赖氨大黄酸对氧化应激引起的小鼠动脉血管损伤的保护作用及其机制

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doi:10.13481/j.1671-587x.20150614

摘要：

目的:观察赖氨大黄酸(RHL)对氧化应激引起的小鼠动脉血管损伤的保护作用,并探讨其作用机制。方法:采用腹腔注射百草枯方法建立活性氧引起血管损伤小鼠模型。将30只C57小鼠随机分为对照组(n=10)、百草枯模型组(n=10)和RHL干预组(n=10)。RHL干预组小鼠在造模前1周灌胃给予RHL,对照组和百草枯模型组灌胃给予等体积蒸馏水。百草枯模型组和RHL干预组腹腔注射百草枯,对照组腹腔注射等体积生理盐水,每周注射1次,持续2周。造模2周后检测血清丙二醛(MDA)水平、超氧化物歧化酶(SOD)和谷胱甘肽过氧化物酶(GSH-Px)活性;DCFH-DA染色观察血管活性氧水平;HE染色观察腹主动脉病理表现;Western blotting 法检测血管损伤相关基因的表达。结果:与对照组比较,百草枯模型组小鼠血管组织小鼠血清SOD和GSH-Px活性下降(P<0.05),MDA水平升高(P<0.05);与百草枯模型组比较,RHL干预组MDA水平下降(P<0.05),SOD和GSH-Px活性升高(P<0.05);DCFH-DA和HE染色,百草枯模型组小鼠血管组织血管组织活性氧水平升高(P<0.05),血管结构破坏, RHL干预组血管活性氧水平下降(P<0.05),病理变化明显减轻。Western blotting法检测,与对照组比较,百草枯模型组小鼠血管组织内皮型一氧化氮合酶(eNOS)和caspase-3表达水平降低(P<0.05),caspase-3裂解片段表达水平升高(P<0.05);与模型组比较,RHL干预组小鼠血管组织 eNOS和caspase-3 表达水平升高 (P<0.05),caspase-3裂解片段表达水平降低(P<0.05)。结论:百草枯能够诱导体内活性氧水平升高引起血管细胞损伤,RHL通过清除活性氧,上调eNOS表达,下调caspase-3裂解片段表达来拮抗百草枯的作用。

关键词: 赖氨大黄酸, 百草枯, 活性氧, 细胞凋亡

Abstract:

Objective To investigate the protective effects of rhein lysinate (RHL) on the blood vessel damage induced by oxidative stress in the mice,and to explore its mechanism. Methods The mouse models of oxidative damage were established by intraperitoneal injection of paraquat.30 C57 mice were randomly divided into control,paraquat model,and RHL prevention groups.The mice in RHL prevention group were given RHL by gavage for one week before performing model.The mice in other two groups were given equal volume of distilled water.For making model,paraquat was intraperitoneally injected in the mice in paraquat model and RHL prevention groups once a week for two weeks.The activities of superoxide dismutase (SOD) and glutathione peroxidase(GSH-Px) and the content of serum malonaldehyde (MDA) of the mice were detected 2 weeks after modeling.The pathological profile of blood vessel was observed by hematoxylin and eosin (HE) staining and the level of reactive oxygen species was observed by DCFH-DA staining.The expressions of genes related to blood vessel damage were detected by Western blotting method. Results Compared with control group,the activities of SOD and GSH-Px were decreased and the content of MDA was increased in paraquat model group (P<0.05).Compared with paraquat model group,the activities of SOD and GSH-Px were increased and the content of MDA was decreased in RHL prevention group (P<0.05).The pathological examination indicated the structure of blood vessel of the mice was damaged and the level of reactive oxygen species of blood vessel was increased (P<0.05) in paraquat model group.The pathological changes were significantly improved and the level of reactive oxygen species of blood vessel of the mice was decreased (P<0.05) in RHL prevention group.The Western blotting analysis showed that compared with control group,the expression levels of nitric oxide endothelial synthase (eNOS) and caspase-3 of the mice in paraquat model group were decreased (P<0.05),however the expression level of cleaved fragment of caspase-3 was increased (P<0.05).Compared with paraquat model group,the expression levels of eNOS and caspase-3 of the mice in RHL prevention group were increased (P<0.05) and the expression level of cleaved fragment of caspase-3 was decreased (P<0.05). Conclusion Paraquat could induce vascular cell damage in vivo through increasing the levels of reactive oxygen species,and RHL could antagonize the effects of paraquat by scavenging reactive oxygen species,and up-regulating the eNOS expression and reducing the expression of the cleaved fragment of caspase-3.

Key words: rhein lysinate, paraquat, reactive oxygen species, apoptosis

中图分类号:

R285.5

冯秋生, 阚泉, 吕翠平, 李冉, 魏静波, 赵毓芳, 甄永占. 赖氨大黄酸对氧化应激引起的小鼠动脉血管损伤的保护作用及其机制[J]. 吉林大学学报(医学版), 2015, 41(06): 1171-1175.

FENG Qiusheng, KAN Quan, LYU Cuiping, LI Ran, WEI Jingbo, ZHAO Yufang, ZHEN Yongzhan. Protective effect of rhein lysinate on blood vessel damage induced by oxidative stress in mice and its mechanism[J]. Journal of Jilin University Medicine Edition, 2015, 41(06): 1171-1175.

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