笃斯越桔花青素提取物对3T3-L1前脂肪细胞生长抑制作用

目的：研究笃斯越橘花青素提取物对3T3-L1 前脂肪细胞生长抑制作用及其作用机理。方法：采用MTT法观察细胞生长抑制并确定半数抑制浓度(IC50),LDH(lactate dehydrogenase)法评价提取物引起的细胞膜损伤,流式细胞计法测定细胞周期,荧光显微镜观察凋亡细胞的形态学特征。结果：笃斯越橘花青素提取物能够有效地抑制3T3-L1 前脂肪细胞的生长且IC50 约为214μg/mL;LDH 实验表明细胞LDH 释放率在24、48、72h 分别为89.0%、85.2%、67.8%,72h 时相对24h 或48h 明显降低(P ＜ 0.05),提取物对3T3-L1 前脂肪细胞膜的通透性未见明显增加;笃斯越橘花青素提取物能够有效诱导细胞凋亡,同时在S 或G0/G1 期产生阻滞,但是对细胞的G2/M 期未见明显影响,荧光显微镜观察细胞出现典型凋亡特征。结论：笃斯越橘花青素提取物能有效抑制3T3-L1 前脂肪细胞生长的作用,具有开发为天然抗肥胖功能因子的潜在可能。     

Effects of casein- and whey protein–dextran conjugates on the stability of bog bilberry anthocyanin extract

Bog bilberry ( Vaccinium uliginosum ) contains a high concentration of anthocyanins. Anthocyanin stability depends on many factors, including pH, temperature and composition of food matrices in a product, such as protein and polysaccharide. The effect of casein–dextran and whey protein–dextran conjugates on the stability of bog bilberry anthocyanin extract (BBAE) was investigated in an accelerated model system. Cyanidin-3-glucoside was the main anthocyanin (79%) found in BBAE. The cyanidin-3-glucoside content, absorbance and ABTS [2, 2-Azinobis (3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt] radical scavenging activity of BBAE significantly ( P  < 0.05) decreased after heating at pH 8.0 and 80 °C for 60 min. The addition of casein–dextran and whey protein–dextran conjugates significantly ( P  < 0.05) prevented cyanidin-3-glucoside from degradation, and increased the absorbance and ABTS radical scavenging activity of heated BBAE. The protection of casein–dextran conjugate on BBAE was significantly higher ( P  < 0.05) than that of whey protein–dextran conjugate. The results suggest that protein–dextran conjugates have the ability to stabilise BBAE.     

Kinetics of Thermal Degradation of the Anthocyanins Delphinidin-3-rutinoside and Malvidin-3-glucoside

The kinetics of the thermal degradation of the two anthocyanins delphinidin-3-rutinoside and malvidin-3-glucoside at 78, 88, 98 and 108 °C, depending on the pH and the type of solvent has been investigated. The first order reaction of delphinidin-3-rutinoside and malvidin-3-glucoside degradation has been established. The rate constants (K ± 10^?5s^?1), the activation parameters, factor Q₁₀ and the half-decay period have been calculated. The rate of delphinidin-3-rutinoside degradation is higher than that of malvidin-3-glucoside.     

基于Caco-2单层细胞模型的兜唇石斛多糖的转运机制研究

以兜唇石斛多糖(DAP)为研究对象,借助人小肠上皮细胞Caco-2(the human colon carcinoma cell line)单层细胞模型,体外模拟DAP在小肠中的转运情况。在前期得到的DAP为基础,探究DAP的粘度对其转运情况的影响,同时建立Caco-2细胞的模型,采用MTT法测定DAP对Caco-2细胞存活率的影响,并考察了DAP在小肠的吸收转运的特征。实验表明:DAP的粘度与其在Caco-2细胞中的转运能力呈负相关趋势,粘度越大,转运速率越慢;在测定TEER值及荧光素钠渗透情况,观察Caco-2细胞的形态,建立了良好的细胞模型;DAP的存在下,Caco-2细胞的存活率为91.8%以上,因此DAP对Caco-2细胞存活无影响;DAP在Caco-2不同单边层的转移率分别为0.35%±0.02%和0.32%±0.02%,表观渗透率分别为(0.42±0.02)×10-6 cm/s和(0.19±0.18)×10-6 cm/s,表明DAP在小肠中不易被吸收。本研究所得结果为进一步研究DAP在肠道中的吸收机制奠定基础。     

Copigmentation between malvidin-3-glucoside and some wine constituents and its importance to colour expression in red wine

Thermodynamic parameters for intermolecular copigmentation interactions involving malvidin-3-glucoside were determined by UV/visible spectroscopy at wine pH (pH 3.6). These included association constants, enthalpy and entropy changes, which were measured for chlorogenic acid, caffeic acid, quercetin, quercetin-3-glucoside, (−)-epicatechin, (+)-catechin, procyanidin dimer and seed tannin. Quercetin produced the strongest copigment (K_CP = 2900 ± 1300), whilst the addition of glucose at position 3 (quercetin-3-glucoside) reduced its effect by almost 10-fold. Malvidin-3-glucoside self-association (K_D = 3300 ± 300 mol⁻¹ l) was thermodynamically favoured over intermolecular interaction with any of the copigments tested. No colour enhancement due to self-association was observed for malvidin-3-glucoside-derived pigments that cannot enter hydration reactions. In addition, malvidin-3-(6-O-p-coumaryl)glucoside did not show colour enhancement suggesting that the p-coumaryl group prevents self-association. The malvidin-3-glucoside circular dichroism (CD) spectrum was not affected by indicated changes in malvidin-3-glucoside concentration. These observations demonstrate that self-association of malvidin-3-glucoside is more important than copigmentation in young red wine.     

柚叶总黄酮对LPS所致Caco-2结肠上皮细胞间高通透性的保护作用

探讨柚叶总黄酮(PLTFE)对脂多糖(LPS,2 μg/mL)诱发Caco-2高通透性细胞模型的保护作用。Caco-2模型细胞以不同浓度的PLTFE(0、10、50、100、150 μg/mL)处理培养24 h进行后续实验。MTT法测定细胞生存率,细胞乳酸脱氢酶(lactate dehydrogenase,LDH)水平依说明书使用试剂盒测定。酶联法(enzyme linked immunosorbent assay,ELISA)测定白介素(interlukin-1β,IL-1β)、IL-8和肿瘤坏死因子(tumor necrosis factor-α,TNF-α)分泌水平。跨上皮细胞电阻(trans epithelial electrical resistance,TEER)值和异硫氰酸荧光素-右旋糖酐(FD40)透过度用于评估细胞通透性水平。实时定量PCR(Quantitative real-time PCR,qRT-PCR)检测细胞IL-1β、IL-8、TNF-α、闭锁蛋白(Occludin)、紧密连接蛋白-1(claudin-1)、封闭小带蛋白(ZO-1)和肌球蛋白轻链激酶(myosin light chain kinase,MLCK)的mRNA表达。结果表明,柚叶总黄酮能显著提高受损细胞生存率至86.1%,抑制受损细胞中LDH的溢出(p<0.05)。同时还能有效抑制受损细胞中炎性细胞因子(IL-1β、IL-8、TNF-α)的分泌及mRNA转录。此外,柚叶总黄酮可增强细胞紧密连接因子(Occludin、claudin-1、ZO-1)的mRNA转录,抑制MLCK的mRNA转录,改善细胞间高通透性qRT-PCR法检测细胞中相关因子的mRNA转录水平。结果提示,柚叶总黄酮具有较强的抗炎活性,能通过上调细胞内细胞紧密连接相关因子的mRNA转录显著改善LPS造成的Caco-2细胞间高通透性的发生(p<0.05)。     

三种茶叶中多酚提取物的抑菌活性及其对致病菌膜渗透性的比较分析

目的:研究3种茶叶中茶多酚的抑菌活性及对菌株细胞膜渗透性的影响。方法:以西湖龙井、铁观音和普洱茶为材料提取茶多酚,以滤纸片法测定茶多酚的抑菌圈直径,二倍肉汤稀释法测定其最小抑菌浓度(MIC)和最小杀菌浓度(MBC),比较其抑菌效果。通过DNA渗透性实验、DNA损伤实验评价茶多酚对细胞膜完整性及对DNA损伤的影响。结果:相同条件下,绿茶(西湖龙井)中茶多酚含量最高为(48.12±3.22) mg/g,青茶(铁观音)次之为(37.36±2.64) mg/g,黑茶(普洱茶)最低为(31.61±1.92) mg/g。绿茶对金黄色葡萄球菌的抑菌作用最强,抑菌圈直径为(1.80±0.06) cm,MIC值为0.125 mg/mL、MBC值为0.25 mg/mL。随着茶多酚提取物浓度的增加,260 nm下的细菌渗透性物质逐渐增多,并且基因组DNA的损伤程度逐渐增大。结论:3种茶叶中绿茶具有更好的抑菌效果,并通过增强细菌细胞膜渗透性,进而对DNA产生一定的损伤。     

苹果模拟胃肠消化后的多酚释放和抗HepG2细胞增殖活性研究

分别采用80%丙酮提取以及模拟胃肠消化法提取苹果中的多酚,测定提取物的多酚释放量以及抗HepG2细胞增殖的活性。结果显示,在水晶富士、嘎啦果和青苹果的80%丙酮提取物中,绿原酸的释放量最高,分别为30.72±1.54、22.37±1.12、8.70±0.44 mg/kg FW(Fresh weight,FW)。模拟胃消化后释放量较高的为原儿茶酸、对羟基苯甲酸、没食子酸和绿原酸等酚酸;模拟肠消化后释放量较高的为槲皮苷、槲皮素-3-葡萄糖苷、扁蓄苷等黄酮醇。水晶富士、嘎啦果和青苹果的模拟胃消化及模拟肠消化后的提取物(浓度分别为297、99、255 mg/mL和198、154、106 mg/mL)作用于HepG2细胞72 h后,细胞的存活率分别为26%、42%、44%和23%、28%、28%。这表明,模拟胃消化主要促进酚酸的释放,模拟肠消化主要促进黄酮醇的释放;在一定浓度范围内,苹果模拟胃肠消化提取物对HepG2细胞有较强的抗增殖活性。     

中国主要水果抑制肝癌HepG2细胞和结肠腺癌Caco-2细胞增殖活性评价

分别采用Folin-Ciocalteu法测定了水果提取物的总酚含量,采用亚甲基蓝法测定了其抗人肝癌细胞Hep G2和人结肠腺癌细胞Caco-2增殖的活性,分析了总酚含量与抗Hep G2和Caco-2细胞增殖活性之间的相关性。结果显示,25种水果中李子的总酚含量最高(1686.08±96.94μmol GAE/100鲜果),西瓜(83.54±2.10μmol GAE/100 g鲜果)和哈密瓜(79.35±0.76μmol GAE/100 g鲜果)的总酚含量最低;在可测出抗增殖EC50值的水果中,李子(18.99±0.06 mg/m L)和番石榴(20.94±0.09 mg/m L)抗Hep G2细胞增殖的活性最强,梨的活性最弱(389.63±10.82 mg/m L)。李子抗Caco-2细胞增殖的活性最强(8.73±0.11 mg/m L),火龙果的活性最弱(388.07±7.04 mg/m L)。水果的抗Hep G2和Caco-2细胞增殖活性与其总酚含量相关性显著(R2=0.4147,p0.01;R2=0.4071,p0.05),该相关性表明水果中的多酚具有良好的抗肿瘤细胞增值的活性。     

Production and transepithelial transportation of angiotensin-I-converting enzyme (ACE)-inhibitory peptides from whey protein hydrolyzed by immobilized Lactobacillus helveticus proteinase

Lactobacillus helveticus LB 10 proteinases immobilized with sodium alginate were used to hydrolyze whey protein to produce angiotensin-I-converting enzyme (ACE)-inhibitory peptides. The generated hydrolysates were tested for ACE-inhibitory activity and for their ability to be transported across Caco-2 cell monolayers. Using a response surface method, we determined that a proteinase concentration of 7.55 mg/mL, sodium alginate concentration of 2.03 g/100 mL, and glutaraldehyde concentration of 0.39% were found to be the optimal immobilization conditions. Compared with free proteinase, the immobilized proteinase had significantly higher pH, thermal and storage stability, and reusability. Whey protein hydrolysates were fractionated by gel filtration chromatography and ACE-inhibitory peptide mixtures were transported across Caco-2 cell monolayers in a human intestinal-absorption model. The di- and tripeptides KA, EN, DIS, EVD, LF, AIV, and VFK (half-maximal inhibitory concentrations (mean ± standard deviation) of 1.24 ± 0.01, 1.43 ± 0.04, 1.59 ± 0.27, 1.32 ± 0.05, 1.60 ± 0.39, 2.66 ± 0.02, and 1.76 ± 0.09 mmol/L, respectively) were detected on the basolateral side of the Caco-2 cell monolayer using ultra-performance liquid chromatography-tandem mass spectrometry. These results highlight that ACE-inhibitory peptides are present on the basolateral side of the Caco-2 cell model after transportation of whey protein hydrolysate across the Caco-2 cell membrane.     

Interactions of Oligomeric Procyanidins in Model Wine Solutions Containing Malvidin-3-Glucoside and Acetaldehyde

Interactions of grape seed flavanols, dimeric, galloylated dimeric and trimeric procyanidins with malvidin-3-glucoside and acetaldehyde were studied in wine-like model solutions. Malvidin-3-glucoside and oligomeric procyanidins reacted very slowly, with eventual formation of yellow xanthylium salts. The concentrations of malvidin-3-glucoside and oligomeric procyanidins decreased faster in the presence of acetaldehyde and the colour augmentation with shift towards violet was attributed to the formation of new coloured compounds, detectable by HPLC and linked at different positions by CH₃CH bridges. The rate of loss of procyanidins in the presence of malvidin-3-glucoside and acetaldehyde varied according to the complexity of the components. Monomeric (+)-catechin and (-)-epicatechin reacted more slowly than dimer procyanidins B₁, B₂ and B₃ while trimer C₁ (epicatechin 4β→8 epicatechin 4β→8 epicatechin) presented the highest degradative reaction rate.     

Bioavailability of a bilberry anthocyanin extract and its impact on plasma antioxidant capacity in rats

Anthocyanins are secondary metabolites from the flavonoid family, frequently found in fruits and vegetables. They have been shown to possess beneficial health effects and various in vitro assays have highlighted their powerful antioxidant capacity. However, little is known about their metabolism after digestion and their antioxidant potency in vivo. The aim of this work was to evaluate anthocyanin bioavailability and to determine the impact of an anthocyanin‐rich diet on plasma antioxidant status in rats. Animals were fed for 8 days with a vitamin E‐deficient diet supplemented with a bilberry (Vaccinium myrtillus L.) anthocyanin extract providing daily 1.43 mmol anthocyanins kg^?1 body weight. An anthocyanin‐enriched diet significantly enhanced the plasma antioxidant capacity compared with a control diet (P < 0.0001). Moreover, anthocyanins were recovered in urine in the intact glycosidic forms in addition to unknown metabolites. Urinary excretion of bilberry anthocyanins and their metabolites was 0.71 ± 0.08 µmol per 24 h (ie 0.22% of the ingested dose). Anthocyanins and their corresponding aglycones were detected in caecal contents. After a single oral administration of the bilberry extract, native anthocyanins quickly (30 min) appeared in plasma. Hence, in spite of a low bioavailability, bilberry anthocyanin extract consumption has a positive effect on plasma antioxidant capacity. Copyright © 2005 Society of Chemical Industry     

苦荞麸皮多酚抑制人结肠癌细胞Caco-2增殖活性研究

研究了产自重庆酉阳的苦荞(Fagopyrum tartaricum L.Gaerth,FG1)麸皮和四川西昌的苦荞(Fagopyyrum tartaricum L.Gaerth,FG2)麸皮中酚类化合物的含量、抗氧化和抗增殖活性。结果表明:FG1和FG2 2种苦荞麸皮总酚含量分别为(26.40±0.41)和(27.00±0.72)mg GAE/g DW,且游离态是其酚类化合物的主要存在形式(约占其总酚含量的93%);2种苦荞麸皮酚提取物均表现出一定的抗氧化性,并且FG2的抗氧化能力强于FG1(P0.05),FG1和FG2的DPPH自由基清除能力总IC50值分别为(39.56±2.76)和(24.69±0.33)μg·mL~(-1);还原力总EC_(50)值分别为(97.92±1.4)和(73.18±4.32)μg·mL~(-1);在抗增殖试验中,与对照组相比,FG1和FG2游离酚提取物均能明显抑制人结肠癌细胞Caco-2增殖(P0.05)。然而,在不产生细胞毒性的浓度范围内,其结合酚提取物却表现出极弱的抗增殖作用。     

Anthocyanin composition in aged Chilean Cabernet Sauvignon red wines

Anthocyanins in aged Cabernet Sauvignon red wines were studied by HPLC–MS. The major anthocyanin in 6, 7, and 8 year old red wine extracts was the dimer vitisin A produced by condensation of malvidin-3-O-glucoside mediated by pyruvic acid. In aged wine, the content of malvidin-3-O-glucoside decreased with age with a concomitant increase of vitisin A. The latter is accompanied by several condensation products with molecular weight up to 1500 m/z. Differential pulse voltammetry indicated that aged wines have lower antioxidant capacity than young wine (400 mV), but higher than malvidin-3-O-glucoside (483 mV). Tafel’s plots showed that the electrochemical process occurring in aged wines is different from young wines. Six, seven and eight year old wines show similar behaviour with plots of 234, 177 and 188 mV/dec, respectively. These values are higher than the expected 120 mV/dec corresponding to a first electronic transfer but smaller compared to the 523 mV/dec corresponding to young wine.     

On the bioavailability of flavanols and anthocyanins: flavanol-anthocyanin dimers

The bioavailability of flavanols, anthocyanins and anthocyanin-derived pigments like flavanol-anthocyanin dimers already reported to occur in food products is a major unsolved issue. The absorption of the flavanol-anthocyanin dimer (+)-catechin-(4,8)-malvidin-3-O-glucoside (Cat-Mv3glc) through Caco-2 cells was assessed by performing transepithelial transport assays. The ability of Cat-Mv3glc to cross Caco-2 cells was compared with that of malvidin-3-glucoside (Mv3glc), (+)-catechin (Cat) and procyanidin B3 (Cat-Cat), in order to evaluate the influence of some structural features on the transport efficiency. The flavanol-anthocyanin dimer was absorbed in this intestinal model although with a lower efficiency than the monomers Cat and Mv3glc. On the other hand, Cat-Mv3glc was found to cross the intestinal barrier model more significantly than Cat-Cat. This feature may be related to the presence of the glucose moiety in its structure. Overall, this study brings more insights into the bioavailability of anthocyanins and flavanols and represents the first report on the bioavailability of flavanol-anthocyanins.     

Determination of anthocyanins,flavonoids and phenolic acids in potatoes. I: Coloured cultivars of Solanum tuberosum L

The major anthocyanins, flavonoids and phenolic acids in the tubers (skin and flesh), flowers and leaves of 26 cultivars of Solanum tuberosum L with coloured skins and/or flesh have been identified and quantified using analytical HPLC. Red tubers contained mostly pelagonidin-3-(p-coumaroyl-rutinoside)-5-glucoside (200–2000 μg g⁻¹ FW) plus lesser amounts of peonidin-3-(p-coumaroyl-rutinoside)-5-glucoside (20–400 μg g⁻¹ FW). Light to medium purple tubers contained petunidin-3-(p-coumaroyl-rutinoside)-5-glucoside (1000–2000 μg g⁻¹ FW) plus small amounts of malvidin-3-(p-coumaroyl-rutinoside)-5-glucoside (20–200 μg g⁻¹ FW) whilst dark purple–black tubers contained similar levels of petunidin-3-(p-coumaroyl-rutinoside)-5-glucoside together with much higher concentrations of malvidin-3-(p-coumaroyl-rutinoside)-5-glucoside (2000–5000 μg g⁻¹ FW). Tuber flesh also contained chlorogenic acid (30–900 μg g⁻¹ FW) and other phenolic acids plus low concentrations of flavonoids (0–30 μg g⁻¹ FW). Tuber skins showed much higher levels (1000–4000 μg g⁻¹ FW) of chlorogenic acid. The major anthocyanins in flowers were present as the rutinosides or other glycosides of pelargonidin, petunidin and malvidin whilst glycosides of cyanidin and delphinidin were found in some flowers, together with many of the same phenolic acids as found in tubers. The commonest flavonoids included rutin, kaempferol-3-rutinoside and two quercetin-rhamnose-glucosides. Flowers and leaves contained higher concentrations of flavonoids which fell into two patterns, with some cultivars containing high concentrations of quercetin-3-glycosides, whilst others had much lower concentrations. © 1998 SCI.     

Phytosterols: lack of cytotoxicity but interference with beta-carotene uptake in Caco-2 cells in culture.

Ingestion of phytosterols has been shown to reduce plasma cholesterol in both animals and humans. The esterified forms of phytosterols are increasingly being incorporated into margarine and fat spreads, which are then marketed as functional foods. The aim was to assess the cytotoxicity and uptake of four phytosterols, beta-sitosterol, campesterol, stigmasterol and stigmastanol, in human intestinal cells in culture. Another aim was to determine if phytosterols would interfere with alpha-tocopherol or beta-carotene uptake by these cells. Human adenocarcinoma Caco-2 cells were supplemented for 24 h with increasing concentrations (0-12.5 microM) of each phytosterol. Cytotoxicity was assessed by neutral red uptake (NRU), lactate dehydrogenase release (LDH) and fluorescein diacetate/ethidium bromide (FDA/EtBr) assays. The phytosterols had no significant effects on Caco-2 cell viability assessed using LDH and FDA/EtBr assays. The highest concentrations of beta-sitosterol and campesterol tested (12.5 microM) resulted in decreased cell viability assessed using the NRU assay. All phytosterols were taken up by Caco-2 cells in culture. The results demonstrate a reduction in the uptake of beta-carotene when Caco-2 cells were supplemented with 20 microM beta-sitosterol. beta-Sitosterol did not interfere with alpha-tocopherol uptake by the cells. In conclusion, Caco-2 cells are a useful model system to study potential interactive effects of phytosterols with fat-soluble dietary components.     

Changes of flavonoid content and antioxidant capacity in blueberries after illumination with UV-C

The levels of flavonoids in blueberries (Vaccinium corymbosum L.) were found to increase after illumination with UV-C. Phytochemicals affected included resveratrol, myricetin-3-arabinoside, quercetin-3-galactoside, quercetin-3-glucoside, kaempferol-3-glucuronide, delphinidin-3-galactoside, cyanidin-3-galactoside, delphinidin-3-arabinoside, petunidin-3-galactoside, petunidin-3-glucoside, petunidin-3-arabinoside, malvidin-3-galactoside, malvidin-3-arabinoside, and chlorogenic acid as analyzed by HPLC. Significantly higher antioxidant capacity was detected in fruit treated with 2.15, 4.30, or 6.45 kJ m⁻² compared to the control fruit. UV-C dosage of 0.43 kJ m⁻² also increased phenolics and anthocyanins, but to a lesser extent. The optimum doses of UV-C for enhancing phytochemical content in blueberries were 2.15 and 4.30 kJ m⁻². These data suggest that proper use of UV-C illumination is capable of modifying the phytochemical content of blueberries. Time course measurements of the effects of UV-C revealed that the strongest responses of fruit to UV-C treatment occurred instantly after the illumination and the effects diminished with time. Therefore, even though residual effects were evident following UV-C exposure, the best results were obtained immediately after the treatment.     

Bilberry and blueberry anthocyanins act as powerful intracellular antioxidants in mammalian cells

Berry anthocyanins have pronounced health effects, even though they have a low bioavailability. The common mechanism underlying health protection is believed to relate to antioxidant activity. Berry extracts, chemically characterised for their phenolic content, were prepared from bilberries (Vaccinium myrtillus L.) and blueberries (Vaccinium corymbosum L.); the bilberry extract was further purified to obtain the anthocyanin fraction. The antioxidant activity of each extract was examined at the cellular level. For this purpose a specific assay, known as cellular antioxidant activity assay (CAA), was implemented in different cell lines: human colon cancer (Caco-2), human hepatocarcinoma (HepG2), human endothelial (EA.hy926) and rat vascular smooth muscle (A7r5). Here we show for the first time that anthocyanins had intracellular antioxidant activity if applied at very low concentrations (<1 μg/l; nM range), thereby providing a long-sought rationale for their health protecting effects in spite of their unfavorable pharmacokinetic properties.     

姜黄素抑制Cu~(2+)诱导的转APP_(695)基因SH-SY5Y细胞氧化损伤和凋亡作用

目的:研究Cu~(2+)诱导的转β-淀粉样前体蛋白(amyloid-β precursor protein,APP)基因SH-SY5Y(SHSY5Y-APP_(695))细胞氧化损伤、凋亡及姜黄素的抑制作用。方法:在姜黄素预保护和无姜黄素预保护条件下,50 μmol/L Cu~(2+)处理细胞24 h,测定细胞存活率、胞外乳酸脱氢酶(lactate dehydrogenase,LDH)水平、胞内活性氧(reactive oxygen,ROS)水平、线粒体膜电位、半胱氨酸天冬氨酸蛋白酶(caspase)-3、caspase-8和caspase-9活力,蛋白免疫印迹法测定核转录NF-E2相关因子2(NF-E2-related factor 2,Nrf2)Ser40位点磷酸化(p Ser40-Nrf2)水平以及血红素加氧酶(heme oxygenase,HO)-1蛋白表达水平。结果:与空白对照相比,Cu~(2+)损伤组细胞存活率降低,胞内ROS水平和胞外LDH活性升高,线粒体膜电位下降,caspase-9和caspase-3活性明显升高,p Ser40-Nrf2和HO-1含量增加,而姜黄素保护组细胞存活率升高,胞内ROS水平和LDH释放水平降低,线粒体膜电位恢复升高,caspase-9和caspase-3活性明显降低,p Ser40-Nrf2和HO-1表达量减少。结论:姜黄素在一定程度上减弱了Cu~(2+)诱导的氧化损伤和细胞凋亡作用。