本文關(guān)鍵詞： 二十碳五烯酸 白蛋白 過氧化物酶體增殖物激活受體-γ 整合素β1 整合素連接激酶　出處：《山東大學(xué)》2014年碩士論文　論文類型：學(xué)位論文

【摘要】：研究背景與目的 在各種慢性腎臟病進(jìn)展中,腎臟纖維化是其共同的病理基礎(chǔ),也是導(dǎo)致終末期腎病的主要原因。。腎臟纖維化的病理改變主要包括腎小球硬化和小管間質(zhì)纖維化。腎小管上皮細(xì)胞(human proximal tubular cells, HK-2)損傷并分泌大量致炎因子、促纖維形成因子和基質(zhì)蛋白等,形成小管間質(zhì)損傷的惡性循環(huán),是引起腎間質(zhì)纖維化的主要原因。整合素連接激酶(integrin-linked kinase, ILK)、整合素β1(integrin β1)、過氧化物酶體增殖物激活受體γ(peroxisome proliferators-activated receptor gamma, PPARy)等與腎臟纖維化的進(jìn)展有著十分密切的關(guān)系。多不飽和脂肪酸(polyunsaturated Fatty Acids,PUFA)是哺乳動物體內(nèi)的必需脂肪酸。但是其體內(nèi)不能自身合成PUFA,只能從食物中攝取。食品來源的多不飽和脂肪酸有Ω-3(n-3PUFA和Ω-6(n-6)PUFA兩種類型。從深海魚類中提取的魚油,含豐富的n-3PUFA,二十二碳五烯酸(eicosapentaenoic acid, EPA)和二十二碳六烯酸(docosahexaenoic acid, DHA)是其主要活性成分。研究發(fā)現(xiàn)魚油除具有如抗血栓、調(diào)血脂等作用外,還對腎臟有一定的保護(hù)作用,延緩腎臟疾病的進(jìn)展。 本實(shí)驗(yàn)通過白蛋白致腎小管上皮細(xì)胞損傷模型觀察EPA對HK-2ILK,整合素β1、PPARy表達(dá)的影響,探究EPA對腎臟的保護(hù)作用機(jī)制,為臨床應(yīng)用提供理論依據(jù)。 方法 取對數(shù)生長期HK-2,常規(guī)培養(yǎng)24h后,換無血清培養(yǎng)24h,使細(xì)胞同步于GO期。分為：正常對照組；白蛋白組(模型組,白蛋白20.0mg/L); EPA干預(yù)組(白蛋白20.0mg/L+EPA10μmol/L或100μmol/L)。各組分別培養(yǎng)24h、48h,72h后,進(jìn)行指標(biāo)測定。 1采用Western Blot法觀察EPA對白蛋白刺激HK-2PPARγ、ILK和整合素β1表達(dá)的影響； 2采用RT-PCR法觀察EPA對白蛋白刺激HK-2PPARγ、ILK和整合素p1mRNA表達(dá)的影響。 結(jié)果 1.EPA對白蛋白刺激HK-2細(xì)胞PPARy表達(dá)的影響 白蛋白可使HK-2細(xì)胞PPARy mRNA和蛋白表達(dá)下降；EPA可增加白蛋白誘導(dǎo)的HK-2細(xì)胞PPARy mRNA和蛋白的表達(dá)。 2.EPA對白蛋白刺激HK-2細(xì)胞ILK表達(dá)的影響 白蛋白可增加HK-2細(xì)胞ILK mRNA和蛋白的表達(dá)；EPA可使白蛋白誘導(dǎo)的HK-2細(xì)胞ILK mRNA和蛋白表達(dá)下降。 3.EPA對白蛋白刺激HK-2細(xì)胞整合素β1mRNA表達(dá)的影響 白蛋白可使HK-2細(xì)胞整合素β1mRNA的表達(dá)下降；EPA可增加白蛋白誘導(dǎo)的HK-2細(xì)胞整合素β1mRNA表達(dá)。 結(jié)論 1.EPA能夠促進(jìn)白蛋白誘導(dǎo)的HK-2細(xì)胞PPARy蛋白和基因的表達(dá)。 2.EPA可以抑制白蛋白誘導(dǎo)的HK-2細(xì)胞ILK蛋白和基因的表達(dá)。 3.EPA可以促進(jìn)白蛋白誘導(dǎo)的HK-2細(xì)胞整合素β1蛋白和基因的表達(dá)。
[Abstract]:Research background and purpose. Among the progress of chronic kidney disease, renal fibrosis is the common pathological basis. The pathological changes of renal fibrosis mainly include glomerulosclerosis and tubulointerstitial fibrosis. Fiber forming factors and matrix proteins form a vicious circle of tubulointerstitial injury. Integrin linked kinase (ILKN), integrin 尾 1 integrin 尾 1 (integrin 尾 1), peroxisome proliferators-activated receptor gamma (PPA Ryma) are closely related to the progression of renal fibrosis. Polyunsaturated fatty acids (PUFAs) are essential fatty acids in mammals, but they cannot synthesize PUFAs by themselves and can only be ingested from foods. There are two types of polyunsaturated fatty acids from food sources: 惟 -3n -3PUFA and 惟 -6n -6 PUFA. Fish oil extracted from deep-sea fish, Rich n-3 PUFAA, 22 carbapentaenoic acid acidand 22 carbohexaenoic acid acid. it is found that fish oil not only has some protective effects on kidney, but also has antithrombotic and lipid effects. Delay the progression of kidney disease. In this study, we observed the effects of EPA on the expression of HK-2 ILK and integrin 尾 1 PPARy in renal tubular epithelial cells induced by albumin, and explored the protective mechanism of EPA on kidney, which provided theoretical basis for clinical application. Method. The logarithmic growth phase HK-2 was taken for 24 hours after conventional culture, and the cells were cultured in go phase for 24 hours. The cells were divided into three groups: normal control group, albumin 20.0 mg / L group (model group), EPA intervention group (20.0 mg / L EPA10 渭 mol/L or 100 渭 mol / L mol/L or 100 渭 mol / L EPA) for 24 h and 48 h, respectively. The index was measured. 1the effect of EPA on the expression of HK-2PPAR 緯 -ilk and integrin 尾 1 stimulated by albumin was observed by Western Blot assay. 2the effect of EPA on the expression of HK-2PPAR 緯 -ilk and integrin p1 mRNA was observed by RT-PCR assay. Results. 1. The effect of EPA on the expression of PPARy in HK-2 cells stimulated by albumin. Albumin can decrease the expression of PPARy mRNA and protein in HK-2 cells. It can increase the expression of PPARy mRNA and protein in HK-2 cells induced by albumin. 2. The effect of EPA on the expression of ILK in HK-2 cells stimulated by Albumin. Albumin could increase the expression of ILK mRNA and protein in HK-2 cells. EPA could decrease the expression of ILK mRNA and protein in HK-2 cells induced by albumin. 3. Effect of EPA on integrin 尾 1 mRNA expression in HK-2 cells stimulated by albumin. Albumin can decrease the expression of integrin 尾 1 mRNA in HK-2 cells. EPA can increase the expression of integrin 尾 1 mRNA in HK-2 cells induced by albumin. Conclusion. 1. EPA could promote the expression of PPARy protein and gene in HK-2 cells induced by albumin. 2. EPA could inhibit the expression of ILK protein and gene in HK-2 cells induced by albumin. 3. EPA could promote the expression of integrin 尾 1 protein and gene in HK-2 cells induced by albumin.
【學(xué)位授予單位】：山東大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2014
【分類號】：R692

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