本文選題：紊流　切入點：震蕩剪切力　出處：《天津醫(yī)科大學(xué)》2017年碩士論文

【摘要】：目的:動靜脈內(nèi)瘺(Arteriovenous fistulae,AVF)是維持性血液透析(Maintenance hemodialysis,MHD)患者的生命線,AVF吻合口處的靜脈段狹窄是內(nèi)瘺失功的主要原因,其病理機制為內(nèi)膜增生,與特殊部位的壁面剪切力及血流模式改變有關(guān)。且透析患者體內(nèi)炎癥介質(zhì)與氧化應(yīng)激密切相關(guān),兩者之間相互作用,共同損傷血管內(nèi)皮細(xì)胞。有研究表明紊流促進血管內(nèi)皮細(xì)胞增殖。小窩蛋白-1(caveolin-1,Cav-1)大量存在于血管內(nèi)皮細(xì)胞中,是分布于其表面的力學(xué)感受器,參與細(xì)胞內(nèi)多種信號通路的調(diào)節(jié),內(nèi)皮細(xì)胞受到剪切力作用時,膜表面蛋白表達(dá)及分布會發(fā)生改變。轉(zhuǎn)化生長因子-β1(transforming growth factor-β1,TGF-β1)不僅具有誘導(dǎo)細(xì)胞增殖和遷移的作用,還能促進細(xì)胞外基質(zhì)積聚。本研究旨在研究體外生理環(huán)境中以及尿素癥環(huán)境中紊流作用下Cav-1、TGF-β1在血管內(nèi)皮的分布及功能的表達(dá)情況。方法:應(yīng)用體外洄流模型模擬體內(nèi)紊流狀態(tài),分別用平行平板流動小室來模擬血管管腔,蠕動泵來控制循環(huán)流量,注射泵制造洄流血流,含血清的培養(yǎng)基模擬血液,以臍靜脈內(nèi)皮細(xì)胞(HUVEC)模擬靜脈血管內(nèi)皮細(xì)胞,尿毒癥患者血清按比例加入到培養(yǎng)基中模擬尿毒癥環(huán)境。洄流作用不同時間(分別為0小時、2小時、6小時和12小時)對內(nèi)皮細(xì)胞的影響。以及用低于生理水平的層流剪切力(6dynes/cm2)、生理水平的層流剪切力(12dynes/cm2)和超生理水平的層流剪切力(20dynes/cm2)以及洄流產(chǎn)生的震蕩剪切力(0±5dynes/cm2)分別作用于HUVEC 12h,對照組不受剪切力作用。應(yīng)用Western blotting和免疫熒光技術(shù)檢測洄流產(chǎn)生的震蕩剪切力作用不同時間以及不同形式的剪切力作用下Cav-1、TGF-β1的分布及功能的表達(dá)情況。結(jié)果:在洄流產(chǎn)生的震蕩剪切力作用下,Cav-1的表達(dá)隨時間的延長呈逐漸下降趨勢,且其分布表現(xiàn)為由質(zhì)膜向胞質(zhì)轉(zhuǎn)移。而TGF-β1的表達(dá)則恰恰相反。對比洄流和不同大小的層流剪切力作用12h,發(fā)現(xiàn)Cav-1在洄流產(chǎn)生的震蕩剪切力作用下以及低于生理水平的層流剪切力作用下表達(dá)下降。Cav-1在高于生理水平的層流剪切力作用下其表達(dá)增多。而TGF-β1在洄流作用和低于生理水平的層流剪切力表達(dá)增多。在高于生理水平的層流剪切力作用下其表達(dá)有所下降。在尿毒癥環(huán)境中,內(nèi)皮細(xì)胞的活性比正常血清環(huán)境下有所減低。各組差異均具有統(tǒng)計學(xué)意義。結(jié)論:血流形式影響細(xì)胞因子的表達(dá)。紊流、低于生理水平的層流剪切力和長時間的作用使內(nèi)皮細(xì)胞的結(jié)構(gòu)和功能發(fā)生改變,在動靜脈內(nèi)瘺失功中,起到關(guān)鍵性作用。尿毒癥環(huán)境影響血管內(nèi)皮細(xì)胞中的細(xì)胞因子的表達(dá)。
[Abstract]:Objective: Arteriovenous fistulaeus AVFis is the main cause of arteriovenous fistula failure in maintenance hemodialysis patients with maintenance hemodialysis and maintenance maintenance hemodialysis (MHD), and its pathological mechanism is intimal hyperplasia. And the inflammatory mediators in dialysis patients are closely related to oxidative stress, and they interact with each other. Some studies have shown that turbulence promotes the proliferation of vascular endothelial cells. Fossa protein -1caveolin-1 (Cav-1) is abundant in vascular endothelial cells and is a mechanical receptor distributed on its surface, which is involved in the regulation of various signal pathways in cells. When endothelial cells were subjected to shear stress, the expression and distribution of membrane proteins changed. Transforming growth factor-尾 1(transforming growth factor- 尾 1 (TGF- 尾 1) could not only induce cell proliferation and migration, but also induce cell proliferation and migration. The aim of this study was to study the distribution and function of Cav-1 TGF- 尾 1 in vascular endothelium in vitro and in uremic environment. Parallel flat flow chamber was used to simulate vascular lumen, peristaltic pump to control circulation flow, injection pump to make blood flow, serum-containing medium to simulate blood, and umbilical vein endothelial cell (HUVECs) to simulate venous endothelial cells. The serum of uremic patients was proportionally added to the medium to simulate the uremic environment. The effects of whirl on endothelial cells at different times (0 hours / 2 hours / 6 hours and 12 hours respectively). Laminar shear stress (6dynes / cm2), physiological laminar shear stress (12dynes / cm2), superphysiological laminar shear force (20dynes / cm2) and whirlwind shear force (0 鹵5dynes / cm2) were applied to HUVEC for 12 h, respectively. The control group was not subjected to shear stress. Western blotting and immunofluorescence were used. The distribution of Cav-1 TGF- 尾 1 and the expression of TGF- 尾 1 under different time and different forms of shear force were detected by the technique. Results: the expression of Cav-1 was expressed over time under the shear force generated by the whirl. The prolongation showed a decreasing trend. The expression of TGF- 尾 1 was the opposite. Compared with the swirl current and laminar shear force of different sizes for 12 h, it was found that Cav-1 was subjected to oscillatory shear force generated by whirl flow and was lower than physiological level. The expression of Cav-1 increased under laminar shear stress, while TGF- 尾 1 increased under whirl flow and laminar shear stress below physiological level, while TGF- 尾 1 increased under laminar shear stress above physiological level. The expression of fluid shear stress decreased. In uremic environment, The activity of endothelial cells was lower than that of normal serum environment. The differences were statistically significant. Conclusion: the form of blood flow affects the expression of cytokines. Laminar shear stress at lower than physiological level and long-term action changed the structure and function of endothelial cells and played a key role in arteriovenous fistula dysfunction. Uremia environment affected the expression of cytokines in vascular endothelial cells.
【學(xué)位授予單位】：天津醫(yī)科大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2017
【分類號】：R692.5

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