双脱甲氧基姜黄素滴丸的质量研究

目的将双脱甲氧基姜黄素制备成滴丸剂型,对滴丸进行质量评价和双脱甲氧基姜黄素的含量测定。方法采用自制滴丸机制备双脱甲氧基姜黄素滴丸,并对滴丸进行外观检查、重量差异、重复性、溶出度等体外研究。结果所制的双脱甲氧基姜黄素滴丸各项规定均符合要求,体外测定符合药典规定。结论该批滴丸的制备工艺稳定,重现性好,为开发双脱甲氧基姜黄素的新型给药系统提供实验依据。     

HPLC法测定姜黄提取物中有效成分含量

目的：建立高效液相色谱法同时测定姜黄提取物中姜黄素、一脱甲氧基姜黄素、二脱甲氧基姜黄素的含量.方法：采用C18色谱柱（150 mm×6 mm,5μm）,流动相为乙腈-水-冰乙酸（39∶57∶4）,流速为1.4 ml· min-1;检测波长：428 nm;柱温：35℃;进样量：20μl.结果：同时测定了姜黄素、一脱甲氧基姜黄素、二脱甲氧基姜黄素的含量,三者的线性范围分别为81.31～650.50 μg ·ml-1,26.93 ～215.42μg· ml-1,16.32～130.54 μg ·ml-1.平均回收率分别为100.9％,100.9％,100.6％,RSD分别为1.7％,1.2％,1.5％（n=9）.结论：该方法简便、准确、重复性好,可用于姜黄提取物中有效成分的测定.     

高效液相色谱法同时测定协日嘎四味汤胶囊中姜黄素、脱甲氧基姜黄素、二脱甲氧基姜黄素的含量

目的建立同时测定协日嘎四味汤胶囊中姜黄素、脱甲氧基姜黄素、二脱甲氧基姜黄素含量的方法。方法采用高效液相色谱法。应用Agilent ZORBAX SB-C18(4.6 mm×150 mm,5μm)色谱柱;以乙腈-0.4%冰醋酸溶液(48∶52)为流动相;流速为0.8 mL.min-1;柱温为30℃;检测波长为430 nm。结果姜黄素、脱甲氧基姜黄素、二脱甲氧基姜黄素基线分离,线性范围分别为0.508~4.572、0.516~4.644、0.253~2.277μg.mL-1;平均回收率依次为99.5%、99.0%、98.8%,RSD分别为1.5%、1.7%、2.3%。结论本试验方法简便、快捷,结果准确、重复性好,可用于该制剂中姜黄素类成分的含量测定。     

HPLC法测定通络胶囊中姜黄素、脱甲氧基姜黄素、双脱甲氧基姜黄素的含量

目的 建立通络胶囊中多种有效成分的含量测定方法。方法 采用HPLC法测定制剂中姜黄素、脱甲氧基姜黄索、双脱甲氧基姜黄素等有效成分的含量。结果 姜黄素在0．02048-0．2048μg范围内线性良好，加样回收率为98．89％，RSD为1．04％；脱甲氧基姜黄素在0．00968-0．0968μg范围内线性良好，加样回收率为97．86％，RSD为0．51％；双脱甲氧基姜黄素在0．00992-0．0992μg范围内线性良好，加样回收率为98．66％，RSD为1．82％。结论 本测定方法简便可行、重复性好，可用于本制剂的质量控制。     

HPLC法测定中药莪术中3种姜黄素的含量

目的：用高效液相色谱法测定中药莪术中3种姜黄素成分即姜黄素(curcumin, I)、脱甲氧基姜黄素(demethoxycurcumin, II)和双脱甲氧基姜黄素(bisdemethoxycurcumin, III)。方法：采用Inertsil ODS-3色谱柱,流动相为甲醇—异丙醇—水—冰乙酸(20∶27∶48∶5),流速为0.5 ml.min^-1,检测波长为420 nm。结果：3种姜黄素在0.1～1.9 μg呈良好线性关系,方法回收率为98.2%～101.1%,曾用本法测定了8种莪术样品。结论：本法准确,快速,重现性好,可用于考察研究中药莪术的不同品种以及同属其它中药的质量。     

双脱甲氧基姜黄素的提取、纯化及鉴定

目的 由中药姜黄中提取双脱甲氧基姜黄素。方法 采用柱层析法由姜黄醇提物中分离双脱甲氧基姜黄素，然后采用重结晶法对其进行纯化；分别采用薄层层析法和高效液相色谱法(HPIC)对分离产物进行纯度鉴定，最后采用质谱法测定其分子量。结果 (1)薄层层析法检测结果表明，经柱层析法获取的双脱甲氧基姜黄素中无其他色素成分；但HPLC法的检测结果则表明，分离产物中的杂质峰总面积约占8．2％。采用结晶法纯化后，其中的杂质峰总面积降至3．4％。(2)质谱法鉴定结果表明，分离产物的分子量为308，与双脱甲氧基姜黄素的分子量完全一致，证明所获取的产物即双脱甲氧基姜黄素。结论 柱层析法与结晶纯化法相结合是分离、纯化双脱甲氧基姜黄素的有效方法。     

双脱甲氧基姜黄素脂质体对体外培养人肺腺癌A549细胞增殖活性的影响研究

目的:考察双脱甲氧基姜黄素脂质体对体外培养人肺腺癌A549细胞增殖活性的影响。方法:将双脱甲氧基姜黄素脂质体作用于体外培养人肺腺癌A549细胞,分别应用形态学观察、MTT检测、流式细胞仪检测观察其生长形态、细胞周期等生物学特性变化。结果:显微镜下可见A549细胞形态明显改变;双脱甲氧基姜黄素脂质体对A549细胞增殖有抑制作用,其IC50为12.108μg/ml;流式细胞仪显示细胞周期阻滞在S期。结论:双脱甲氧基姜黄素脂质体可抑制人肺腺癌A549细胞的增殖,在肺腺癌治疗中有一定应用前景。     

脱甲氧基姜黄素纳米脂质载体的制备及处方工艺优化

摘 要 目的：优选脱甲氧基姜黄素纳米脂质载体（CurⅡ-NLC）的最佳处方工艺。方法: 以包封率为主要考察指标,采用单因素考察和正交试验法研究制备工艺中影响纳米粒质量的主要因素,筛选出较理想的处方和工艺。结果: 优选的最佳处方工艺为：药脂比为1∶40,液态脂质占总脂质的比例为10%,卵磷脂的用量为2%,表面活性剂用量为4%。优化工艺所制备的纳米粒为球形或类球形实体,粒径在110nm左右,PDI为0.199,粒径分布均匀。结论:由正交试验法优选处方工艺稳定可行,为脱甲氧基姜黄素进一步研究提供参考依据。     

去甲氧基姜黄素Mannich碱衍生物的合成及体外抗肿瘤活性研究

目的设计并合成去甲氧基姜黄素的曼尼希碱衍生物,评价其体外抗肿瘤增殖活性。方法以乙酰丙酮为起始原料,与香草醛及对羟基苯甲醛经两次Adol缩合制得去甲氧基姜黄素,乙酰化保护后再经亲核取代反应引入双苄基,然后经脱保护及Mannich反应合成目标化合物。采用MTT法测定了目标化合物对人宫颈癌细胞、人乳腺癌细胞、人肝癌细胞、人纤维肉瘤细胞及人慢性髓原白血病细胞等5种细胞的抗增殖活性。结果与结论合成了14个未见文献报道的化合物,其中11个为去甲氧基姜黄素的Mannich碱类化合物,结构经1H-NMR和HR-MS确证;初步药理结果显示活性大多高于去甲氧基姜黄素。     

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     

姜黄素对尤文氏肉瘤RD-ES细胞的诱导凋亡研究

目的 探讨姜黄素对体外培养的人尤文氏肉瘤RD-ES细胞的影响. 方法 体外培养人尤文氏肉瘤RD-ES细胞,姜黄素组加入终浓度分别为1,10,100 mg.mL^-1的姜黄素处理RD-ES细胞,对照组不加姜黄素,24～72 h后观察细胞的形态变化,分别用噻唑蓝(MTT)法、原位末端转移酶标记技术(TUNEL)染色、逆转录 聚合酶链反应(RT-PCR)及图像分析技术检测各组细胞生长、凋亡发生、Bcl-2 基因表达水平的变化. 结果1,10,100 mg.mL^-1姜黄素组细胞生长抑制率分别为(11.8±2.0)%,(56.4±5.0)%,(67.6±6.0)%,24 h半数抑制浓度为10.74 mg.mL^-1;1,10,100 mg.mL^-1姜黄素作用于RD-ES细胞48 h,随着浓度的增强,细胞凋亡率增高;Bcl-2表达减少. 结论 姜黄素可诱导RD-ES细胞凋亡,并与Bcl-2有关.     

莪术的质量研究

目的　研究莪术的化学成分及其药理作用,寻找与中医用药功能主治相吻合的有效成分,并比较不同品种及产地的莪术在质量上的差异。方法　用柱色谱的方法分离提取单体化合物,确定结构,并分别进行药效学实验,确定有效成分,通过HPLC比较不同品种中有效成分含量,评价莪术的质量。结果　从中提取分离了7个单体化合物,姜黄素显出较强的药理活性,但在广西莪术中未检出;吉马酮的含量较高,有一定的活性,在3种莪术中均有分布;挥发油作为一个提取部位,显出较强的药理活性。结论　莪术中姜黄素是主要有效成分之一,吉马酮可选作特征成分,挥发油可为有效部位。莪术不同品种所含成分有显著差异,以此3个指标作为莪术质量评价指标。     

姜黄提取工艺研究

目的：优选姜黄提取工艺.方法：采用单因素法,以姜黄挥发油收率为评价指标,优选姜黄挥发油提取工艺条件;采用正交试验法,以姜黄素、干膏率为评价指标,优选姜黄醇提工艺条件.结果：姜黄挥发油提取工艺条件为：粉碎过一号筛,加8倍水,蒸馏提取7 h;姜黄醇提的工艺条件为：加80%乙醇,提取3次,每次加药材8倍量乙醇,提取60 min.结论：优选的姜黄提取工艺条件稳定可行.     

姜黄素对皮层神经元氧化损伤的保护作用

目的　观察姜黄素对第三丁基过氧化氢 (tert butylhy droperoxide,t BHP ,tBHP)诱导的大鼠皮层神经元氧化损伤的影响 ,探讨可能的机制。方法　培养胚胎鼠皮层神经元 ,MTT法测定细胞活力 ,DNA断裂评价细胞凋亡 ,流式细胞术测定线粒体膜电位和细胞内活性氧水平 ,分光光度法测定细胞内谷胱甘肽 (GSH)水平 ,Westernblot法测定Bcl 2和Bax蛋白和胞浆细胞色素C以及活化型半胱氨酸天冬氨酸蛋白酶 3(caspase 3)和多聚 (ADP 核糖 )聚合酶 [poly (ADP ribose)poly merase,PARP]水平。结果　姜黄素 (2 5～ 2 0 μmol·L-1)可有效减少tBHP对神经元的氧化损伤和tBHP引起的细胞内GSH水平降低 ,降低细胞内的活性氧水平 ,增加线粒体膜电位和细胞内GSH以及Bcl 2蛋白水平 ,减少线粒体内细胞色素C向胞浆释放和Bax蛋白表达水平 ,最终明显减少cas pase 3和PARP活化和tBHP引起的神经元凋亡。结论　姜黄素可减弱tBHP对原代皮层神经元的氧化损伤作用 ,其作用可能与降低细胞内的活性氧水平 ,保护线粒体功能有关     

Evaluation in vitro of synthetic curcumins as agents promoting monocytic gene expression related to β-amyloid clearance

Accumulation of amyloid-beta (Aβ) is one of the hallmarks of Alzheimer's disease (AD), and efficient clearance of Aβ by cells of the innate immune system may be an important mechanism for controlling or preventing disease onset. It was reported that peripheral blood mononuclear cells (PBMCs) of most AD patients are defective in the phagocytosis of soluble Aβ. Natural curcumins were shown to restore Aβ phagocytosis by AD PBMCs and to up-regulate the expression of key genes including MGAT3 and those encoding Toll-like receptors (TLRs). Bisdemethoxycurcumin (BDC), a minor component of natural curcumin, was shown to have the greatest potency for stimulating AD PBMCs. Because natural curcumins have inherent limitations with regard to physicochemical properties, synthetic curcumin analogues were developed that showed improved solubility, stability, and bioavailability. An in vitro system using human monocytic cell lines (U-937, THP-1) was used to evaluate analogues for the potency of innate immune cell stimulation. These cell lines showed responses to curcuminoids and to 1α,25-dihydroxyvitamin D3 (VD3) resembling those seen in human PBMCs. From more than 45 curcuminoids analyzed, the most potent compounds possessing enhanced pharmaceutical properties were identified. The most promising candidates included prodrug versions containing water solubility-enhancing amino acids and stability-increasing modifications near the central diketone. In vivo studies showed compound (5) substantially increased bioavailability by combining several promising structural modifications. Studies examining ex vivo phagocytosis of Aβ and bead particles in mouse microglia showed that BDC and several water-soluble analogues were quite effective compared to curcumin or an unnatural analogue. In vitro studies using monocytic cell lines reported herein complement those using human PBMCs and represent a routinely accessible and uniform cellular resource allowing direct comparisons between compounds.     

Inhibitory effect of curcuminoids on acetylcholinesterase activity and attenuation of scopolamine-induced amnesia may explain medicinal use of turmeric in Alzheimer's disease

Curcuminoids (a mixture of curcumin, bisdemethoxycurcumin and demethoxycurcumin) share vital pharmacological properties possessed by turmeric, a well known curry spice, considered useful in Alzheimer's disease (AD). The aim of this study was to evaluate if curcuminoids possess acetylcholinesterase (AChE) inhibitory and memory enhancing activities. The in-vitro and ex-vivo models of AChE inhibitory activity were used along with Morris water maze test to study the effect on memory in rats. Curcuminoids inhibited AChE in the in-vitro assay with IC₅₀ value of 19.67, bisdemethoxycurcumin 16.84, demethoxycurcumin 33.14 and curcumin 67.69 µM. In the ex-vivo AChE assay, curcuminoids and its individual components except curcumin showed dose-dependent (3-10 mg/kg) inhibition in frontal cortex and hippocampus. When studied for their effect on memory at a fixed dose (10 mg/kg), all compounds showed significant (p < 0.001) and comparable effect in scopolamine-induced amnesia. These data indicate that curcuminoids and all individual components except curcumin possess pronounced AChE inhibitory activity. Curcumin was relatively weak in the in-vitro assay and without effect in the ex-vivo AChE model, while equally effective in memory enhancing effect, suggestive of additional mechanism(s) involved. Thus curcuminoids mixture might possess better therapeutic profile than curcumin for its medicinal use in AD.     

Nitric Oxide Scavenging by Curcuminoids

Because curcumin, a compound with anti-inflammatory and anticancer activity, inhibits induction of nitric oxide synthase in activated macrophages and has been shown to be a potent scavenger of free radicals we have investigated whether it can scavenge nitric oxide directly. Curcumin reduced the amount of nitrite formed by the reaction between oxygen and nitric oxide generated from sodium nitroprusside. Other related compounds, e.g. demethoxycurcumin, bisdemethoxycurcumin and diacetylcurcumin were as active as curcumin, indicating that the methoxy and the phenolic groups are not essential for the scavenging activity. The results indicate curcumin to be a scavenger of nitric oxide. Because this compound is implicated in inflammation and cancer, the therapeutic properties of curcumin against these conditions might be at least partly explained by its free-radical scavenging properties, including those toward nitric oxide.     

姜黄素-3脂质体对小鼠四氯化碳急性肝损伤的保护作用及其机制

目的：研究姜黄素-3脂质体对肝损伤的保护作用及机制。方法：取昆明种小鼠40只,随机分成4组,每组10只：正常对照组、四氯化碳（CCl4）肝损伤模型组、姜黄素-3注射剂组、姜黄素-3脂质体组。采用腹腔注射CCl4致小鼠肝损伤模型,测定动物血清ALT、AST的活性和肝组织脂质过氧化产物丙二醛（MDA）的含量;研究肝组织病理变化。结果：模型组血清ALT、AST及肝组织MDA较正常对照组明显增高,肝组织切片出现肝小叶内炎细胞浸润;治疗组小鼠各生化指标及炎症程度较模型组均明显减轻,脂质体组对肝损伤的药效明显优于注射剂组。结论：姜黄素-3脂质体对小鼠四氯化碳性肝损伤有显著的保护作用,可通过降低肝内脂质的过氧化反应而增加其对肝脏内皮细胞的保护作用。     

The hypomethylation agent bisdemethoxycurcumin acts on the WIF-1 promoter, inhibits the canonical Wnt pathway and induces apoptosis in human non-small-cell lung cancer

Hypermethylation of the Wnt inhibitory factor-1 (WIF-1) promoter has been implicated in the overactivation of the Wnt pathway in human lung cancer. Curcuminoids exert anti-cancer effects and have been reported to act as hypomethylating agents. Previously, we have investigated and compared the demethylation effects of three curcuminoids and observed that bisdemethoxycurcumin exhibited the strongest demethylation potency. In this study, we used lung cancer cell lines with WIF-1 promoter hypermethylated as a model to study the demethylating effect of bisdemethoxycurcumin on WIF-1 restoration, Wnt signaling activity and cell death. Bisdemethoxycurcumin directly suppressed the activity of DNA methyltransferase-1 (DNMT1) but did not influence DNMT1 expression. In addition, it induced WIF-1 promoter demethylation and protein re-expression. WIF-1 restoration in lung cancer cells down-regulated nuclear β-catenin and the canonical Wnt cascade. Furthermore, we also showed that down-regulation of Wnt signaling by WIF-1 was required for bisdemethoxycurcumin-induced apoptosis in certain lung cancer cell types. This report is the first to show that bisdemethoxycurcumin induces apoptosis by reactivating WIF-1 from a silenced state. Our results provide new insights into the anti-cancer actions of bisdemethoxycurcumin.