Effect of genistein analogs on oxygen radical production in leukocytes stimulated by unopsonized zymosan

Effects of genistein analogs on oxygen radical production have been analyzed in human neutrophils, human monocytes or murine macrophages Raw264.7 stimulated with unopsonized zymosan by lucigenin- and luminol-enhanced chemiluminescence assays. Genistein exhibited IC50 values of 10.7-11.5 microM on the oxygen radical production in human neutrophils, 10.9-11.0 microM in human monocytes, and 14.8-27.3 microM in Raw264.7 cells. Orobol, a genistein analog with an additional hydroxy group at the 3' position, exhibited IC50 values of 3.0-3.3 microM on the oxygen radical production in human neutrophils, 2.8-3.1 microM in human monocytes, and 1.5-3.9 microM in Raw264.7 cells. Genistin and sophoricoside are genistein glycosides with a glucose moiety at 7 or 4' position, respectively. The genistein glycosides exhibited 23-37% inhibitory effects at 100 microM on the oxygen radical production.     

白藜芦醇对人胚肺成纤维细胞株MRC-5生长的抑制作用

目的探讨白藜芦醇(Res)对人胚肺成纤维细胞生长的抑制作用及相关机制。方法以人胚肺成纤维细胞MRC-5为研究对象,分别予20μL二甲基亚砜(DMSO)及0、12.5、25、50、100、200μmol·L-1Res处理细胞24、48、72 h,通过MTT法分析细胞增殖抑制率。另外,以20μL DMSO(溶媒组)及50、100μmol·L-1Res孵育细胞48 h,采用流式细胞术检测细胞周期和凋亡率,行原位缺口末端标记法(TUNEL)测定细胞凋亡指数(AI),应用荧光实时定量PCR和Western blot分别检测细胞周期蛋白D1(cell cycle protein D1,Cyclin D1)、细胞周期蛋白依赖激酶4(cyclin-dependent kinase 4,CDK4)mRNA与蛋白表达,Western blot检测Bcl-2、Bax蛋白表达。结果随着Res浓度的升高和处理时间的延长,细胞增殖抑制率逐渐增加(P<0.01)。在共同培养48h后,50、100μmol·L-1Res处理组S、G2/M期DNA比例及Cyclin D1、CDK4 mRNA与蛋白表达水平、Bcl-2蛋白表达水平降低,而G0/G1期DNA比例、AI、凋亡率及Bax蛋白表达水平增加,与溶媒组比较,差异均有显著性(P<0.01),并且100μmol·L-1Res效果强于50μmol·L-1(P<0.01)。结论Res能抑制MRC-5细胞增殖,其机制可能与阻碍细胞周期进展及促进细胞凋亡有关。     

Protective effects of resveratrol and its analogs on age-related macular degeneration in vitro

Damage of retinal pigment epithelial (RPE) cells by A2E may be critical for age-related macular degeneration (AMD) management. Accumulation and photooxidation of A2E are known to be one of the critical causes in AMD. Here, we evaluated the protective effect of resveratrol (RES), piceatannol (PIC) and RES glycones on blue-light-induced RPE cell death caused by A2E photooxidation. A2E treatment followed by blue light exposure caused significant damages on human RPE cells (ARPE-19). But the damages were attenuated by post- and pre-treatment of RES and PIC in our in vitro models. The results of cell free system and FAB-MS analysis clearly showed that the reduction of A2E by blue light exposure was significantly rescued, and that oxidized forms of A2E were significantly reduced by RES or PIC treatment. Besides, RES or PIC inhibited the intracellular accumulation of A2E. Not only RES and PIC but RES glycones showed protection of ARPE-19 cells against A2E and blue-light-induced photo-damage. These findings demonstrate that RES and its analogs may have protective effects against A2E and blue-light-induced ARPE-19 cell death through regulation of A2E accumulation as well as photooxidation of A2E. Thus RES and its analogs may be beneficial for AMD treatment.     

Inhibitory effect of a novel resveratrol derivative on nitric oxide production in lipopolysaccharide-activated microglia

Excessive nitric oxide (NO) production by activated microglial cells has been implicated in various neurodegenerative diseases. In the present study, we found that a new resveratrol derivative, (E)-5-(3-nitrostyryl)benzene-1,3-diol (RV06), has a more potential inhibitory effect on the production of NO in LPS-activated N9 microglial cells, and the result was confirmed on primary rat microglial cells. Further studies showed that RV06 inhibited LPS-induced iNOS expression in N9 microglial cells, with no activity on direct scavenging nitric oxide radical in a cell-free environment. The results suggest that RV06 might be a potential anti-inflammatory agent or leading compound which can inhibit inflammatory responses of microglia.     

Inhibitory effects of maesanin and analogs on arachidonic acid metabolizing enzymes.

The substituted 1,4-benzoquinone, maesanin (1), is a potent 5-lipoxygenase (5-LO) inhibitor present in the fruit of Maesa lanceolata Forssk. Thirteen natural, synthetic, semisynthetic, and microbially transformed analogs of 1 were tested for their in vitro inhibition of 5-lipoxygenase (5-LO) and cyclooxygenase-1 (COX-1). Maesanin was the most active 5-LO inhibitor. All other analogs were inactive or less active than the natural products as 5-LO inhibitors. None of the tested compounds was strongly active in the COX-1 inhibition assay.     

Inhibitory effects of novel tetrahydropyridine derivatives on nitric oxide and reactive oxygen species production in glioma cells

The immune response in the central nervous system involves the degeneration of neurological tissue. The therapeutic use of nonsteroidal anti‐inflammatory agents (NSAIDs) can be effective in reducing inflammation and associated neurodegeneration. The current study was designed to examine the effects of previously synthesized N‐(substituted benzoylamino)‐4‐ethyl‐1,2,3,6‐tetrahydropyridines (THPs) on free radical and nitric oxide (NO) generation in C6 glioma cells activated with bacterial endotoxin [lipopolysaccharide (LPS): Escherichia coli 0111:B4] and interferon‐γ. Stimulated C6 cells exhibited elevated iNOS protein expression, an increase of reactive oxygen species and NO₂^?. All parameters were attenuated significantly by indomethacin and various THPs (4‐Ome, 3‐CF3, 4‐Me, 4‐Et, 4‐t‐butyl, and 4‐n‐butyl). The results indicate a possible role for THP derivatives as anti‐inflammatory agents. Drug Dev. Res. 61:189–196, 2004. © 2004 Wiley‐Liss, Inc.     

Inhibitory effects of resveratrol on MCP-1, IL-6, and IL-8 production in human coronary artery smooth muscle cells

Resveratrol, a representative polyphenol compound, is known to have antiatherogenic effects through its various actions including an anti-inflammatory action. The processes of initiation and progression of atherosclerosis are mediated by proinflammatory cytokines. The aim of this study was to determine whether resveratrol affects cytokine production in human coronary artery smooth muscle cells (HCASMCs). Each cytokine concentration in the culture medium of HCASMCs was measured by flow cytometry using the cytometric bead array system, and extracellular signal-regulated kinase (ERK) activity was evaluated by Western blotting. Basal levels of monocyte chemoattractant protein-1 (MCP-1), interleukin (IL)-6, and IL-8 were significantly decreased in the presence of resveratrol at 1–50 μM in a concentration-dependent manner and were significantly decreased in the presence of U0126, an ERK inhibitor. Resveratrol significantly decreased both basal and interferon-γ (IFN-γ) (200 ng/ml)-stimulated levels of MCP-1, IL-6, and IL-8 and significantly attenuated both basal and IFN-γ-stimulated activity of ERK. TNF-α, IL-1β, IL-10, and IL-12p70 were detected only as trace levels in the culture medium with or without IFN-γ. Therefore, resveratrol is thought to inhibit production of MCP-1, IL-6, and IL-8 in HCASMCs through attenuating ERK activity. Inhibition of cytokine production in coronary artery smooth muscle cells may in part explain antiatherogenic action of resveratrol.     

白藜芦醇类似物的合成及其抗氧化活性

目的:合成白藜芦醇类似物,以期获得抗氧化活性更好的活性分子。方法:将白藜芦醇的A环分别用活性杂环川芎嗪和4-氨基喹啉替换。考察目标合物9,12及中间体8,11对DPPH自由基的清除作用。结果与结论:设计合成了2个未见文献报道的白藜芦醇类似物,其结构经MS和1H-NMR图谱确证。化合物9,11和12对DPPH自由基的清除作用较白藜芦醇减弱,化合物8在低浓度时具有比白藜芦醇更强的DPPH自由基清除能力,具有潜在的抗氧化作用。     

Inhibitory effect of resveratrol on angiotensin II-induced cardiomyocyte hypertrophy

Resveratrol is proposed to account in part for the protective effect of red wine on the cardiovascular system. Angiotensin II (Ang II) is a potent hypertrophic stimulus in cardiomyocytes. In this study, we determined the effect of resveratrol on Ang II-induced cardiomyocyte hypertrophy. Cultured neonatal rat cardiomyocytes were stimulated with Ang II, and [³H]leucine incorporation and -myosin heavy chain (-MyHC) promoter activity were examined. Intracellular reactive oxygen species (ROS) were measured by a redox-sensitive fluorescent dye, 2 7-dichlorofluorescin diacetate, and the extracellular signal-regulated kinase (ERK) phosphorylation was examined by Western blotting. Resveratrol inhibited Ang II-increased intracellular ROS levels. Furthermore, resveratrol, as well as the antioxidant N-acetyl-cysteine, decreased Ang II- or H₂O₂-increased protein synthesis, -MyHC promoter activity, and ERK phosphorylation. In summary, we demonstrate for the first time that resveratrol inhibits Ang II-induced cardiomyocyte hypertrophy via attenuation of ROS generation.Abbreviations Ang II Angiotensin II - MAPKs Mitogen-activated protein kinases - ERK Extracellular signal-regulated kinase - JNK c-Jun N-terminal kinase - p38 MAPK p38 mitogen-activated protein kinases - MEK MAPK or ERK kinase - NAC N-acetylcysteine - DCF-DA Dichlorodihydrofluorescein diacetate - DCF Dichlorofluorescein     

Effect of resveratrol, a natural polyphenolic compound, on reactive oxygen species and prostaglandin production

Resveratrol is a natural molecule with antioxidant action. Moreover, resveratrol is also considered to be a molecule with anti-inflammatory action, an effect attributed to suppression of prostaglandin (PG) biosynthesis. The aim of the present study was to investigate the effects of resveratrol, a polyphenol present in most red wines, on reactive oxygen species formation as well as on arachidonic acid (AA) release, cyclooxygenase expression, and PG synthesis in murine resident peritoneal macrophages. Results show that resveratrol exerted a strong inhibitory effect on superoxide radical (O2-) and hydrogen peroxide (H2O2) produced by macrophages stimulated by lipopolysaccharides (LPS) or phorbol esters (PMA). Resveratrol also significantly decreased [3H]AA release induced by LPS and PMA or by exposure to O2- or H2O2. Resveratrol treatment caused a significant impairment of cyclooxygenase-2 (COX-2) induction stimulated by LPS and PMA or by O2- or H2O2 exposure. These effects of resveratrol on [3H]AA release and COX-2 overexpression were correlated with a marked reduction of PG synthesis. Our results indicate that the antioxidant action of resveratrol affects AA mobilization and COX-2 induction.     

绞股兰总皂苷对小鼠脑缺血的保护作用及对自由基生成的抑制作用

目的 :观察绞股兰总皂苷对小鼠脑缺血的保护作用及对自由基生成的抑制作用。方法 :不完全结扎小鼠一侧颈总动脉及迷走神经 ,另侧完全结扎造成脑缺血 ,观察 6h卒中指数 ,12 h存活时间和死亡率 ;体外生成 O·2 。结果 :与对照组比较 ,绞股兰总皂苷 17.5 ,3 5和 5 2 .5 mg· kg-1 ip可以降低卒中指数 ,分别降低 2 5 .6% (P<0 .0 5 ) ,3 6.7% (P<0 .0 1)和 5 2 .4% (P<0 .0 1) ;延长存活时间 ,分别延长 165 % (P<0 .0 1) ,189% (P<0 .0 1)和 2 5 2 % (P<0 .0 1) ;降低死亡率 ,分别降低 3 0 % (P<0 .0 1) ,48%(P<0 .0 1)和 64 % (P<0 .0 1) ;均呈一定的剂量依赖性趋势。绞股兰总皂苷 5 0 ,10 0和 15 0 mg· L-1 对体外生成的 O·2 均有一定的抑制作用。结论 :绞股兰总皂苷对小鼠脑缺血具有保护作用 ,且可能与其对自由基生成具有抑制作用有关     

Pleiotropic effects of resveratrol

Resveratrol, a phytoalexin found in red wine, has several pharmacological properties which include inhibition of arachidonate metabolism in leukocytes and platelets, modulation of lipid metabolism, inhibition of platelet aggregation and lipid peroxidation, reduction of expression and activity of cyclooxygenase-2. Resveratrol induces cell cycle arrest at S/G(2) phase transition and a pro-apoptotic cell death in several types of cancer cells. All these biological activities help to explain the vasorelaxing, anticancer and antiinflammatory activity of resveratrol. This article summarizes the wide range of biological activities of resveratrol in three disease states: cancer, coronary heart disease and pain. In addition, the mechanisms underlying these promising properties of resveratrol are also reviewed.     

Inhibitory effects of radical scavengers on diacylglycerol-promoted transformation in BALB/3T3 cells

To discover the relationship between the activation of protein kinase C and the generation of reactive oxygen in the tumor promotion process, we investigated the effects of radical scavengers on diacylglycerol-promoted transformation in BALB/3T3 cells. Superoxide dismutase (SOD) showed inhibitory effects on both 1-oleoyl-2-acetylglycerol (OAG)-promoted and diolein-promoted transformation. Catalase (CT) suppressed the promoting effects of diolein by up to 70%. Mannitol (MT), a hydroxyl radical (.OH) scavenger, inhibited diacylglycerol-promoted transformation dose-dependently. These results suggest that activation of protein kinase C alone is insufficient and that generation of reactive oxygen accompanied by activation of the enzyme is essential to the promotion process in BALB/3T3 cells.     

白藜芦醇对人结肠癌细胞上皮间质转化的抑制作用

目的观察白藜芦醇(Resveratrol,Res)对人结肠癌细胞HCT-8上皮间-质转化(EMT)的影响,初步探讨其可能存在的机制。方法取对数生长期的人结肠癌HCT-8细胞,采用不同浓度的白藜芦醇(0、40、80、120μmol/L)对细胞分别进行干预24、48、72 h;MTT法检测不同时间Res对HCT-8细胞增殖活性的影响;细胞黏附试验检测Res对HCT-8细胞黏附能力的影响;Transwell小室试验检测Res对HCT-8细胞迁移和侵袭能力的影响;进一步采用蛋白免疫印迹法和qRT-PCR分别检测Res对HCT-8细胞中蛋白激酶B(AKT)、磷酸化蛋白激酶B(P-AKT)、E-钙黏蛋白(E-cadherin)、N-钙黏蛋白(N-cadherin)、波形蛋白(Vimentin)和mRNA表达的影响。结果 Res(浓度为40、80、120μmol/L)在干预后24、48、72 h均可以明显抑制HCT-8细胞的增殖活性(P<0.05)。与空白对照组相比,细胞的同种黏附能力增强,异种黏附能力降低,差异有统计学意义(P<0.05)。与空白对照组相比,Res 120μmol/L对细胞迁移和侵袭能力均具有明显的抑制作用(P<0.05),Res 120μmol/L作用HCT-8细胞48 h后,HCT-8细胞中上皮间质标志性蛋白E-钙黏蛋白(E-cadherin)和mRNA的相对表达量增加(P<0.05),而AKT蛋白的表达无明显变化(P>0.05),P-AKT蛋白的表达明显减少(P<0.05);N-钙黏蛋白(N-cadherin)、波形蛋白(Vimentin)和mRNA的相对表达量显著减少,差异有统计学意义(P<0.05)。结论白藜芦醇通过调控细胞EMT抑制人结肠癌HCT-8细胞的迁移和侵袭能力,其作用机制可能受PI3K/AKT信号通路的调控。     

Inhibitory effects of a novel synthetic protease inhibitor, FUT-175, on the paw edema in rats and zymosan-induced complement activation in vitro

FUT-175 inhibited the zymosan-induced rat paw edema in a dose-dependent manner, while indomethacin exhibited no significant activities in this model. FUT-175 also inhibited the decrease in hemolytic complement (CH50) induced by zymosan in vitro, and indomethacin was inactive. These results suggest that FUT-175 has potent in vitro and in vivo inhibitory activity against the activation of the complement system induced by zymosan.     

基于雌激素代谢调控的白藜芦醇对MNNG诱导子宫内膜癌变的抑制作用

目的考察白藜芦醇(Res)是否可能通过调控雌激素内稳态,进而抑制MNNG诱导小鼠子宫内膜的癌变。方法采用MNNG诱导EC小鼠模型,将小鼠随机分为Control组、Res组、MNNG组、MNNG+Res组。HE染色观察组织病理情况;q PCR检测Ccnd1、CK-19、Erbb2 mRNA的表达;LC-MS/MS检测小鼠血清和子宫中E_1、E_2、2-MeOE_1、4-MeOE_1、2-MeOE_2、4-MeOE_2、16α-OHE_1、2-OHE_1、4-OHE_1、2-OHE_2和4-OHE_2的含量。结果 MNNG组小鼠子宫内膜腺体增多、拥挤,局灶形成筛孔;MNNG+Res组小鼠子宫内膜腺体减少,趋于正常。与Control组相比,MNNG组Ccnd1、Erbb2mRNA表达明显升高,MNNG+Res组明显降低。MNNG组小鼠的血清和子宫组织中,2-MeOE_2、2-MeOE_1和4-MeOE_1含量明显下降,4-OHE_2含量明显增加,给予Res治疗后,血清中4-MeOE_1含量明显升高,子宫组织中2-MeOE_2和2-MeOE_1含量明显升高,4-OHE_2在血清和子宫组织中含量均明显降低。结论 EC小鼠体内雌激素内稳态失衡,Res可以上调雌激素代谢产物2-MeOE_2的含量,降低4-OHE_2的含量,抑制EC的发生发展。     

埃必定对人血白细胞呼吸爆发和氧自由基的抑制作用

目的观察埃必定对人血白细胞呼吸爆发和氧自由基的影响，初步探讨埃必定的抗炎作用机制。方法用化学发光法测定人血白细胞受调理的酵母多糖刺激发生呼吸爆发的活性氧、碱性二甲基亚砜－鲁米诺体系产生的超氧阴离子自由基（Ｏ·２）、ＶｉｔＣ－Ｃｕ２＋酵母多糖产生的羟自由基（·ＯＨ）及过氧化氢（Ｈ２Ｏ２）的释放，观察了埃必定对上述体系产生的自由基的影响。结果埃必定对白细胞的呼吸爆发有显著的抑制作用；对Ｏ·２、·ＯＨ有清除作用，呈剂量依赖性，对Ｈ２Ｏ２无清除作用。结论埃必定对人血白细胞呼吸爆发具有抑制作用，对氧自由基具有清除作用