10种中药提取物的抗氧化作用及ESR波谱研究
|
摘要
氧自由基(oxygen free radicals, OFR)又称活性氧,包括超氧阴离子自由基(O2·)、羟基自由基(OH·)、过氧化氢(H2O2)、单线态氧(1O2·)、脂氧自由基(RO·)、脂过氧自由基(ROO·)、NO等。氧自由基与多种疾病密切相关,因此,预防或减少体内的氧自由基损伤,从而减少疾病的发生是人们一直期待解决的问题。传统合成的抗氧化剂虽然抗氧化能力比较强,但长期食用有潜在的毒性,有的甚至会产生致畸、致癌作用,因此愈来愈受到人们的排斥。因而寻找天然、高效、低毒的中草药抗氧化剂成为了一种必然趋势。因此,本项研究从体内和体外两个层次,以抗氧化为切入点,采用不同的氧化损伤模型,观察10种中药(黄芩、红花、茶叶、贯叶金丝桃、槐米、木蝴蝶、金荞麦、金银花、山楂叶、山楂鲜果)的抗氧化作用,为临床应用提供科学依据。
1.黄芩、红花水提取物对快速老化小鼠衰老模型的抗氧化作用的影响
选取快速老化模型小鼠P/8 (SAM-P/8),观察黄芩、红花水提物对快速老化小鼠的保护作用。实验共分5组：空白对照组,阳性药对照组,黄芩水提物组(7.6,3.8,1.9g生药·kg-1)、红花水提物组(7.6,3.8,1.9g生药.kg-1)剂量组,连续给药4周,末次给药1小时后取胸腺和脑称重测脏器系数；分别测定脑匀浆和血清中MDA, SOD, CAT和GSH-Px水平。实验结果表明：黄芩、红花水提物显著降低衰老小鼠的胸腺系数和脑系数,还可不同程度的提高衰老小鼠体内SOD、GSH-Px、CAT活性,抑制MDA含量的升高,提示黄芩、红花水提取物可能是通过提高机体的抗氧化能力达到抗衰老的作用。
2.黄芩等10种中药醇提物对Fe2+-ADP-NADPH所致肝微粒体脂质过氧化的影响
低温条件下制备大鼠肝微粒体,观察黄芩等10种中药醇提物对Fe2+-ADP-NADPH氧化体系诱发的脂质过氧化的抑制作用,通过计算半数抑制浓度评价上述10种中药醇提物的抗氧化作用。结果表明,10种中药提取物及标准品(水溶性的维生素E—Trolox)在一定浓度时可明显抑制体外Fe2+-ADP-NADPH氧化体系所致大鼠肝微粒体MDA的生成,且在一定浓度范围内呈现剂量-效应关系,标准品、黄芩、红花、茶叶、贯叶金丝桃、槐米冷浸、槐米回流、木蝴蝶、金荞麦、金银花、山楂叶、山楂鲜果的IC50分别为0.014、0.066、0.598、0.030、0.041、0.041、0.058、0.098、0.324、0.458、1.069、6.156 mg·mL-1,抗脂质过氧化活性由大到小依次为标准品>茶叶>槐米冷浸>贯叶金丝桃>槐米回流>黄芩>木蝴蝶>金荞麦>金银花>红花>山楂叶>山楂果；其中,Trolox有较强的抑制脂质过氧化作用,山楂果的脂质过氧化抑制作用较弱。
3.黄芩等10种中药醇提物对ox-LDL所致心肌微血管内皮细胞损伤的影响
采用第三代大鼠乳鼠心肌微血管内皮细胞,观察黄芩等10种中药醇提物及Trolox预处理对ox-LDL诱导损伤的心肌微血管内皮细胞的保护作用。实验分5组,空白对照组：正常完全培养基培养；模型对照组：加入ox-LDL (100μg·mL-1)孵育24h；提取物大、中、小剂量组：分别加入不同浓度的提取物预处理24h,然后换为100μg·mL-1的ox-LDL溶液孵育24h。所有细胞均在37℃,95%O2+5%CO2,饱和湿度条件下的二氧化碳培养箱中培养。实验结束后分别收集细胞上清液测定MDA, SOD和GSH-Px活性。实验结果表明：10种中药醇提物及Trolox预处理能明显提高ox-LDL损伤的内皮细胞存活率,减少MDA含量,提高SOD,GSH-Px活性。结果提示10种中药醇提物对ox-LDL损伤的心脏微血管内皮细胞具有不同程度的保护作用,其机制可能与直接减少自由基生成,增强其清除以及增强内源性抗氧化酶的活性有关。
4.黄芩等10种中药醇提物对黄嘌呤／黄嘌呤氧化酶所致心肌细胞氧化损伤的影响
采用原代培养的乳鼠心肌细胞,建立心肌细胞黄嘌呤／黄嘌呤氧化酶氧化损伤模型,观察10种中药醇提物及Trolox对心肌细胞黄嘌呤／黄嘌呤氧化酶氧化损伤的保护作用。实验分5组,空白对照组：正常普通培养基培养;模型对照组：换为4.2mmol·L-1的黄嘌呤与280U·L-1的黄嘌呤氧化酶孵育6h；提取物大、中、小剂量组：分别加入不同浓度的提取物的同时加4.2mmol·L-1的黄嘌呤与280U·L-1的黄嘌呤氧化酶作用6h。所有细胞均在37℃,95%02+5%CO2,饱和湿度条件下的二氧化碳培养箱中培养。实验结束后分别收集细胞上清液测定LDH,MDA,SOD和GSH-Px活性。实验结果表明：10种中药醇提物及Trolox对能明显提高黄嘌呤／黄嘌呤氧化酶氧化损伤的心肌细胞存活率,减少LDH, MDA含量,提高SOD, GSH-Px活性。结果提示10种中药醇提物对心肌细胞黄嘌呤／黄嘌呤氧化酶氧化损伤具有保护作用,其机制可能与直接减少自由基生成,增强其清除以及增强内源性抗氧化酶的活性有关。
5.茶叶、槐米冷浸、木蝴蝶醇提物对ox-LDL损伤心肌微血管内皮细胞和黄嘌呤／黄嘌呤氧化酶损伤心肌细胞中自由基信号的影响
采用原代培养的心肌微血管内皮细胞和乳鼠心肌细胞,建立心肌微血管内皮细胞ox-LDL损伤模型和心肌细胞黄嘌呤／黄嘌呤氧化酶氧化损伤模型,观察茶叶、槐米冷浸、木蝴蝶醇提物对内皮细胞和心肌细胞中自由基信号的影响。实验分组同前,所有细胞均在37℃,95%O2+5%CO2,饱和湿度条件下的二氧化碳培养箱中培养。实验结束后收集细胞悬液应用电子自旋波谱仪测定活性氧自由基信号。实验结果表明,茶叶、槐米冷浸、木蝴蝶醇提物能够不同程度的降低细胞悬液中自由基信号的峰值。结果提示,茶叶、槐米冷浸、木蝴蝶醇提物减轻细胞氧化损伤的作用可能也是通过减少自由基产生,增强其清除而实现的,与前期实验结果有一致性。
综上,黄芩、红花水提物在整体动物模型,对快速老化小鼠具有一定的抗衰老作用,黄芩等10种中药醇提物在Fe2+-ADP-NADPH所致肝微粒体脂质过氧化模型、微血管内皮细胞ox-LDL损伤模型、心肌细胞黄嘌呤／黄嘌呤氧化酶损伤模型等3个模型上,均可提高细胞上清液中的SOD, GSH-Px活性,降低MDA的含量,通过ESR检测,发现茶叶、槐米冷浸、木蝴蝶还可直接减少自由基的产生,增强其清除而达到抗氧化的作用。结果提示黄芩等10种中药提取物具有抗氧化作用,其机制与减少自由基生成,增强其清除,增强内源性抗氧化酶的活性,减少氧化产物生成等有关。
Oxygen free radicals (OFR) also known as reactive oxygen species, including superoxide anion radical (O2·), hydroxyl radical (OH·), hydrogen peroxide (H2O2), singlet oxygen (1O2·), lipid oxygen free radicals (RO·), lipid peroxy radicals (ROO·), NO and so on. Oxygen free radicals are closely related with many diseases. Therefore, what people have been looking forward to solving is preventing or reducing the body's oxygen free radicals. Although the traditional synthetic antioxidants have strong antioxidant capacity, they also have long-term of potentially toxic, some even inducing teratogenic and carcinogenic effect. Thus finding natural, efficient, low toxicity anti-oxidants from Chinese herbs has become an inevitable trend. Relevant experiments in vivo and vitro showed the 10 kinds of chinese herbs including Scutellaria baicalensis Georgi,Carthamus tinctorius L., Camellia sinensis(L.)O.Ktze,Hypericum perforatum L.,Sophora japonica L., Oroxylum indicum (L.) Vent., Fagopyrum dibotrys(D.Don), Lonicera japonica Thunb., leaves and fruits of Crataegus pinnatifida Bge. have antioxidant activity by different degrees.Therefore, this study was to investigate the effect of 10 kinds of extracts from Chinese Herbs on the oxidative injury in order to provide the scientific evidence for wide clinical application.
1. Antioxidant effects of water extracts of Scutellaria and Carthamus tinctorius in senescence accelerated mouse(SAM-P/8)
Antioxidant effects of aqueous extract of Scutellaria and Carthamus tinctorius were observed by selecting senescence accelerated mouse(SAM-P/8). SAM-P/8 were randomly divided into seven groups:control group with saline only, aqueous extract of Scutellaria dose groups (7.6,3.8,1.9 g crude drug g·kg-1)and aqueous extract of Carthamus tinctorius dose groups (7.6,3.8,1.9 g crude drug g·kg-1), which had been consecutively received by intragastric administration for 4 weeks. Thymus and brain were collected to weigh and calculate organ coefficient.The activities of superoxide dismutase (SOD),glutathione peroxidase(GSH-Px), catalase(CAT) and the contents of malondialdehyde(MDA) in serum and brain homogenate of all groups were determined by kit. The results demonstrated that they could elevate the brain and thymus coefficient. They can also improve the activities of SOD, CAT, GSH-Px in different degree and decrease the contents of MDA.
2. The effects of 10 kinds of ethanol extracts from Chinese Herbs on Fe2+-ADP-NADPH induced lipid peroxidation(LPO) in rat liver microsomes
The effects of 10 kinds of ethanol extracts from Chinese Herbs on Fe2+-ADP-NADPH induced LPO were evaluated by calculating IC50 which repsents the concentration for 50% inhibition of LPO. The results demonstrated that ethanol extracts from Chinese Herbs and Trolox could significantly inhibit the generation of MDA induced by Fe2+-ADP-NADPH oxidation system and presented dose-response relationship in a certain range of concentration. IC50 of Trolox, Scutellaria baicalensis Georgi,Carthamus tinctorius L.,Camellia sinensis(L.)O.Ktze, Hypericum perforatum L., cold leaching of Sophora japonica L.,refluxing of Sophora japonica L. Oroxylum indicum (L.) Vent., Fagopyrum dibotrys(D.Don), Lonicera japonica Thunb., leaves and fruits of Crataegus pinnatifida Bge. are 0.014、0.066、0.598、0.030、0.041、0.041、0.058、0.098、0.324、0.458、1.069、6.156 mg·mL-1.
3. Antioxidant effects of 10 kinds of ethanol extracts from Chinese Herbs on ox-LDL induced injury in cardiac microvascular endothelial cells
The protection of 10 kinds of ethanol extracts from Chinese Herbs and Trolox were investigated after ox-LDL (100μg·mL-1) induced injury on the third generation of rat cardiac microvascular endothelial cells (rCMEC). The supernatant was collected for detecting MDA, SOD, GSH-Px activity. The results demonstrated that 10 kinds of ethanol extracts from Chinese Herbs increased the rCMEC survival rate, reduced MDA contents and improved SOD, GSH-Px activity.
4. The effects of 10 kinds of ethanol extracts from Chinese Herbs on xanthine-xanthine oxidase system injury in cultured neonatal rat cardiomyocytes
Antioxidant effects of 10 kinds of ethanol extracts from Chinese Herbs and Trolox were studied in the cultured neonatal rat cardiomyocytes which were injured by xanthine-xanthine oxidase system injury. The supernatant was collected for detecting lactate dehydrogenase(LDH), MDA, SOD, GSH-Px activity. The results demonstrated that 10 kinds of ethanol extracts from Chinese Herbs and Trolox increased the cardiomyocytes survival rate, and reduced LDH, MDA levels, and improved SOD, GSH-Px activity.
5. The effects of ethanol extracts from Camellia sinensis (L.)O.Ktze, cold leaching of Sophora japonica L., Oroxylum indicum (L.) Vent. on oxygen derived free radicals (OFR) signal of cardiac microvascular endothelial cells injured by ox-LDL and cardi-omyocytes injured by xanthine-xanthine oxidase system
Antioxidant effects of ethanol extracts from Camellia sinensis (L.)O.Ktze, cold leaching of Sophora japonica L., Oroxylum indicum (L.) Vent. on OFR signal were investigated by selecting cardiac microvascular endothelial cells injured by ox-LDL and cardiomyocytes injured by xanthine-xanthine oxidase system. cell suspension was collected for detecting OFR singal by electron spin resonance (ESR). The results demonstrated that OFR signal of the cell suspension decreased significantly.
Therefore, the results were suggested that 10 kinds of extracts from Chinese Herbs had anti-oxidation. The mechanism may be related with scavenging of free radicals, enhancing its clearance, enhancing endogenous antioxidant activity.
1.黄芩、红花水提取物对快速老化小鼠衰老模型的抗氧化作用的影响
选取快速老化模型小鼠P/8 (SAM-P/8),观察黄芩、红花水提物对快速老化小鼠的保护作用。实验共分5组：空白对照组,阳性药对照组,黄芩水提物组(7.6,3.8,1.9g生药·kg-1)、红花水提物组(7.6,3.8,1.9g生药.kg-1)剂量组,连续给药4周,末次给药1小时后取胸腺和脑称重测脏器系数；分别测定脑匀浆和血清中MDA, SOD, CAT和GSH-Px水平。实验结果表明：黄芩、红花水提物显著降低衰老小鼠的胸腺系数和脑系数,还可不同程度的提高衰老小鼠体内SOD、GSH-Px、CAT活性,抑制MDA含量的升高,提示黄芩、红花水提取物可能是通过提高机体的抗氧化能力达到抗衰老的作用。
2.黄芩等10种中药醇提物对Fe2+-ADP-NADPH所致肝微粒体脂质过氧化的影响
低温条件下制备大鼠肝微粒体,观察黄芩等10种中药醇提物对Fe2+-ADP-NADPH氧化体系诱发的脂质过氧化的抑制作用,通过计算半数抑制浓度评价上述10种中药醇提物的抗氧化作用。结果表明,10种中药提取物及标准品(水溶性的维生素E—Trolox)在一定浓度时可明显抑制体外Fe2+-ADP-NADPH氧化体系所致大鼠肝微粒体MDA的生成,且在一定浓度范围内呈现剂量-效应关系,标准品、黄芩、红花、茶叶、贯叶金丝桃、槐米冷浸、槐米回流、木蝴蝶、金荞麦、金银花、山楂叶、山楂鲜果的IC50分别为0.014、0.066、0.598、0.030、0.041、0.041、0.058、0.098、0.324、0.458、1.069、6.156 mg·mL-1,抗脂质过氧化活性由大到小依次为标准品>茶叶>槐米冷浸>贯叶金丝桃>槐米回流>黄芩>木蝴蝶>金荞麦>金银花>红花>山楂叶>山楂果；其中,Trolox有较强的抑制脂质过氧化作用,山楂果的脂质过氧化抑制作用较弱。
3.黄芩等10种中药醇提物对ox-LDL所致心肌微血管内皮细胞损伤的影响
采用第三代大鼠乳鼠心肌微血管内皮细胞,观察黄芩等10种中药醇提物及Trolox预处理对ox-LDL诱导损伤的心肌微血管内皮细胞的保护作用。实验分5组,空白对照组：正常完全培养基培养；模型对照组：加入ox-LDL (100μg·mL-1)孵育24h；提取物大、中、小剂量组：分别加入不同浓度的提取物预处理24h,然后换为100μg·mL-1的ox-LDL溶液孵育24h。所有细胞均在37℃,95%O2+5%CO2,饱和湿度条件下的二氧化碳培养箱中培养。实验结束后分别收集细胞上清液测定MDA, SOD和GSH-Px活性。实验结果表明：10种中药醇提物及Trolox预处理能明显提高ox-LDL损伤的内皮细胞存活率,减少MDA含量,提高SOD,GSH-Px活性。结果提示10种中药醇提物对ox-LDL损伤的心脏微血管内皮细胞具有不同程度的保护作用,其机制可能与直接减少自由基生成,增强其清除以及增强内源性抗氧化酶的活性有关。
4.黄芩等10种中药醇提物对黄嘌呤／黄嘌呤氧化酶所致心肌细胞氧化损伤的影响
采用原代培养的乳鼠心肌细胞,建立心肌细胞黄嘌呤／黄嘌呤氧化酶氧化损伤模型,观察10种中药醇提物及Trolox对心肌细胞黄嘌呤／黄嘌呤氧化酶氧化损伤的保护作用。实验分5组,空白对照组：正常普通培养基培养;模型对照组：换为4.2mmol·L-1的黄嘌呤与280U·L-1的黄嘌呤氧化酶孵育6h；提取物大、中、小剂量组：分别加入不同浓度的提取物的同时加4.2mmol·L-1的黄嘌呤与280U·L-1的黄嘌呤氧化酶作用6h。所有细胞均在37℃,95%02+5%CO2,饱和湿度条件下的二氧化碳培养箱中培养。实验结束后分别收集细胞上清液测定LDH,MDA,SOD和GSH-Px活性。实验结果表明：10种中药醇提物及Trolox对能明显提高黄嘌呤／黄嘌呤氧化酶氧化损伤的心肌细胞存活率,减少LDH, MDA含量,提高SOD, GSH-Px活性。结果提示10种中药醇提物对心肌细胞黄嘌呤／黄嘌呤氧化酶氧化损伤具有保护作用,其机制可能与直接减少自由基生成,增强其清除以及增强内源性抗氧化酶的活性有关。
5.茶叶、槐米冷浸、木蝴蝶醇提物对ox-LDL损伤心肌微血管内皮细胞和黄嘌呤／黄嘌呤氧化酶损伤心肌细胞中自由基信号的影响
采用原代培养的心肌微血管内皮细胞和乳鼠心肌细胞,建立心肌微血管内皮细胞ox-LDL损伤模型和心肌细胞黄嘌呤／黄嘌呤氧化酶氧化损伤模型,观察茶叶、槐米冷浸、木蝴蝶醇提物对内皮细胞和心肌细胞中自由基信号的影响。实验分组同前,所有细胞均在37℃,95%O2+5%CO2,饱和湿度条件下的二氧化碳培养箱中培养。实验结束后收集细胞悬液应用电子自旋波谱仪测定活性氧自由基信号。实验结果表明,茶叶、槐米冷浸、木蝴蝶醇提物能够不同程度的降低细胞悬液中自由基信号的峰值。结果提示,茶叶、槐米冷浸、木蝴蝶醇提物减轻细胞氧化损伤的作用可能也是通过减少自由基产生,增强其清除而实现的,与前期实验结果有一致性。
综上,黄芩、红花水提物在整体动物模型,对快速老化小鼠具有一定的抗衰老作用,黄芩等10种中药醇提物在Fe2+-ADP-NADPH所致肝微粒体脂质过氧化模型、微血管内皮细胞ox-LDL损伤模型、心肌细胞黄嘌呤／黄嘌呤氧化酶损伤模型等3个模型上,均可提高细胞上清液中的SOD, GSH-Px活性,降低MDA的含量,通过ESR检测,发现茶叶、槐米冷浸、木蝴蝶还可直接减少自由基的产生,增强其清除而达到抗氧化的作用。结果提示黄芩等10种中药提取物具有抗氧化作用,其机制与减少自由基生成,增强其清除,增强内源性抗氧化酶的活性,减少氧化产物生成等有关。
Oxygen free radicals (OFR) also known as reactive oxygen species, including superoxide anion radical (O2·), hydroxyl radical (OH·), hydrogen peroxide (H2O2), singlet oxygen (1O2·), lipid oxygen free radicals (RO·), lipid peroxy radicals (ROO·), NO and so on. Oxygen free radicals are closely related with many diseases. Therefore, what people have been looking forward to solving is preventing or reducing the body's oxygen free radicals. Although the traditional synthetic antioxidants have strong antioxidant capacity, they also have long-term of potentially toxic, some even inducing teratogenic and carcinogenic effect. Thus finding natural, efficient, low toxicity anti-oxidants from Chinese herbs has become an inevitable trend. Relevant experiments in vivo and vitro showed the 10 kinds of chinese herbs including Scutellaria baicalensis Georgi,Carthamus tinctorius L., Camellia sinensis(L.)O.Ktze,Hypericum perforatum L.,Sophora japonica L., Oroxylum indicum (L.) Vent., Fagopyrum dibotrys(D.Don), Lonicera japonica Thunb., leaves and fruits of Crataegus pinnatifida Bge. have antioxidant activity by different degrees.Therefore, this study was to investigate the effect of 10 kinds of extracts from Chinese Herbs on the oxidative injury in order to provide the scientific evidence for wide clinical application.
1. Antioxidant effects of water extracts of Scutellaria and Carthamus tinctorius in senescence accelerated mouse(SAM-P/8)
Antioxidant effects of aqueous extract of Scutellaria and Carthamus tinctorius were observed by selecting senescence accelerated mouse(SAM-P/8). SAM-P/8 were randomly divided into seven groups:control group with saline only, aqueous extract of Scutellaria dose groups (7.6,3.8,1.9 g crude drug g·kg-1)and aqueous extract of Carthamus tinctorius dose groups (7.6,3.8,1.9 g crude drug g·kg-1), which had been consecutively received by intragastric administration for 4 weeks. Thymus and brain were collected to weigh and calculate organ coefficient.The activities of superoxide dismutase (SOD),glutathione peroxidase(GSH-Px), catalase(CAT) and the contents of malondialdehyde(MDA) in serum and brain homogenate of all groups were determined by kit. The results demonstrated that they could elevate the brain and thymus coefficient. They can also improve the activities of SOD, CAT, GSH-Px in different degree and decrease the contents of MDA.
2. The effects of 10 kinds of ethanol extracts from Chinese Herbs on Fe2+-ADP-NADPH induced lipid peroxidation(LPO) in rat liver microsomes
The effects of 10 kinds of ethanol extracts from Chinese Herbs on Fe2+-ADP-NADPH induced LPO were evaluated by calculating IC50 which repsents the concentration for 50% inhibition of LPO. The results demonstrated that ethanol extracts from Chinese Herbs and Trolox could significantly inhibit the generation of MDA induced by Fe2+-ADP-NADPH oxidation system and presented dose-response relationship in a certain range of concentration. IC50 of Trolox, Scutellaria baicalensis Georgi,Carthamus tinctorius L.,Camellia sinensis(L.)O.Ktze, Hypericum perforatum L., cold leaching of Sophora japonica L.,refluxing of Sophora japonica L. Oroxylum indicum (L.) Vent., Fagopyrum dibotrys(D.Don), Lonicera japonica Thunb., leaves and fruits of Crataegus pinnatifida Bge. are 0.014、0.066、0.598、0.030、0.041、0.041、0.058、0.098、0.324、0.458、1.069、6.156 mg·mL-1.
3. Antioxidant effects of 10 kinds of ethanol extracts from Chinese Herbs on ox-LDL induced injury in cardiac microvascular endothelial cells
The protection of 10 kinds of ethanol extracts from Chinese Herbs and Trolox were investigated after ox-LDL (100μg·mL-1) induced injury on the third generation of rat cardiac microvascular endothelial cells (rCMEC). The supernatant was collected for detecting MDA, SOD, GSH-Px activity. The results demonstrated that 10 kinds of ethanol extracts from Chinese Herbs increased the rCMEC survival rate, reduced MDA contents and improved SOD, GSH-Px activity.
4. The effects of 10 kinds of ethanol extracts from Chinese Herbs on xanthine-xanthine oxidase system injury in cultured neonatal rat cardiomyocytes
Antioxidant effects of 10 kinds of ethanol extracts from Chinese Herbs and Trolox were studied in the cultured neonatal rat cardiomyocytes which were injured by xanthine-xanthine oxidase system injury. The supernatant was collected for detecting lactate dehydrogenase(LDH), MDA, SOD, GSH-Px activity. The results demonstrated that 10 kinds of ethanol extracts from Chinese Herbs and Trolox increased the cardiomyocytes survival rate, and reduced LDH, MDA levels, and improved SOD, GSH-Px activity.
5. The effects of ethanol extracts from Camellia sinensis (L.)O.Ktze, cold leaching of Sophora japonica L., Oroxylum indicum (L.) Vent. on oxygen derived free radicals (OFR) signal of cardiac microvascular endothelial cells injured by ox-LDL and cardi-omyocytes injured by xanthine-xanthine oxidase system
Antioxidant effects of ethanol extracts from Camellia sinensis (L.)O.Ktze, cold leaching of Sophora japonica L., Oroxylum indicum (L.) Vent. on OFR signal were investigated by selecting cardiac microvascular endothelial cells injured by ox-LDL and cardiomyocytes injured by xanthine-xanthine oxidase system. cell suspension was collected for detecting OFR singal by electron spin resonance (ESR). The results demonstrated that OFR signal of the cell suspension decreased significantly.
Therefore, the results were suggested that 10 kinds of extracts from Chinese Herbs had anti-oxidation. The mechanism may be related with scavenging of free radicals, enhancing its clearance, enhancing endogenous antioxidant activity.
引文
[1]Ralf P,Brandes, Kreuzer J. Vascular NADPH oxidases:molecular mechanisms of activation[J]. Cardiovasc Res,2005,65:16-27.
[2]Vaziri ND, Ni Z. Expression of NOX-I, gp91phox, p47phox and P67phox in the aorta segments above and below coarctation[J]. Biochim Biophys Acta,2005,1723(1-3):321-327.
[3]杨万松.高血压与动脉粥样硬化.见：张鸿修,黄体钢主编.实用冠心病学.第4版.天津：天津科技翻译出版公司。2005.181-185.
[4]魏文青,王荣杰,丛建波,等.自由基与细胞信号转导[J].国外医学生理、病理科学与临床分册,1999,19(5)：329-332.
[5]Raha S, Robinson BH. Mitochondria, oxygen free radicals, and apoptosis[J]. Amj Med Genet,2001,106(1):62-70.
[6]HuangH,Joazeiro CA,Bonfoco E,et al. The inhibitor of apoptosis CIAP2, functions as a ubiquitin-protein ligase and promotes in vitro monoubiquitination of caspase 3 and 7[J]. J Biol Chem,2002,275(35):26661-26664.
[7]兰希发,姚秀文,郭阳.脑缺血再灌注后神经细胞凋亡的机制[J].中国临床康复,2003,7(19)：2726-2727.
[8]张春阳,卫涛涛,马辉,等.活性氧参与一氧化氮诱导的神经细胞凋亡[J].生物化字与生物物理进展,2001,28(1)：81-85.
[9]Ruiz Gines JA, Lopez Ongil S, Gonzalez Rubio M,et al.Reactive oxygen species induce proliferation of bovine aortic endothelial cells [J]. J Cardiovasc Pharmacol,2000,35(1):109-113.
[10]Lum H, Roebuck KA. Oxidant stress and endothelial cell dysfunction[J]. Am J Physiol Cell Physiol,2001,280(4):C719～C741.
[11]Nystrom T. The free-radical hypothesis of aging goes prokaryotic[J].Cell Mol Life Sci,2003,60(7):1333-1341.
[12]Droge W.Oxidative stress and aging[J]. Adv Exp Med Biol,2003,543:191-200.
[13]李国林,印大中.衰老分子生物化学中的羰基应激[J].生命科学研究,2003,7(2)：116-122.
[14]Bensi G,Moretti A. Age peroxidative stress related modifications of the cerebral enzymatic activities linked to mitochondria and the glutathione system [J].Free Radic Biol Med,1995,19:77.
[15]Nordlie MA,Wold LE, Simkhovich BZ, et al. Molecular aspects of ischemic heart disease: ischemia/reperfusion-induced genetic changes and potential applications of gene and RNA interference therapy[J]. J Cardiovasc Pharmacol Ther,2006,11:17-30.
[16]Zhao ZQ. Oxidative stress-elicited myocardial apoptosis during reperfusion[J]. Curr Opin Pharmacol,2004,4:159-165.
[17]Paradies G, Petrosillo G, PistoleseM, et al. Decrease in mitochondrial complex activity in ischemic/reperfused ratheart:involvement of reactive oxygen species and cardiolipin. Circ Res,2004,94:53-59.
[18]PchejetskiD, Kunduzova O, Dayon A, et al. Oxidative stress dependent sphinosine kinase-1 inhibition mediates monoamine oxidase A-associated cardiac cell apoptosis. Circ Res.2007,100:41-49.
[19]Yang JC, CortopassiGA. Induction of the mitochondrial perme ability transition causes release of the apoptogenic factor cytochrome c. Free Radic BiolMed,1998,24:624-631.
[20]谷天祥,张显清,谷春久,等.心肌缺血再灌注损伤亚细胞Ca2+反常与ATP酶泵功能抑制[J].中华心血管病杂志,2001,29(7)：420-423.
[21]NordlieMA,Wold LE, Simkhovich BZ, et al. Molecular aspects of ischemic heartdisease: ischemia/reperfusion-induced genetic changes and potential applica- tions of gene and RNA interference therapy[J].J Cardiovasc Pharmacol Ther,2006, 11(1):17-30.
[22]Kevin LG, Novalija E, Stowe DF. Reactive oxygen species asmediators of cardiac injury and protection:the relevance to anesthesia practice[J].Anesth Analg,2005,101:1275-1287.
[23]Selzner M, Rudiger HA, Selzner N. Transgenicmice overexpressing human Bcl-2 are resistant to hepatic ischemia and reperfusion[J]. J Hepatol,2002,36:218-225.
[24]Liao F, Andalibi A, Qiao JH, et al. Genetic evidence for a common pathway mediating oxidative stress, inflammatory gene induction, and aortic fatty streak formation inmice[J]. J Clin Invest,1994,94(2):877-884.
[25]Cathcart MK. Regulation of superoxide anion production by NADPH oxides in monocytes/macrophages:contributions to atherosclerosis[J]. Arterioscler Thromb Vasc Biol,2004,24(1):23-28.
[26]Steinberg D. Low density lipoprotein oxidation its pathobiological significance [J]. J Biol Chem,1997,272(34):20963-20966.
[27]Cecile M, Martin A, Mojgan DM, et al. Oxidized low density Lipoprotem induces activation of the transeriptional factor NF-KB in fibroblasts, endothelial and smooth muscle cells[J]. Biochem Mol Biol Int,1996,39(6):1201-1207.
[28]Cominacini L, Ulisse G, Pasini AF, et al. Antioxidants inhibit the expression of intercellular cell adhension molecula-1 and vascular cell adhension molecule-1 induced by oxidized LDL on human umbilical endothelial cells[J]. Free Radic Biol Med,1997,22(1-2):117-127.
[29]Kunsch C, Medford RM. Oxidative stress as a regulator of gene expression in the vasculature[J]. Circ Res,1999,85(8):753-766.
[30]Galis ZS, Muszynski M, Sukhova GK, et al. Enhanced expression of vascular matrix metalloproteinases induced in vitro by cytokines and in regions of human atherosclerotic lesions[J]. Ann N Y Acad Sci,1995,748:501-507.
[31]王晓雯,蒋晓燕,邬梨娅·伊明,等.肉苁蓉总甙体外清除自由基对·OH引起DNA的氧化损伤保护[J].中国药学杂志,2001,36(1)：29.
[32]金鸣,李金荣,吴伟.红花黄色素抗氧化作用的研究[J].中国中药杂志,2004,29(5)：447-449.
[33]陈新.黄芪对急性心肌粳塞患者心功能和氧自由基的影响及强心机制探讨[J].中国中西医结合杂志,1999,15(3)：141.
[34]何桂霞,杜方麓,杨伟丽,等.藤茶总黄酮清除氧自由基与抗脂质过氧化作用[J].中药材,2003,26(5)：338-340.
[35]朱建明.人参皂苷的抗衰老作用研究进展[J].中医药信息,1998,23(2)：18.
[36]JiXY,Tan Benny Kg,Zhu YC,et al.Comparison of cardioprotective effects using ramipril and DanShen for the treatment of acute myocardial infarction in rats[J]. Life Sci,2003,73:1413-1426.
[37]叶锦霞,梁日欣,王岚,等.红花水提物对ox-LDL损伤心肌微血管内皮细胞的保护作用及ESR谱研究[J].中国中药杂志,2008,33(21)：2513-2517.
[38]张启春,卞慧敏,喻丽珍,等.六味地黄方含药血清对氧化低密度脂蛋白损伤血管内皮细胞的保护作用[J].中国实验方剂学杂志,2007,13(7)：31-34.
[39]李志刚,蓝荣芳,关丽,等.人参皂苷Rgl抗O-LDL诱导内皮细胞凋亡及分子机制[J].中国组织化学与细胞化学杂志,2002,11(3)：248-253.
[40]宋瑞霞,余静,梁连生,等.甘肃黄芪异黄酮化合物对血管内皮细胞凋亡的影响[J].兰州大学学报,2007,33(1)：43-47
[41]高伟,梁日欣,肖永庆,等.川芎内酯A预处理对心肌微血管内皮细胞低氧／复氧损伤保护作用及机制研究[J].中国中药杂志,2007,32(2)：133-137.
[42]孙连胜,李妍,马佐英,等.葛根素对损伤内皮细胞白细胞介素-8释放的影响[J].天津医药,2006,23(2)：147-148.
[43]山丽梅,张锦超,赵艳玲,等.槟榔碱抗动脉粥样硬化分子机制的研究[J].中国药理学通报,2004,20(2)：146.
[44]Zhao B, Zhang Y, Lin B, et al.Endothelial Cells injured oxidized low density lipoproteins.[J].A mJ H-tol,1995,49(3):251.
[45]Celemaje DS. Endothelial dysfunction:does is matter?Is it reversible[J]? Am Coll Cardiol,1997,30:325.
[46]庞荣清,潘兴华,吴亚玲,等.三七总皂苷对兔血管平滑肌细胞核因子kappa B和细胞周期的影响[J].中国微循环,2004,8(3)：154.
[47]崔华,何作云.白藜三醇对黄嘌呤／黄嘌呤氧化酶所致平滑肌细胞增殖的干预作用[J].中国微循环,2003,7(4)：218-220.
[48]崔华,何作云,高玉琪.白藜三醇对氧自由基致兔血管平滑肌细胞NF-KB活化及MCP-1表达的影响[J].中国病理生理杂志,2004,20(10)：1837-1841.
[49]干蓓,陈可冀.血府逐瘀浓缩九防治43例冠心病内支架植入术后再狭窄的临床研究[J].中国中西医结合杂志,1998,18(10)：585.
[50]吴宗贵.炎症与动脉粥样硬化：挑战远未结束[J].上海医学,2004,27(4)：211.
[51]王毅,马志强,黄波,等.粉防己碱对高脂饮食兔血清白细胞介素-1p、肿瘤坏死因子-α及血管壁细胞核因子-κB表达的影响[J].中国临床康复,2004,8(25)：5045.
[52]浦奎,祝善俊,姚智.心肌缺血-再灌注时体内细胞因子活性动态变化的实验研究[J].中国心血管病杂志,1997,25(4)：305-307.
[53]常天辉,刘晓阳,章新华,等.白花前胡及前胡甲素对心肌缺血再灌注大鼠IL-6水平及Fas、Bax、BCL-2蛋白表达的影响[J].中国医科大学学报,2003,32(1)：1-3.
[54]徐强,司良毅,赵小兰,等.Caspase-3活性对老年大鼠心肌缺血再灌注损伤的影响[J].实用老年医学,2004,18(2)：71-74.
[55]王秀华,李红,魏征人,等.蒺藜皂苷对NaCN诱导大鼠乳鼠心肌细胞凋亡的抑制作用及机制[J].吉林大学学报(医学版),2005,31(1)：5-9.
[56]季凤清,李宝红,孙海梅,等.槲皮素对急性心肌缺血大鼠心肌caspase3基因蛋白表达的影响[J].首都医科大学学报,2004,25(4)：435-437
[57]黎玉,万福生,李少华.川芎嗪对大鼠缺血心肌线粒体NO及自由基的影响[J].中医药学报,2004,32(2)：47-48.
[58]Jenningds RB,Reimer KA.Lethal myocardial ischemic injury.Am J Pathol, 1981,102:241.
[59]黄起壬,何明,李萍,等.油茶皂苷抗心肌缺血大鼠氧自由基和脂质过氧化作用[J].中国药理学通报,2003,19(9)：1034-1036.
[60]李立中,吴基良,胡宗礼,等.茶多酚对大鼠心肌缺血再灌注损伤的影响[J],中药药理与临床,1997,13(4)：21-23.
[61]Silverman HS,Stern MD.Ionic basis of ischemic cardiac injurying:insights from cellular studies[J].Cardiovasc Res,1994,28(5):581～597.
[62]杨远友,刘宁,莫正纪,等.淫羊藿对大鼠内脏器官PDC钙通道及其心肌缺血性损伤的影响[J].四川大学学报(自然科学版),2005,42(1)：122-127.
[1]陈瑗,周玫.自由基医学.第1版,北京：人民军医出版社,1991,142-161.
[2]张明霞,李效忠.红花抗衰老作用的实验研究[J].中草药,2001,32(1)：52-54
[3]金鸣,李金荣,蔡亚欣,等.红花水溶性成分抗氧化作用的研究[J].心肺血管病杂志,1998,17(4)：227-279.
[4]陈桂枝,罗德生,郑红花,等.四氯化碳肝损伤与脂质过氧化时黄芩煎剂的保护作用[J].咸宁学院学报(医学版),2005,19(1)：31-33.
[5]张永钦,周井炎,徐辉碧.黄芩甙的抗氧化作用[J].华中理工大学学报,1999,27(4)：111-11.
[6]陈瑾歆.自由基与衰老关系的研究进展[J].川北医学院学报,2004,19(1)：207-209.
[7]李磷,丁安伟,等.衰老机理研究进展[J].西北药学杂志2000,15(4)：177-178.
[8]Takeda T. Senescence-acceleratedmouse (SAM):a biogerontological resource in aging research[J]. Neurobiol Aging,1999,20(2):105-10.
[9]聂伟,张永祥.快速老化小鼠-研究衰老及衰老相关疾病的动物模型[J].中国药理学通报,,2000,16(2)：132-137.
[10]李国星,刘克明,何宁,等.快速老化鼠和自然衰老鼠中超氧化物歧化酶的比较研究[J].中国老年学杂志,2007,27(9)：820-822.
[11]Boldyrev AA, Yuneva MO, Sorokina EV, et al. Antioxidant systems in tissues of senescence accelerated mice.Biochemistry (Mosc)[J].2001,66 (10):1157-63.
[12]Yokozawa T, Satoh A, Cho EJ. Ginsenoside Rd attenuates oxidative damage related to aging in senescence accelerated mice. Pharm Pharmaco [J].2004,56(1):107-13.
[13]陈志蓉.抗衰老药物的研究进展[J].海峡药学,2008,20(1)：8-11.
[14]Bin Yang, Akira Kotani, Kensuke Arai, et al. Relationship of Electrochemical Oxidation of Catechins on Their Antioxidant Activity in Microsomal Lipid Peroxidation[J].Chem,Pharm, Bull,2001,49(6):747-751.
[15]张均田.现代药理实验方法(下册)[M].北京：北京医科大学、中国协和医科大学联合出版社。1997.1220-123.
[16]赵保路编著.氧自由基和天然抗氧化剂[M].北京：科学出版社,1999.
[17]Nishida M, Carley WW, Gerritsen ME, et al. Isolation and characterization of human and rat cardiac microvascular endotheliall cells. Am J Physiol,1993,264:H639-52.
[18]Anderson JW, Diwadkar VA, Bridges SR. Selective effects of different antioxidants on oxidation of lipoproteins from rats. Proc Soc Exp Biol Med,1998:218(4):376-81.
[19]司徒镇强,吴军正主编.细胞培养[M].西安：世界图书出版公司,2000年
[20]曹莹,梁日欣,王岚.升麻甙与5-O甲基维斯阿米醇甙对血管内皮细胞分泌细胞因子的影响[J].中药药理与临床,2007,23(3)：13-15.
[21]Deshpande NN, Sorescu D, Seshiah P, et al. Mechanism of hydrogen peroxide-induced cell cycle arrest in vascular smooth muscle [J]. Antioxid Redox Signal,2002,4(5):845-54
[22]Forman HJ,Torres M,Fukuto J.Redox signaling [J].Mol Cell Biochem,2002,234-235(1-2): 49-62
[23]Kuzkaya N, Weissmann N, Harrison DG, et al. Interactions of peroxynitrite tetrahydrobiopterin, ascorbic acid, and thiols:implications for uncoupling endothelial nitric-oxide synthase [J]. J Biol Chem,2003,278(25):22546-54
[24]Seifried HE, McDonald SS, Anderson DE, et al. The antioxidant conundrumin cancer[J].Cancer Res.2003,63(15):4295-4298.
[25]Diplock AT, Charleux JL, Crozier-Willi G, et al. Functional food scienceand defence against reactive oxidative species[J].Br JNutr.1998,80 Suppl 1:S77-112.
[26]Chang MY, Lees AM, Lees RS. Low-density lipoprotein modification and arterial wall accumulation in a rabbit model of atherosclerosis[J].Biochemistry.1993,32:8518-8524.
[27]Hein TW, KuoL. LDLs impair vasomotor function of the coronary microcirculation:role of superoxide anions[J].Circ Res.1998,83:404-414.
[28]黄从刚,魏劲波.自由基清除剂对动脉粥样硬化的影响[J].医学综述,1998,(4)：166.
[29]汪长华.心肌细胞的培养及注意事项[J].中国心脏起搏与心电生理杂志,2003,17(3)：230-232.
[30]Webster KA, Discher DJ, Bishopric NH. Cardioprotection in an in vitro model of hypoxic preconditioning. J Mol Cell Cardiol,1995,27(1):453-8.
[31]王达理,曾爱萍,南柏松.红花水提物抗自由基保护心肌细胞的作用[J].中国中医基础医学杂志,2001,7(2)：34-35.
[32]张小郁,李文广,高明堂等.葡萄籽中原花青素对心肌细胞的保护作用[J].中药药理与临床,2001,17(6)：14-16.
[33]工兴祥,周利龙,冯义柏等.超氧阴离子白由基对心肌细胞的损伤作用[J].临床心血管病杂志,2003,19(7)：410-412.
[34]曹莹,梁日欣,杨滨等(-)-儿茶素没食子酸酯和(+)-表儿茶素对心肌细胞保护作用研究[J].中国实验方剂学杂志,2006,12(10)：36-38.
[35]叶锦霞,梁日欣,王岚,等.红花水提物对ox-LDL损伤心肌微血管内皮细胞的保护作用及ESR谱研究[J].中国中药杂志,2008,33(21)：2513-2517.
[36]Hausenloy DJ, Duchen MR, Yellon DM. Inhibiting mitochondrial permeability transition poreopening at reperfusion protects against ischaemia-reperfusion injury [J]. Cardiovasc Res, 2003,60(3):617-25
[37]Yang XM, Downey JM, Cohen MW. Multiple brief coronary occlusions during early reperfusion protect rabbit hearts by activation of ERK and production of nitric oxide [J]. Circulation,2003,108(supplIV):158
[38]孙成文,赵春燕,钟国赣.人参二醇单体对大鼠心肌作用的单钙通道分析及ESR谱研究[J].基础医学与临床,1994,14(4)：286.
[39]Tatiana D, Galina T, Vilhelmina J.Characterization of the Radical Scavenging Activity of Lignins- natural Antioxidants[J].Biores Tech,2004,95:309-317.
[2]Vaziri ND, Ni Z. Expression of NOX-I, gp91phox, p47phox and P67phox in the aorta segments above and below coarctation[J]. Biochim Biophys Acta,2005,1723(1-3):321-327.
[3]杨万松.高血压与动脉粥样硬化.见：张鸿修,黄体钢主编.实用冠心病学.第4版.天津：天津科技翻译出版公司。2005.181-185.
[4]魏文青,王荣杰,丛建波,等.自由基与细胞信号转导[J].国外医学生理、病理科学与临床分册,1999,19(5)：329-332.
[5]Raha S, Robinson BH. Mitochondria, oxygen free radicals, and apoptosis[J]. Amj Med Genet,2001,106(1):62-70.
[6]HuangH,Joazeiro CA,Bonfoco E,et al. The inhibitor of apoptosis CIAP2, functions as a ubiquitin-protein ligase and promotes in vitro monoubiquitination of caspase 3 and 7[J]. J Biol Chem,2002,275(35):26661-26664.
[7]兰希发,姚秀文,郭阳.脑缺血再灌注后神经细胞凋亡的机制[J].中国临床康复,2003,7(19)：2726-2727.
[8]张春阳,卫涛涛,马辉,等.活性氧参与一氧化氮诱导的神经细胞凋亡[J].生物化字与生物物理进展,2001,28(1)：81-85.
[9]Ruiz Gines JA, Lopez Ongil S, Gonzalez Rubio M,et al.Reactive oxygen species induce proliferation of bovine aortic endothelial cells [J]. J Cardiovasc Pharmacol,2000,35(1):109-113.
[10]Lum H, Roebuck KA. Oxidant stress and endothelial cell dysfunction[J]. Am J Physiol Cell Physiol,2001,280(4):C719～C741.
[11]Nystrom T. The free-radical hypothesis of aging goes prokaryotic[J].Cell Mol Life Sci,2003,60(7):1333-1341.
[12]Droge W.Oxidative stress and aging[J]. Adv Exp Med Biol,2003,543:191-200.
[13]李国林,印大中.衰老分子生物化学中的羰基应激[J].生命科学研究,2003,7(2)：116-122.
[14]Bensi G,Moretti A. Age peroxidative stress related modifications of the cerebral enzymatic activities linked to mitochondria and the glutathione system [J].Free Radic Biol Med,1995,19:77.
[15]Nordlie MA,Wold LE, Simkhovich BZ, et al. Molecular aspects of ischemic heart disease: ischemia/reperfusion-induced genetic changes and potential applications of gene and RNA interference therapy[J]. J Cardiovasc Pharmacol Ther,2006,11:17-30.
[16]Zhao ZQ. Oxidative stress-elicited myocardial apoptosis during reperfusion[J]. Curr Opin Pharmacol,2004,4:159-165.
[17]Paradies G, Petrosillo G, PistoleseM, et al. Decrease in mitochondrial complex activity in ischemic/reperfused ratheart:involvement of reactive oxygen species and cardiolipin. Circ Res,2004,94:53-59.
[18]PchejetskiD, Kunduzova O, Dayon A, et al. Oxidative stress dependent sphinosine kinase-1 inhibition mediates monoamine oxidase A-associated cardiac cell apoptosis. Circ Res.2007,100:41-49.
[19]Yang JC, CortopassiGA. Induction of the mitochondrial perme ability transition causes release of the apoptogenic factor cytochrome c. Free Radic BiolMed,1998,24:624-631.
[20]谷天祥,张显清,谷春久,等.心肌缺血再灌注损伤亚细胞Ca2+反常与ATP酶泵功能抑制[J].中华心血管病杂志,2001,29(7)：420-423.
[21]NordlieMA,Wold LE, Simkhovich BZ, et al. Molecular aspects of ischemic heartdisease: ischemia/reperfusion-induced genetic changes and potential applica- tions of gene and RNA interference therapy[J].J Cardiovasc Pharmacol Ther,2006, 11(1):17-30.
[22]Kevin LG, Novalija E, Stowe DF. Reactive oxygen species asmediators of cardiac injury and protection:the relevance to anesthesia practice[J].Anesth Analg,2005,101:1275-1287.
[23]Selzner M, Rudiger HA, Selzner N. Transgenicmice overexpressing human Bcl-2 are resistant to hepatic ischemia and reperfusion[J]. J Hepatol,2002,36:218-225.
[24]Liao F, Andalibi A, Qiao JH, et al. Genetic evidence for a common pathway mediating oxidative stress, inflammatory gene induction, and aortic fatty streak formation inmice[J]. J Clin Invest,1994,94(2):877-884.
[25]Cathcart MK. Regulation of superoxide anion production by NADPH oxides in monocytes/macrophages:contributions to atherosclerosis[J]. Arterioscler Thromb Vasc Biol,2004,24(1):23-28.
[26]Steinberg D. Low density lipoprotein oxidation its pathobiological significance [J]. J Biol Chem,1997,272(34):20963-20966.
[27]Cecile M, Martin A, Mojgan DM, et al. Oxidized low density Lipoprotem induces activation of the transeriptional factor NF-KB in fibroblasts, endothelial and smooth muscle cells[J]. Biochem Mol Biol Int,1996,39(6):1201-1207.
[28]Cominacini L, Ulisse G, Pasini AF, et al. Antioxidants inhibit the expression of intercellular cell adhension molecula-1 and vascular cell adhension molecule-1 induced by oxidized LDL on human umbilical endothelial cells[J]. Free Radic Biol Med,1997,22(1-2):117-127.
[29]Kunsch C, Medford RM. Oxidative stress as a regulator of gene expression in the vasculature[J]. Circ Res,1999,85(8):753-766.
[30]Galis ZS, Muszynski M, Sukhova GK, et al. Enhanced expression of vascular matrix metalloproteinases induced in vitro by cytokines and in regions of human atherosclerotic lesions[J]. Ann N Y Acad Sci,1995,748:501-507.
[31]王晓雯,蒋晓燕,邬梨娅·伊明,等.肉苁蓉总甙体外清除自由基对·OH引起DNA的氧化损伤保护[J].中国药学杂志,2001,36(1)：29.
[32]金鸣,李金荣,吴伟.红花黄色素抗氧化作用的研究[J].中国中药杂志,2004,29(5)：447-449.
[33]陈新.黄芪对急性心肌粳塞患者心功能和氧自由基的影响及强心机制探讨[J].中国中西医结合杂志,1999,15(3)：141.
[34]何桂霞,杜方麓,杨伟丽,等.藤茶总黄酮清除氧自由基与抗脂质过氧化作用[J].中药材,2003,26(5)：338-340.
[35]朱建明.人参皂苷的抗衰老作用研究进展[J].中医药信息,1998,23(2)：18.
[36]JiXY,Tan Benny Kg,Zhu YC,et al.Comparison of cardioprotective effects using ramipril and DanShen for the treatment of acute myocardial infarction in rats[J]. Life Sci,2003,73:1413-1426.
[37]叶锦霞,梁日欣,王岚,等.红花水提物对ox-LDL损伤心肌微血管内皮细胞的保护作用及ESR谱研究[J].中国中药杂志,2008,33(21)：2513-2517.
[38]张启春,卞慧敏,喻丽珍,等.六味地黄方含药血清对氧化低密度脂蛋白损伤血管内皮细胞的保护作用[J].中国实验方剂学杂志,2007,13(7)：31-34.
[39]李志刚,蓝荣芳,关丽,等.人参皂苷Rgl抗O-LDL诱导内皮细胞凋亡及分子机制[J].中国组织化学与细胞化学杂志,2002,11(3)：248-253.
[40]宋瑞霞,余静,梁连生,等.甘肃黄芪异黄酮化合物对血管内皮细胞凋亡的影响[J].兰州大学学报,2007,33(1)：43-47
[41]高伟,梁日欣,肖永庆,等.川芎内酯A预处理对心肌微血管内皮细胞低氧／复氧损伤保护作用及机制研究[J].中国中药杂志,2007,32(2)：133-137.
[42]孙连胜,李妍,马佐英,等.葛根素对损伤内皮细胞白细胞介素-8释放的影响[J].天津医药,2006,23(2)：147-148.
[43]山丽梅,张锦超,赵艳玲,等.槟榔碱抗动脉粥样硬化分子机制的研究[J].中国药理学通报,2004,20(2)：146.
[44]Zhao B, Zhang Y, Lin B, et al.Endothelial Cells injured oxidized low density lipoproteins.[J].A mJ H-tol,1995,49(3):251.
[45]Celemaje DS. Endothelial dysfunction:does is matter?Is it reversible[J]? Am Coll Cardiol,1997,30:325.
[46]庞荣清,潘兴华,吴亚玲,等.三七总皂苷对兔血管平滑肌细胞核因子kappa B和细胞周期的影响[J].中国微循环,2004,8(3)：154.
[47]崔华,何作云.白藜三醇对黄嘌呤／黄嘌呤氧化酶所致平滑肌细胞增殖的干预作用[J].中国微循环,2003,7(4)：218-220.
[48]崔华,何作云,高玉琪.白藜三醇对氧自由基致兔血管平滑肌细胞NF-KB活化及MCP-1表达的影响[J].中国病理生理杂志,2004,20(10)：1837-1841.
[49]干蓓,陈可冀.血府逐瘀浓缩九防治43例冠心病内支架植入术后再狭窄的临床研究[J].中国中西医结合杂志,1998,18(10)：585.
[50]吴宗贵.炎症与动脉粥样硬化：挑战远未结束[J].上海医学,2004,27(4)：211.
[51]王毅,马志强,黄波,等.粉防己碱对高脂饮食兔血清白细胞介素-1p、肿瘤坏死因子-α及血管壁细胞核因子-κB表达的影响[J].中国临床康复,2004,8(25)：5045.
[52]浦奎,祝善俊,姚智.心肌缺血-再灌注时体内细胞因子活性动态变化的实验研究[J].中国心血管病杂志,1997,25(4)：305-307.
[53]常天辉,刘晓阳,章新华,等.白花前胡及前胡甲素对心肌缺血再灌注大鼠IL-6水平及Fas、Bax、BCL-2蛋白表达的影响[J].中国医科大学学报,2003,32(1)：1-3.
[54]徐强,司良毅,赵小兰,等.Caspase-3活性对老年大鼠心肌缺血再灌注损伤的影响[J].实用老年医学,2004,18(2)：71-74.
[55]王秀华,李红,魏征人,等.蒺藜皂苷对NaCN诱导大鼠乳鼠心肌细胞凋亡的抑制作用及机制[J].吉林大学学报(医学版),2005,31(1)：5-9.
[56]季凤清,李宝红,孙海梅,等.槲皮素对急性心肌缺血大鼠心肌caspase3基因蛋白表达的影响[J].首都医科大学学报,2004,25(4)：435-437
[57]黎玉,万福生,李少华.川芎嗪对大鼠缺血心肌线粒体NO及自由基的影响[J].中医药学报,2004,32(2)：47-48.
[58]Jenningds RB,Reimer KA.Lethal myocardial ischemic injury.Am J Pathol, 1981,102:241.
[59]黄起壬,何明,李萍,等.油茶皂苷抗心肌缺血大鼠氧自由基和脂质过氧化作用[J].中国药理学通报,2003,19(9)：1034-1036.
[60]李立中,吴基良,胡宗礼,等.茶多酚对大鼠心肌缺血再灌注损伤的影响[J],中药药理与临床,1997,13(4)：21-23.
[61]Silverman HS,Stern MD.Ionic basis of ischemic cardiac injurying:insights from cellular studies[J].Cardiovasc Res,1994,28(5):581～597.
[62]杨远友,刘宁,莫正纪,等.淫羊藿对大鼠内脏器官PDC钙通道及其心肌缺血性损伤的影响[J].四川大学学报(自然科学版),2005,42(1)：122-127.
[1]陈瑗,周玫.自由基医学.第1版,北京：人民军医出版社,1991,142-161.
[2]张明霞,李效忠.红花抗衰老作用的实验研究[J].中草药,2001,32(1)：52-54
[3]金鸣,李金荣,蔡亚欣,等.红花水溶性成分抗氧化作用的研究[J].心肺血管病杂志,1998,17(4)：227-279.
[4]陈桂枝,罗德生,郑红花,等.四氯化碳肝损伤与脂质过氧化时黄芩煎剂的保护作用[J].咸宁学院学报(医学版),2005,19(1)：31-33.
[5]张永钦,周井炎,徐辉碧.黄芩甙的抗氧化作用[J].华中理工大学学报,1999,27(4)：111-11.
[6]陈瑾歆.自由基与衰老关系的研究进展[J].川北医学院学报,2004,19(1)：207-209.
[7]李磷,丁安伟,等.衰老机理研究进展[J].西北药学杂志2000,15(4)：177-178.
[8]Takeda T. Senescence-acceleratedmouse (SAM):a biogerontological resource in aging research[J]. Neurobiol Aging,1999,20(2):105-10.
[9]聂伟,张永祥.快速老化小鼠-研究衰老及衰老相关疾病的动物模型[J].中国药理学通报,,2000,16(2)：132-137.
[10]李国星,刘克明,何宁,等.快速老化鼠和自然衰老鼠中超氧化物歧化酶的比较研究[J].中国老年学杂志,2007,27(9)：820-822.
[11]Boldyrev AA, Yuneva MO, Sorokina EV, et al. Antioxidant systems in tissues of senescence accelerated mice.Biochemistry (Mosc)[J].2001,66 (10):1157-63.
[12]Yokozawa T, Satoh A, Cho EJ. Ginsenoside Rd attenuates oxidative damage related to aging in senescence accelerated mice. Pharm Pharmaco [J].2004,56(1):107-13.
[13]陈志蓉.抗衰老药物的研究进展[J].海峡药学,2008,20(1)：8-11.
[14]Bin Yang, Akira Kotani, Kensuke Arai, et al. Relationship of Electrochemical Oxidation of Catechins on Their Antioxidant Activity in Microsomal Lipid Peroxidation[J].Chem,Pharm, Bull,2001,49(6):747-751.
[15]张均田.现代药理实验方法(下册)[M].北京：北京医科大学、中国协和医科大学联合出版社。1997.1220-123.
[16]赵保路编著.氧自由基和天然抗氧化剂[M].北京：科学出版社,1999.
[17]Nishida M, Carley WW, Gerritsen ME, et al. Isolation and characterization of human and rat cardiac microvascular endotheliall cells. Am J Physiol,1993,264:H639-52.
[18]Anderson JW, Diwadkar VA, Bridges SR. Selective effects of different antioxidants on oxidation of lipoproteins from rats. Proc Soc Exp Biol Med,1998:218(4):376-81.
[19]司徒镇强,吴军正主编.细胞培养[M].西安：世界图书出版公司,2000年
[20]曹莹,梁日欣,王岚.升麻甙与5-O甲基维斯阿米醇甙对血管内皮细胞分泌细胞因子的影响[J].中药药理与临床,2007,23(3)：13-15.
[21]Deshpande NN, Sorescu D, Seshiah P, et al. Mechanism of hydrogen peroxide-induced cell cycle arrest in vascular smooth muscle [J]. Antioxid Redox Signal,2002,4(5):845-54
[22]Forman HJ,Torres M,Fukuto J.Redox signaling [J].Mol Cell Biochem,2002,234-235(1-2): 49-62
[23]Kuzkaya N, Weissmann N, Harrison DG, et al. Interactions of peroxynitrite tetrahydrobiopterin, ascorbic acid, and thiols:implications for uncoupling endothelial nitric-oxide synthase [J]. J Biol Chem,2003,278(25):22546-54
[24]Seifried HE, McDonald SS, Anderson DE, et al. The antioxidant conundrumin cancer[J].Cancer Res.2003,63(15):4295-4298.
[25]Diplock AT, Charleux JL, Crozier-Willi G, et al. Functional food scienceand defence against reactive oxidative species[J].Br JNutr.1998,80 Suppl 1:S77-112.
[26]Chang MY, Lees AM, Lees RS. Low-density lipoprotein modification and arterial wall accumulation in a rabbit model of atherosclerosis[J].Biochemistry.1993,32:8518-8524.
[27]Hein TW, KuoL. LDLs impair vasomotor function of the coronary microcirculation:role of superoxide anions[J].Circ Res.1998,83:404-414.
[28]黄从刚,魏劲波.自由基清除剂对动脉粥样硬化的影响[J].医学综述,1998,(4)：166.
[29]汪长华.心肌细胞的培养及注意事项[J].中国心脏起搏与心电生理杂志,2003,17(3)：230-232.
[30]Webster KA, Discher DJ, Bishopric NH. Cardioprotection in an in vitro model of hypoxic preconditioning. J Mol Cell Cardiol,1995,27(1):453-8.
[31]王达理,曾爱萍,南柏松.红花水提物抗自由基保护心肌细胞的作用[J].中国中医基础医学杂志,2001,7(2)：34-35.
[32]张小郁,李文广,高明堂等.葡萄籽中原花青素对心肌细胞的保护作用[J].中药药理与临床,2001,17(6)：14-16.
[33]工兴祥,周利龙,冯义柏等.超氧阴离子白由基对心肌细胞的损伤作用[J].临床心血管病杂志,2003,19(7)：410-412.
[34]曹莹,梁日欣,杨滨等(-)-儿茶素没食子酸酯和(+)-表儿茶素对心肌细胞保护作用研究[J].中国实验方剂学杂志,2006,12(10)：36-38.
[35]叶锦霞,梁日欣,王岚,等.红花水提物对ox-LDL损伤心肌微血管内皮细胞的保护作用及ESR谱研究[J].中国中药杂志,2008,33(21)：2513-2517.
[36]Hausenloy DJ, Duchen MR, Yellon DM. Inhibiting mitochondrial permeability transition poreopening at reperfusion protects against ischaemia-reperfusion injury [J]. Cardiovasc Res, 2003,60(3):617-25
[37]Yang XM, Downey JM, Cohen MW. Multiple brief coronary occlusions during early reperfusion protect rabbit hearts by activation of ERK and production of nitric oxide [J]. Circulation,2003,108(supplIV):158
[38]孙成文,赵春燕,钟国赣.人参二醇单体对大鼠心肌作用的单钙通道分析及ESR谱研究[J].基础医学与临床,1994,14(4)：286.
[39]Tatiana D, Galina T, Vilhelmina J.Characterization of the Radical Scavenging Activity of Lignins- natural Antioxidants[J].Biores Tech,2004,95:309-317.