本文選題：終末期腎臟病 + 心血管疾病�。� 參考：《蘭州大學》2014年碩士論文

【摘要】：目的：體外建立終末期腎臟病(End stage renal disease, ESRD)患者內(nèi)環(huán)境模型,誘導人臍靜脈血管內(nèi)皮細胞(Human umbilical vein endothelial cells, HUVECs)凋亡,觀察黃芪總黃酮(Total flavonoids of astragalus, TFA)干預后氧化應激(Oxidative stress, OS)終產(chǎn)物4-羥基壬烯醛(4-hydroxy nonene aldehyde,4-HNE)及內(nèi)質(zhì)網(wǎng)應激(Endoplasmic reticulum stress, ERS)信號分子C/EBP同源蛋白(C/EBPhomologous protein, CHOP)勺表達變化,探討TFA減少ESRD患者血清誘導的血管內(nèi)皮細胞(Vein endothelial cell, VEC)凋亡的分子機制,為ESRD患者防治心血管疾病(Cardiovascular disease, CVD)提供新的理論依據(jù)和治療靶點。 方法：HUVECs培養(yǎng)于含10%胎牛血清、100U/mL青霉素和10Oμg/mL鏈霉素的DMEM普通培養(yǎng)基中,置于37℃、5%CO2環(huán)境的培養(yǎng)箱中培養(yǎng),根據(jù)細胞生長情況擇期更換新鮮培養(yǎng)基,應用0.25%濃度的胰酶對貼壁生長的內(nèi)皮細胞進行消化傳代。于實驗前24h更換為無血清培養(yǎng)基,使細胞完成同步化。隨機將細胞分為6組：空白對照組(10%胎牛血清)、正常對照組(10%健康志愿者血清)、終末期腎臟病組(10%終末期腎臟病患者血清)、低劑量(TFA1)、中劑量(TFA2)、高劑量(TFA3)黃芪總黃酮組(在終末期腎臟病組基礎上分別加入0.5mg/mL、1.0mg/mL、2.0mg/mL黃芪總黃酮)。培養(yǎng)24h后倒置顯微鏡下觀察HUVECs形態(tài),四甲基偶氮畔鹽(Methyl thiazolyl tetrazolium, MTT)試驗檢測細胞生存率,彗星實驗檢測細胞凋亡率,酶聯(lián)免疫吸附測定(Enzyme-linked immuno sorbent assay, ELISA)法檢測4-HNE濃度,免疫細胞化學法檢測CHOP的表達量。 結果：正常對照組與空白對照組比較,細胞生存率、凋亡率、4-HNE濃度和CHOP表達量,差異無統(tǒng)計學意義(P0.05)。ESRD患者血清誘導HUVECs培養(yǎng)24h后出現(xiàn)細胞體積縮小,結構緊密,胞漿濃縮,細胞器聚集,胞膜出芽脫落,細胞核固縮呈均一的致密物等凋亡形態(tài)學變化。與正常對照組比較,終末期腎臟病組HUVECs生存率降低(P0.01),凋亡率增高(P0.01),4-HNE濃度和CHOP表達量均升高(P0.05)。與終末期腎臟病組比較,TFA干預后各組HUVECs生存率提高(P0.05),凋亡率減少(P0.05),4-HNE濃度降低,呈劑量依賴性(P0.05),CHOP的表達量降低(P0.05),中劑量組優(yōu)于低劑量組(P0.01),但高劑量組與中劑量組比較差異無統(tǒng)計學意義(P0.05)。相關性分析示：4-HNE濃度與CHOP的表達量呈正相關(r=0.913,P0.01)。 結論：ESD患者血清可誘導HUVECs凋亡；TFA對ESRD患者血清誘導的HUVECs凋亡具有抑制作用,其可能通過減輕OS和ESR發(fā)揮抗凋亡作用。
[Abstract]:Objective: to establish an in vitro model of end-stage renal disease (End stage renal disease, ESRD) patients and induce apoptosis of human umbilical vein endothelial cells (HUVECs) in vitro. To observe the changes of oxidative stress (OS) end product 4-hydroxy nonene aldehydededededededededededededededededededededededededededededededededededededededededededededededehne4-HNE( 4-HNE4) after the intervention of Total flavonoids of astragalus, TFA) of Astragalus membranaceus Total reticulum stress, ERS) and the expression of C/EBP homologous protein (CHOPP) in the endoplasmic reticulum (ER) signal molecule Endoplastic reticulum stress, ERS). To explore the molecular mechanism of TFA reducing the apoptosis of vascular endothelial cells (VEC) induced by serum in patients with ESRD, and to provide a new theoretical basis and therapeutic target for the prevention and treatment of cardiovascular disease (CVDs) in patients with ESRD. Methods 1: HUVECs were cultured on the DMEM medium containing 10% fetal bovine serum 100 U / mL penicillin and 10 O 渭 g/mL streptomycin, and cultured in a incubator with CO 2 at 37 鈩，

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