不同来源莪术中姜黄素类成分的含量差异研究

目的：用高效液相色谱法测定不同来源的莪术中3种姜黄素成分,比较姜黄素（cur_cumin,Ⅰ）、去甲氧基姜黄素（demethoxycurcumin,Ⅱ）和双去甲氧基姜黄素（bisdemethoxycurcum_in,Ⅲ）的含量差异。方法：采用Kromasil色谱柱,流动相为乙腈-水（0.5%冰乙酸）,流速为1 mL· min-1,检测波长为425 nm。结果：姜黄素、去甲氧基姜黄素、双去甲氧基姜黄素的线性范围分别为0.04~0.16μg （r=0.9991）、0.037~0.150μg （r=0.9990）、0.00015~0.00062μg （r=0.9996）;平均回收率（n=9）分别为103.32%、100.32%、100.06%。结论：不同来源的莪术中姜黄素类成分的含量存在着明显的差异。     

中国姜黄属植物根茎中姜黄素类化合物的含量测定

在硅胶G板上,以正丁醉-26%氨水-无水乙醇(30:3:1)为展开剂,用薄层扫描法分离测定了中国姜黄属植物毛郁金Curcuma aromatica Salisb.,温郁金C. wenyujin Y. H. Chen et C.Ling(or C. aromatica cv. Wenyujin),莪术C. aeruginosa Roxb.,黄莪术C. zedoaria (Christm.)Rosc.,桂莪术C. kwangsiensis S. G. Lee et C. F. Liang,川郁金C. chuanyujin C. K. Hsich et H.Zhang,姜黄C. longa L.根茎中姜黄素(curcumin),去甲氧基姜黄素(demethoxycurcumin),双去甲氧基姜黄素(bisdemethoxycurcumin)和总姜黄素(curcuminoids)的含量,回收率97.07%～102.0%(n=6),变异系数为3.1%～5.0%。     

脱甲氧基姜黄素对血管生成影响的实验研究

目的：从脱甲氧基姜黄素对内皮细胞增殖和迁移的影响角度，探讨脱甲氧基姜黄素抗血管生成的作用机制。方法：用MTT法检测脱甲氧基姜黄素对牛主动脉内皮细胞及人胃腺癌细胞系SGC-7901细胞增殖的影响；用琼脂糖刮除法检测脱甲氧基姜黄素对牛内皮细胞迁移的影响。结果：脱甲氧基姜黄素能明显抑制牛内皮细胞增殖，对非内皮细胞与肿瘤细胞也有抑制作用，但与前者相比，差异有统计学意义。脱甲氧基姜黄素能显著抑制牛内皮细胞迁移，在实验浓度范围内，其抑制作用呈明显量效关系。结论：脱甲氧基姜黄素是一种特异性血管生成抑制剂。抑制内皮细胞增殖和迁移可能是脱甲氧基姜黄素抑制血管生成的机制之一。     

姜黄素-3单体脂质体的研制及评价

目的制备包封率高、粒径均匀、稳定性好的姜黄素-3(双脱甲氧基姜黄素,demethoxycurcumin)单体脂质体。方法通过均匀设计筛选工艺,以乙醇注入法等制备姜黄素-3脂质体。结果制备姜黄素-3脂质体平均粒径为130.5nm,分布均匀,平均包封率为87.89%,稳定性好。结论以脂质体技术包裹姜黄素-3单体形成脂质体制剂可行,为进一步研究在肿瘤治疗中的应用提供了基础。     

Microarray-based Analysis of Anti-angiogenic Activity of Demethoxycurcumin on Human Umbilical Vein Endothelial Cells: Crucial Involvement of the Down-regulation of Matrix Metalloproteinase

cDNA microarray-based gene expression analysis has been successfully employed to explore the action mechanism and to validate the targets of several drugs. In the present study, we evaluated anti-angiogenic activity of demethoxycurcumin (DC), a structural analog of curcumin, isolated from Curcuma aromatica , and investigated the effect of DC on genetic reprogramming in cultured human umbilical vein endothelial cells (HUVECs) using cDNA microarray analysis. Of 1024 human cancer-focused genes arrayed, 187 genes were up-regulated and 72 genes were down-regulated at least 2-fold by DC. Interestingly, 9 angiogenesis-related genes were down-regulated over 5-fold in response to DC, suggesting that the genetic reprogramming was crucially involved in anti-angiogenesis by the compound. To verify the results obtained from cDNA microarray analysis, matrix metalloproteinase-9 (MMP-9), the product of one of the angiogenesis-related genes down-regulated over 5-fold by DC, was investigated using gelatin zymography. DC potently inhibited the expression of MMP-9, yet showed no direct effect on its activity. These data show that gene expressional change of MMP-9 is a major mediator for angiogenesis inhibition by DC. All genes identified and microarray data are available on the web at http://dasan.sejong.ac.kr/bioprobe/ .     

天然姜黄素类化合物的合成

目的以化学合成法制备天然姜黄素类化合物。方法以对羟基苯甲醛或香草醛和乙酰丙酮为原料经硼络合保护,分别合成姜黄素、一脱甲氧基姜黄素和二脱甲氧基姜黄素,测定熔点,并以氢谱进行结构解析。结果姜黄素、一脱甲氧基姜黄素和二脱甲氧基姜黄素合成品分别为亮黄色、橙黄色和橙红色粉末,氢谱所确证的结构与已知结构一致。结论本合成法操作简单,条件温和,可在短时间内制备大量天然姜黄素类化合物,效率较传统的提取分离法大大提高。     

Comparison of inhibitory potency of three different curcuminoid pigments on nitric oxide and tumor necrosis factor production of rat primary microglia induced by lipopolysaccharide

Microglia are the resident innate immune cells in the central nervous system. Evidence supports that the unregulated activation of microglia results in the production of pro-inflammatory cytokines and chemokines that propagate neuronal injury and finally cause neurodegenerative diseases. Curcuminn (Cur), demethoxycurcumin (DMC), and bisdemethoxycurcumin (BDMC) are curcuminoid pigments extracted from turmeric (Curcuma longa L.). Cur has been reported to suppress the activation of microglia by reducing toxic factors production, but little is known about whether the two natural demethoxy derivatives of Cur, DMC and BDMC, have the similar effects as Cur. In the present study, we found that all of the three curcuminoid pigments significantly suppressed nitric oxide (NO) production by LPS-activated microglia and the relative potency was DMC > BDMC > Cur. This result was verified by RT-PCR analysis of iNOS mRNA. The NO-scavenging abilities of three curcuminoid pigments are very weak, which suggested that the indirect effect may not be critical in inhibiting NO production by LPS-activated microglia. Moreover, these three curcuminoid pigments attenuated the expression of mRNA and proteins of tumor necrosis factor-alpha (TNF-α) in a concentration-dependent manner and the relative potency was also DMC > BDMC > Cur. In conclusion, Cur, DMC and BDMC were found as potent microglia-activation inhibitors, and DMC exhibited the strongest inhibitory activity on NO and TNF-α production. These results provided an interesting clue for designing new compounds which could have better potential therapeutic implications for various neurodegenerative diseases.     

Curcumin and its demethoxy derivatives possess p300 HAT inhibitory activity and suppress hypertrophic responses in cardiomyocytes

The natural compound, curcumin (CUR), possesses several pharmacological properties, including p300-specific histone acetyltransferase (HAT) inhibitory activity. In our previous study, we demonstrated that CUR could prevent the development of cardiac hypertrophy by inhibiting p300-HAT activity. Other major curcuminoids isolated from Curcuma longa including demethoxycurcumin (DMC) and bisdemethoxycurcumin (BDMC) are structural analogs of CUR. In present study, we first confirmed the effect of these three curcuminoid analogs on p300-HAT activity and cardiomyocyte hypertrophy.Our results showed that DMC and BDMC inhibited p300-HAT activity and cardiomyocyte hypertrophy to almost the same extent as CUR. As the three compounds have structural differences in methoxy groups at the 3-position of their phenol rings, our results suggest that these methoxy groups are not involved in the inhibitory effects on p300-HAT activity and cardiac hypertrophy. These findings provide useful insights into the structure–activity relationship and biological activity of curcuminoids for p300-HAT activity and cardiomyocyte hypertrophy.     

Metabolites Identification of Curcumin, Demethoxycurcumin and Bisdemethoxycurcumin in Rats After Oral Administration of Nanoparticle Formulations by Liquid Chromatography Coupled with Mass Spectrometry

Background: Curcuminoids are promising cancer chemopreventive agents. Curcumin, demethoxycurcumin(DMC) and bisdemethoxycurcumin(BDMC) are the major bioactive curcuminoids in turmeric. However, comprehensive metabolic studies of these three curcuminoids are still limited.Objective: To identify the metabolites of curcumin, DMC and BDMC in rats after oral administration of solid lipid nanoparticles(SLNs).Methods: Male Sprague-Dawley rats(250 ± 20 g, body weight) were randomly divided into 4 groups(n=3), and were orally administered with curcumin-SLN, DMC-SLN, BDMC-SLN, or blank-SLN, respectively. Plasma samples(500 μL) via the angular vein were collected at 1, 2 and 4 h post dosing, and the urine and feces samples were collected at 0–12 h and 12–24 h post-intake. An HPLC-DAD-ESI-MSnmethod was developed to identify the metabolites. The structures of phase II metabolites were further confirmed by enzyme hydrolysis.Results: A total of 34 metabolites were identified in rats plasma, urine, and feces. Most of them were phase II metabolites, including glucuronide conjugates and sulfate conjugates. Among them, the glucuronide conjugates were the major metabolites in rats plasma. In the meanwhile, the three parent curcuminoids were detected in high amounts in the urine and feces samples.Conclusion: The possible metabolic pathways of curcuminoids in rats were proposed.     

姜黄及其制剂中姜黄素类化合物的高效液相色谱分离与测定

姜黄为姜料植物Curcuma longa L.的根茎,为一常用中药。近年来用于治疗高脂血症有较好疗效,且无毒副作用。其主要化学成分为姜黄素、去甲氧基姜黄素及二去甲氧基姜黄素。实验证实姜黄素为其降血脂的主要有效成分。姜黄素类化合物过去多采用在420 nm附近测定黄色素的方法,此类方法不够稳定,易受其它黄色素的干扰。利用姜黄素与硼生成红色络合物而进行测定,比较专一而灵敏。对