姜黄素对胃癌细胞株凋亡抑制蛋白家族及Smac基因表达的影响

目的　观察姜黄素(curcumin) 对人胃癌细胞株凋亡抑制蛋白(IAP) 家族和Smac基因表达的影响, 探讨姜黄素诱导胃癌细胞凋亡的分子机制。方法　10~40μmol/L 姜黄素分别处理胃癌细胞株MKN -45 6~24 h后, MTT比色法检测癌细胞生长活性, 末端TdT酶标记技术和DNA Ladder检测细胞凋亡; 逆转录聚合酶链反应(RT PCR)和Western blot法检测IAP家族(Survivin、XIAP) 和Smac基因表达; 比色法检测癌细胞Caspase 3活性改变。结果　同对照组比较, 不同浓度姜黄素作用后癌细胞生长明显减慢, 抑制率为12. 18%~68. 15% (P<0. 01), 部分细胞呈现典型凋亡形态学改变, 凝胶电泳可见“梯形”条带, 凋亡率为9 .24%~28. 12% (P<0. 01); Survivin、XIAP基因mRNA和蛋白水平显著下调(均P<0. 01), 而Smac基因表达增强(P<0. 01), 癌细胞Caspase- 3 活性增强2 27~6 67倍(P<0 .01)。结论　姜黄素可显著诱导胃癌MKN- 45 细胞凋亡, 通过上调Smac、下调Survivin和XIAP基因表达, 进而活化Caspase- 3是其作用机制之一。     

姜黄素对人肺癌细胞凋亡及相关信号的影响

目的研究姜黄素对人肺癌细胞内凋亡相关信号的影响。方法取对数生长期SPC-A1细胞,用20μmol/L姜黄素作用SPC-A1细胞0、1、2、3、4h,用荧光探测的办法测定细胞内Ca2 浓度。用原位杂交的方法检测人肺癌细胞caspase8mRNA表达。结果20μmol/L姜黄素处理SPC-A1细胞后,细胞内Ca2 浓度随作用时间增高而升高;20μmol/L姜黄素作用于癌细胞24h后,使人肺癌细胞caspase8mRNA表达明显增高。结论姜黄素诱导SPC-A1细胞的凋亡可能与细胞内Ca2 浓度升高以及caspase8mRNA表达增高有关。     

Effect of curcumin on the gene expression of low density lipoprotein receptors

Elevated level of low density lipoprotein-cho-lesterol(LDL-C)in plasma is one of the majorcauses for atherosclerosis and coronary heart dis-ease.3-hydroxy-3-methylglutaryl-coenzyme A(HMG-CoA)reductaseinhibitors,such as statins,alreadyin used clinically to reduce cholesterol lev-els,are effective in patients with atherosclerosis.They show a good safety profile in patients withhigh cholesterol levels and/or cardiovascular dis-ease,however,statins may be potentially associat-ed with developmen…     

Preparation of Curcumin Prodrugs and Their in Vitro Anti-tumor Activities

The curcumin prodrugs, which could be selectively activated in tumor cells, were prepared to establish a basis for the targeted chemotherapy for cancer. On the basis of the molecular structure of curcumin, the N-maleoyl-L-valine-curcumin （NVC）, N-maleoyl- glycine-curcumin （NGC） were chemically synthesized and identified by IR and NMR spectroscopy. After treatment with these two prodrugs for 6--24 h, the rates of growth inhibition on human bladder cancer EJ cells and renal tubular epithelial （HKC） cells were detected by MTT colorimetry. Our results showed that after the treatment with 20 μmol/L--40 μmol/L NVC and NGC for 6--24 h, the growth inhibitory effects on EJ cells were 6.71%-65.13 % （P〈0.05）, 10. 96 %-73.01 % （p〈0. 05）, respectively, in both dose- and time-dependent manners. When compared with the curcumin of same concentrations, the growth inhibitory effects of these two prodrugs on HKC cells were significantly decreased （P〈0.01）. It is concluded that activation of curcumin prodrugs via hydrolysis functions of cellular esterase could inhibit the growth activities of tumor cells, and reduce the side effects on normal diploid cells. This provided a novel strategy for further exploration of tumor targeted chemotherapeutic drugs.     

莪术中姜黄素在大鼠体内的药动学研究

目的研究莪术中姜黄素在大鼠体内的药动学。方法采用高效液相色谱法测定姜黄素的血药浓度。色谱柱:L ichrospher-5-C18(250 mm×4.6 mm,5μm);柱温:25℃;流动相:乙腈-5%醋酸水(45∶55);体积流量:1 mL/m in;检测波长:420 nm。结果姜黄素在6.5~104μg/mL与峰面积呈良好的线性关系(r=0.999 5)。平均回收率为98.5%,RSD为2.41%(n=5)。采用药动学计算程序处理,姜黄素的药动学参数:ka为0.53/h、ke为0.10/h、t1/2ka为1.32 h、t1/2k为6.89 h、tpeak为3.89 h、Cm ax为93.15 ng/mL、AUC为1 369.38 ng/mL。结论本法稳定、简单、可靠,可用于姜黄素的血药浓度分析及其药动学研究。     

姜黄素对非酒精性脂肪肝小鼠代谢组学的调控研究

目的 探讨姜黄素（CUR）对非酒精性脂肪肝病（NAFLD）的代谢调节作用。方法 小鼠适应性饲养3 d后,随机分为3组：对照（C）组、高脂饮食模型（HFD）组、CUR （50 mg/kg）组。C组饲喂普通饲料,HFD和CUR组饲喂高脂饲料,CUR同时ig给药10周,C组和HFD组ig给予0.5% CMC-Na溶液,给药体积均为10 mL/kg。给药结束后,取血和肝脏,肝组织切片后进行HE染色和油红O染色,光学显微镜观察病理变化;采用基于气相色谱-质谱联用（GC/MS）的代谢组学方法检测血清和肝脏组织样品,数据通过Simca-P13.0及Metaboanalysis网络工具进行分析,找出代谢组变化趋势和相关差异化合物。此外,制备小鼠肝原代细胞,棕榈酸（PA）100 mmol/L诱导细胞NAFLD体外模型,观察CUR （10 μmol/L）对代谢组变化的影响。结果 油红O染色结果显示,HFD小鼠肝脏组织存在大量红染油滴,CUR组小鼠肝脏组织中的脂质油滴明显减少。HE染色分析显示,与C组比较,HFD组小鼠肝脏组织有空泡状,CUR组小鼠肝脏组织与C组比较没有明显区别。对血清样和肝组织样品的GC-MS数据进行PLS-DA分析发现C组与HFD组偏离较远,而CUR组与HFD组相比出现向C组偏移的趋势,进一步S-plot分析发现影响较大的代谢为胆固醇的合成、酮体的生成以及氨基酸代谢,细胞实验得到了类似的结果;经通路MetPA分析和代谢产物富集,血清和细胞中各组间差异化合物代谢主要集中在氨基酸代谢,肝脏中代谢差异主要集中在氨基酸代谢和部分脂质代谢通路;进一步进行差异代谢物的筛选,HFD组生糖氨基酸和生酮氨基酸较对照组均有不同程度的升高,CUR能够降低这些氨基酸的水平。结论 高脂饮食会造成小鼠三大代谢的异常,姜黄素能够一定程度上改善小鼠代谢紊乱。     

Preparation of Curcuma Longa L. Extract Nanoparticles Using Supercritical Solution Expansion

Nanoparticles of Curcuma longa Linn (turmeric) rhizome extract were prepared using supercritical carbon dioxide (SC-CO₂). The SC-CO₂ was used for sample pre-treatment, including lipophilic compounds removal and extraction, as well as particle production. The particle formation process was based on the expansion of supercritical solution of plant extract into a secondary chamber. In the course of the expansion of supercritical solution process, the herbal extract changed from dissolved mode at higher pressures to precipitated mode at lower pressures, as long as the pressures were higher than the critical pressure. The particle growth via coagulation was limited by a large number of unsuccessful collisions between CO₂ molecules and primary nuclei due to the presence of pressurized CO₂ molecules where the particle formation occurs. The presence of curcumin derivatives in nanoparticles was confirmed by liquid chromatography-mass spectrometry results. Irregular to quasi-spherical particles with average diameter of 47 ± 20 nm (n = 300) were prepared at a pre-expansion pressure of 35 MPa, pressure drop of 24 MPa, temperature of 50°C, equilibration time of 30 min, collection time of 60 min, extract volume of 30 μL, and feeding solution concentration of 2 mg mL^?1.     

Curcumin entrapped hyaluronan containing niosomes: preparation,characterisation and in vitro/in vivo evaluation

Curcumin, a natural polyphenolic compound, has numerous pharmacological activities; while it faces several bioavailability problems, due to its poor solubility and stability. So, many nanostructures have been designed to overcome these drawbacks. The aim of this study was to prepare a polymeric niosomal structure by incorporating hyaluronan to improve curcumin efficiencies. Hyaluronan containing niosomes were prepared by thin film hydration medium with slight modifications. In the formulation of hyaluronan containing niosomes size and zeta potential studies, Atomic Force Microscopy (AFM), Transmission Electron Microscopy (TEM), Scanning Electron Microscopy (SEM), Differential Scanning Calorimetry (DSC), X-Ray Diffraction (XRD), in-vitro release test, DPPH antioxidant assay and in-vivo anti-inflammatory test were investigated. The results showed that hyaluronan containing niosomes were 249.83?±?6.38?nm and the entrapment of curcumin was 98.28?±?0.278% (w/w). In addition, the shape of the hyaluronan containing niosomes was spherical. 500?µl of the prepared formulation with 4.002?×?10^?7 moles of curcumin showed 100% antioxidant effect. Moreover, the anti-inflammatory effect of the hyaluronan containing niosomes was higher than the anti-inflammatory effect of the simple suspension of curcumin.     

姜黄素对宫颈癌小鼠的抗肿瘤活性及免疫功能的影响

目的 探讨姜黄素对宫颈癌小鼠的抗肿瘤活性及免疫功能的影响。方法 采用腹腔及前肢腋下接种U14细胞建立宫颈癌小鼠模型,随机分为模型组、姜黄素组和阳性对照组,每组10只。模型组给予0.9% NaCl 0.2 mL,姜黄素组给予100 mg·mL^-1的姜黄素0.2 mL,连续灌胃14 d,阳性对照组给予顺铂3 mg·kg^-1·d^-1,每隔3 d腹腔注射1次,共5次。比较各组移植瘤形态,检测各组肿瘤体积和重量并计算抑瘤率,ELISA法检测血清中白细胞介素-2（IL-2）、肿瘤坏死因子（TNF-α）、干扰素-γ（IFN-γ）、骨桥蛋白（OPN）、癌胚抗原（CEA）和鳞状细胞癌抗原（SCC-Ag）水平,流式细胞仪检测CD3⁺、CD4⁺、CD8⁺细胞及NKT细胞含量。结果 模型组肿瘤细胞核大、深染,核质增加,核分裂多,姜黄素组和阳性对照组肿瘤细胞明显皱缩,核固缩或碎裂,病理性核分裂减少。姜黄素组和阳性对照组肿瘤体积、质量、IL-2、TNF-α、IFN-γ、OPN、CEA、SCC-Ag、CD8⁺水平显著低于模型组,而抑瘤率、CD3⁺、CD4⁺和NKT细胞含量显著高于模型组,差异均有统计学意义（P<0.05）。结论 姜黄素可抑制宫颈癌小鼠的抗肿瘤活性,其机制可能为增强免疫功能,降低炎症因子和肿瘤标志物水平。     

Protective effects of maternal administration of curcumin and hesperidin in the rat offspring following repeated febrile seizure: Role of inflammation and TLR4

Neuroinflammation has a key role in seizure generation and perpetuation in the neonatal period, and toll-like receptor 4 (TLR4) pathway has a prominent role in neuroinflammatory diseases. Administration of antioxidants and targeting TLR4 in the embryonic period may protect rat offspring against the next incidence of febrile seizure and its harmful effects. Curcumin and hesperidin are natural compounds with anti-inflammatory and antioxidant properties and have an inhibitory action on TLR4 receptors. We evaluated the effect of maternal administration of curcumin and hesperidin on infantile febrile seizure and subsequent memory dysfunction in adulthood.Hyperthermia febrile seizure was induced on postnatal days 9–11 on male rat pups with 24 h intervals, in a Plexiglas box that was heated to ~45 °C by a heat lamp. We used enzyme-linked immunosorbent assay, Western blotting, malondialdehyde (MDA), and glutathione (GSH) assessment for evaluation of inflammatory cytokine levels, TLR4 protein expression, and oxidative responses in the hippocampal tissues. For assessing working memory and long-term potentiation, the double Y-maze test and Schaffer collateral-CA1 in vivo electrophysiological recording were performed, respectivelyOur results showed that curcumin and hesperidin decreased TNF-α, IL-10, and TLR4 protein expression and reversed memory dysfunction. However, they did not provoke a significant effect on GSH content or amplitude and slope of recorded fEPSPs in the hippocampus. In addition, curcumin, but not hesperidin, decreased interleukin-1β (IL-1β) and MDA levels.These findings imply that curcumin and hesperidin induced significant protective effects on febrile seizures, possibly via their anti-inflammatory and antioxidant properties and downregulation of TLR4.