Kaemtakols A-D,highly oxidized pimarane diterpenoids with potent anti-inflammatory activity from Kaempferia takensis

Four highly oxidized pimarane diterpenoids were isolated from Kaempferia takensis rhizomes.Kaemtakols A-C pos-sess a tetracyclic ring with either a fused tetrahydropyran or tetrahydrofuran motif.Kaemtakol D has an unusual rearranged A/B ring spiro-bridged pimarane framework with a C-10 spirocyclic junction and an adjacent 1-methyltri-cyclo[3.2.1.02,7]octene ring.Structural characterization was achieved using spectroscopic analysis,DP4+and ECD calculations,as well as X-ray crystallography,and their putative biosynthetic pathways have been proposed.Kaemtakol B showed significant potency in inhibiting nitric oxide production with an IC50 value of 0.69μM.Molecular docking provided some perspectives on the action of kaemtakol B on iNOS protein.

Improved dissolution of Kaempferia parviflora extract for oral administration by preparing solid dispersion via solvent evaporation

Kaempferia parviflora, a plant in the family Zingiberaceae, has been used in Thai traditional medicines for treating hypertension and promoting longevity with good health and wellbeing. However, its limited aqueous solubility and low dissolution restrict its bioavailability.The aim of the study was therefore to improve the dissolution rate of K. parviflora extracted with dichloromethane(KPD) by solid dispersions. Different water-soluble polymers were applied to improve dissolution of KPD. The solid dispersions in different ratios were prepared by solvent evaporation method. Only hydroxypropyl methylcellulose(HPMC) and polyvinyl alcohol-polyethylene glycol grafted copolymer(PVA-co-PEG) could be used to produce homogeneous, powdered solid dispersions. Physical characterization by scanning electron microscopy, hot stage microscopy, differential scanning calorimetry and powder X-ray diffractometry, in comparison with corresponding physical mixtures, showed the changes in solid state during the formation of solid dispersions. Dissolution of a selected marker,5,7,4′-trimethoxyflavone(TMF), from KPD/HPMC and KPD/PVA-co-PEG solid dispersions was significantly improved, compared with pure KPD. The dissolution enhancement by solid dispersion was influenced by both type and content of polymers. The stability of KPD/HPMC and KPD/PVA-co-PEG solid dispersions was also good after 6-month storage in both longterm and accelerated conditions. These results identified that the KPD/HPMC and KPD/PVAco-PEG solid dispersions were an effective new approach for pharmaceutical application of K. parviflora.

Establishment and Optimization Growth of Shoot Buds-Derived Callus and Suspension Cell Cultures of <i>Kaempferia parviflora</i>

Callus and suspension cells culture of Kaempferia parviflora was successfully established. Meristematic shoots can be used for utilization of plant cell biosynthetic capabilities for obtaining useful products from valuable medicinal plant to meet out the pharmaceutical demand and also for studying the metabolism. The medium containing combination of 0.2 mg/L 2,4-dichlorophenoxyacetic acid (2,4-D) and 0.2 mg/L napthyleneacetic acid (NAA) promoted the highest callus induction at 20%. Transferring the initiated callus on the medium with 1 mg/L 2,4-D enhanced the proliferation rate up to maximum fresh weight of 6.71 gm. Growth curve of cultured cells revealed that the cells continued to grow until 50 days of culture and showed the highest peak (fresh weight) at 40 days in all different initial weight tested ( 0.2, 0.5 and 1.0 gram). Isolated embryogenic callus was found to produce the highest in weight when suspended in liquid medium supplemented with 1 mg/L 2,4-D at 110 rpm resulted 13.5 gram fresh weight and 1080 mg dry weight.

Optimization of Extraction Conditions for Total Phenolics and Total Flavonoids from <i>Kaempferia parviflora</i>Rhizomes

Kaempferia parviflora plants derived from in vitro culture were grown in the glasshouse. A comparison of the yield of total phenolics and total flavonoids under varying extraction conditions from rhizomes harvested from plants of different ages was undertaken. The results showed that phenolic and flavonoid contents in the rhizomes were highest 8 months after planting. Another study found that 2 g rhizomes extracted in 50 ml of water at 90°C for 120 minutes gave the best yield of phenolics and flavonoids. Under these conditions, an average of 210 mg GAE/g dry weight of total phenolics and 81 μg QCE/g dry weight of total flavonoids were obtained.

Micropropagation of Kaempferia angustifolia Roscoe via Direct Regeneration

This study has established an efficient and reproducible protocol to micropropagation Kaempferia angustifolia Roscoe via direct regeneration. The use of young shoots as explants showed the best results compared to rhizome shoots, where the young shoots showed a low percentage of contamination of 10% - 30% (agar 6 g/L) and 45% - 55% (agar 3 g/L), respectively, compared to the use of rhizome shoots, where the contamination rate exceeded 80%. For shoot initiation, the combination of BAP (6 Benzylaminopurine) and NAA (1-Naphthaleneacetic acid) showed higher results for the percentage of initial shoots and number of micro shoots/explants compared to BAP with Kin (Kinetin). The highest concentration of BAP (5 mg/L) combined with the lowest concentration of NAA (0.5 mg/L) resulted in 90% of initial shoots and a number of shoots/explants of 5.8. The highest number of shoots for micropropagation was in treatment with 30 g/L sucrose that was segmented with 3 mg/L BAP + 0.5 mg/L NAA. For the number of roots, the highest number of roots was 11.8 recorded at sucrose (45) with only BAP (1 mg/L) used as the plant growth regulator, while the longest length of roots was 7 - 8 cm, recorded both at sucrose with the combination of BAP and NAA.

Kaempferia parviflora ethanol extract improves self-assessed sexual health in men:a pilot study

Background： Sexual health positively correlates with overall wellbeing. Existing therapeutics to enhance male sexual health are limited by factors that include responsiveness, adherence and adverse effects. As the population ages, safe and effective interventions that preserve male sexual function are needed. Published research suggests that various preparations of Kaempferia parviflora, a plant in the Zingiberaceae （ginger） family, support cardiovascular health and may ameliorate erectile function. Objective： The aim of this study was to examine the effects of KaempMaxTM, an ethanol extract of the K. parvi]lora rhizome, on erectile function in healthy middle-aged and older men. Design, setting, participants and interventions： We conducted an open-label, one-arm study on 14 generally healthy males aged 50-68 years with self-reported mild erectile dysfunction, who were not using pre- scription treatments. Participants took 100 mg KaempMaxTM daily for 30 days. Main outcome measures： Evaluations were conducted at baseline and on the final study assessment. Primary efficacy analyses included the International Index of Erectile Function （IIEF）; secondary efficacy analyses included the Global Assessment Question about erectile function. Results： Thirteen participants completed the 30-day study. Supplementation with KaempMaxTM resulted in statistically significant improvements in erectile function, intercourse satisfaction and total scores on the IIEF questionnaire. KaempMaxTM was well tolerated and exhibited an excellent safety profile. Conclusion： Our results suggest that KaempMaxTM may improve erectile function in healthy middle-aged and older men. While the effects were not as pronounced as what might be seen with prescription med- ication, most participants found them satisfactory. Additional, longer and placebo-controlled clinical tri- als will be needed. Trial registration： Clinicaltrials.gov identifier Ncr03389867.

In vitro cytotoxic and toxicological activities of ethanolic extract of Kaempferia galanga Linn. and its active component,ethyl-p-methoxycinnamate, against cholangiocarcinoma

Objective:To evaluate the cytotoxic,apoptotic,mutagenic and immunomodulatory activities of Kaempferia galanga Linn.(KG)extract and ethyl-p-methoxycinnamate(EPMC)in vitro.Methods:The present study investigated the cytotoxic[using the 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphe nyl-2-H-tetrazolium bromide test],apoptotic(using a mitochondrial membrane potential assay),mutagenic(using a micronucleus test)and immunomodulatory(using flow cytometry)activities of the ethanolic extract of KG and its bioactive component,EPMC,against two cholangiocarcinoma(CCA)cell lines,CL-6 and HuCCT1,and one normal human cell line,OUMS-36 T-1 F.Results:Both KG extract and EPMC exhibited moderate cytotoxic activity against both CCA cells.The cytotoxic activity was supported by their concentration-dependent induction of apoptosis.CL-6 was most sensitive(3–4 fold)and selective to 5-fluorouracil(5-FU),compared with KG extract and EPMC[median half inhibiting concentration(IC50)and selectivity index(SI)were 23.01 lg/mL and 17.32;78.41 lg/mL and 4.44;100.76 lg/mL and 2.20,respectively for 5-FU vs.KG extract vs.EPMC].HuCCT1 was relatively more sensitive and selective to 5-FU and EPMC than KG extract[median IC50 and SI were 66.03 lg/mL and6.04;60.90 lg/mL and 3.65;156.60 lg/mL and 2.23,respectively for 5-FU vs.EPMC vs.KG extract].EPMC produced relatively potent cytotoxic activity against polymorphonuclear cells(IC50=92.20 lg/mL).KG extract and EPMC exhibited concentration-dependent mutagenic activity,as well as inhibition of tumor necrosis factor-a and interleukin-6.Conclusion:Considering cytotoxic,apoptotic,immunomodulatory and mutagenic activities,further development of KG as a drug candidate is likely to focus on the oral pharmaceutical formulation of a standardized KG extract rather than isolated compounds.

不同遮阴处理对小花山奈生长的影响

为探究小花山奈的光照需求特性,为其在华南地区的栽培提供理论依据。在小花山奈块茎繁殖期间进行遮阴栽培试验,测量不同遮阴处理下小花山奈地下部分、地上部分的鲜重和干重,并得到月生长量、含水量和产量等数据。结果表明:在小花山奈生长初期(7~8月),不同遮阴处理对小花山奈地上、地下部分的生长无显著影响;在小花山奈生长中期(9~10月),30%的遮阴处理有利于促进小花山奈地上、地下部分的生长;在小花山奈生长后期(11月~次年1月),30%的遮阴处理有利于促进小花山奈地上部分的生长,全光照有利于促进小花山奈地下部分的生长。小花山奈的地上部分迅速增长期集中在8~11月,地下部分迅速增长期集中在10~12月;适度的遮阴(30%遮阴度)处理有利于提高小花山奈的产量,但遮阴度太高(50%遮阴度)反而会大大降低小花山奈的产量。因此,在夏季给予30%遮阴处理、秋冬季给予全光照是小花山奈在华南地区种植的最佳光照模式。

小花山柰根状茎挥发油和营养成分及其抗植物病原菌活性分析

【目的】阐明小花山柰根状茎挥发油的化学组成和相对含量,明确根状茎中多种营养成分含量,分析根状茎不同提取物的抗菌活性,从而为小花山柰资源的综合开发利用提供理论依据。【方法】采用水蒸汽蒸馏法提取小花山柰挥发油,通过气相色谱—质谱联用仪(GC-MS)对挥发油进行化学成分分析;采用凯氏定氮法、酸—苯酚比色法和电感耦合等离子体—质谱法(ICP-MS)半定量分析法分别测定小花山柰根状茎中蛋白质、粗多糖和微量元素含量,采用茚三酮柱后衍生离子交换色谱法测定氨基酸组成与含量;进一步采用抑菌圈法和菌丝生长速率法测定小花山柰根状茎不同提取物的抗细菌和抗真菌活性。【结果】小花山柰根状茎中挥发油得率为0.14‰;从根状茎挥发油中共鉴定出66种成分,占挥发性成分总量的94.51%,主要有烯烃类、醇类、醛类、酮类和酯类,主要成分为冰片(13.02%)、芳樟醇(12.27%)和大根香叶烯D(5.42%)等。小花山柰根状茎中蛋白质含量为3.83 g/100 g,粗多糖含量为3.33 g/100 g;16种氨基酸总量为2.94 g/100 g,其中精氨酸含量最高(0.61 g/100 g),其次是天冬氨酸(0.46 g/100 g)。小花山柰根状茎中共检测出28种微量元素,其中钾元素含量最高(3270 mg/kg),镁(813 mg/kg)、钙(289 mg/kg)、锰(146 mg/kg)和铝(135 mg/kg)的含量也较高,其他微量元素的含量均在80 mg/kg以下。小花山柰根状茎乙酸乙酯层提取物的抗细菌活性最强,石油醚层提取物对杜英生假隐丛赤壳菌的抑制活性最强,半最大效应浓度(EC50)为14.11μg/mL。【结论】小花山柰根状茎中含有种类丰富的挥发油,以及蛋白质、氨基酸和微量元素等多种营养成分;根状茎乙酸乙酯层提取物具有较强的抗细菌活性,石油醚层提取物对杜英生假隐丛赤壳菌具有明显的抑制作用,小花山柰作为林下新资源植物具有重要的开发和应用价值。

小花山柰高效快繁技术研究

通过研究不同激素配比对小花山柰不定芽的诱导、不定芽增殖及生根的影响,筛选出小花山柰组培快繁最佳的激素配比,建立小花山柰芽基部的高效组培快繁体系。结果表明:小花山柰最佳的不定芽诱导培养基为MS+30g/L蔗糖+7g/L琼脂+2mg/L 6-BA+0.1mg/L NAA;最佳的不定芽增殖培养基为MS+30g/L蔗糖+8g/L琼脂+2mg/L 6-BA+0.5mg/L KT+0.1mg/L NAA;最佳的生根培养基为MS+30g/L蔗糖+7g/L琼脂+0.2mg/L NAA;炼苗移栽15d后成活率达100%,缩短了小花山柰繁殖时间并提升了繁殖系数,降低了小花山柰的育苗成本,为小花山柰组培苗工业化生产提供了技术参考。

中药材山柰的X衍射Fourier图分析

每种中药材中均含有数十种乃至上百种成分，是一个多组分系统。Ｘ衍射将提供一种既能反映中药材整体固有结构特征，又能表现来自其局部成分变化的图谱化与数值化的方法。它将在衍射空间中实现中药材的鉴定。考虑到中药材的品种、产地、生长年限、采集季节等物候因素的影响，本研究选择了不同产地和采集时间的植物药山柰与苦山柰的６个样品进行Ｘ衍射Ｆｏｕｒｉｅｒ图分析。获得了可用于山柰与苦山柰鉴别的特征标记峰与衍射模糊图形。研究结果表明，Ｘ衍射Ｆｏｕｒｉｅｒ图分析法尚可用于性状与组织结构相似。

GC/MS法分析广东阳春沙姜精油的化学成分

沙姜精油是沙姜中的主要呈味成分。本文选用广东阳春所产鲜沙姜用酶解法配合水汽蒸镏提取其中精油,并对此精油进行气相色谱—质谱分析,以确定广东阳春沙姜精油的成分及含量。实验表明;沙姜精油中含30种主要成分,其中反式—对甲氧基肉桂酸乙酯的含量最高(59.24%),其它肉桂酸系列物质如顺式—对甲氧基肉桂酸乙酯(6.14%)、肉桂酸乙酯(5.27%)的含量也较高。这些数据为进一步开发利用沙姜资源提供了可靠的依据。

超临界CO_2流体萃取法和水蒸汽蒸馏法提取山柰挥发油化学成分的研究

采用超临界CO2流体萃取法(SFE)与水蒸气蒸馏法(SD)从山柰中提取挥发油.用气相色谱-质谱联用技术对化学成分进行分析,用归一化法测定其相对百分含量.超临界CO2流体萃取法提取的挥发油共鉴定了39种成分,占挥发油总成分的86%以上;水蒸气蒸馏法提取的挥发油共鉴定了51种成分,占挥发油总成分的89%以上.研究表明超临界CO2流体萃取法提取的挥发油能更真实、全面地反映药材中的化学成分.

山奈提取物抗氧化性能研究

分别利用DPPH.法和碘量法测试山奈(Kaempferia galanga L.)提取物的抗氧化活性。结果表明,山奈的乙酸乙酯提取物(EAK)和正丁醇提取物(BK)具有良好的抗氧化活性。在碘量法测试中,0.04%的EAK抗氧化作用最强,且超过0.02%的BHT;在对DPPH自由基清除试验中,BK的效果最好,清除DPPH自由基的EC50值为1.13 mg/mL,是BHT的61%。

海南三七(姜科)的食源性欺骗传粉

有花植物为繁殖成功,进化出各种各样的花部特征来吸引传粉者,如为传粉者提供花蜜、花粉、栖息地等,然而在33科146属的被子植物中也存在着不提供任何报酬而欺骗昆虫为其传粉的现象。这种欺骗性传粉模式主要出现在高度进化的具有多样化传粉模式的兰科植物中。报道了在姜科植物中首次发现的食源性欺骗传粉模式。对姜科山柰属海南三七进行连续2年的传粉生物学观察和研究发现,海南三七的花在早上5:30～6:00之间开放,下午17:00～18:00左右闭合萎蔫,持续大约11～12h。开花过程中花粉活性与柱头可受性均保持较高水平(>90%)。花粉/胚珠比率(P/O)为82.20±47.89(n=20)。木蜂是其主要的访花和传粉昆虫,访花目的是吸取花蜜。海南三七虽有细长线形的蜜腺,但并不分泌花蜜作为传粉昆虫访花的报酬,采用食源性欺骗的方式欺骗木蜂为其传粉。繁育系统的研究表明广西弄化的海南三七居群主要是通过根茎进行无性繁殖。

山萘杀线虫活性成分的分离及鉴定

采用超声波提取法,用甲醇作溶剂对山萘进行粗提,分别测定了山萘提取物对松材线虫和南方根结线虫的毒性,结果显示,1 000 mg/L山萘甲醇粗提取物处理松材线虫和南方根结线虫,24 h校正死亡率均达到100%。采用活性追踪和柱层析法,以5种植物病原线虫为对象,提取、分离、纯化出山萘根中的活性成分,结果显示,在正己烷萃取组分中,得到活性较高的化合物A、B;用化合物A处理南方根结线虫、松材线虫、马铃薯腐烂茎线虫、海南根结线虫、象耳豆根结线虫72 h后,其LC50分别为17.79、29.70、43.21、57.64和36.94 mg/L;用化合物B处理72 h后LC50分别为1.49、2.81、10.09、26.67和14.47 mg/L。化合物A和B经1H-NMR1、3C-NMR、GC-MS分析鉴定表明,A为肉桂酸乙酯,B为对甲氧基肉桂酸乙酯。

索氏法和水蒸汽蒸馏法提取山柰(鲜品)挥发油化学成分的比较研究

采用以石油醚为溶剂的索氏提取法及水蒸汽从山柰(鲜品)的根茎中提取挥发油,用气相-质谱联联用技术对化学成分进行分析,以峰面积归一化法测定其相对含量,比较了不同的加工办法对挥发油的主要化学成分和相对含量的影响。研究表明:这两种途径获得的挥发油中其主要成分都为反式对甲氧基肉桂酸乙酯、反式肉桂酸乙酯、十五烷、顺式对甲氧基肉桂酸乙酯、蓝桉醇等,但化学成份的种类及相对含量随加工方法的不同而有较大差异。

沙姜中肉桂酸乙酯类化合物抗促癌作用的研究

采用EB病毒早期抗原 (EBV EA)诱导抑制实验、小鼠耳肿胀实验、表皮组织鸟氨酸脱羧酶活性 (ODC)实验及小鼠皮肤乳头状瘤诱导实验等方法对沙姜中肉桂酸乙酯类化合物进行抗促癌活性研究。结果显示 ,沙姜中的顺式 和反式 对甲氧基肉桂酸乙酯具有抗促癌活性 ,其IC5 0分别为 5 5和 9 5 μmol L。经进一步经典的小鼠皮肤乳头状瘤两阶段致癌实验、ODC实验及小鼠耳肿胀实验对该化合物的验证 ,该成分可有效地抑制由DMBA TPA诱导的小鼠皮肤乳头状瘤的生成 ,使荷瘤鼠比例及鼠荷瘤数降低 ,并可抑制小鼠皮肤ODC活性和小鼠的耳肿胀程度。

沙姜挥发油化学成分的GC/MS分析

采用超临界流体萃取法对沙姜挥发油的化学成分进行了分离提取研究,利用气相-质谱联机技术,对其中的28种组分进行了鉴定,并测定了相对含量.结果表明,质量分数在0.5%以上的组分有:对甲氧基桂酸乙酯(67.88%),桂皮酸乙酯(4.36%)、十七烷(5.11%)、1,8桉油素(4.83%)、乙酯(4.36%)、十五烷(2.16%)、δ蒈烯(1.37%)、二甲基苏合香烯(1.34%)、龙脑(1.31%)、樟烯(0.57%)、α蒎烯(0.56%)、β蒎烯(0.55%)等,占挥发油总量的94.33%.

对甲氧基肉桂酸乙酯的制备及其抑菌活性测定

采用水蒸气蒸馏法提取沙姜挥发油,然后以溶剂结晶法制得对甲氧基肉桂酸乙酯。通过气-质联用色谱法对其进行了结构鉴定,通过体外抑菌实验,利用纸片法和试管法分别测定了其对金黄色葡萄球菌、大肠杆菌、绿脓杆菌、白色念珠菌和黑曲霉菌的抑菌活性和最低抑菌浓度(M IC)。结果表明,对甲氧基肉桂酸乙酯对这5种常见皮肤致病菌均有良好的抑菌活性,对绿脓杆菌、大肠杆菌、金黄色葡萄球菌、黑曲霉菌和白色念珠菌的最低抑菌浓度分别为0.625,1.25,2.5,2.5和10.0 mg/mL。