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  本文選題：載脂蛋白A-I模擬肽D-4F + 氧化型低密度脂蛋白(ox-LDL)��； 參考：《福建醫(yī)科大學》2015年碩士論文

【摘要】：目的:探討載脂蛋白A-I模擬肽D-4F拮抗血管平滑肌細胞(vascular smooth muscle cells,VSMCs)增殖及遷移的機制。方法:大鼠胸主動脈VSMCs分離與培養(yǎng)。分別給予D-4F及氧化型低密度脂蛋白(oxidized low-density lipoprotein,ox-LDL)處理VSMCs,CCK-8(Cell counting kit-8)試劑盒來測定細胞增殖,transwell小室和劃痕實驗來評估細胞遷移,而活性氧探針,即雙氫乙酰乙酸一二氯熒光黃(dihydrochloride acetyl acid dichloride fluorescent yellow,DCFH-DA)測定VSMCs中活性氧簇(reactive oxygen species,ROS)。利用western blotting檢測D-4F對VSMCs中P13K/Akt信號通路磷酸化激活和血紅素加氧酶-1(heme oxygenase-1,HO-1)蛋白表達。利用P13K/Akt抑制劑LY294002預孵育VSMCs,western blotting檢測HO-1蛋白表達。利用HO-1抑制劑Znpp拮抗VSMCs中HO-1的活性,再次檢測D-4F對ox-LDL誘導的ROS釋放以及VSMCs增殖和遷移的抑制作用。結果:D-4F在體外能夠抑制ox-LDL誘導的血管平滑肌細胞(VSMCs)的增殖和遷移。D-4F能夠誘導VSMCs中P13K/Akt磷酸化激活,D-4F上調VSMCs中HO-1蛋白的表達,PI3K/Akt抑制劑LY294002能夠拮抗D-4F誘導的VSMCs中HO-1蛋白表達。此外D-4F能夠有效的抑制ox-LDL誘導的VSMCs中ROS的釋放,進而抑制ox-LDL誘導的VSMCs的增殖和遷移。HO-1抑制劑Znpp能夠拮抗D-4F的抗氧化與抗增殖和抗遷移活性。結論:載脂蛋白A-I模擬肽D-4F通過激活PI3K/Akt通路上調VSMCs中HO-1蛋白的表達,以此來抑制ox-LDL誘導的ROS釋放,拮抗VSMCs增殖和遷移。
[Abstract]:Aim: to investigate the mechanism of antagonistic effect of apolipoprotein A-I mimic peptide D-4F on proliferation and migration of vascular smooth muscle cells in vascular smooth muscle cells (VSMCs). Methods: VSMCs was isolated and cultured from rat thoracic aorta. D-4F and oxidized low density lipoprotein (LDL) -treated VSMCsCCK-8 cell counting kit-8 were used to measure cell proliferation, transwell chamber and scratch test to evaluate cell migration, and reactive oxygen probe (Ros). That is, dihydroacetoacetic acid, dichlorofluorescein dihydrochloride acetyl acid dichloride fluorescent yellow.DCFH-DAA was used to determine reactive oxygen species, oxygen speciesrossuch as Ros in VSMCs. Western blotting was used to detect the activation of phosphorylation of P13K/Akt signaling pathway and the expression of hemhemoxygenase-1 (HO-1) protein in VSMCs by D-4F. P13K/Akt inhibitor LY294002 preincubated VSMCswestern blotting to detect the expression of HO-1 protein. HO-1 inhibitor Znpp was used to antagonize the activity of HO-1 in VSMCs. The inhibitory effects of D-4F on ROS release induced by ox-LDL and VSMCs proliferation and migration were detected again. Results D-4F could inhibit the proliferation and migration of vascular smooth muscle cells induced by ox-LDL. D-4F could induce P13K/Akt phosphorylation in VSMCs to activate the expression of HO-1 protein in VSMCs. PI3K / Akt inhibitor LY294002 could antagonize the expression of HO-1 protein in VSMCs induced by D-4F. In addition, D-4F could effectively inhibit the release of ROS in VSMCs induced by ox-LDL, and then inhibit the proliferation and migration of VSMCs induced by ox-LDL. Znpp, an inhibitor of HO-1, could antagonize the antioxidant, anti-proliferation and anti-migration activities of D-4F. Conclusion: apolipoprotein A-I mimic peptide D-4F up-regulates the expression of HO-1 protein in VSMCs by activating the PI3K/Akt pathway, thereby inhibiting the ROS release induced by ox-LDL and antagonizing the proliferation and migration of VSMCs.
【學位授予單位】：福建醫(yī)科大學
【學位級別】：碩士
【學位授予年份】：2015
【分類號】：R541.4

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本文編號：1873901