腹腔內(nèi)一次性注射(ip)鏈脲佐菌素(Streptozotocin,STZ)建立DN大鼠模型。3周后造模成功后,隨機將DN大鼠(56只)分為模型對照組和TWP (4.5,9.0、18.0mg·kg-1)藥物治療組,每組14只,TWP治療組給予灌胃(ig),每天1次,連續(xù)8周,等體積飲用水灌胃給予正常組和模型對照組。實驗過程中觀察大鼠的飲食、尿量、體重、毛發(fā)、精神狀況等一般情況。實驗結(jié)束前2d將大鼠置代謝籠,收集24h尿液,用于檢測24h尿白蛋白排泄率。用藥8周末,大鼠稱重,10%水合氯醛麻醉大鼠,心臟取血離心,檢測肌酐清除率(creatinine clearance, Ccr)、尿素氮(blood urea nitrogen,BUN),可見光方法檢測血清CAT活性,比色法測定血清O2-水平。取雙腎,稱重,左腎制成10%勻漿,比色法測定GSH-Px活性,硫代巴比妥酸縮合法檢MDA含量；右腎切成8mm3小塊,甲醛固定,切片HE染色觀察病理學改變。 結(jié)果 1.成功建立了DN大鼠模型。 2.TWP對DN大鼠一般狀況的影響用藥4周后,TWP9.0和18.0mg·kg-1組大鼠進食、飲水、尿量、精神等一般狀況有所改善,用藥8周后上述現(xiàn)象逐漸恢復正常。 3.TWP對各組大鼠體質(zhì)量、腎質(zhì)量、腎質(zhì)量/體質(zhì)量的影響與正常組相比,模型對照組大鼠腎質(zhì)量、腎質(zhì)量/體質(zhì)量顯著升高(P0.01),體質(zhì)量顯著降低(P0.01)；與模型對照組相比,TWP9.0和18.0mg·kg-1治療組大鼠腎質(zhì)量、腎質(zhì)量/體質(zhì)量顯著降低(P0.01),體質(zhì)量顯著升高(P0.01)。 4.TWP對DN大鼠血糖、血BUN、Ccr和24hUAER變化的影響與正常組相比,模型對照組大鼠血糖、24hUAER、血BUN較其同時期顯著升高(P0.01),Ccr顯著降低(P0.01)；與模型對照組相比,TWP9.0和18.0mg·kg-1組大鼠24hUAER、血BUN、血糖較其同時期顯著下降(P0.01),血Ccr顯著升高(P0.01)。 5.TWP對DN大鼠血清CAT活性和O2-含量及腎組織GSH-Px活性和MDA含量的影響與正常組相比,模型對照組腎組織中GSH-Px和血清CAT活性顯著降低(p0.01),血清O2-和腎組織MDA水平顯著上升(p0.01)；與模型對照組相比,TWP9.0和18.0mg·kg-1組血清CAT活性和腎組織中GSH-Px活性明顯升高(p0.01),各組間CAT活性和GSH-Px活性差異均有統(tǒng)計學意義(p0.05)；與模型對照組相比,TWP9.0和18.0mg·kg-1組血清O2-水平和腎組織MDA含量明顯降低(p0.01)。 6.TWP對DN大鼠腎臟病理學改變的影響與正常組相比,模型組大鼠腎小球肥大、系膜細胞增生、腎小球變形；與模型對照組相比,TWP4.5、9.0、18.0mg·kg-1治療組上述病理狀況明顯改善。 結(jié)論 1.TWP9.0和18.0mg·kg-1可使DN大鼠BUN、腎重/體重及UAER均顯著降低,Ccr明顯升高,其病理改變也明顯減輕,表明TWP對DN大鼠腎臟有一定保護作用。 2.TWP對DN大鼠腎臟的保護機制之一可能與其增高血清CAT活性和腎組織GSH-Px活性,降低血清O2-水平和腎組織MDA含量,抑制氧化應激、增強機體抗氧化能力有關(guān)。
[Abstract]:Background diabetic nephropathy (diabetic nephropathy DN) is one of the most common chronic complications of diabetes, is a major cause of end-stage renal failure in developed countries, glomerular extracellular matrix (extracelluar, matrix, ECM) hyperplasia, basement membrane thickening, renal tubulointerstitial fibrosis is the basic pathologic feature, may eventually lead to kidney failure. Confirmed by clinical and research in recent years, DN not only increased the incidence in adults, and the overall incidence in children has increased significantly, so the children of diabetic nephropathy has become a serious threat to children's physical and mental health of metabolic diseases. But the mechanism is not clear, recent studies have found that oxidative stress (oxitative stress OS), in DN, plays an important role in the development of tripterygium glycosides (Tripterygium wilfordii, polyglycoside, TWP) is extracted from Tripterygium Glycosides in plants, the main Divided into two epoxy terpenoid lactone. The results show that TWP can inhibit cellular and humoral immunity, inhibit the immune response from many aspects. Li Junhua [[24] reported that TWP had antioxidant effect on the intestinal mucosa, then whether TWP has antioxidant on renal tissue for the investigation of new is not reported in China. Based on this as a starting point, based on the establishment of DN rat model, to analyze the effect of TWP on renal oxidative stress response in DN rats and protective effects of TWP on the kidney of DN rats, and provide experimental data and theoretical basis for TWPDN.
Purpose of this study is to establish the rat model of DN, the effect of TWP on DN rat diet, drinking water, hair, body weight, general condition and mental state of blood glucose, creatinine clearance rate (creatinine, clearance, Ccr), blood urea nitrogen (blood urea nitrogen, BUN 24h), urinary albumin excretion rate (urinary albumin excretion rate, UAER) of biochemical indicators, analysis of TWP on the content of renal tissue in DN rats (Malondialdehyde, MDA) content and glutathione peroxidase (glutathione peroxidase, GSH-Px) and the activity of serum catalase (catalase, CAT) and the activity of serum superoxide anion (superoxide oxygen, anion, O2-) level, the effect of TWP the renal pathological changes in DN rats, to investigate the renoprotective effects of TWP in DN rats, provide experimental data and theoretical basis for the treatment of DN TWP.
鏂規(guī)硶85鍙仴搴烽泟鎬，

本文編號：1550729