本文選題：冠狀動脈旁路移植術(shù) + 靜脈橋��； 參考：《河北醫(yī)科大學(xué)》2015年碩士論文

【摘要】：目的:冠狀動脈旁路移植術(shù)(coronary artery bypass graft,CABG)作為一種穩(wěn)定而有效的血管重建方法[1]被廣泛地應(yīng)用于治療冠狀動脈粥樣硬化性心臟病,但其遠(yuǎn)期療效不甚理想。血管內(nèi)膜增生是導(dǎo)致血管重建術(shù)后自體靜脈橋管腔狹窄或閉塞的主要原因,已被大量的研究所證實。引起靜脈移植再狹窄的主要原因是血管內(nèi)皮損傷,其主要是由于血管內(nèi)膜增生,血管平滑肌細(xì)胞(vascular smooth muscle,VSMCs)過度增殖及分泌大量細(xì)胞外基質(zhì)。即再狹窄是一種由于手術(shù)的物理性損傷導(dǎo)致的缺血再灌注損傷后的局部血管修復(fù)反應(yīng),是血管重塑的過程。因此,阻斷血管平滑肌的過度增殖是預(yù)防CABG術(shù)后靜脈橋再狹窄的有效策略。大量的研究證實,阿托伐他汀能夠抑制血管平滑肌的增殖,據(jù)此推測,阿托伐汀可能阻斷CABG術(shù)后發(fā)生靜脈橋再狹窄,但其發(fā)生機(jī)制至今尚未闡明。NF-κB是從B淋巴細(xì)胞核中檢測到的一種能與免疫球蛋к輕鏈基因的增強(qiáng)子κB序列特異結(jié)合的核蛋白因子。非活化狀態(tài)下NF-κB以與IκB聚合的三聚體形式或與前體蛋白聚合的二聚體形式存在于胞漿中,在多種細(xì)胞因子的作用下,通過多種信號途徑IκB發(fā)生磷酸化并與NF-κB解聚,NF-κB被激活,將信息由胞漿傳遞至胞核并啟動相關(guān)靶基因的轉(zhuǎn)錄,參與自由基損傷、細(xì)胞凋亡、免疫調(diào)節(jié)、炎癥反應(yīng)等病理生理過程[2]。此外,還有一些研究發(fā)現(xiàn),當(dāng)血管內(nèi)皮細(xì)胞損傷發(fā)生時,多種炎癥介質(zhì)和細(xì)胞因子釋放,激活NF-κB,NF-κB二聚體被活化,NF-κB激活后上調(diào)平滑肌細(xì)胞及內(nèi)皮細(xì)胞等基因表達(dá),引起血管內(nèi)膜增生而誘導(dǎo)管腔狹窄。但CABG術(shù)后靜脈橋血管內(nèi)皮細(xì)胞增殖是否通過NF-κB信號轉(zhuǎn)導(dǎo)途徑誘導(dǎo)再狹窄,尚有待闡明。為此,本實驗旨在觀察CABG術(shù)后靜脈橋血管內(nèi)皮細(xì)胞增殖是否通過NF-κB信號轉(zhuǎn)導(dǎo)通路誘導(dǎo)再狹窄,探討阿托伐他汀是否通過NF-κB信號通路發(fā)揮對靜脈橋的保護(hù)作用。方法:健康雄性新西蘭大白兔(3±0.5Kg)40只,建立自體靜脈移植動物模型。1阿托伐他汀鈣對靜脈橋血管壁內(nèi)膜厚度、平滑肌細(xì)胞數(shù)及超微結(jié)構(gòu)的影響。具體分組如下:①假手術(shù)組(sham)組(n=10):僅分離左側(cè)頸外靜脈,不行血管移植,假手術(shù)1d起,經(jīng)胃管灌注生理鹽水;②移植對照組(control)組(n=10):靜脈移植術(shù)后1d起,經(jīng)胃管灌注生理鹽水;③阿托伐他汀鈣(atorvastatin)5mg組(n=10):靜脈移植術(shù)后1d起,經(jīng)胃管灌注阿托伐他汀,劑量為5mg/kg.d;④阿托伐他汀鈣(atorvastatin)10mg組(n=10):靜脈移植術(shù)后1d起,經(jīng)胃管灌注阿托伐他汀,劑量為10mg/kg.d。每組共處理4w,在靜脈移植后或假手術(shù)后4w取靜脈橋,分別用HE染色法、免疫組化、電鏡觀察阿托伐他汀對靜脈橋血管壁內(nèi)膜厚度、平滑肌細(xì)胞數(shù)及超微結(jié)構(gòu)的影響。2阿托伐他汀鈣對NF-κB蛋白及NF-κB p50/p65二聚體復(fù)合物表達(dá)的影響。具體分組如下:①假手術(shù)組(sham)組(n=10):僅分離左側(cè)頸外靜脈,不行血管移植,假手術(shù)1d起,經(jīng)胃管灌注生理鹽水;②移植對照組(control)組(n=10):靜脈移植術(shù)后1d起,經(jīng)胃管灌注生理鹽水;③阿托伐他汀鈣(atorvastatin)5mg組(n=10):靜脈移植術(shù)后1d起,經(jīng)胃管灌注阿托伐他汀,劑量為5mg/kg.d;④阿托伐他汀鈣(atorvastatin)10mg組(n=10):靜脈移植術(shù)后1d起,經(jīng)胃管灌注阿托伐他汀,劑量為10mg/kg.d。每組共處理4w,在靜脈移植術(shù)后或假手術(shù)后4w取靜脈橋,應(yīng)用Western blot方法觀察NF-κB蛋白表達(dá)的變化,免疫共沉淀(Co-immunoprecipitation)方法觀察NF-κB p50/p65二聚體復(fù)合物表達(dá)的變化。探討阿托伐他汀對NF-κB表達(dá)的影響。結(jié)果:1阿托伐他汀鈣對靜脈橋血管壁內(nèi)膜厚度的影響采用HE染色方法,在光鏡下觀察靜脈橋移植術(shù)后,靜脈橋血管壁內(nèi)膜厚度的變化。Control組(Fig.2)、atoravastatin 5mg組(Fig.3)、atoravastatin 10mg組(Fig.4)靜脈橋血管管壁與sham組(Fig.1)相比,均有不同程度的增厚(P0.05)。與Sham組(Fig.1)相比,Control組(Fig.2)靜脈橋血管壁明顯增厚(P0.01)。與Control組(Fig.2)相比,atoravastatin 5 mg組(Fig.3)無顯著性差異(P0.05);atoravastatin 10 mg組(Fig.4)靜脈橋血管管壁平滑,內(nèi)膜增生程度明顯減少(P0.05)。2阿托伐他汀鈣對靜脈橋血管壁內(nèi)膜平滑肌數(shù)目的影響采用免疫組化方法,在光鏡下觀察靜脈橋移植術(shù)后,靜脈橋血管壁內(nèi)膜平滑肌細(xì)胞數(shù)目的變化。Control組(Fig.6)、atoravastatin 5 mg組(Fig.7)、atoravastatin 10 mg組(Fig.8)靜脈橋血管內(nèi)皮及平滑肌層與sham組(Fig.5)相比,均有不同程度的增殖(P0.05)。與Sham組(Fig.5)相比,Control組(Fig.6)有大量增殖細(xì)胞深染(P0.01);atoravastatin 5 mg組(Fig.7)深染的增殖細(xì)胞雖較Control組有所減少,但無明顯變化(P0.05);atoravastatin 10 mg組(Fig.8)深染的增殖細(xì)胞明顯減少(P0.05)。3阿托伐他汀鈣對靜脈橋血管壁超微結(jié)構(gòu)的影響在電鏡下觀察靜脈橋移植術(shù)后,靜脈橋血管壁超微結(jié)構(gòu)的變化。Control組(Fig.9)可見靜脈橋血管壁內(nèi)膜明顯增厚。與Control組(Fig.9)相比,atoravastatin 5 mg組(Fig.10),靜脈橋血管壁內(nèi)膜有一定程度的增厚,雖較Control組(Fig.9)少,但無顯著差異(P0.05);atoravastatin 10 mg組(Fig.11)內(nèi)膜連續(xù)性好(P0.05)。4阿托伐他汀鈣對NF-κB蛋白表達(dá)的影響應(yīng)用Western blot方法觀察靜脈橋移植術(shù)后,阿托伐他汀鈣對NF-κB蛋白表達(dá)的影響NF-κB p50(Fig.12)在Sham組、control組、atoravastatin 5 mg組、atoravastatin 10 mg均有一定的表達(dá)量。Sham表達(dá)量較少。與Sham組相比,control組、atoravastatin 5 mg組、atoravastatin 10 mg組其蛋白表達(dá)均有所上調(diào)(P0.05)。與control組相比,atoravastatin 5 mg組其蛋白表達(dá)雖有所下調(diào),但無顯著性差異(P0.05),atoravastatin 10 mg組此蛋白表達(dá)明顯下調(diào)(P0.05)。上述結(jié)果表明,阿托伐他汀鈣能夠顯著下調(diào)NF-κB p50的表達(dá)。5阿托伐他汀鈣對NF-κB p50/p65二聚體復(fù)合物表達(dá)的影響通過免疫共沉淀(Co-immunoprecipitation)的方法研究了靜脈橋移植術(shù)后,阿托伐他汀鈣對NF-κB p50/p65二聚體復(fù)合物表達(dá)的影響。NF-κB p50/p65二聚體復(fù)合物(Fig.12)在Sham組、control組、atoravastatin 5 mg組、atoravastatin 10 mg組均有表達(dá),其中control組此二聚體復(fù)合物表達(dá)最多。與Sham組相比,control組其表達(dá)顯著上調(diào)(P0.01);atoravastatin 5 mg組此二聚體復(fù)合物表達(dá)雖有所下調(diào),但明顯高于Sham組(P0.05)。與control組相比,atoravastatin 10 mg組其表達(dá)明顯下調(diào),差異有顯著性(P0.01)。上述結(jié)果表明,阿托伐他汀鈣能夠有效阻斷靜脈橋移植術(shù)后NF-κB p50/p65二聚體復(fù)合物數(shù)量的上調(diào)。小結(jié):1靜脈橋移植術(shù)后,NF-κB的蛋白表達(dá)顯著上調(diào);NF-κB被激活,即表現(xiàn)為NF-κB發(fā)生二聚體化,NF-κB二聚體復(fù)合物明顯上調(diào),并由胞漿轉(zhuǎn)移至胞核。2靜脈橋移植術(shù)后,經(jīng)10 mg atoravastatin處理,下調(diào)NF-κB p50蛋白的表達(dá),下調(diào)NF-κB p50/p65的表達(dá)二聚體復(fù)合物的表達(dá)。結(jié)論:靜脈橋移植術(shù)后,靜脈橋血管內(nèi)皮細(xì)胞增殖通過NF-κB信號通路誘導(dǎo)再狹窄;阿托伐他汀鈣通過NF-κB信號通路發(fā)揮對靜脈橋的保護(hù)作用。
[Abstract]:Objective: coronary artery bypass graft (CABG), as a stable and effective vascular reconstruction method, is widely used in the treatment of coronary atherosclerotic heart disease, but its long-term effect is not satisfactory. Vascular intima hyperplasia is the cause of autogenous vena cava stenosis or occlusion after the reconstruction of blood tube. The main cause has been confirmed by a large number of studies. The main cause of restenosis is vascular endothelial damage, mainly due to vascular intima hyperplasia, vascular smooth muscle cells (vascular smooth muscle, VSMCs) excessively proliferation and secretion of a large number of extracellular matrix. Restenosis is a result of physical damage caused by surgery. The local vascular repair reaction after ischemia-reperfusion injury is a process of vascular remodeling. Therefore, blocking the excessive proliferation of vascular smooth muscle is an effective strategy to prevent the restenosis after CABG. A large number of studies have confirmed that atorvastatin can inhibit the proliferation of vascular smooth muscle. Accordingly, it is presumed that opavastin may block the post operation of CABG. However, the mechanism of.NF- kappa B is a nuclear protein factor that is specific to the enhancer kappa B sequence that can be detected from the B lymphocyte nucleus. NF- kappa B is in the form of polymerization with I kappa B or in the form of two polymer polymerization with the precursor protein in the non activated state. In the cytoplasm, I kappa B is phosphorylated and depolymerization with NF- kappa B through a variety of signaling pathways, and NF- kappa B is deactivated. NF- kappa B is activated. The information is transferred from the cytoplasm to the nucleus and activates the transcription of the related target genes, and participates in the pathological and physiological processes, such as free radical damage, cell apoptosis, immunoregulation, and inflammatory reaction, [2]., and some other studies. It was found that when vascular endothelial cell injury occurred, many inflammatory mediators and cytokines were released, activated NF- kappa B, NF- kappa B two polymer was activated. After activation of NF- kappa B, the gene expression of smooth muscle cells and endothelial cells was up-regulated, causing intima hyperplasia to induce the stenosis of the lumen, but whether the proliferation of vascular endothelial cells in vein bridge after CABG has passed NF-. In this experiment, the aim of this experiment was to observe whether the proliferation of vascular endothelial cells in vein bridge after CABG was restenosis through the NF- kappa B signal transduction pathway, and to explore the protective effect of atorvastatin on the vein bridge through the NF- kappa B signaling pathway. Methods: healthy male New Zealand white rabbits (B). 3 + 0.5Kg) 40 rats, the effect of autologous vein transplantation animal model.1 atorvastatin calcium on the intima thickness, the number of smooth muscle cells and the ultrastructure of the vascular wall of the vein bridge were established as follows: (1) the sham operation group (sham) group (n=10): only the left external jugular vein, the blood tube transplantation, the false operation 1D, the saline infusion through the gastric tube, and the transplantation Control group (control) group (n=10): 1D after intravenous transplantation, saline infusion through gastric tube, and group of atorvastatin calcium (atorvastatin) 5mg (n=10) group (n=10): intravenous infusion of atorvastatin after intravenous transplantation, perfusion of atorvastatin through gastric tube, dosage of 5mg/kg.d; (n=10) atorvastatin calcium (atorvastatin) 10mg group (n=10): after vein transplantation, 1D, perfusion of atorvastatin through gastric tube The statins were treated with a dose of 10mg/kg.d., each group was treated with a total of 4W. After the vein graft or after the sham operation, the intravenous bridge was taken by 4W. HE staining, immunohistochemistry, and electron microscopy were used to observe the effect of atorvastatin on the intima thickness, the number of smooth muscle cells and the ultrastructure of the vascular wall of the vein bridge,.2 alfavastine calcium to NF- kappa B protein and NF- kappa B p50/p65 two polymer complex. The effects of the expression were as follows: (1) the sham operation group (sham) group (n=10): only the left external jugular vein, no vascular transplantation, the sham operation 1D, the saline infusion through the gastric tube, and the control group (control) group (n=10): 1D after the vein transplantation, the saline infusion through the gastric tube, and the atorvastatin calcium (atorvastatin) 5mg group (n=10): vein (n=10): vein After 1D, perfusion of atorvastatin through gastric tube, dosage of 5mg/kg.d, and atorvastatin calcium (atorvastatin) 10mg group (n=10): 1D after intravenous transplantation, perfusion of atorvastatin through gastric tube, the dose of 10mg/kg.d. in each group was treated with 4W, and the venous bridge was taken 4W after the vein graft or after the false operation, and Western blot method was used to observe NF- kappa eggs. The changes in the expression of white expression and immunoprecipitation (Co-immunoprecipitation) method were used to observe the changes in the expression of NF- kappa B p50/p65 two polymer complex. The effect of atorvastatin on the expression of NF- kappa B was investigated. Results: the effect of 1 atorvastatin calcium on the intima thickness of vascular wall intima of venous bridge was observed by HE staining, and the vein bridge transplantation was observed under the light microscope. .Control group (Fig.2), atoravastatin 5mg group (Fig.3), atoravastatin 10mg group (Fig.4), atoravastatin 10mg group (Fig.4), the vascular wall of venous bridge was thickened in varying degrees (P0.05). Compared with the Sham group, the vascular wall of the vein bridge was thickened significantly. The Vastatin 5 mg group (Fig.3) had no significant difference (P0.05), atoravastatin 10 mg group (Fig.4), the vascular wall of the vein bridge was smooth and the degree of intimal hyperplasia decreased significantly (P0.05). The effect of.2 atorvastatin calcium on the number of intima smooth muscle of vascular wall of the vein bridge was adopted by immunohistochemical method. The vascular wall of the vein bridge was observed under the light microscope. The number of intimal smooth muscle cells in group.Control (Fig.6), atoravastatin 5 mg group (Fig.7), atoravastatin 10 mg group (Fig.8), vascular endothelium and smooth muscle layer of the vein bridge were different to the sham group (P0.05). Compared with the Sham group, there were a large number of proliferating cell deep staining; 5 The proliferating cells of group Mg (Fig.7) were less than those in the Control group, but there was no significant change (P0.05). The proliferation of atoravastatin 10 mg group (Fig.8) decreased significantly (P0.05) the effect of.3 atorvastatin calcium on the ultrastructure of vein bridge wall ultrastructure under electron microscope observation of vein bridge transplantation, the changes of vascular wall ultrastructure of vein bridge In group.Control (Fig.9), the intima of vascular wall of vein bridge was obviously thickened. Compared with group Control (Fig.9), atoravastatin 5 mg group (Fig.10), the intima of vascular wall of vein bridge was thickened to a certain extent, although less than Control group (Fig.9), but there was no significant difference (P0.05). The effect of F- kappa B protein expression was used to observe the effect of Western blot method on the expression of NF- kappa B protein after vein bridge transplantation. NF- kappa B P50 (Fig.12) was in Sham group, control group and 5 groups. The protein expression of group atoravastatin 10 mg was up to up (P0.05). Compared with group control, the protein expression of group atoravastatin 5 mg decreased, but there was no significant difference (P0.05), and the expression of this protein in group atoravastatin 10 mg decreased significantly (P0.05). The above results showed that atorvastatin calcium could significantly reduce the expression of NF- kappa B. The effect of atorvastatin calcium on the expression of NF- kappa B p50/p65 two polymer complex was studied by immunoprecipitation (Co-immunoprecipitation) method to study the effect of atorvastatin calcium on the expression of NF- kappa B p50/p65 two polymer complex (.NF- kappa B p50/p65 two) complex (Fig.12) in Sham, 5 Group mg was expressed in group atoravastatin 10 mg, and the expression of this two polymer complex was the most in group control. Compared with Sham group, the expression of control group was significantly up (P0.01). The expression of this two polymer complex in atoravastatin 5 mg group was down, but obviously higher than Sham group (P0.05). Compared with control group, the expression of 10 groups was obviously expressed. Down regulation, the difference was significant (P0.01). The above results showed that atorvastatin calcium could effectively block the increase in the number of NF- kappa B p50/p65 two polymer complex after vein bridge transplantation. Conclusion: after 1 vein bridge transplantation, the protein expression of NF- kappa B was significantly up-regulated, NF- kappa B was activated, that is, NF- kappa B two polycondensation, NF- kappa B two polymer complex. After 10 mg atoravastatin treatment, the expression of NF- kappa B P50 protein was downregulated and the expression of NF- kappa B p50/p65 was down regulated by 10 mg atoravastatin treatment. Conclusion: the vascular endothelial cell proliferation of vein bridge was restenosis by NF- kappa B signaling pathway after vein bridge transplantation; atorvastatin. Calcium plays a protective role in the vein bridge through the NF- B signaling pathway.

【學(xué)位授予單位】：河北醫(yī)科大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2015
【分類號】：R654.2

【參考文獻(xiàn)】

相關(guān)期刊論文 前3條

1 魏爾清，，王瑤塵，辛小華，卞如濂;白三烯拮抗劑ONO-1078對抗原誘導(dǎo)的豚鼠皮膚微血管滲漏的抑制作用[J];現(xiàn)代應(yīng)用藥學(xué);1994年06期

2 張世紅,魏爾清,朱朝陽,陳忠,張松法;白三烯受體拮抗劑ONO-1078對內(nèi)皮素-1誘導(dǎo)的大鼠局灶性腦缺血的保護(hù)作用[J];藥學(xué)學(xué)報;2004年01期

3 張緯萍,魏爾清,梅如煥,朱朝陽,趙孟輝;白三烯受體拮抗劑0N0-1078對大鼠局灶性腦缺血的神經(jīng)保護(hù)作用(英文)[J];Acta Pharmacologica Sinica;2002年10期

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