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《溫莪術(shù)對(duì)tgf-β1誘導(dǎo)腎小管上皮細(xì)胞轉(zhuǎn)分化的干預(yù)作用》由會(huì)員上傳分享,免費(fèi)在線閱讀,更多相關(guān)內(nèi)容在學(xué)術(shù)論文-天天文庫。
1、論文題目:漁董苤盟!魚星:墾:!透昱豎叢!笪土廑細(xì)胞裝盆絲的王亟佳昱答辯委員會(huì)主席:衄童登塾援答辯委員會(huì)成員:衄童登熬援塞4毅主焦醫(yī)婭董玉迭主任蟲醫(yī)垣2量攪醫(yī)堂瞳亟±堂焦迨塞lI哪10哪帥㈣㈣吣刪100IY2397504目錄縮略詞表???···??···???????·??軍中文摘要???····???·??????··??多Abstract????··??????????··?·7前言??....????....???..???·??彤剛吾??一一????一一???一???一?一刀材料與方法?????··???????·??··侈實(shí)驗(yàn)結(jié)果???.···????????··??
2、?·功分析與討論???????????????··力結(jié)論???????????????··??.形參考文獻(xiàn)??????????????·??韶附錄?????????????????··彩致謝?????????????????··衫綜述?????????????????··彩獨(dú)創(chuàng)性聲明?????????·???·····?··彭全文縮略詞表AGEadvancedglycationendproducts終末期糖基化產(chǎn)物AngIIAngiotensinII血管緊張素UCKDChronicKidneyDisease慢性腎臟病Co卜ICollagenII型膠原蛋白DMSODimethyl
3、Sulfoxide二甲基亞砜ESRDendstagerenaldisease終末期腎病ECMExtracellularmatrix細(xì)胞外基質(zhì)EMTEpithelial—myofibroblast上皮一間充質(zhì)細(xì)胞轉(zhuǎn)分化transdifferentiationE-cadE-cadherinE一鈣粘素Fsplfibroblastspecificproteinl成纖維細(xì)胞特異性蛋白lFNFibronectin纖維連接蛋白HGFhepatocytegrowthfactor肝細(xì)胞生長因子IL一1Interleukin1白細(xì)胞介素一1ILKIntegrin—linkedkinase整合素連接
4、激酶MFMyofibroblast肌成纖維細(xì)胞MMP一2Matrixmetalloproteinase一2基質(zhì)金屬蛋白酶一2MMP一9Matrixmetalloproteinase-9基質(zhì)金屬蛋白酶一9NRK一52ENormalratkidneycell大鼠腎小管上皮細(xì)胞ODOpticaldensity光密度PKCproteinkinaseC蛋白激酶CPBSPhosphatebuffersolution磷酸鹽緩沖液RASRenin—angiotensinsystem腎素一血管緊張素系統(tǒng)RIFRenalinterstitialfibrosis腎小管間質(zhì)纖維化TGF—pTransf
5、orminggrowthfactor—B轉(zhuǎn)化生長因子一BTBMTubularbasementmembrane腎小管基底膜UUOUnilateralureteralobstruction單側(cè)輸尿管梗阻Zo‘1Zonaoccludensprotein1緊密連接蛋白一1按首字母順序排列4溫莪術(shù)對(duì)TGF.13,誘導(dǎo)腎小管上皮細(xì)胞轉(zhuǎn)分化的干預(yù)作用中文摘要目的:本研究在體外建立大鼠腎小管上皮細(xì)胞轉(zhuǎn)分化(Epithelial-myofibroblasttransdifferentiation)模型,通過檢測(cè)細(xì)胞骨架蛋白岱.a(chǎn)ctin,上皮細(xì)胞黏附分子E.cad,纖維化標(biāo)志a.SMA、ILK、
6、FN、C01.I等蛋白及基因的表達(dá),探討溫莪術(shù)對(duì)EMT的干預(yù)作用及可能的作用機(jī)制。方法:通過MTr法選取合適的溫莪術(shù)含藥血清給藥濃度后,將體外培養(yǎng)的正常大鼠腎小管上皮細(xì)胞(NRK.52E)隨機(jī)分為6組:正常對(duì)照組、TGF.p1誘導(dǎo)模型組、溫莪術(shù)低濃度組、溫莪術(shù)中濃度組、溫莪術(shù)高濃度組、鹽酸貝那普利組,其中溫莪術(shù)及鹽酸貝那普利均以體外制作的含藥血清方式給藥。除正常對(duì)照組外,各組轉(zhuǎn)化生長因子.131(Transforminggrowthfactor-13l,TGF.131)以10ng/mL的濃度誘導(dǎo)24h后,溫莪術(shù)組及鹽酸貝那普利組再加入藥物作用48h。運(yùn)用倒置顯微鏡觀察各組細(xì)胞形
7、態(tài)學(xué)的改變,細(xì)胞免疫熒光法檢測(cè)各組細(xì)胞漿內(nèi)骨架成分B.肌動(dòng)蛋白(B.a(chǎn)ctin),纖維化標(biāo)志物a.平滑肌肌動(dòng)蛋白(a.smoothmuscleactin,a—SMA)的分布;用實(shí)時(shí)熒光定量PCR法檢測(cè)上皮細(xì)胞黏附分子E鈣黏蛋白(E.cadherin,E.cad)mRNA、及細(xì)胞內(nèi)a.SMA、整合素連接激酶(Integrin.1inkedkinase,ILK)、纖維黏連蛋白(fibronectin,F(xiàn)N)、I型膠原蛋白(CollagenI,C01.I)的mRNA的表達(dá);用ELISA法檢測(cè)各組細(xì)