芹菜黄酮提取及其抗氧化与降血脂作用研究
摘要
本文采用比色法测定了不同品种芹菜在不同生长时期及不同部位中黄酮类化合物的含量， 并采用响应面分析法（RSA）优化出芹菜黄酮提取工艺条件。针对芹菜黄酮提取物，分别采用比色法和化学发光法研究其体外清除自由基能力，利用高脂饲料诱导大鼠高脂血症模型，研究其降血脂及体内抗氧化作用。研究结果表明：
1.芹菜中黄酮类化合物随着生长时期的延长，其含量逐渐下降。芹菜不同部位黄酮类化合物含量不同，叶子中的含量比茎中高，高约为5倍。
2.采用RSA法优化出芹菜黄酮提取的最佳工艺条件为：以芹菜干粉为材料，以40％乙醇为溶剂，料液比1:15，提取温度80℃，提取时间2.5h。在此条件下，芹菜黄酮含量为0.27％。
3.芹菜黄酮提取物具有较强的清除羟自由基和1,1-二苯基苦基苯肼（DPPH）自由基能力，其IC50分别为10.1和8mg/L。芹菜黄酮提取物清除自由基能力强于VC和槲皮素。
4.芹菜黄酮提取物各极性分部呈现出一定的清除超氧阴离子自由基、羟自由基、过氧化氢和DPPH自由基的能力，其中，清除超氧阴离子自由基能力最强，清除过氧化氢次之，清除DPPH自由基能力最弱。在这四种自由基体系中，芹菜各部提取物清除自由基能力表现出一致性，都是乙酸乙酯部强于正丁醇部，正丁醇部强于水部。芹菜提取物清除自由基能力与其黄酮含量有关。
5.芹菜黄酮提取物能显著降低高脂血症大鼠血清中总胆固醇（TC）、甘油三酯(TG)、低密度脂蛋白胆固醇(LDL-C)、载脂蛋白B(ApoB)含量，显著升高高密度脂蛋白胆固醇(HDL-C)和载脂蛋白A(ApoA)含量，有降血脂作用。芹菜黄酮提取物还能显著降低动脉粥样硬化指数AI1和AI2，有预防动脉粥样硬化的作用。
6.芹菜黄酮提取物能显著降低高脂血症大鼠血清和肝脏脂质过氧化产物丙二醛（MDA）含量，显著提高血清和肝脏谷胱甘肽过氧化物酶(GSH-Px)活性，显著增强肝脏中超氧化物歧化酶(SOD)活性，有提高血清中超氧化物歧化酶（SOD）活性的趋势，但达不到显著性水平。
In this paper, flavoniod content of different celery varieties in different growing phase and different parts was measured, and the processing conditions of leaching flavoniod from celery were optimized with response surface analysis (RSA)method. The free radical scavenging effects in vitro of celery flavoniod extracts was studied by using colorimetry and chemiluminescence method respectively. Celery flavoniod extracts had hypocholesterolaemia and antioxidation in vivo in hypercholesterolaemic rats induced by high fat diet. The results were showed as follow:
1. Flavoniod content of celery reduced gradually as the growth of celery, and was different in different celery parts. The flavoniod content of celery leaf was about quintuple higher than stem.
2. The optimal processing conditions of leaching celery flavoniod with response surface analysis method were 40% ethanol, ratio of solid to liquid 1:15, extracting time 2.5h, extracting temperature 80℃.Under this condition, flavoniod content was 0.27%.
3. The ethanol extracts of celery flavoniod showed stronger scavenging effects on hydroxyl free radical(·OH) and 1,1-diphenyl-2-picrylhyrazyl free radical(DPPH·), which IC50 valued at the levels of 10.1mg/L and 8mg/L respectively(flavoniod content), and its activities on scavenging free radicals were weaker than quercetin and stronger than vitamin C.
4. Different solvent extracts of celery flavoniod had significant scavenging effects on superoxide anion free radical(O2－·), hydroxyl free radical(·OH), hydrogen peroxide(H2O2) and 1,1-diphenyl-2-picrylhyrazyl free radical(DPPH·), and its scavenging effect on O2－·was the strongest , the second on scavenging H2O2 ,and the weakest on scavenging DPPH·. In four free radical systems, scavenging free radicals effects of different solvent extracts of celery showed consistency, The ethyl acetate extract was stronger than the buthonal extract, buthonal extract was stronger than the aqueous extract. The scavenging free radical effects of celery extract related to its flavoniod content.
5. Celery flavoniod extracts decreased significantly triglyceride(TG), total cholesterol(TC), low density lipoprotein cholesterol(LDL-C), apolipoprotein B(APOB),
increased markedly high density lipoprotein cholesterol(HDL-C) and apolipoprotein A(APOA) in serum in hypercholesterolaemic rats, and showed hypocholesterolaemia effect. Celery flavoniod extracts decreased significantly atherosclerosis index(AI1, AI2), and prevented atherosclerosis effect.
6. Celery flavoniod extracts reduced effectively lipid peroxidate malony dialdehyde(MDA) content in serum and liver in hypercholesterolaemic rats, improved markedly glutathione peroxidase(GSH-Px) activity, and enhanced significantly superoxide dismutase(SOD) activity in liver in hypercholesterolaemic rats, and but had no significant level in serum. Celery flavoniod extracts had antioxidation effect.
1.芹菜中黄酮类化合物随着生长时期的延长，其含量逐渐下降。芹菜不同部位黄酮类化合物含量不同，叶子中的含量比茎中高，高约为5倍。
2.采用RSA法优化出芹菜黄酮提取的最佳工艺条件为：以芹菜干粉为材料，以40％乙醇为溶剂，料液比1:15，提取温度80℃，提取时间2.5h。在此条件下，芹菜黄酮含量为0.27％。
3.芹菜黄酮提取物具有较强的清除羟自由基和1,1-二苯基苦基苯肼（DPPH）自由基能力，其IC50分别为10.1和8mg/L。芹菜黄酮提取物清除自由基能力强于VC和槲皮素。
4.芹菜黄酮提取物各极性分部呈现出一定的清除超氧阴离子自由基、羟自由基、过氧化氢和DPPH自由基的能力，其中，清除超氧阴离子自由基能力最强，清除过氧化氢次之，清除DPPH自由基能力最弱。在这四种自由基体系中，芹菜各部提取物清除自由基能力表现出一致性，都是乙酸乙酯部强于正丁醇部，正丁醇部强于水部。芹菜提取物清除自由基能力与其黄酮含量有关。
5.芹菜黄酮提取物能显著降低高脂血症大鼠血清中总胆固醇（TC）、甘油三酯(TG)、低密度脂蛋白胆固醇(LDL-C)、载脂蛋白B(ApoB)含量，显著升高高密度脂蛋白胆固醇(HDL-C)和载脂蛋白A(ApoA)含量，有降血脂作用。芹菜黄酮提取物还能显著降低动脉粥样硬化指数AI1和AI2，有预防动脉粥样硬化的作用。
6.芹菜黄酮提取物能显著降低高脂血症大鼠血清和肝脏脂质过氧化产物丙二醛（MDA）含量，显著提高血清和肝脏谷胱甘肽过氧化物酶(GSH-Px)活性，显著增强肝脏中超氧化物歧化酶(SOD)活性，有提高血清中超氧化物歧化酶（SOD）活性的趋势，但达不到显著性水平。
In this paper, flavoniod content of different celery varieties in different growing phase and different parts was measured, and the processing conditions of leaching flavoniod from celery were optimized with response surface analysis (RSA)method. The free radical scavenging effects in vitro of celery flavoniod extracts was studied by using colorimetry and chemiluminescence method respectively. Celery flavoniod extracts had hypocholesterolaemia and antioxidation in vivo in hypercholesterolaemic rats induced by high fat diet. The results were showed as follow:
1. Flavoniod content of celery reduced gradually as the growth of celery, and was different in different celery parts. The flavoniod content of celery leaf was about quintuple higher than stem.
2. The optimal processing conditions of leaching celery flavoniod with response surface analysis method were 40% ethanol, ratio of solid to liquid 1:15, extracting time 2.5h, extracting temperature 80℃.Under this condition, flavoniod content was 0.27%.
3. The ethanol extracts of celery flavoniod showed stronger scavenging effects on hydroxyl free radical(·OH) and 1,1-diphenyl-2-picrylhyrazyl free radical(DPPH·), which IC50 valued at the levels of 10.1mg/L and 8mg/L respectively(flavoniod content), and its activities on scavenging free radicals were weaker than quercetin and stronger than vitamin C.
4. Different solvent extracts of celery flavoniod had significant scavenging effects on superoxide anion free radical(O2－·), hydroxyl free radical(·OH), hydrogen peroxide(H2O2) and 1,1-diphenyl-2-picrylhyrazyl free radical(DPPH·), and its scavenging effect on O2－·was the strongest , the second on scavenging H2O2 ,and the weakest on scavenging DPPH·. In four free radical systems, scavenging free radicals effects of different solvent extracts of celery showed consistency, The ethyl acetate extract was stronger than the buthonal extract, buthonal extract was stronger than the aqueous extract. The scavenging free radical effects of celery extract related to its flavoniod content.
5. Celery flavoniod extracts decreased significantly triglyceride(TG), total cholesterol(TC), low density lipoprotein cholesterol(LDL-C), apolipoprotein B(APOB),
increased markedly high density lipoprotein cholesterol(HDL-C) and apolipoprotein A(APOA) in serum in hypercholesterolaemic rats, and showed hypocholesterolaemia effect. Celery flavoniod extracts decreased significantly atherosclerosis index(AI1, AI2), and prevented atherosclerosis effect.
6. Celery flavoniod extracts reduced effectively lipid peroxidate malony dialdehyde(MDA) content in serum and liver in hypercholesterolaemic rats, improved markedly glutathione peroxidase(GSH-Px) activity, and enhanced significantly superoxide dismutase(SOD) activity in liver in hypercholesterolaemic rats, and but had no significant level in serum. Celery flavoniod extracts had antioxidation effect.
引文
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