本文選題：動脈粥樣硬化 + 血管緊張素Ⅱ��； 參考：《西南醫(yī)科大學(xué)》2017年碩士論文

【摘要】：目的:建立ApoE基因敲除大鼠(Apolipoprotein E-deficient Rats,ApoE~(-/-)Rats)動脈粥樣硬化模型及培養(yǎng)大鼠巨噬細胞,探討鈣庫操作性Ca~(2+)內(nèi)流調(diào)節(jié)因子(Store-Operated Calcium Entry Associated Regulatory Factor,SARAF)在動脈粥樣硬化中的作用及機制。方法:第一部分:建立ApoE基因敲除大鼠動脈粥樣硬化模型將8周齡ApoE基因敲除大鼠共16只,分為實驗組和對照組,予高脂飼料喂養(yǎng)12周。實驗組在干預(yù)的前4周予2004型Alzet滲透泵植入大鼠皮下,內(nèi)部填充血管緊張素Ⅱ(AngiotensinⅡ,AngⅡ),濃度為25mg/ml,以0.25μl/hr的速率穩(wěn)定釋放28天。對照組僅予等量生理鹽水。檢測血壓、心率、體重,12周后麻醉取血并處死,檢測炎癥因子白細胞介素-1β(Interleukin-1β,IL-1β)、腫瘤壞死因子-α、(Tumor necrosis factor-α,TNF-α),血脂TC、TG、HDL、LDL;取心臟及主動脈組織制備病理切片,作HE、油紅O、Masson染色,評估動脈粥樣硬化斑塊情況;采用實時定量基因擴增熒光檢測系統(tǒng)(q PCR)以及蛋白免疫印跡法(western-blotting)檢測主動脈SARAF、TNF-α、IL-1β表達。第二部分:SARAF蛋白對巨噬細胞炎癥因子的調(diào)節(jié)作用將NR8383巨噬細胞分為對照組(Blank),ox-LDL組,空病毒干擾組(NC+ox-LDL),SARAF過表達慢病毒載體轉(zhuǎn)染組(SARAF+ox-LDL)。分別以空載體慢病毒和過表達SARAF載體慢病毒,轉(zhuǎn)染NC+ox-LDL組和SARAF+ox-LDL組各24 h,ox-LDL(終濃度100μg/ml)干預(yù)ox-LDL組、NC+ox-LDL組和SARAF+ox-LDL組各24 h,采用western blotting技術(shù)檢測各組SARAF、TNF-α、IL-1β表達。再將NR8383巨噬細胞分為對照組(Blank),ox-LDL組,空病毒載體干擾組(NC+ox-LDL),si-SARAF慢病毒干擾載體組(si-SARAF+ox-LDL)。分別以空載體慢病毒和si-SARAF干擾載體慢病毒,轉(zhuǎn)染NC+ox-LDL組和si-SARAF+ox-LDL組各24 h,ox-LDL(終濃度100μg/ml)干預(yù)ox-LDL組、NC+ox-LDL組和si-SARAF+ox-LDL組各24 h,western blotting檢測SARAF、TNF-α、IL-1β表達。結(jié)果:1.成功建立ApoE基因敲除大鼠動脈粥樣硬化模型,大鼠主動脈根部粥樣斑塊明顯;2.實驗組ApoE基因敲除大鼠的主動脈SARAF蛋白及炎癥因子TNF-α、IL-1β表達升高;3.SARAF過表達組NR8383巨噬細胞炎癥因子IL-1β、TNF-α表達增加;4.SARAF基因干擾組NR8383巨噬細胞IL-1β、TNF-α表達降低。結(jié)論:1.血管緊張素Ⅱ誘導(dǎo)可成功建立ApoE基因敲除大鼠動脈粥樣硬化模型,SARAF可能與動脈粥樣硬化有關(guān);2.SARAF可能通過增加巨噬細胞炎癥因子IL-1β、TNF-α表達,促進動脈粥樣硬化斑塊形成。
[Abstract]:Aim: to establish the atherosclerosis model of Apolipoprotein E-deficient ratssis-ratsl in ApoE knockout rats and to investigate the role and mechanism of Store-Operated Calcium Entry Associated Regulatory factorial SARAF in atherosclerosis, and to investigate the role of Store-Operated Calcium Entry Associated Regulatory Regulatory SARAF in atherosclerosis. Methods: in the first part, 16 8-week-old ApoE knockout rats were divided into experimental group and control group, and fed with high-fat diet for 12 weeks. The experimental group was subcutaneously implanted with 2004 Alzet osmotic pump in the first four weeks of intervention, and was filled with angiotensin 鈪，

本文編號：1785528