二至丸提取物对体外肝细胞损伤的保护作用

 目的：研究二至丸乙酸乙酯提取物(ethyl acetate extractsof Erzhi pill,EAEP)对体外肝细胞损伤的保护作用及其机理。方法：培养L-O2型肝细胞,采用CCl4和H2O2体外分别诱导肝细胞损伤,检测培养上清液中天门冬氨酸转换酶(AST)和丙氨酸氨基转换酶(ALT)水平,测定上清液中丙二醛(MDA)的含量和过氧化物岐化酶(SOD)活力及谷胱甘肽过氧化物酶(GSH-Px)活性,MTT法检测细胞存活和增殖活性。结果：① EAEP(0.32～40 μg?mL-1)剂量组可明显降低由CCl4所致肝细胞培养上清液中AST和ALT水平及MDA含量的升高,还可提高由H2O2所致肝细胞存活率和SOD活力及GSH-Px活性的降低。② EAEP(0.32～40 μg?mL-1)剂量组可使H2O2升高的肝细胞培养上清液中ALT和ALT水平及MDA含量明显降低或恢复,还可提高CCl4降低的肝细胞存活率和SOD活力及GSH-Px活性。结论：提示AEEP对体外肝细胞损伤有直接保护作用,该作用可能与其抗氧化作用有关。     

金丝桃苷对四氯化碳诱导肝细胞损伤的保护作用

目的研究金丝桃苷(hyperfine,HP)对四氯化碳(CCl4)诱导肝细胞损伤的保护作用及其机制。方法用CCl4体外诱导人正常肝细胞L-02损伤,检测培养上清液中天门冬酸氨基转移酶(AST)和丙氨酸氨基转移酶(ALT)水平,测定肝细胞中丙二醛(MDA)和谷胱甘肽(GSH)的含量,四甲基偶氮唑盐微量酶反应比色(MTT)法检测肝细胞存活率。结果金丝桃苷可明显降低由CCl4升高的肝细胞培养上清液中ALT和AST水平及肝细胞MDA含量,还可提高由CCl4降低的肝细胞存活率和GSH含量。结论金丝桃苷对人肝细胞氧化性损伤有直接保护作用,这可能与金丝桃苷抑制抗氧化酶活性和抗自由基活性有关。     

黄芪总提物对体外肝细胞损伤的保护作用

目的研究黄芪总提物 (TEA)对体外肝细胞损伤的保护作用及其机理。方法采用Ⅳ型胶原酶灌流法分离大鼠肝细胞进行原代培养 ,用CCl4 和H2O2 体外分别诱导肝细胞损伤 ,检测肝细胞丙二醛 (MDA)、谷胱甘肽 (GSH)含量和谷胱甘肽过氧化物酶 (GSHpx)活性以及培养上清液中天门冬氨酸转换酶 (AST)和/或丙氨酸氨基转换酶 (ALT)水平。结果 (1)TEA(5～80mg·L-1)可明显降低或恢复由CCl4 升高的肝细胞MDA含量及肝细胞培养上清液中AST水平 ,还可使CCl4 降低的肝细胞GSH含量和GSHpx活性升高或恢复 ;(2)TEA(5～80mg·L-1)可使H2O2升高的肝细胞培养上清液中ALT水平和肝细胞MDA含量明显降低或恢复 ,还可使H2O2降低的肝细胞GHS含量和GSHpx活性明显升高或恢复。结论提示TEA对体外肝细胞损伤有直接保护作用 ,该作用可能与其抗氧化作用有关     

白藜芦醇与白藜芦醇苷抗氧化活性比较

目的：比较白藜芦醇与白藜芦醇苷清除自由基的能力以及对H202造成人脐静脉血管内皮细胞ECV304损伤的保护作用。方法：以邻二氮菲．Fe2＋体系产生羟基自由基（·OH）的方法研究白藜芦醇与白藜芦醇苷清除自由基的能力;采用H2O2诱导人脐静脉血管内皮细胞（ECV304）氧化损伤模型,四甲基偶氮噻唑蓝（MTF）比色法检测两种药物对H202致氧化损伤细胞的保护作用。结果：白藜芦醇与白藜芦醇苷清除·OH的能力呈剂量依赖性,其中白藜芦醇苷的作用比白藜芦醇强,但均弱于维生素C;白藜芦醇对H2O2致氧化损伤的ECV304细胞的保护作用强于白藜芦醇苷。结论：白藜芦醇对H202损伤的ECV304细胞具有明显的保护作用。     

玄参中苯丙素苷对肝细胞损伤保护作用的研究

目的：研究苯丙素苷对肝细胞损伤的保护作用。方法：通过D－氨基半乳糖造成体外和体内肝组织损伤，观察苯丙素苷对肝细胞存活率、LDH、ALT和AST的影响。结果：苯丙素苷在体外能提高肝原代培养细胞的存活率，降低LDH水平。在体内能降低衰竭大鼠ALT和AST水平。结论：苯丙素苷对D－氨基半乳糖造成肝细胞损伤具有明显的保护作用。     

二至丸保肝活性成分群对体外肝细胞损伤的保护作用

<正>二至丸出自明代吴旻辑的《扶寿精方》[1],由女贞子(蒸)和墨旱莲等量组成,用于治疗肝肾阴虚、眩晕耳鸣等症。通过前期二至丸乙酸乙酯部位对体外肝细胞损伤保护作用的研究表明了该部位的保肝活性[2],本实验进一步通过聚酰胺柱层析对乙酸乙酯部位分离得到50%乙醇洗脱活性成分群(AIEP),采用体外培养L-O2型肝细胞,用四氯化碳(CCl4)和过氧化氢(H2O2)分别诱导肝细胞损伤模型。观察AIEP对体外肝细胞损伤的保护作用及其机制。     

生姜油提取物对过氧化氢致大鼠肝细胞损伤的保护作用

目的 比较=三个不同生姜油提取物对过氧化氢(H2O2)损伤培养的大鼠肝细胞的保护作用.方法 用硅胶柱层析分离生姜油后获取A(主要为萜烯类)、B(姜油酮类)、C(姜酚类和姜烯酚类)三个不同提取部位溶液.6只SD大鼠分为6组,4组分别以生姜油A、B、C三个不同部位和联苯双酯制备含药血清干预,另2组给溶媒制备对照血清.利用H2O2体外诱导大鼠肝细胞损伤模型,检测肝细胞经含药血清处理后上清液中ALT、AST、丙二醛(MDA)和谷胱甘肽过氧化物酶(GSH-Px)活性的变化.结果 生姜油A部位含药血清能显著降低ALT和AST水平,抑制MDA的产生,升高GSH-Px活性;B部位亦能明显降低ALT和AST水平,抑制MDA的产生,升高GSH-Px活性,但降低ALT作用不如A部位;C部位能降低ALT和AST水平,升高GSH-Px活性,但对MDA含量无明显影响且降低AST作用不如A部位.结论 不同成份的生姜油提取物含药血清对H2O2引起的大鼠肝细胞损伤均有不同程度的保护作用,但以A部位效果较好,其机制可能与其抗氧化作用有关.     

二至丸水提物对体外肝细胞损伤的保护作用

目的研究二至丸水提物（aqueous extract of Erzhi Pill,AEEP）对体外肝细胞损伤的保护作用及其机制。方法培养L-O2型肝细胞,采用H2O2和CCl4体外分别诱导肝细胞损伤,检测培养上清液中天门冬氨酸转换酶（AST）和丙氨酸氨基转换酶（ALT）水平,测定上清液中丙二醛（MDA）的含量和过氧化物岐化酶（SOD）活力,MTT法检测细胞存活和增殖活性。结果①AEEP（0.32~40μg/ml）剂量组可明显降低由H2O2升高的肝细胞培养上清液中AST和ALT水平及MDA含量,还可提高H2O2降低的肝细胞存活率和SOD活力;②AEEP（0.32~40μg/ml）剂量组可使CCl4升高的肝细胞培养上清液中ALT和ALT水平及MDA含量明显降低或恢复,还可提高CCl4降低的肝细胞存活率和SOD活力。结论提示AEEP对体外肝细胞损伤有直接保护作用,该作用可能与其抗氧化作用有关。     

卡维地洛对过氧化氢致血管内皮细胞氧化应激损伤的保护作用

目的观察卡维地洛(carved ilol)对过氧化氢(hydrogenperoxide,H2O2)致内皮细胞损伤及表面粘附分子表达的影响。方法采用H2O2作为外源性自由基生成系统,模拟内皮细胞的脂质过氧化损伤,建立离体培养的ECV-304细胞氧化应激损伤模型,观察卡维地洛对H2O2致内皮细胞损伤及表面粘附分子表达的影响。结果卡维地洛各浓度组均明显改善H2O2(1.0×10-6mol.L-1)所致ECV-304细胞形态学损伤,提高细胞生存率,降低LDH释放,并可使细胞内及细胞培养液中MDA含量降低,SOD活性升高,亦可下调ICAM-1蛋白及细胞内ICAM-1mRNA表达水平,上述作用随药物浓度增加呈增强趋势。结论卡维地洛可保护内皮细胞结构和功能的完整性,提高内皮细胞抗氧化能力,并从转录水平抑制脂质过氧化诱导的粘附分子表达增加,降低单核-内皮细胞粘附,有利于减少动脉粥样硬化的始动环节和早期事件的发生。     

肝细胞生长因子预处理对过氧化氢诱导大鼠神经干细胞凋亡的保护作用

目的研究肝细胞生长因子(HGF)对神经干细胞(NSC)凋亡的保护作用及其作用机制,为HGF用于NSC移植提供实验基础。方法分离培养大鼠NSC。细胞分为正常对照、模型(H2O2100μmo.lL-1)、HGF+H2O2(HGF15,30及60μg.L-1预处理24h后,再加入H2O2100μmol.L-1处理4h),LY294002(PI3K/Akt通路抑制剂)+HGF+H2O2(先加入LY29400220μmol.L-1处理30min,再加入HGF60μg.L-1处理24h,最后再加入100μmo.lL-1H2O2培养4h)组。MTT法检测细胞存活率;TUNEL法检测细胞凋亡率;比色法检测半胱氨酸天冬氨酸蛋白酶(caspase)-3活性;Western印迹分析Bcl-2,Bax蛋白表达。结果MTT检测发现,随着HGF浓度的增加,NSC细胞的存活率也增加。与模型组〔(63.5±2.4)%〕比较,HGF15,30及60μg.L-1预处理组细胞存活率明显升高〔(79.1±7.5)%,(83.8±6.1)%和(86.6±8.2)%;n=3,P<0.05〕。TUNEL法检测发现,HGF预处理组凋亡细胞明显减少,模型组的凋亡率为(43.5±6.2)%,HGF预处理组则分别为(34.2±8.6)%,(21.7±3.8)%及(19.4±4.0)%。Caspase-3活性检测表明,与模型组相比,HGF预处理组细胞caspase-3活性降低。Western印迹分析结果显示,与模型组比较,HGF预处理使细胞的Bcl-2蛋白表达升高,但Bax蛋白表达不受影响;HGF的抗凋亡效应可被PI3K/Akt通道阻滞剂LY294002阻断。结论HGF可减轻H2O2所诱导的大鼠NSC凋亡,且呈一定的浓度依赖关系,其作用机制可能与NSC的PI3K/Akt通路激活和Bcl-2表达增强有关。     

虎杖苷对CCl_4损伤原代培养大鼠肝细胞的保护作用

目的研究虎杖苷（ＰＤ）对ＣＣｌ４损伤原代培养大鼠肝细胞的保护作用。方法原位灌流法分离大鼠肝细胞，培养２４ｈ后加入ＰＤ，同时造成ＣＣｌ４损伤，分别于损伤后６，１２，２４和４８ｈ测培养液中谷丙转氨酶（ＡＬＴ）、丙二醛（ＭＤＡ）和还原型谷胱甘肽（ＧＳＨ）活性，４８ｈ后用ＭＴＴ比色法测肝细胞存活率。结果ＰＤ１×１０－７～１×１０－４ｍｏｌ·Ｌ－１能不同程度地抑制ＡＬＴ和ＭＤＡ的上升，提高ＧＳＨ水平，显著提高细胞存活率，以１×１０－５ｍｏｌ·Ｌ－１效果最好。结论ＰＤ１×１０－７～１×１０－４ｍｏｌ·Ｌ－１能有效保护ＣＣｌ４造成的原代培养大鼠肝细胞损伤     

尼莫地平对过氧化氢诱导猪脑基底动脉氧化应激损伤的保护作用

目的：探讨尼莫地平对过氧化氢（H2O2）诱导猪脑基底动脉损伤的保护作用。方法：采用去内皮离体血管环灌流的方法，建立H2O2损伤模型。比较正常对照组，H2O2（2×10^-4mol／L）损伤组，维生素C（10^-4mol／L）组，尼莫地平高、中、低剂量（5×10^-6、5×10^-7、5×10^-8mol／L）组血管环对KCl、苯肾上腺素的张力；检测各组血管组织匀浆中的超氧化物歧化酶（SOD）、过氧化氢酶（CAT）、谷胱甘肽过氧化物酶（GSH—PX）活性及丙二醛（MDA）含量。结果：（1）H2O2损伤组与正常对照组比较，血管环对KCl、苯肾上腺素的收缩反应明显增加；尼莫地平高、中、低剂量组呈剂量依赖性抑制KCl、苯肾上腺素对血管环的收缩反应。（2）H2O2损伤组MDA含量升高，SOD、CAT、GSH—PX活性降低，与正常对照组比较，差异有统计学意义（P〈0．05）。尼莫地平高、中、低剂量均能降低组织中MDA含量，增强SOD、CAT、GSH-PX，的活性，与H2O2损伤组比较，差异有统计学意义（P〈0．05）。结论：钙通道阻断剂尼莫地平具有抗氧化应激作用，能预防H2O2对脑基底动脉血管的氧化损伤。     

Protective effects of prostaglandin E1 on human umbilical vein endothelial cell injury induced by hydrogen peroxide

Aim:

To investigate the protective effects of prostaglandin E₁ (PGE₁) against H₂O₂-induced oxidative damage on human umbilical vein endothelial cells (HUVECs).

Methods:

HUVECs were pretreated with PGE₁ (0.25, 0.50, and 1.00 μmol/L) for 24 h and exposed to H₂O₂ (200 μmol/L) for 12 h, and cell viability was measured by the MTT assay. LDH, NO, SOD, GSH-Px, MDA, ROS, and apoptotic percentage were determined. eNOS expression was measured by Western blotting and real-time PCR.

Results:

PGE₁ (0.25−1.00 μmol/L) was able to markedly restore the viability of HUVECs under oxidative stress, and scavenged intracellular reactive oxygen species induced by H₂O₂. PGE₁ also suppressed the production of lipid peroxides, such as MDA, restored the activities of endogenous antioxidants including SOD and GSH-Px, and inhibited cell apoptosis. In addition, PGE₁ significantly increased NO content, eNOS protein, and mRNA expression.

Conclusion:

PGE₁ effectively protected endothelial cells against oxidative stress induced by H₂O₂, an activity that might depend on the up-regulation of NO expression.     

过氧化氢诱导肺微血管内皮损伤的实验研究

目的　探讨H2 O2 诱导培养肺微血管内皮细胞损伤的机制。方法　观察H2 O2 对大鼠肺微血管内皮细胞 (RP MVEC)的形态、单层通透性、F 肌动蛋白和 β AR的影响。结果　H2 O2 浓度大于 1mmol·L-1作用 48h内观察到细胞脱落和破裂。 10mmol·L-1H2 O2 90min内可使RPMVEC单层通透性增高、F 肌动蛋白发生明显解聚和 β AR显著下调 ;FOR、山莨菪碱和CTX可抑制上述变化。结论　H2 O2引起的RPMVEC脱落与破裂呈浓度和时间依赖性。H2 O2引起RPMVEC单层通透性增高的机制与F 肌动蛋白解聚密切相关 ;β AR参与内皮通透性调节。FOR、山莨菪碱和CTX对H2 O2 引起RPMVEC单层通透性增高有一定的保护作用。     

Evidence of crocin against endothelial injury induced by hydrogen peroxide in vitro

Crocin, the digentiobiosyl ester of crocetin, was investigated for its cytoprotective effect on hydrogen peroxide-induced injury in bovine aortic endothelial cells (BAECs). The morphology of BAECs was observed by inverted phase contrast and electron microscopy. The MTT assay was used to measure cell viability. Cell apoptosis was evaluated by DNA argarose gel electrophoresis. The cells treated with H₂O₂ (200 μM) showed apoptotic changes as revealed by cell shrinkage, condensation of nuclei, membrane blebbing and formation of apoptotic body. A concentration-dependent inhibition of cell injury was seen in cultures treated with crocin at dosages ranging from 1 to 10 μM. Furthermore, in the H₂O₂-treated group, agarose gel electrophoresis displayed a “DNA ladder”. Whereas in the 10 μM crocin-pretreated group, cells remained intact and no “DNA ladder” was observed in agarose gel electrophoresis. Only very little DNA debris appeared on DNA-fragmentation analysis in the 1 μM crocin-pretreated group. Our data demonstrated that crocin has preventive effects on the cell apoptosis induced by H₂O₂, which may contribute to its utilisation for cardiovascular diseases (e.g., atherosclerosis and hypertension).     

Evidence of crocin against endothelial injury induced by hydrogen peroxide in vitro

Crocin, the digentiobiosyl ester of crocetin, was investigated for its cytoprotective effect on hydrogen peroxide-induced injury in bovine aortic endothelial cells (BAECs). The morphology of BAECs was observed by inverted phase contrast and electron microscopy. The MTT assay was used to measure cell viability. Cell apoptosis was evaluated by DNA argarose gel electrophoresis. The cells treated with H₂O₂ (200 μM) showed apoptotic changes as revealed by cell shrinkage, condensation of nuclei, membrane blebbing and formation of apoptotic body. A concentration-dependent inhibition of cell injury was seen in cultures treated with crocin at dosages ranging from 1 to 10 μM. Furthermore, in the H₂O₂-treated group, agarose gel electrophoresis displayed a “DNA ladder”. Whereas in the 10 μM crocin-pretreated group, cells remained intact and no “DNA ladder” was observed in agarose gel electrophoresis. Only very little DNA debris appeared on DNA-fragmentation analysis in the 1 μM crocin-pretreated group. Our data demonstrated that crocin has preventive effects on the cell apoptosis induced by H₂O₂, which may contribute to its utilisation for cardiovascular diseases (e.g., atherosclerosis and hypertension).     

依达拉奉对过氧化氢致血管内皮细胞氧化损伤的保护作用

目的探讨依达拉奉对过氧化氢(H2O2)致内皮细胞氧化损伤的影响。方法体外培养人脐静脉内皮细胞(HUVEC),采用H2O2作为外源性自由基生成系统,模拟血管内皮细胞氧化应激损伤模型,观察不同剂量依达拉奉对H2O2所致内皮细胞的影响。通过检测各组丙二醛(MDA)的含量反应细胞的损伤程度,检测超氧化物歧化酶(SOD)的活性及总抗氧化能力(T-AOC)反应细胞的抗氧化能力。结果依达拉奉各浓度组均明显降低MDA的含量,提高SOD的活性及总T-AOC,上述作用随药物浓度增加呈增强趋势。结论依达拉奉能降低内皮细胞的氧化损伤程度,提高内皮细胞抗氧化能力。     

Protective effects of raw and cooked blackcurrant extract on DNA damage induced by hydrogen peroxide in human lymphoblastoid cells

Context: Blackcurrant (Ribes nigrum L.) is a classical fruit that has long been used to make juice, liqueur and sometimes medicines in Europe. The beneficial effects of blackcurrant, which are inhibition of lipopolysaccharide-stimulated inflammatory, anticarcinogenesis and other health effects, have been reported.

Objective: Previously, we reported the antimutagenic activities of blackcurrant using a yeast gene mutation assay. In this study, we investigated whether this antimutagenicity of blackcurrant was confirmed in human cells.

Materials and methods: We prepared four types of aqueous blackcurrant extracts (BCE) from mature and premature with or without heat treatment by microwave. Antioxidant activities of BCE were measured by the DPPH radical scavenger assay. In the DPPH radical scavenger assay, the maximum concentration of BCE was 1.6?mg/reaction. We investigated the antigenotoxic activities of BCE by the comet assay and micronucleus test using the human lymphoblastoid cell line TK6. In the comet assay, TK6 was treated with 300?μM?H₂O₂ without or with BCE at concentrations of 0.5,?1.0,?2.0 and 3.0?mg/mL. In the micronucleus test, TK6 was treated with 1?mg/mL BCE without or with H₂O₂.

Results: All BCEs exhibited more than 90% of inhibition rates of DPPH radicals at the maximum concentration of BCE. DNA damage and micronuclei induced by H₂O₂ significantly decreased in the each BCE-treated condition.

Conclusion: The results suggest that BCE treatment can reduce the genomic instability induced by H₂O₂ in human cells. We consider that these antigenotoxic effects are related to polyphenols, l-ascorbic acid and other antioxidant compounds.