姜黄素对人增生性瘢痕成纤维细胞生长及功能的影响

目的 探讨姜黄素对人增生性瘢痕(hyperplastic scar)成纤维细胞（fibroblast, FB）增殖及胶原合成的影响,以寻找治疗人增生性瘢痕的有效药物。方法 应用组织块法分离培养瘢痕和正常真皮组织的FB,在倒置相差显微镜下观察细胞形态,应用细胞计数方法绘制细胞生长曲线,测定细胞生长速度;采用不同浓度的姜黄素（0、12.5、25、50和100 μmol/L）刺激瘢痕FB,采用四氮甲基唑盐比色法（MTT法）检测姜黄素对FB生长的抑制效应,采用RT PCR检测姜黄素对FBα1（Ⅰ）前胶原基因的转录活性的影响。结果 细胞生长曲线显示:增生性瘢痕FB倍增时间约为5天,正常皮肤FB后细胞倍增时间为4天,两者比较,差异有统计学意义(P<0.05);MTT检测结果显示：12.5 μmol/L姜黄素作用于增生性瘢痕FB,表现为增殖倾向,其光吸收值（A值）显著地高于正常对照组。而在25～100 μmol/L 的浓度范围内,随着姜黄素药物浓度的增加,其抑制作用逐渐增强,50 μmol/L和100 μmol/L姜黄素能显著抑制瘢痕FB增殖（P<0.05）;此外,50 μmol/L和100 μmol/L姜黄素能够显著抑制瘢痕FBⅠ型胶原合成（P<0.01）。结论 较高浓度的姜黄素能有效抑制增生性瘢痕FB增殖和Ⅰ型胶原合成,对增生性瘢痕可能具有治疗作用。     

Protection of liver cells from ethanol cytotoxicity by curcumin in liver slice culture in vitro

Curcumin, a widely used spice and colouring agent in food has been shown to have a broad spectrum of biological activities such as anti-inflammatory, anti-neoplastic, antimutagenic and antioxidant. We have used liver slice culture model to demonstrate hepatoprotective activity of curcumin in vitro. Ethanol has been used as a hepatotoxin and the cytotoxicity of ethanol is estimated by quantitating the release of LDH. Ethanol induces 3.5 times more release of LDH from the liver cells and twice the amount of lipid peroxidation as compared to the cells from untreated liver tissue and this was significantly reduced in presence of curcumin (5 microM). We measured the activity of antioxidant enzymes (AOEs) namely superoxide dismutase, catalase and peroxidase and found that in ethanol treated cells activity of all three enzymes was elevated. However, when curcumin was added along with ethanol their levels were kept low. The fact that release of LDH is significantly reduced along with lipid peroxidation and the activity of AOEs is kept low indicates that curcumin by its antioxidant activity reduced the oxidative stress induced by ethanol and protected the liver cells in vitro.     

姜黄素对动脉粥样硬化家兔血管内皮功能的影响

目的: 观察姜黄素对实验性动脉粥样硬化(atherosclerosis,AS)家兔血浆一氧化氮(NO)水平,结构型一氧化氮合酶(constitutive NOS, cNOS)活性和非对称性二甲基精氨酸(asymmetric dimethylarginine,ADMA)的影响,探讨姜黄素抗AS作用及与ADMA的相关性. 方法: 38只雄性日本大耳白兔随机分为4组,即对照组(8只,喂食普通饲料)、模型组(10只,喂食高脂饲料)、低剂量姜黄素组(10只,喂食高脂饲料+100 mg·kg^-1·d^-1姜黄素)和高剂量姜黄素组(10只,喂食高脂饲料+200 mg·kg^-1·d^-1姜黄素).于实验的第12周末测定血浆TC,LDL-C,NO,内皮素(endothelin,ET),ADMA水平,主动脉组织cNOS活力,同时留取主动脉组织行病理检查. 结果: 喂食高脂饲料的3组较对照组家兔血浆ADMA和ET水平浓度均有升高(P<0.01),血浆NO浓度、动脉组织cNOS活性降低(P<0.01).姜黄素组家兔血浆ADMA和ET水平较模型组低(P<0.05),血浆NO浓度、动脉组织cNOS活性则要高出于模型组(P<0.01),2个姜黄素组之间未见显著性差异. 结论: 姜黄素有可能通过降低血浆ADMA水平,在AS进程中发挥重要的内皮保护作用.     

Neuroprotective role of curcumin from curcuma longa on ethanol-induced brain damage.

In the present study, curcumin from Curcuma longa was screened for neuroprotective activity using ethanol as a model of brain injury. Oral administration of curcumin to rats caused a significant reversal in lipid peroxidation, brain lipids and produced enhancement of glutathione, a non-enzymic antioxidant in ethanol intoxicated rats, revealing that the antioxidative and hypolipidaemic action of curcumin is-responsible for its protective role against ethanol induced brain injury.     

Study on antifibrotic effects of curcumin in rat hepatic stellate cells

Suppression of activation or fibrogenesis and induction of apoptosis, in hepatic stellate cells (HSCs) have been proposed as therapeutic strategies against liver fibrosis. Curcumin, an active compound isolated from yellow curry pigment of turmeric (Curcuma longa Linn), has been demonstrated to be an effective anti‐inflammatory and antioxidant compound. In this study, we investigated the in vitro antifibrogenic effects of curcumin on HSCs at the concentration range of (1–40 µM). A cell line of rat HSCs (HSC‐T6) was stimulated with transforming growth factor‐β1 (TGF‐β1). The inhibitory effects of curcumin (1.25～10 µM) on fibrosis‐related markers including α‐smooth muscle actin (α‐SMA) and collagen were assessed. In addition, the induction effects of curcumin (20～40 µM) on apoptosis in HSC‐T6 cells were also assessed by Hoechst and propidium iodide stains. Curcumin (1.25～10 µM) concentration‐dependently suppressed TGF‐β1‐induced α‐SMA expression and collagen deposition in HSC‐T6 cells, without cytotoxicity. Whereas, higher concentrations of curcumin (20～40 µM) induced cell apoptosis and cytochrome c release in HSC‐T6 cells. Our results suggest that curcumin exerted antifibrotic effects, possibly through two different mechanisms depending on its concentrations. At lower concentrations (1.25～10 µM), curcumin exerted antifibrogenic effects, whereas at higher concentrations (20～40 µM), curcumin exerted induction of apoptosis in HSCs. Copyright © 2009 John Wiley & Sons, Ltd.     

芎芍胶囊对兔实验性血管再狭窄血管胶原酶基因表达的影响

目的 观察芎芍胶囊对兔实验性血管再狭窄（RS）血管胶原酶基因表达的影响,探讨芎芍胶囊干预RS的机制。方法 采用球囊剥脱兔腹主动脉内皮结合高脂饲料方法建立实验性RS的模型,80只兔随机分层分组,分为8组,即正常对照组,单纯内皮损伤组,模型3天、2周、6周组,普罗布考对照组,芎芍胶囊小剂量、大剂量组,每只10只。采用RT-PCR方法、计算机图像分析仪,观察芎芍胶囊对再狭窄过程中血管胶原酶基因表达的影响,结合病理形态学和胶原的变化探讨芎芍胶囊改善血管重构预防RS的机理。结果 模型2周时管腔出现代偿性扩张,至6周时管腔却明显缩小,增殖指数明显升高。各用药组在抑制内膜增殖方面以芎芍大剂量组及普罗布考对照组尤为明显。模型2周时内膜胶原堆积不明显,但胶原总量增加,并达到高峰。6周时,内膜胶原逐渐堆积达到高峰。各用药组内膜胶原减少,中、外膜胶原总量减少,以芎芍大剂量组最明显。在正常对照及模型各组,基质金属蛋白酶-1（MMP-1） mRNA有弱表达,芎芍大剂量组与模型6周组比较差异有统计学意义（P＜0.05）。结论 芎芍胶囊可明显增强MMP 1mRNA在损伤血管部位的表达,提示芎芍胶囊预防RS可能与上调MMP-1mRNA的表达,增加胶原的降解,减少胶原在血管壁的沉积有关。     

姜黄素与阿米洛利联用对大鼠肝星状细胞氧应激所致纤维化的抑制作用

目的：探讨钠氢交换抑制剂阿米洛利(Amibride)与抗氧化剂姜黄素(curcumin，Cur)联用对大鼠肝星状细胞(hepatic stellate cells，HSC)氧应激所致纤维化的作用。方法：HSC与三醋酸亚硝酸铁(FeNTA)共同培养。以MTY法观察阿米洛利、姜黄素对HSC增殖的效应。LDH法观察阿米洛利、姜黄素对HSC的毒性作用。以：ELISA法测定细胞培养液中Ⅰ型胶原的水平。以试剂盒推荐方法分别测定细胞培养液中丙二醛(MDA)、谷胱苷肽(GSH)、谷胱甘肽过氧化物酶(GSH-Px)、超氧化物歧化酶(SOD)水平。结果：HSC与FeNTA共同培养，细胞内MDA、GSH水平明显升高，GSH-Px、SOD活力明显降低。FeNTA与HSC共同培养，能明显促进：HSC增殖和提高细胞培养上清中Ⅰ型胶原的水平。阿米洛利、姜黄素可以逆转上述所有FeNTA所致效应，且联用组优于单用组。结论：作用于HSC不同靶点的抗纤维化药物姜黄素、阿米洛利合用优于单用。     

姜黄素对人胚肺成纤维细胞外基质影响的试验研究

目的：探讨姜黄素对经转化生长因子β（TGF-β1）诱导后人胚肺成纤维细胞一型胶原与c-jun,在人胚肺成纤维细胞中的表达的影响及其与特发性肺纤维化的关系。方法：细胞分为正常组、姜黄素组、模型组、治疗组采用采用RT-PCR,Western bloting法检测、I型胶原与c-jun在人胚肺成纤维细胞的变化。结果：治疗组与正常组、模型组相比,I型胶原与c-jun,表达明显减弱。结论：姜黄素可抑制人胚肺成纤维细胞的胶原沉积,在延缓肺纤维化形成中起一定作用,其作用机制可能是通过抑制核转录因子c-jun表达而实现的。     

姜黄素上调博来霉素诱导肺纤维化C57BL/6小鼠Cathepsin K表达

目的:研究姜黄素对博来霉素诱导肺纤维化C57BL/6小鼠Cathepsin K表达的影响。方法:博来霉素气管注射造小鼠肺纤维化模型,随机分为对照组(气管内注入0.1mL生理盐水)、模型组(气管内注入0.025U博来霉素)、姜黄素组(气管内注入0.025U博来霉素);造模后第2d,对照组和模型组给予0.5%羧甲基纤维素钠生理盐水灌胃,姜黄素组给予200 mg.kg-1.d-1姜黄素灌胃,分别于造模后第7、14、28d取材。采用H&E及Mallory染色观察肺组织病理学变化;应用免疫组学Cathepsin K蛋白的表达,经图像分析各组之间的差异。结果:姜黄素组、模型组与对照组比较都有不同程度的炎症及纤维化病灶,但是姜黄素组与对照组比较炎症及纤维化程度明显减轻;姜黄素组、模型组与对照组比较Cathepsin K表达都有不同程度增加,但是姜黄素组与对照组比较Cathep-sin K表达显著增加。结论:姜黄素可能是通过增加Cathepsin K的表达从而降解胶原而实现抗纤维化作用。     

姜黄抗前列腺增生作用有效成分研究

目的:观察姜黄75%乙醇提取物对良性前列腺增生症模型小鼠前列腺增生的抑制效果,探讨姜黄抗前列腺增生作用的活性成分。方法:测定75%乙醇提取的姜黄不同溶剂萃取物中姜黄素的含量,并观察不同溶剂萃取物对良性前列腺增生症模型小鼠前列腺系数的影响。结果:姜黄素含量最高的乙醚萃取物与姜黄素含量最低的正丁醇萃取物均能降低小鼠前列腺系数,且两者治疗效果无明显差异(P 0.05)。结论:姜黄75%乙醇提取物具有较好的治疗良性前列腺增生症的作用,除姜黄素外,其他成分也具有与姜黄素相似的抑制前列腺增生的作用。     

姜黄素激活过氧化物酶体增殖因子活化受体γ信号对大鼠肝星状细胞基质金属蛋白酶2、9活性和胞核核因子-κBp65表达的影响

目的 研究姜黄素激活过氧化物酶体增殖因子活化受体γ（PPARγ）信号对大鼠肝星状细胞（HSC）活化、基质金属蛋白酶（MMPs）活性和胞核核因子-κBp65（RelA）表达的影响。方法 采用肝脏原位灌流酶消化、Nycodenz密度梯度离心法分离大鼠HSC。药物处理后收集裂解细胞,Western blot检测PPARγ、α平滑肌肌动蛋白（αSMA）、Ⅰ型胶原、RelA。收集细胞培养上清,明胶酶谱法检测MMP2、9的活性。结果 随着HSC活化程度增加PPAR7表达水平不断下降,姜黄素上调其表达水平（P〈0．01）,拮抗剂GW9662显著阻断这种作用（P〈0．01）。姜黄素抑制αSMA的表达、Ⅰ型胶原的生成以及胞核内活化RelA的表达（P〈0．01）,显著升高MMP2、9的活性（P〈0．01）。结论 姜黄素激活PPARγ信号途径抑制HSC活化,升高MMP2、9活性,抑制／干扰NFκB的核转位。     

聚乙烯吡咯烷酮K30对姜黄素的增溶作用研究

目的评价聚乙烯吡咯烷酮K30(PVP K30)对姜黄素的增溶作用.方法选择PVP K30作为载体,用溶剂法制备姜黄素固体分散体;通过差示扫描量热分析和X射线粉末衍射鉴别姜黄素在载体中的存在状态以及与载体之间的相互作用,并采用紫外-可见分光光度法测定姜黄素固体分散体的溶解度以及体外溶出度.结果紫外-可见分光光度法测定姜黄素的溶出量,利用PVP K30制备姜黄素固体分散体,与姜黄素本身相比,固体分散体中姜黄素溶解度提高了880倍以上,溶出速度明显增大.结论PVP K30可作为载体以提高姜黄素溶解度及溶出速度.     

Protective role of curcumin in ethanol toxicity

The effect of curcumin on ethanol induced liver toxicity was evaluated. The increased levels of aspartate transaminase and alkaline phosphatase induced by ethanol were significantly lowered by curcumin. Elevated serum cholesterol, phospholipids and free fatty acids were observed in ethanol fed rats, but on curcumin treatment they decreased. We have also observed a marked decrease in the levels of thiobarbituric acid reactive substances in serum of alcoholic rats fed curcumin. Thus this study shows that curcumin offers protection against ethanol induced toxicity. © 1998 John Wiley & Sons, Ltd.     

姜黄素诱导结肠癌LoVo细胞凋亡的作用及机制研究

目的: 探讨姜黄素促人结肠癌LoVo细胞凋亡的生物学作用及其调控机制。 方法: 体外培养人结肠癌LoVo细胞,0~20 mg·L^-1不同浓度姜黄素处理后,采用MTT 比色法测定姜黄素对细胞的增殖抑制作用,利用透射电镜观察细胞的超微结构,采用PI单标流式细胞仪测定细胞周期分布,Annexin V-FITC/PI双标法检测细胞凋亡率; 采用 RT-PCR检测Bax,Bcl-2,Caspase-3,Bcl-xL mRNA表达水平。 结果: MTT检测显示姜黄素能显著抑制人结肠癌LoVo细胞的生长、増殖,呈现浓度和时间效应关系;透射电镜结果显示姜黄素能使LoVo细胞发生形态学改变,呈现典型凋亡细胞特征;流式细胞仪分析显示姜黄素能阻滞细胞周期于S期,诱导细胞发生凋亡。RT-PCR检测显示姜黄素可促进细胞中Bax,Caspase-3 mRNA 表达,抑制Bcl-2,Bcl-xL mRNA 表达。 结论: 姜黄素能显著抑制人结肠癌LoVo细胞的增殖并促进其凋亡,这种生物学效应可能与激活Bax表达、抑制Bcl-2和Bcl-xL表达而活化Caspase-3的信号通路有关。     

姜黄素对A549人肺腺癌细胞增殖抑制作用及其机制的研究

目的:探讨姜黄素对人肺腺癌细胞(A549)抗癌作用及其分子机制.方法:采用荧光显微镜、四甲基偶氮唑蓝(MTT)比色法、流式细胞仪(FCM)技术结合PI及AnnexinV-FITC双标记染色、Western blot法.结果:(1)姜黄素作用于癌细胞后,光镜下可见有细胞脱壁,悬浮培养液中;荧光镜下可见细胞核破碎,裂解成大小不等的凋亡小体.(2)MTT法测得不同浓度姜黄素作用A549细胞72 h后,可产生显著的细胞增殖抑制作用.将量效关系进行直线回归,得IC50值18 μmol/L.(3)姜黄素诱导凋亡作用呈剂量依赖性.随着药物浓度从5 μmo/L增加至30 μmol/L,Annexix-FITC单阳性细胞(早期凋亡细胞)由3.4%增加到59.1%;当姜黄素浓度增至40 μmol/L时,PI及AnnexinV-FITC双阳性细胞(凋亡继发性坏死细胞)成为主要的细胞组成.同时发现G2期细胞比例明显增多,出现了G2期阻滞.(4)Western blot法观察到,随着姜黄素浓度增至10 μmol/L,116 kD裂解成为89 kD片段的多聚ADP-核糖聚合酶的表达同步增加.结论:姜黄素能干扰细胞周期进程,对A549细胞有生长抑制作用,且呈剂量依赖性,其抑制作用不仅是由于非特异性的细胞毒性造成的,还与诱导肿瘤细胞发生凋亡有关.     

Negative Effects of Curcumin on Liver Injury Induced by Alcohol

Curcumin is known for its antiinflammatory and antifibrotic properties in liver damage. However, the negative effects of curcumin on alcoholic liver damage are seldom reported. In this study, both advantageous and disadvantageous functions of curcumin on alcoholic liver injury were observed. In order to determine the effects of curcumin on liver fibrosis induced by alcohol, 5% ethanol and/or curcumin (1 × 10^–3 or 1 × 10^–4 m ) were injected intravenously in mice. Hematoxylin‐eosin staining was performed to determine the value of liver injury by optical density analyses. Liver histology was evaluated by an experienced hepatopathologist blinded to the type of treatment received by the animals. Ethanol accelerates serum levels of serum aspartate aminotransferase (AST) and alanine aminotransferase (ALT), liver injury, production of tumor necrosis factor‐alpha (TNF‐α), transforming growth factor‐beta (TGF‐β) and nuclear factor‐kappa B (NF‐κB) during 5% ethanol‐induced liver injury. 1 × 10^–3 m curcumin accelerates liver injury and liver cellular edema during only 5% ethanol‐induced liver injury evolution, whereas 1 × 10^–4 m curcumin does not lead to (or protect) alcoholic liver injury. Therefore, it is suggested that curcumin may have dual effects on alcoholic liver injury depending on its concentration. Copyright © 2012 John Wiley & Sons, Ltd.     

姜黄素对人肝癌细胞Sk-hep-1抗癌作用的研究

目的:研究姜黄素对人肝癌细胞Sk-hep-1体外抗癌作用,探讨其抗癌作用的分子机制.方法:应用MTT法、DAPI核染色法和细胞周期分析法,检测姜黄素对Sk-hep-1细胞生长和凋亡的影响,并通过RT-PCR方法检测姜黄素对Sk-hep-1细胞抗凋亡基因(Survivin和Bcl-xL)和耐药基因(DRG2和MDR1)mRNA表达的影响.结果:姜黄素抑制Sk-hep-1细胞增殖呈量效关系.姜黄素处理组肝癌细胞Sk-hep-1,HepG2和Hep3B细胞发生不同程度的凋亡,正常肝细胞Chang's liver无明显凋亡.姜黄素处理组Sk-hep-1细胞周期发生变化,G_0/G_1或G_2/M期细胞比例增多.姜黄素处理组Sk-hep-1细胞耐药基因MDR1 mRNA水平显著下降,而抗凋亡基因的表达无明显变化.结论:姜黄素能够抑制Sk-hep-1细胞增殖,并诱导其凋亡,推测可能是通过下调MDR1 mRNA的表达而起作用.     

姜黄素类化合物的分离及其抗肝癌和子宫内膜癌的研究

目的从中药姜黄提取3种姜黄素类化合物并对它们进行抗肝癌细胞和子宫内膜癌作用研究。方法采用柱层析法从姜黄提取物中分离出3种姜黄素类化合物,用MTT法测定其抗肝癌细胞和子宫内膜癌作用。结果3种姜黄素化合物分别鉴定为姜黄素、脱甲氧基姜黄素和双脱甲氧基姜黄素。0.01 mg/m l双脱甲氧基姜黄素,0.01 mg/m l和0.001 mg/m l脱甲氧基姜黄素能明显抑制肝癌细胞的生长;0.01 mg/m l和0.001 mg/m l双脱甲氧基姜黄素,0.01 mg/m l脱甲氧基姜黄素,0.01 mg/m l和0.001 mg/m l姜黄素能明显抑制子宫内膜癌细胞的生长。结论一定浓度的姜黄素化合物能明显抑制肝癌和子宫内膜癌细胞的生长。     

姜黄素防治慢性心力衰竭大鼠左心室胶原网络重构的实验研究

目的 观察姜黄素对慢性心力衰竭（CHF）大鼠左室胶原网络重构的影响。方法 采用缩窄大鼠腹主动脉的方法，将大鼠随机分成假手术组、手术对照组、卡托普利组和姜黄素组，姜黄素组又分为高、中、低3个剂量组，造模后每周测定鼠尾动脉血压，分别于造模后第12、20周观察左室质量指数、心脏血流动力学变化，采用VG染色方法检测心肌胶原含量。结果 12周末手术对照组LVEDP、LVMI、CVF-V、CVF—NV均明显强于其余各组，20周末与手术对照组相比，姜黄素各剂量组和卡托普利组大鼠LVSP均增加，LVEDP、LVMI以及CVF-V、CVF-NV的表达显著降低，姜黄素高剂量组尤为明显；不同时间段，假手术组LVEDP、LVMI、CVF-V、CVF-NV的前后表达相近，而手术对照组、卡托普利组和姜黄素各剂量组前后差别显著。结论 姜黄素具有抑制胶原网络重构的作用，可在一定程度改善心功能，延缓CHF的进程。     

姜黄素抑制人胃癌BGC-823细胞生长并诱导其凋亡的机制研究

目的:探讨姜黄素抑制人胃癌BGC-823细胞生长并促人胃癌BGC-823细胞凋亡的生物学作用及其调控机制。方法:常规体外培养对数生长期胃癌BGC-823细胞,细胞分为对照组,低、中、高姜黄素处理组四组,姜黄素浓度分别为0mg/L5,mg/L1,0mg/L,20mg/L。姜黄素处理24h后,采用甲基噻唑(MTT)比色法及流式细胞仪测定细胞增殖水平及细胞凋亡率;采用免疫组化法测定细胞内Bax、Bcl-2蛋白表达;采用PCR检测Caspase-3的mRNA表达水平。结果:MTT检测显示姜黄素能抑制人胃癌BGC-823细胞増殖,呈现浓度依赖性;流式细胞仪显示姜黄素能有效诱导细胞的凋亡,呈现浓度依赖性,其中20mg/L姜黄素处理24h后细胞凋亡率为48.3%;免疫组化试验表明姜黄素处理使人胃癌BGC-823细胞中Bax表达水平上调,同时Bcl-2蛋白表达水平下调;且细胞中Caspase-3的mRNA表达水平受姜黄素诱导而增高。结论:姜黄素对人胃癌BGC-823细胞的增殖具有明显抑制作用,呈浓度依赖性促进细胞凋亡,这种生物学效应可能与激活Bax蛋白表达、抑制Bcl-2蛋白表达而活化Caspase-3的信号通路有关。该研究为深入探讨姜黄素诱导人胃癌BGC-823细胞凋亡的机制提供了重要依据。