丁香酚体内外抗氧化作用及其机制研究

发布时间：2019-01-04 22:12

【摘要】：丁香酚是丁香挥发精油的主要成分，目前对丁香酚的报道多见于其抑菌能力、对鱼类的麻醉作用，其在机体内抗氧化活性、对活细胞的抗氧化活性方面的研究鲜有报道。实验通过对丁香酚体外抗氧化能力的测定、对急性肝损伤小鼠的保护作用以及对细胞氧化损伤保护作用的研究，综合探讨丁香酚抗氧化活性及作用机制，旨在为丁香酚在食品添加剂和功能性食品中的应用提供一定的理论参考。通过对丁香酚还原能力、清除羟自由基能力、清除超氧阴离子自由基能力、清除DPPH能力和抗脂质氧化能力的测定，实验结果表明，100~250μg/mL各浓度下丁香酚的还原能力高于BHT（P0.05）；丁香酚对·OH自由基IC50剂量为152.52μg/mL，，低于BHT；250μg/mL的丁香酚对DPPH的清除率达到92.5%，显著高于BHT（P0.05）；对O2-·的IC50为215.99μg/mL；50~200μg/mL范围各浓度下丁香酚与Vc对卵黄脂蛋白体系氧化抑制作用差异不显著（P0.05），250μg/mL浓度下丁香酚对卵黄脂蛋白体系氧化抑制作用显著高于Vc（P0.05）。50~250μg/mL浓度范围内丁香酚的各项抗氧化指标均随浓度的增加而增大，差异显著（P0.05）。采用CCl4作用于健康Balb/c小鼠建立急性肝损伤模型：测定实验小鼠肝指数；血清AST、ALT酶活力；肝脏匀浆T-AOC能力、MDA含量、SOD酶活力、CAT酶活力、GSH-Px酶活力；肝组织做切片，并在显微镜下观察病理学变化。结果表明，丁香酚各剂量组不同程度地抑制了CCl4致肝损伤导致的血清AST、ALT活性的升高，对CCl4引起的肝脏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)组各项指标与模型组差异显著（P0.01），小鼠肝脏病理学显微镜检验结果表明丁香酚5mg/(kg·d)、10mg/(kg·d)和15mg/(kg·d)的给药量对小鼠肝脏结构、细胞完整性均具有保护作用，15mg/(kg·d)的给药量对小鼠肝脏的保护效果最好。采用H2O2诱导建立人肝细胞HL7702氧化损伤模型，测定丁香酚对细胞增值的影响、细胞抗氧化酶活性、细胞裂解液丙二醛的含量，结果表明10mmol/L的丁香酚对细胞的增值最为有利，0.1~10mg/L的丁香酚可显著提高H2O2损伤HL7702细胞的SOD、CAT、GSH-Px酶活性（P0.05），并降低细胞裂解液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).
【学位授予单位】：河南科技大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：TS202.3

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