本文選題：α-硫辛酸 + 硫酸吲哚酚��； 參考：《河北醫(yī)科大學(xué)》2014年碩士論文

【摘要】：目的:慢性腎臟疾病(chronic kidney disease,CKD)患者尤其是終末期腎衰患者的心血管發(fā)病率和死亡率遠(yuǎn)高于普通人群。加速性動(dòng)脈粥樣硬化是尿毒癥患者心血管并發(fā)癥的主要病理機(jī)制之一，起始于血管內(nèi)皮細(xì)胞損傷。CKD患者隨著腎小球?yàn)V過率降低，腎毒性物質(zhì)在體內(nèi)蓄積，可誘導(dǎo)氧化應(yīng)激反應(yīng),促進(jìn)動(dòng)脈血管產(chǎn)生粥樣硬化病變。硫酸吲哚酚（indoxylsulfate,IS）即是其中一種代表性的蛋白結(jié)合毒素，誘發(fā)氧化應(yīng)激使心血管系統(tǒng)受損并加速慢性腎臟病進(jìn)程。α-硫辛酸(α-lipoic acid,LA)是天然存在的最強(qiáng)抗氧化劑，目前在糖尿病、糖尿病并發(fā)癥、缺血再灌注損傷等疾病的防治方面應(yīng)用較廣泛，而在CKD患者應(yīng)用較少。本研究以人臍靜脈內(nèi)皮細(xì)胞(human umbilical vein endothelial cell，HUVEC)為對(duì)象,探討α-硫辛酸能否對(duì)硫酸吲哚酚致血管內(nèi)皮細(xì)胞產(chǎn)生的氧化損傷起到保護(hù)作用及其可能的作用機(jī)制，，為α-硫辛酸在慢性腎臟病中應(yīng)用提供實(shí)驗(yàn)依據(jù)。 方法:培養(yǎng)人臍靜脈內(nèi)皮細(xì)胞,將其分成對(duì)照組（CONTROL組），α-硫辛酸組（LA組），硫酸吲哚酚組（IS組）和硫酸吲哚酚+α-硫辛酸組（IS+LA組）。用MTS法檢測細(xì)胞存活率；硫代巴比妥酸法測定丙二醛（MDA）的含量；用流式細(xì)胞術(shù)檢測細(xì)胞內(nèi)活性氧自由基(reactiveoxygen species,ROS)的變化；應(yīng)用RT-PCR法測定NADPH氧化酶4（NOX4）mRNA的表達(dá)。 統(tǒng)計(jì)學(xué)處理采用GraphPad Prism5軟件，實(shí)驗(yàn)數(shù)據(jù)中計(jì)量資料用均數(shù)±標(biāo)準(zhǔn)差表示。多組間均數(shù)比較采用單因素方差分析（ANOVA）,均數(shù)兩兩比較采用Bonferroni法檢驗(yàn)，P＜0.05具有統(tǒng)計(jì)學(xué)意義。各組實(shí)驗(yàn)至少重復(fù)3次。 結(jié)果: 1IS呈濃度（100、200、500μm/L）依賴性的降低細(xì)胞存活率，大于等于200μm/L時(shí)與對(duì)照組相比具有統(tǒng)計(jì)學(xué)意義（P0.001）；而LA呈濃度（20、50、100、250、500μm/L）依賴性的提高細(xì)胞存活率，大于等于250μm/L時(shí)與對(duì)照組有顯著性差異（P0.01）；250μm/L LA預(yù)處理細(xì)胞再與500μm/L IS共培養(yǎng)，與IS組（500μm/L）相比，細(xì)胞存活率顯著提高（P0.01）。選定500μm/L和250μm/L分別作為IS和LA繼續(xù)實(shí)驗(yàn)濃度。 2與對(duì)照組（1.29±0.40nmol/ml）相比，IS組（2.87±0.66nmol/ml）細(xì)胞MDA含量升高（P0.001）；細(xì)胞經(jīng)LA預(yù)處理，IS+LA組（2.04±0.46nmol/ml）較IS組（2.87±0.66nmol/ml） MDA含量降低（P0.05）。 3與對(duì)照組(62.07±1.60%)相比，IS組(76.17±2.26%)ROS陽性細(xì)胞率增加(P0.001)；細(xì)胞經(jīng)LA預(yù)處理，IS+LA組(69.70±3.01%)較IS組(76.17±2.26%)ROS陽性細(xì)胞率降低（P0.05）。 4與對(duì)照組（1.00±0.24）相比，IS組（1.65±0.18）細(xì)胞NOX4mRNA相對(duì)表達(dá)量升高（P0.05）；細(xì)胞經(jīng)LA預(yù)處理, IS+LA組（1.11±0.41）較IS組（1.65±0.18）NOX4mRNA相對(duì)表達(dá)量降低（P0.05）。 結(jié)論： 1硫酸吲哚酚誘導(dǎo)氧化應(yīng)激使內(nèi)皮細(xì)胞受損 2α-硫辛酸可能通過下調(diào)NADPH氧化酶，降低ROS水平，改善硫酸吲哚酚對(duì)內(nèi)皮細(xì)胞的氧化損傷
[Abstract]:Objective: cardiovascular morbidity and mortality in patients with chronic kidney disease, especially in patients with end-stage renal failure, were significantly higher than those in the general population. Accelerated atherosclerosis is one of the main pathological mechanisms of cardiovascular complications in uremic patients. It begins with the decrease of glomerular filtration rate and accumulation of nephrotoxic substances in patients with vascular endothelial cell injury. Promote atherosclerotic lesions in the arteries. Indole sulfate indoxyl sulfate is one of the typical protein-binding toxins, which induces oxidative stress to damage the cardiovascular system and accelerates the progression of chronic kidney disease. 偽 -lipoic acid (偽 -lipoic acid) is the strongest natural antioxidant in diabetes. Diabetes complications, ischemia reperfusion injury and other diseases are widely used in the prevention and treatment, but less in patients with CKD. In this study, human umbilical vein endothelial cells (HUVECs) were used to investigate whether 偽 -lipoic acid could protect endothelial cells from oxidative damage induced by indole sulfate and its possible mechanism. To provide experimental basis for the application of 偽-lipoic acid in chronic kidney disease. Methods: human umbilical vein endothelial cells were cultured and divided into control group (control group), 偽 -lipoic acid group (LA group), indoleol sulfate group (is group) and indolol sulfate 偽 -lipoxylic acid group (ISLA group). MTS assay was used to detect cell survival rate, thiobarbituric acid method to determine the content of malondialdehyde (MDA), flow cytometry to detect the changes of reactive oxygen species (Ros), and RT-PCR method to detect the expression of NADPH oxidase 4(NOX4)mRNA. GraphPad Prism5 software was used to process statistics, and the measurement data in experimental data were expressed as mean 鹵standard deviation. Single factor analysis of variance (ANOVAX) was used to compare the mean of the two groups, and the Bonferroni method was used to test the difference between the two groups (P < 0. 05, P < 0. 05). The experiments in each group were repeated at least 3 times. Results: 1IS decreased the cell survival rate in a concentration-dependent manner (> 200 渭 m / L) and increased the cell survival rate in a dose-dependent manner compared with the control group (P 0.001 / L), while LA increased the cell survival rate in a concentration-dependent manner. Compared with the control group, there was a significant difference when the concentration of LA was greater than or equal to 250 渭 m / L, and the cell survival rate was significantly higher than that in the is group (P < 0.01), and then co-cultured with 500 渭 m 路L ~ (-1) L 路L 路L ~ (-1) LA, compared with 500 渭 m 路L ~ (L) in the is group (P _ (0.01) 路L ~ (-1)). 500 渭 m / L and 250 渭 m / L were selected as the continued concentration of is and LA, respectively. 2Compared with the control group (1.29 鹵0.40 nmol / ml), the MDA content of the cells in the is group increased (2.87 鹵0.66 nmol / ml), and the MDA content in the ISLA group decreased by 2.04 鹵0.46 nmol / ml (2.87 鹵0.66 nmol / ml) after LA preconditioning, compared with the control group (2.87 鹵0.66 nmol / ml). 3Compared with the control group (62.07 鹵1.60), the percentage of positive cells of 76.17 鹵2.26%)ROS was increased in the is group, and the percentage of positive cells in the LA group (69.70 鹵3.01) was significantly lower than that in the is group (76.17 鹵3.01). 4Compared with the control group (1.00 鹵0.24), the relative expression of NOX4mRNA in the is group (1.65 鹵0.18) was higher than that in the control group (P 0.05), and the relative expression of NOX4mRNA in the is LA group was 1.11 鹵0.41) lower than that in the is group (1.65 鹵0.18). Conclusion: 1. Oxidative stress induced by indole sulfate damaged endothelial cells 2 偽 -lipoic acid may decrease the level of ROS by down-regulating NADPH oxidase, and improve the oxidative damage of endothelial cells induced by indole sulfate.
【學(xué)位授予單位】：河北醫(yī)科大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2014
【分類號(hào)】：R692

【參考文獻(xiàn)】

相關(guān)期刊論文 前1條

1 盧斌;馬健;楊翠華;王堅(jiān);;硫辛酸對(duì)db/db小鼠胰島內(nèi)NADPH氧化酶表達(dá)的影響[J];中西醫(yī)結(jié)合心腦血管病雜志;2013年02期

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