【摘要】：丁香酚是丁香揮發(fā)精油的主要成分，目前對(duì)丁香酚的報(bào)道多見(jiàn)于其抑菌能力、對(duì)魚(yú)類(lèi)的麻醉作用，其在機(jī)體內(nèi)抗氧化活性、對(duì)活細(xì)胞的抗氧化活性方面的研究鮮有報(bào)道。實(shí)驗(yàn)通過(guò)對(duì)丁香酚體外抗氧化能力的測(cè)定、對(duì)急性肝損傷小鼠的保護(hù)作用以及對(duì)細(xì)胞氧化損傷保護(hù)作用的研究，綜合探討丁香酚抗氧化活性及作用機(jī)制，旨在為丁香酚在食品添加劑和功能性食品中的應(yīng)用提供一定的理論參考。 通過(guò)對(duì)丁香酚還原能力、清除羥自由基能力、清除超氧陰離子自由基能力、清除DPPH能力和抗脂質(zhì)氧化能力的測(cè)定，實(shí)驗(yàn)結(jié)果表明，100~250μg/mL各濃度下丁香酚的還原能力高于BHT（P0.05）；丁香酚對(duì)·OH自由基IC50劑量為152.52μg/mL，，低于BHT；250μg/mL的丁香酚對(duì)DPPH的清除率達(dá)到92.5%，顯著高于BHT（P0.05）；對(duì)O2-·的IC50為215.99μg/mL；50~200μg/mL范圍各濃度下丁香酚與Vc對(duì)卵黃脂蛋白體系氧化抑制作用差異不顯著（P0.05），250μg/mL濃度下丁香酚對(duì)卵黃脂蛋白體系氧化抑制作用顯著高于Vc（P0.05）。50~250μg/mL濃度范圍內(nèi)丁香酚的各項(xiàng)抗氧化指標(biāo)均隨濃度的增加而增大，差異顯著（P0.05）。 采用CCl4作用于健康Balb/c小鼠建立急性肝損傷模型：測(cè)定實(shí)驗(yàn)小鼠肝指數(shù)；血清AST、ALT酶活力；肝臟勻漿T-AOC能力、MDA含量、SOD酶活力、CAT酶活力、GSH-Px酶活力；肝組織做切片，并在顯微鏡下觀察病理學(xué)變化。結(jié)果表明，丁香酚各劑量組不同程度地抑制了CCl4致肝損傷導(dǎo)致的血清AST、ALT活性的升高，對(duì)CCl4引起的肝臟T-AOC能力降低、SOD、GSH-Px、CAT活力下降具有拮抗作用，10mg/(kg·d)和15mg/(kg·d)組各項(xiàng)指標(biāo)與模型組差異顯著（P0.01），小鼠肝臟病理學(xué)顯微鏡檢驗(yàn)結(jié)果表明丁香酚5mg/(kg·d)、10mg/(kg·d)和15mg/(kg·d)的給藥量對(duì)小鼠肝臟結(jié)構(gòu)、細(xì)胞完整性均具有保護(hù)作用，15mg/(kg·d)的給藥量對(duì)小鼠肝臟的保護(hù)效果最好。 采用H2O2誘導(dǎo)建立人肝細(xì)胞HL7702氧化損傷模型，測(cè)定丁香酚對(duì)細(xì)胞增值的影響、細(xì)胞抗氧化酶活性、細(xì)胞裂解液丙二醛的含量，結(jié)果表明10mmol/L的丁香酚對(duì)細(xì)胞的增值最為有利，0.1~10mg/L的丁香酚可顯著提高H2O2損傷HL7702細(xì)胞的SOD、CAT、GSH-Px酶活性（P0.05），并降低細(xì)胞裂解液MDA含量（P0.05）。
[Abstract]:Eugenol is the main component of eugenol volatile essential oil. At present, eugenol is mainly reported in its bacteriostatic ability, anaesthesia to fish, antioxidant activity in organism and antioxidant activity in living cells. The protective effect of eugenol on acute liver injury in mice and the protective effect of eugenol on cell oxidative injury in vitro were studied, and the antioxidant activity and mechanism of eugenol were discussed. The purpose is to provide a theoretical reference for the application of eugenol in food additives and functional foods. The ability of reducing eugenol, scavenging hydroxyl radical, scavenging superoxide anion radical, scavenging DPPH and resisting lipid oxidation were determined. The reduction ability of eugenol at 100 渭 g/mL concentration was higher than that of BHT (P0.05). The dosage of eugenol to OH radical IC50 was 152.52 渭 g / mL, and the clearance rate of eugenol to DPPH was 92.5 渭 g 路mL ~ (-1) lower than that of BHT;250 渭 g/mL, which was significantly higher than that of BHT (P0.05), IC50 of O _ 2- was 215.99 渭 g / mL. There was no significant difference between eugenol and Vc in the oxidative inhibition of vitelloprotein in the range of 50 渭 g/mL (P0.05). The inhibitory effect of eugenol on the oxidation of egg yolk lipoprotein system was significantly higher than that of Vc at the concentration of 250 渭 g/mL (P0.05). The antioxidant indexes of eugenol in the range of 50 渭 g/mL increased with the increase of concentration, and the difference was significant (P0.05). The acute liver injury model of healthy Balb/c mice was established by CCl4. The liver index, serum AST,ALT enzyme activity, T-AOC ability, MDA content, SOD enzyme activity, CAT enzyme activity and GSH-Px enzyme activity of liver homogenate were measured. The liver tissue was sectioned and the pathological changes were observed under microscope. The results showed that eugenol inhibited the increase of serum AST,ALT activity induced by liver injury induced by CCl4 to varying degrees, and antagonized the decrease of T-AOC activity and SOD,GSH-Px,CAT activity induced by CCl4. The indexes of 10mg/ (kg d) and 15mg/ (kg d) groups were significantly different from those of model group (P0.01). The pathological examination of mouse liver showed that eugenol 5mg/ (kg d), was detected by microscope. The dosage of 10mg/ (kg d) and 15mg/ (kg d) had protective effect on the liver structure and cell integrity of mice, and the dose of 15mg/ (kg d) had the best protective effect on the liver of mice. The HL7702 oxidative damage model of human hepatocytes induced by H2O2 was established. The effect of eugenol on cell proliferation, the activity of cell antioxidant enzymes and the content of malondialdehyde in cell lysate were measured. The results showed that eugenol of 10mmol/L was the most beneficial to cell proliferation. Eugenol of 0.1~10mg/L significantly increased the activity of SOD,CAT,GSH-Px enzyme in HL7702 cells damaged by H2O2 (P0.05), and decreased the content of MDA in cell lysate (P0.05).
【學(xué)位授予單位】：河南科技大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2014
【分類(lèi)號(hào)】：TS202.3

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