Inhibitory effects of Curcuma aromatica oil on proliferation of hepatoma in mice

AIM To reveal the inhibitory effects ofCurcuma aromatica oil(CAO)on cellproliferation of hepatoma in mice.METHODS Two tumor inhibitory experimentsof CAO on hepatoma in mice were conducted.The inhibitory effects of CAO on proliferation ofhepatoma in mice were evaluated by DNA imagecytometry and immunohistochemical staining ofproliferating cell nuclear antigen(PCNA).RESULTS The tumor inhibitory rates of CAOwere 52% and 51% in two experiments,respectively.Compared with those of the saline-treated control groups,both differences werestatistically significant(P【0.01).In the groupof mice treated with CAO,the cellular nuclearDNA OD value(249±70),areas(623 μm^2±228 μm^2)and DNA(2.38±0.67)index of hepaticcarcinomas were significantly lower than thoseof the control group(430±160,1073 μm^2±101 μm^2 and 4.48±0.71).CAO also couldincrease diploidy cell rates(29.00%±9.34% vs2.97%±5.69%,P【0.01)and decreasepentaploidy cell exceeding rate(30.04%±15.10% vs 70.89%±14.94%,P【0.01).In thegroup of mice treated with CAO,the labelingindexes of proliferating cell nuclear antigen (PCNA-LI)were 30%±4%,which weresignificantly lower than 40%±6% of the controlgroup(P【0.01).CONCLUSION The inhibition of CAO on thegrowth of hepatoma in mice might be associatedwith its depression on cellular proliferativeactivity.

Antimicrobial activity and essential oils of Curcuma aeruginosa,Curcuma mangga,and Zingiber cassumunar from Malaysia

Objective:To analyze the chemical composition of the essential oils of Curcuma aeruginosa (C.aeruginosa),Curcuma mangga(C.mangga),and Zingiber cassumunar(Z.cassumunar). and study their antimicrobial activity.Methods:Essential oils obtained by steam distillation were analyzed by gas chromatography-mass speclrometry(GC-MS).The antimicrobial activity of the essential oils was evaluated against four bacteria:Bacillus cereus(H.cereus).Staphylococcus aureus(S.aureus).Escherichia coli(E.coli).and Pseudomonas aeruginosa(P.aeruginosa);and two fungi:Candida albicans(C.albicans) and Cyptococcus neoformans(C.neoformans),using disc-diffusion and broth microdilution methods.Results:Cycloisolongifolene,8.9-dehydro-9- formyl(35.29%) and dihydrocoslunolide(22.51%) were the major compounds in C.aeruginosa oil; whereas caryophyllene oxide(18.71%) and caryophyllene(12.69%) were the major compounds in C.mangga oil:and 2,6.9,9-tetramethyl-2.6.10-cycloundecatrien-1-one(60.77%) andα-caryophyllene(23.92%) were abundant in Z.cassumunar oil.The essential oils displayed varying degrees of antimicrobial activity against all lested microorganisms.C.mangga oil had the highest and most broad-spectrum activity by inhibiting all microorganisms tested,with C.neoformans being the most sensitive microorganism by having the lowest minimum inhibitory concentration(MIC) and minimum fungicidal concentration(MFC) values of 0.1μL/mL. C.aeruginosa oil showed mild antimicrobial activity,whereas Z.cassumunar had very low or weak activity against the tested microorganisms.Conclusions:The preliminary results suggest promising antimicrobial properties of C.mangga and C.aeruginosa,which may be useful for food preservation,pharmaceutical treatment and natural therapies.

Essential oil of Curcuma wenyujin induces apoptosis in human hepatoma cells

AIM: To investigate the effects of the essential oil of Curcuma wenyujin (CWO) on growth inhibition and on the induction of apoptosis in human HepG2 cancer cells. METHODS: The cytotoxic effect of drugs on HepG2 cells was measured by 3-(4,5-dimethylthiazol-2- yl)-2,5-diphenyltetra-zolium bromide (MTT) assay. DNA fragmentation was visualized by agarose gel electrophoresis. Cell cycle and mitochondrial transmembrane potential (△Ψm) were determined by flow cytometry (FCM). Cytochrome C immunostaining was evaluated by fluorescence microscopy. Caspase-3 enzymatic activity was assayed by the cleavage of Ac-DEVD-R110. Cleaved PARP and active caspase-3 protein levels were measured by FCM using BD? CBA Human Apoptosis Kit. RESULTS: Treatment with CWO inhibited the growth of HepG2 cells in a dose-dependent manner, and the IC50 of CWO was approximately 70 μg/mL. CWO was found to inhibit the growth of HepG2 cells by inducing a cell cycle arrest at S/G2. DNA fragmentation was evidentlyobserved at 70 μg/mL after 72 h of treatment. During the process, cytosolic HepG2 cytochrome C staining showed a markedly stronger green fluorescence than in control cells in a dose-dependent fashion, and CWO also caused mitochondrial transmembrane depolarization. Furthermore, the results clearly demonstrated that both, activity of caspase-3 enzyme and protein levels of cleaved PARP, significantly increased in a dose- dependent manner after treatment with CWO. CONCLUSION: CWO exhibits an antiproliferative effect in HepG2 cells by inducing apoptosis. This growth inhibition is associated with cell cycle arrest, cytochrome C translocation, caspase 3 activation, Poly- ADP-ribose polymerase (PARP) degradation, and loss of mitochondrial membrane potential. This process involves a mitochondria-caspase dependent apoptosis pathway. As apoptosis is an important anti-cancer therapeutic target, these results suggest a potential of CWO as a chemotherapeutic agent.

Chemical profiling and antimicrobial activity of essential oil from Curcuma aeruginosa Roxb.,Curcuma glans K.Larsen&J.Mood and Curcuma cf.xanthorrhiza Roxb.collected in Thailand

Objective: To investigate chemical constituents and new antimicrobial agents among essential oils from the rhizomes of Curcuma aeruginosa(C. aeruginosa) Roxb., Curcuma glans K. Larsen & J. Mood and Curcuma cf. xanthorrhiza Roxb.Methods: The essential oils were obtained by hydro-distillation and analyzed by gas chromatography/mass spectroscopy. Agar-well diffusion assay was used to study the antimicrobial activity and also broth-micro dilution techniques were examined for minimum inhibitory concentration(MIC) against four bacterial strains and yeast.Results: The gas chromatography/mass spectroscopy analysis showed monoterpenes predominantly(88.53%) in the rhizome oil of Curcuma cf. xanthorrhiza. Sesquiterpenes(50.10%) was the most abundant component in the essential oil of C. glans, while monoterpenes(45.55%) and sesquiterpenes(45.81%) were found in C. aeruginosa with a significant amount. The major components of C. aeruginosa were characterized as camphor(29.39%) and germacrone(21.21%). Germacrone(15.76%), b-pinene(9.97%)and camphor(9.96%) were found as major compounds in the rhizome oils of C. glans while a-terpinolene(24.86%) and p-cymen-7-ol(12.17%) were found as major compositions in Curcuma cf. xanthorrhiza. The essential oils were tested against four bacterial strains and yeast. As a result, the rhizome oil of C. aeruginosa exhibited potent activity against Staphylococcus aureus [inhibition zone(21.94 ± 0.24) mm, MIC 125 μg/mL],Bacillus cereus [inhibition zone(20.83 ± 0.36) mm, MIC 125 μg/mL], and Candida albicans [inhibition zone(11.60 ± 0.30) mm, MIC 250 μg/mL].Conclusions: The essential oils from three Curcuma species possessed greater activity against the gram-positive bacteria(Staphylococcus aureus and Bacillus cereus) than gram-negative bacteria(Escherichia coli and Pseudomonas aeruginosa). The results suggest that the essential oils from the fresh rhizome of Curcuma spp. might be a potential source of natural antimicrobial substances.

广西莪术和蓬莪术中药超分子“印迹模板”特征研究

目的基于中药超分子“气析”理论结合化学计量学对广西莪术和蓬莪术挥发油成分“印迹模板”的特征进行分析和研究。方法采用水蒸气蒸馏法提取广西莪术和蓬莪术挥发油,并用气质联用法建立其指纹图谱。应用总量统计矩法对2种莪术“印迹模板”的印迹性进行对比;计算各批莪术挥发油的物芯指数(CI),结合化学计量学对2种莪术“印迹模板”的拓扑结构特征进行对比。结果广西莪术和蓬莪术的挥发油提取率无明显差异;两者的总量零阶矩分别为(1.907±0.177)×10^(8)、(1.979±0.413)×10^(8)μV·s,表明两者挥发油成分总含量无明显差异;总量一阶矩分别为(30.969±0.962)、(33.198±0.409)min,总量二阶矩分别为(56.176±11.368)、(43.891±4.113)min^(2),表明两者挥发油成分含量配比及种类有显著差异;2种莪术总量统计矩相似度大多低于界定值,表明两者挥发油的整体化学成分与其构成比存在差异性;主成分分析和正交偏最小二乘-判别分析可明显将广西莪术和蓬莪术分为两类;通过P值与变量重要性投影值分析,确定广西莪术和蓬莪术的Xvp四阶、Xvpc五阶、Xvpc六阶、Xvpc七阶、Xvc三阶、Xvpc四阶6个阶数的CI值为主要差异性数值。结论通过对中药超分子“印迹模板”进行“印迹性”及“拓扑特征”的表征并结合化学计量学分析可成功区分广西莪术和蓬莪术,并找到2种莪术“印迹模板”差异性的CI值。

莪术挥发油化学成分及其药理作用研究

莪术挥发油为中药莪术发挥药用价值的主要活性部位,总结有关莪术挥发油化学成分的文献报道,发现目前从莪术挥发油中共鉴定出成分130余种,包括萜类(单萜、倍半萜)、酮(醛)类、醇(酚)类、烷(烯)烃类、酸(酯)类、醚类、芳香族等化合物,其中萜类为其主要化合物成分。莪术挥发油具有抗肿瘤、抗炎抑菌、抗病毒、抗血栓、保肝、保护神经、抗氧化等药理作用。目前,莪术挥发油所开发出的莪术油注射液、复方莪术油栓等广泛应用于临床。文章对莪术挥发油的提取工艺、化学成分及药理作用进行文献综述,以期为莪术挥发油在临床用药、制剂等方面提供参考。

不同干燥方式对姜黄中姜黄素和姜黄油的影响

以新鲜姜黄根茎为原料,采用高效液相色谱法、水蒸气蒸馏法及气相色谱质谱联用(GC-MS)等方法研究了晒干、阴干、30℃热风烘干、60℃热风烘干和冷冻干燥5种干燥方式对姜黄素的相对含量和姜黄油组分组成的影响。结果表明,不同干燥方式下,姜黄素的相对含量和姜黄油的含量及组分的变化较大,但姜黄的色度变化较小,其中自然阴干方式下姜黄素和姜黄挥发油含量相对较高,分别为1.84%、6.15%。研究结果为姜黄的开发利用和产品加工提供一定的技术支持。

离线全二维逆流色谱-液相色谱分离莪术油成分

中药挥发油成分复杂,一维色谱分离由于有限的峰容量难以完全分离中药挥发油成分,全二维气相色谱为分离挥发油成分提供了有力的方法,然而气相色谱一般无法用于天然活性成分的筛选。为建立挥发油成分全二维色谱分析新方法,研究建立以液相色谱为基础的全二维色谱分离分析方法。本文主要研究全二维逆流色谱-液相色谱分离莪术油成分的方法,并探讨两种色谱技术之间的正交性,为活性成分筛选提供新的技术支持。通过优化离线全二维逆流色谱-液相色谱分离方法,对全二维色谱峰容量、正交性和空间覆盖率进行度量。优化液相色谱分析条件并筛选逆流色谱分离两相溶剂体系,通过比色法筛选了逆流色谱两相溶剂体系并采用下相为流动相进行梯度洗脱。在290~375 min采用推挤洗脱,莪术油在第一维逆流色谱分离中达到了良好的分离。第二维反相高效液相色谱的流动相组成为乙腈(A)和水(B)。梯度洗脱程序为0~10 min,50%A~65%A;10~14 min,65%A;14~21 min,65%A~85%A;21~25 min,85%A~95%A;25~30 min,95%A~55%A;30~40 min,55%A。在上述条件下进行全二维色谱分析,结果表明该全二维分离体系总峰容量达954个,理论峰容量达到一维色谱的10倍以上,同时具有良好的正交性(线性相关性r=0.17)和空间覆盖率(68.1%)。以上结果表明,离线全二维逆流色谱-液相色谱分离莪术油具有良好的正交性,能显著提高复杂样品的分离效果,为挥发油物质基础研究和活性成分筛选以及二维色谱指纹图谱的建立提供了新的方法与思路。

莪术不同部位挥发油化学成分GC-MS分析

目的通过莪术不同部位,包括根茎、块根以及须根在内的挥发油化学成分GC-MS进行分析,对不同部位挥发性化学成分差异进行评估。方法应用水蒸气蒸馏法对不同部位莪术挥发油进行提取,采取气相色谱-质谱联用仪(GC-MS)对莪术挥发油中有效化学成分进行分析,并经面积归一化法对其化合物所占百分比进行检测。结果莪术样本的根茎、块根、须根部位挥发油获得率分别为0.6%,0.4%以及4.1%,且根茎以及块根颜色分别为为黄色至紫色,而须根挥发油表现为紫色。另选择莪术根茎、块根及须根部位内挥发油在种类以及含量上存在差异性,并鉴定出80种化合物,其中根茎获得化合物共61个,占总量76.25%,从块根中鉴定64种化合物,占挥发油总量80.00%,须根而中仅仅鉴定出化合物共48种化合物,占挥发油总量60.00%。结论表明莪术根茎、块根、须根不同部位的挥发油种类以及所占比均存在差异,并证实根茎部分也存在相当大的开发和应用价值,可为莪术不同部位开发提供参考依据。

油菜素内酯对姜黄次生代谢产物合成的影响

【目的】姜黄是我国的传统中药材,被国家列入药食同源目录。通过喷施不同质量浓度的油菜素内酯(BR),分析外源BR对姜黄代谢产物的影响,为姜黄栽培提供理论依据。【方法】以姜黄盆栽苗为试验材料,采用完全随机分组试验设计,叶面分别喷施0.01、0.05、0.1和0.2 mg/L BR后,检测姜黄根状茎的氨基酸、总黄酮、姜黄素类化合物和挥发油的含量。【结果】喷施0.1 mg/L BR后,姜黄总黄酮、姜黄素、去甲氧基姜黄素和双去甲氧基姜黄素含量显著提高,分别比对照提高128.99%、73.05%、99.56%和97.42%;0.01和0.05 mg/L BR显著提高姜黄挥发油含量;亮氨酸与天冬氨酸含量在0.05 mg/L BR时达到最大值。【结论】喷施0.05~0.1 mg/L外源BR能够促进姜黄次生代谢产物的生物合成,进而提高姜黄的药材品质。

复方莪术油微囊中莪术醇的气相色谱测定

目的 :建立莪术油、干蟾皮水溶性提取物微囊制剂和莪术油原料的定量分析方法。方法 :用气相色谱法使微囊制剂和原料油中挥发性有效成分达到良好分离 ,并对主要抗肿瘤成分莪术醇进行定性定量分析。结果 :组分与内标分离良好 ,莪术醇在 0 .0 5～ 1.6 g/ L 范围内线性关系良好 ,γ=0 .9999,高、低二个浓度的日内与日间精密度以及 3个浓度加样回收率的相对标准误差均 <2 %。结论 :本法定量准确可靠 ,重现性好 。

GC-MS法建立温莪术挥发油指纹图谱研究

目的 :建立温莪术挥发油的指纹图谱测定分析方法 ,为温莪术药材的质量控制提供参考 ,同时也为温莪术中药制剂制定指纹图谱奠定基础。方法 :采用水蒸气蒸馏法提取温莪术挥发油 ,用GC MS联用技术对其进行指纹图谱测定 ,并采用国家药品监督管理局推荐的中药指纹图谱计算软件进行计算 ,建立温莪术挥发油的共有指纹图谱。结果与结论 :建立的GC MS研究温莪术挥发油指纹图谱的方法有较好的重现性、精密度和稳定性 (RSD均小于 5 % ) ,再加上GC MS具有多成分同时定性的优势 ,所建立的温莪术挥发油的GC MS指纹图谱为其质量控制打下基础。

莪术油自乳化制剂处方研究

目的 :寻找莪术油自乳化最佳处方条件。方法 :通过溶解度实验、正交筛选和相图绘制 ,以形成乳剂的乳化程度和乳滴粒径大小为指标 ,对莪术油处方中的油相、非离子表面活性剂、助溶剂进行筛选 ,寻找最佳处方。结果 :莪术油的自乳化的处方中油相为油酸乙酯 ,非离子表面活性剂为吐温 - 85 ,其最佳比例范围为 4∶ 6～ 7∶ 3。 结论 :自乳化莪术油处方比例为莪术油∶油酸乙酯∶吐温 - 85为 6∶ 5∶

莪术油注射液对慢性低氧大鼠学习与记忆的影响

本文旨在探讨莪术油注射液对慢性低氧大鼠学习与记忆的影响及其可能机制。将Sprague-Dawley大鼠随机分为对照组、慢性低氧组、5mg/kg体重莪术油组、10mg/kg体重莪术油组、20mg/kg体重莪术油组,每组14只。慢性低氧处理采用低氧舱内吸入大约10%O2、5%CO2,饲养10h/d,持续饲养28d。莪术油组大鼠低氧处理前腹腔注射相应浓度的莪术油注射液。实验结束次日,通过Morris水迷宫测试各组动物学习和记忆成绩的变化;测定各组大鼠血清和海马组织丙二醛(malonaldehyde,MDA)含量和超氧化物歧化酶(superoxide dismutase,SOD)活性以及海马组织Ca2+浓度([Ca2+i]);通过免疫组织化学和Western blot检测磷酸化Ca2+/钙调蛋白依赖性蛋白激酶II(phosphorylated Ca2+/calmodulin-dependent pro-tein kinase II,p-CaMKII)在海马组织和胞膜上的表达。结果显示,与对照组相比,慢性低氧组大鼠隐蔽平台的逃避潜伏期明显延长(P<0.05),血清和海马组织MDA含量明显增高,SOD活性显著降低(P<0.05,P<0.01),海马组织[Ca2+]i明显增高(P<0.01),海马p-CaMKII表达量显著降低(P<0.01)。与慢性低氧组比较,莪术油各组发生以下变化:10、20mg/kg体重莪术油组大鼠隐蔽平台的逃避潜伏期显著缩短(P<0.05);5、10、20mg/kg体重莪术油组大鼠血清和海马组织MDA含量均显著降低(P<0.05,P<0.01);10、20mg/kg体重莪术油组大鼠血清SOD活性显著增加(P<0.01);5、10、20mg/kg体重莪术油组大鼠海马组织[Ca2+]i明显降低(P<0.01);10、20mg/kg体重莪术油组大鼠海马p-CaMKII表达也明显增加(P<0.05,P<0.01)。以上结果提示,莪术油改善慢性低氧导致的大鼠学习与记忆能力下降,其机制可能与降低血清和海马组织MDA含量、增加SOD活性、降低海马组织[Ca2+]i和/或增加海马组织p-CaMKII表达有关。

广西几个姜黄品种姜黄油GC-MS分析

通过对广西地区栽培的三个姜黄品种:博白姜黄、大新姜黄、市场姜黄的姜黄油GC-MS分析,比较不同姜黄品种姜黄油成分的差异性,为筛选特定姜黄品种提供依据。不同姜黄品种其主要成分含量差异显著,主成份姜黄酮(含芳香-姜黄酮)含量依次为博白姜黄56.07%、大新姜黄47.79%、市场姜黄28.38%,市场姜黄的姜黄酮含量只有博白姜黄的一半。因此从姜黄油主要成份(含量>20%)姜黄酮含量来考察,以种植博白姜黄为最好。

微胶囊化姜黄油对冷藏鲢鱼鱼丸的保鲜效果

以鲢鱼鱼丸为研究对象,将姜黄油及微胶囊化姜黄油添加到鱼丸中,测定分析其在冷藏过程中微生物、理化和感官等品质指标的变化,评价微胶囊化姜黄油对鲢鱼鱼丸的保鲜效果。结果表明:4℃下空白组鱼丸在贮藏5 d后菌落总数(TVC)达到6.2×104 cfu/g,超过国家规定的卫生标准;而添加0.1%微胶囊化姜黄油的鱼丸样品细菌生长受到明显抑制,可使鱼丸货架期延长8 d(TVC值为5.9×104 cfu/g)。微胶囊化姜黄油还可以有效地减缓脂肪氧化,提高了鱼丸的白度,而对弹性影响不大。该生物保鲜剂在鱼糜制品保鲜中具有明显效果,使产品在较长时间内保持良好的食用品质。

莪术油对小鼠肝癌细胞抑制作用的分子机理

目的 :研究莪术油抑制小鼠HepA肝癌细胞的分子机制。方法 :建立小鼠HepA肝癌模型 ,进行莪术油抑癌试验 ,并用免疫组化法检测细胞增殖核抗原 (PCNA)、细胞周期素D1(cyclinD1)的表达 ,探讨莪术油抑制小鼠肝癌的分子机理。结果 :莪术油治疗组的瘤重、PCNA标记指数、cyclinD1标记指数均低于生理盐水组 (P <0 .0 1)。结论 :莪术油可能是通过下调cyclinD1的表达 ,抑制PCNA的阳性表达率 ,进而抑制肿瘤的生长。

莪术油乳膏对大鼠乳腺增生的作用

目的观察外用莪术油乳膏对乳腺增生模型大鼠的影响,并探讨其作用机制。方法将60只雌性Wistar大鼠随机分为正常对照组、模型对照组、散结乳癖贴膏组和莪术油乳膏大、中、小剂量组,每组10只。采用大鼠后腿内侧肌内注射苯甲酸雌二醇联合黄体酮法复制乳腺增生模型,给予不同剂量药物干预,连续4周。酶联免疫吸附法、苏木精-伊红(HE)染色法和免疫组化法探讨外用莪术油对乳腺增生的作用机制。结果莪术油乳膏对乳腺增生有一定防治作用。与模型对照组比较,莪术油大剂量组大鼠乳房直径显著缩小(P<0.01),体质量恢复正常,血清雌二醇(P<0.01)、促卵泡激素(P<0.05)、泌乳素(P<0.01)均明显降低,孕酮、睾酮、促黄体生成素含量升高(P<0.01),小叶内腺泡数量及直径显著减少。结论莪术油乳膏能显著改善大鼠血清激素水平紊乱,抑制乳腺增生发生。

莪术挥发油提取及包合工艺的研究

以莪术挥发油提取率为评价指标 ,利用正交实验L9(34)筛选莪术挥发油提取最佳工艺条件为 :莪术粗粉浸泡 1h ,加 8倍水煎煮 ,提油时间 8h。按优选的最佳工艺条件验证三批 ,测得挥发油平均提取率 2 38% ,相对标准偏差 (RSD) 1 2 3% (n =3)。分别考察了饱和水溶液法 β CD包合莪术挥发油时超声和搅拌对包合的影响 ,测得两种方法包合率分别为 98 77%和 86 98% ,RSD分别为 2 4 2 %和 2 6 5 % (n =3) ,表明用超声法包合挥发油比搅拌法所得包合率高。

莪术挥发油的提取工艺和主要成分测定

用水蒸气蒸馏法提取莪术挥发油 ,通过正交试验设计法优选获取莪术挥发油的技术参数 ,结果表明 ,以药材粗粉为原料 ,按m (药材 )∶m (水 ) =1∶8加水 ,蒸馏 8h ,药材出油得率平均可达2 .5 1% ,该工艺稳定、可靠 .莪术挥发油的GC