本文關(guān)鍵詞：纖維細(xì)胞在高脂飲食致大鼠動(dòng)脈粥樣硬化形成中的作用　出處：《昆明理工大學(xué)》2016年碩士論文　論文類型：學(xué)位論文

  更多相關(guān)文章： 高脂飲食 大鼠 動(dòng)脈粥樣硬化 纖維細(xì)胞 肌成纖維細(xì)胞

【摘要】：研究背景隨著發(fā)展中國(guó)家生活水平的提高以及西方生活方式的普及,動(dòng)脈粥樣硬化(Atherosclerosis, AS)造成的心血管疾病將是世界范圍內(nèi)最主要的死亡原因。因此,研究動(dòng)脈粥樣硬化形成的原因和機(jī)制,對(duì)動(dòng)脈粥樣硬化的有效防治具有重要的臨床價(jià)值和意義。研究發(fā)現(xiàn),肌成纖維細(xì)胞在形成動(dòng)脈粥樣硬化斑塊中數(shù)量增多,產(chǎn)生過量膠原和其他細(xì)胞外基質(zhì)成分,沉積形成纖維帽被認(rèn)為是動(dòng)脈硬化斑塊和纖維帽形成過程中的重要成分。新近研究表明,纖維細(xì)胞是肌成纖維細(xì)胞的一個(gè)重要來(lái)源,參與動(dòng)脈粥樣硬化形成。然而,纖維細(xì)胞在動(dòng)脈粥樣硬化中作用的研究報(bào)道甚少。本研究從纖維細(xì)胞途徑,探討其動(dòng)脈粥樣硬化形成的作用,有助于為動(dòng)脈粥樣硬化的防治提供新的策略和思路。方法動(dòng)脈粥樣硬化SD大鼠模型建立：采用兩種高脂飲食(A：基礎(chǔ)飼料78.8%,豬油12%,膽固醇3.5%,丙基硫氧嘧啶0.2%,白糖5%,膽酸鈉0.5%; B：基礎(chǔ)飼料60%,豬油10%,蛋黃粉10%,白糖20%)前后分段喂養(yǎng)SD大鼠,應(yīng)用全自動(dòng)生化儀定期監(jiān)測(cè)血脂(TC、TG、HDL-C和LDL-C水平)和HE染色判斷腹主動(dòng)脈斑塊形成,確定大鼠動(dòng)脈粥樣硬化模型建成。纖維細(xì)胞在動(dòng)脈粥樣硬化中的作用的研究方法：①采用免疫組織化學(xué)方法評(píng)價(jià)纖維細(xì)胞(白細(xì)胞分化抗原45(cluster of differentiation 45, CD45+ CD34+ Col I+)腹主動(dòng)脈斑塊的存在；②酶聯(lián)免疫吸附法(ELIS A)觀測(cè)纖維細(xì)胞表面趨化因子配體12 (Chemotaxis cytokine ligand-12, CXCL12)和趨化因子受體4(Chemotaxis cytokine receptor-4, CXCR4) 在AS斑塊中的表達(dá),發(fā)現(xiàn)CXCL12/CXCR4軸在纖維細(xì)胞遷移中的作用；③免疫組織化學(xué)和逆轉(zhuǎn)錄聚合酶鏈?zhǔn)椒磻?yīng)(RT-PCR)檢測(cè)平滑肌肌動(dòng)蛋白α(α-smooth muscle actin, α-SMA+)肌成纖維細(xì)胞的表達(dá),以揭示纖維細(xì)胞的分化作用；④RT-PCR和蛋白質(zhì)印跡法(Western Blot)方法,測(cè)量正常組和模型組腹主動(dòng)脈中纖維細(xì)胞分泌的細(xì)胞外基質(zhì)(I型膠原蛋白, collagen type I, Col I)、促血管生成因子(platelet-derived growth factor A, PDGF-A)基礎(chǔ)金屬蛋白酶(matrix metalloproteinase, MMP-2)、促生長(zhǎng)因子(transforming growth factor-β, TGF-β)以及趨化因子(monocyte chemoattractant protein-1, MCP-1)的基因和蛋白表達(dá)水平,探索纖維細(xì)胞在動(dòng)脈粥樣硬化形成過程中發(fā)揮的作用。結(jié)果(1)兩種高脂飲食(聯(lián)合維生素D3)分段喂養(yǎng)成功建立動(dòng)脈粥樣硬化模型；(2)在正常組和模型組大鼠主動(dòng)脈上CD45+、CD34+、Col I+三個(gè)因子的免疫熒光均呈陽(yáng)性反應(yīng),但模型組中熒光信號(hào)強(qiáng)度明顯高于正常組,表明模型組大鼠主動(dòng)脈上纖維細(xì)胞的數(shù)量明顯增加；(3)模型組大鼠主動(dòng)脈上的熒光強(qiáng)度和基因表達(dá)水平明顯高于正常組,提示α-SMA+肌成纖維細(xì)胞存在于大鼠主動(dòng)脈斑塊上；(4)模型組大鼠主動(dòng)脈上Col I+、PDGF-A、MMP-2、TGF-β、MCP-1的蛋白和mRNA表達(dá)明顯高于正常組,表明纖維細(xì)胞分泌的這些因子可能在動(dòng)脈粥樣硬化形成中發(fā)揮了重要作用。結(jié)論(1)兩種高脂飼料(聯(lián)合維生素D3)分段喂養(yǎng)是建立動(dòng)脈粥樣硬化模型一種理想的方法。(2)纖維細(xì)胞遷移、分化成為肌成纖維細(xì)胞、分泌多種細(xì)胞因子(Col I、PDGF-A、MMP-2、TGF-β和MCP-1)在動(dòng)脈粥樣硬化形成中發(fā)揮重要作用。這些發(fā)現(xiàn)有助于為動(dòng)脈粥樣硬化的防治提供新的策略和思路。
[Abstract]:Background: with the life level of the developing countries and the improvement of the popularity of Western lifestyle, atherosclerosis (Atherosclerosis, AS) caused by cardiovascular disease is the leading cause of death in the world. Therefore, studying the causes and mechanism of the formation of atherosclerosis, has important clinical value and significance of effective prevention and treatment of atherosclerosis. The study found that muscle the number of fiber cell formation in atherosclerotic plaque increased, excessive production of collagen and other extracellular matrix deposition, formation of fibrous cap is the artery plaque and fibrous cap formation is an important component in the process. Recent research shows that muscle fiber cells are an important source of fiber cells, participate in the formation of atherosclerosis. However, research reported effects of fiber cells in atherosclerosis is rare. This study from the fiber cell Way, to explore the role of atherosclerosis, contribute to the prevention and treatment of atherosclerosis to provide new methods and ideas. To establish a method for atherosclerosis SD rat model: using two kinds of high fat diet (A: 78.8% basic feed, 12% lard, 3.5% cholesterol, 0.2% propylthiouracil, 5% sugar, 0.5% sodium cholate; B Foundation: feed 60%, 10% lard, 10% yolk powder, sugar, 20%) before and after feeding SD rats, using automatic biochemical analyzer, regular monitoring of lipid (TC, TG, HDL-C and LDL-C level) and HE staining of abdominal aortic plaque formation, set up a rat model of atherosclerosis. Methods the role of fiber cells in atherosclerosis. The evaluation of the fiber cell immunohistochemical method (leukocyte differentiation antigen 45 (cluster of differentiation 45, CD45+ CD34+ Col I+) abdominal aortic plaque; II ELISA Adsorption method (ELIS A) and observation of fiber cell surface chemokine ligand 12 (Chemotaxis cytokine, ligand-12, CXCL12) and chemokine receptor 4 (Chemotaxis cytokine, receptor-4, CXCR4) expression in AS plaque, found in fiber cell migration in CXCL12/CXCR4 axis; chemical and reverse transcriptase polymerase chain reaction and immunohistochemistry (detection of alpha smooth muscle actin (RT-PCR) -smooth muscle alpha actin, alpha -SMA+) muscle fibroblast cells, to reveal the differentiation of fibroblasts; the RT-PCR and Western blotting (Western Blot method), extracellular matrix fiber cell secretion was measured in normal group and model group in the abdominal aorta (type I collagen, collagen type I, Col I), angiogenic factors (platelet-derived growth factor A PDGF-A (matrix) based metalloproteinase metalloproteinase, MMP-2 (trans), growth factor Forming growth factor- TGF- beta, beta chemokine (monocyte) and chemoattractant protein-1, MCP-1) the expression level of gene and protein, explore the fiber cell formation process play a role in atherosclerosis. Results (1) two kinds of high fat diet (combined vitamin D3) segment feeding atherosclerosis model was successfully established; (2) CD45+ in the normal group and model group, rat aortic CD34+, immunofluorescence three factor Col I+ was positive in the model group, but the fluorescence signal intensity was significantly higher than the normal group, model group rats showed that aortic number of fibroblasts increased significantly; (3) the fluorescence intensity and the gene of aorta of rats in the model group the expression level was significantly higher than the normal group, suggesting that alpha -SMA+ fibroblasts in rat aortic plaque; (4) the aorta of rats in the model group Col I+, PDGF-A, MMP-2, TGF- beta, MCP-1. The white and the expression of mRNA was significantly higher than the normal group, showed that the fiber cells secrete these cytokines may play an important role in the formation of atherosclerosis. Conclusion (1) two kinds of high fat diet (vitamin D3) segmented feeding is the model of atherosclerosis was established an ideal method. (2) cell migration, differentiation into muscle fibroblasts secrete a variety of cytokines (Col, I, PDGF-A, MMP-2, TGF- beta and MCP-1) play an important role in the formation of atherosclerosis. These findings provide new methods and ideas in the prevention of atherosclerosis.

【學(xué)位授予單位】：昆明理工大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2016
【分類號(hào)】：R543.5

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