半枝莲提取物对人肺癌A549细胞的凋亡作用及其对凋亡相关基因表达的影响

目的:了解半枝莲提取物对人肺癌A549细胞的凋亡作用及其对凋亡相关基因表达的影响,探讨半枝莲提取物的抗肺癌作用及其分子生物学作用机制。方法:采用70%乙醇水溶液制备半枝莲提取物;实验组的半枝莲提取物浓度为3mg/L、6mg/L、12mg/L,对照组不加药物。采用MTT法测定药物对细胞生长的抑制率;采用流式细胞仪检测细胞凋亡率;采用RT-PCR法测定细胞凋亡相关基因Survivin、Caspase-3和内对照β-actin的表达水平。结果:3mg/L、6mg/L、12mg/L浓度实验组的OD值均低于对照组;实验组的抑制率分别为(23.53±5.84)%、(39.71±6.77)%和(66.18±8.45)%。对照组3mg/L、6mg/L、12mg/L浓度实验组的细胞凋亡率分别是(3.76±0.82)%、(16.61±3.19)%、(21.34±4.52)%、(35.19±5.80)%;3mg/L、6mg/L、12mg/L浓度实验组的细胞凋亡率均高于对照组。3mg/L、6mg/L、12mg/L浓度实验组的Survivin/β-actin均低于对照组;3mg/L、6mg/L、12mg/L浓度实验组的Caspase-3/β-actin均高于对照组。结论:半枝莲提取物可通过诱导细胞凋亡作用抑制人肺癌A549细胞的生长,其分子机制为下调Survivin的表达水平和上调Caspase-3的表达水平。     

草苁蓉苯丙素苷对肺癌细胞周期分布和凋亡的影响

目的:探讨草苁蓉苯丙素苷(PGBR)对肺癌细胞增殖的抑制作用及其可能机制。方法:采用MTT法观察PGBR对A549肺癌细胞增殖的影响,采用流式细胞术检测对A549细胞周期分布和细胞凋亡的影响,采用TUNEL染色法检测对A549细胞凋亡的影响,免疫细胞化学法检测凋亡相关蛋白p53,Fas,Bax,Bc1-2蛋白的表达。结果:PGBR可时间和浓度依赖性地抑制肺癌细胞增殖,200 mg.L-1 PGBR作用于A549细胞12,24,48 h时,抑制率分别为(20.4±2.1)%,(27.1±2.0)%,(30.7±2.2)%。PGBR改变肺癌细胞周期分布,使G0/G1期细胞由(45.3±4.1)%增至(57.4±4.0)%,表现出G0/G1期阻滞;同时诱导肺癌细胞凋亡,凋亡率由(2.4±0.90)%增至(7.8±1.2)%。PGBR明显增加p53和Fas表达,增加Bax表达和降低Bc1-2表达。结论:PGBR可通过诱导细胞周期阻滞和细胞凋亡而发挥抗肺癌作用。     

半枝莲提取物诱导人肺癌SPC-A-1细胞凋亡及对凋亡相关基因表达的影响

目的:探讨半枝莲提取物(Scutellaria barbataextract,SBE)对人肺癌SPC-A-1细胞凋亡及凋亡相关基因survivin和caspase-3表达的影响。方法:体外培养肺癌SPC-A-1细胞,以2.5、5、10 mg/L SBE处理SPC-A-1细胞48h,并以2.5 mg/L顺铂(DDP)作为阳性对照。用MTT法检测细胞抑制率,TUNEL法检测凋亡率,半定量RT-PCR检测survivin和caspase-3 mRNA表达。结果:与正常对照组相比,SBE各组细胞抑制率显著上升(P<0.001),细胞凋亡率明显升高(P<0.01),Caspase-3 mRNA表达显著上升(P<0.05或P<0.01),Survivin mRNA表达明显下降(P<0.05或P<0.01)。结论:SBE具有诱导肺癌SPC-A-1细胞凋亡的作用,其分子机制可能与上调caspase-3及下调survivin基因表达有关。     

蛴螬提取物诱导人肺癌A549细胞凋亡的机制研究

目的:观察蛴螬提取物对人肺癌A549细胞增殖的影响及其机制。方法:采用MTT法检测蛴螬提取物对人肺癌A549细胞的生长抑制率;用HE染色方法观测凋亡细胞的形态学变化;运用免疫组织化学SP法检测用药前后细胞周期蛋白D1(CyclinD1)的表达改变。结果:用蛴螬提取物处理的人肺癌A549细胞生长抑制率明显高于对照组;施药早期细胞出现凋亡形态学变化;A549细胞经蛴螬提取物处理后CyclinD1表达均下降。结论:蛴螬提取物在体外对人肺癌A549细胞株有显著的增殖抑制作用,其机制与下调CyclinD1的表达有关。     

透骨草提取物对人肺癌A549细胞凋亡及其PUMA/Bcl-2/Caspase-3蛋白表达的影响

目的:观察透骨草提取物对人肺癌A549细胞PUMA(P53上调凋亡调空因子)、Bcl-2(B淋巴细胞瘤-2)、Caspase-3(天冬氨酸特异性半胱氨酸蛋白酶-3)蛋白表达的影响,从凋亡诱导的角度探讨透骨草提取物抑制A549细胞增殖的作用机制。方法:将A549细胞分为实验组和对照组;吖啶橙染色观察透骨草提取物对A549细胞形态的影响;免疫组化法检测透骨草提取物对A549细胞PUMA、Bcl-2、Caspase-3蛋白表达的影响。结果:吖啶橙染色结果显示,39.7 mg/L透骨草提取物处理的A549细胞皱缩,胞核固缩、呈边集,出现凋亡小体。免疫组化检测结果显示,39.7 mg/L透骨草提取物处理的A549细胞PUMA蛋白显著高于对照组(P0.05),而Bcl-2蛋白和Caspase-3蛋白表达显著低于对照组(P0.05)。结论:透骨草提取物对A549细胞具有凋亡诱导作用,机制与促进A549细胞PUMA蛋白表达、抑制Bcl-2和Caspase-3蛋白表达有关。     

中药单体汉黄芩素诱导肺癌细胞凋亡及其机制初步研究

目的 考察汉黄芩素对肺癌细胞株A549的增殖、凋亡的影响,并初步探讨其分子机制.方法 MTT法检测汉黄芩素对A549增殖的影响;Hochest染色、流式细胞仪等检测对细胞凋亡的影响;Western-blotting检测汉黄芩素对凋亡相关蛋白的影响.结果 汉黄芩素能抑制A549细胞的生长,其细胞增殖抑制率与药物浓度及作用时间呈依赖关系.形态学上观察汉黄芩素明显诱导肺癌细胞A549凋亡,同时明显抑制了凋亡相关蛋白BCL-2、survivin蛋白的表达,提升Bax蛋白表达.结论 汉黄芩素可以明显诱导肺癌细胞A549的凋亡,其诱导凋亡分子机制可能与下调BCL-2、survivin蛋白表达有关.     

南蛇藤提取物对高表达maspin的人胃癌MGC-803细胞凋亡的影响

目的:利用高表达抑癌基因maspin的人胃癌MGC-803细胞探讨南蛇藤提取物对其凋亡的影响。方法:以人胃癌MGC-803细胞为研究对象,不同质量浓度20,40,80,160,320 mg·L-1南蛇藤提取物作用于MGC-803细胞24,48 h后,MTT法检测南蛇藤提取物对高表达maspin的人胃癌MGC-803细胞增殖的影响;以不同质量浓度COE组(0,20,40,80 mg·L-1)及32 mg·L-1的5-FU阳性对照处理24 h后,TUNEL法检测南蛇藤提取物对细胞凋亡的影响,Western blot检测对maspin,Bcl-2,Bcl-2L12及Bax蛋白表达的影响。结果:南蛇藤提取物(20,40,80,160,320 mg·L-1)能明显抑制高表达maspin的人胃癌MGC-803细胞的增殖,呈现时间及浓度依赖性;与空白组比较,不同质量浓度的南蛇藤提取物(20,40,80 mg·L-1)作用24 h后,镜下显示,随着药物浓度升高,细胞数目减少,形态变圆,胞浆浓缩,出现凋亡小体,TUNEL结果表明南蛇藤提取物(20,40,80 mg·L-1)能促进细胞凋亡,Western blot结果显示南蛇藤提取物(20,40,80,160 mg·L-1)可呈浓度依赖性地增加抑癌基因maspin蛋白的表达,降低Bcl-2,Bcl-2L12的表达,均具有明显的统计学差异(P0.05,P0.01)。结论:南蛇藤提取物能够抑制高表达maspin的人胃癌MGC-803细胞的增殖,促进其凋亡,其机制可能与南蛇藤提取物促进抑癌基因maspin的表达以及抑制Bcl-2,Bcl-2L12的表达有关。     

玉竹高异黄酮抑制人肺癌细胞A549增殖的作用及机制

目的:以人肺癌A549细胞为研究对象,探讨玉竹中提取的高异黄酮对A549细胞增殖的抑制作用及其作用机制。方法:从玉竹中提取高异黄酮,作用于肺癌细胞A549;噻唑蓝(MTT)比色法观察高异黄酮12.5,25,50,100 mg·L~(-1)作用于A549细胞6,12,24 h对细胞存活率的影响。通过流式细胞仪检测细胞凋亡及周期变化,并通过蛋白免疫印迹法(Western blot)分析与细胞凋亡相关半胱氨酸天冬氨酸蛋白酶-3(Caspase-3),B细胞淋巴瘤(白血病)-2(Bcl-2),Bcl-2对抗杀伤性蛋白(Bak)和与细胞周期相关的磷酸化周期素依赖激酶2(p-Cdc 2),细胞周期蛋白依赖性激酶2(Cdc 2)及p38蛋白表达。结果:与空白组比较,高异黄酮呈剂量和时间依赖性抑制A549细胞的增殖(P0.05,P0.01),高异黄酮对A549细胞处理12 h后,细胞周期阻滞于G2/M期(P0.05);与空白组比较,25,50,100 mg·L~(-1)高异黄酮可显著促进凋亡蛋白Caspase-3和Bak表达,抑制Bcl-2表达(P0.01),可显著促进细胞周期蛋白p-Cdc 2和p38表达,抑制Cdc 2表达(P0.01)。结论:玉竹中提取的高异黄酮对A549细胞增殖具有抑制作用,能使A549细胞阻滞于细胞周期G2/M期,其机制与线粒体介导的细胞凋亡和p38 MAPK通路相关。     

纳米雄黄对肺癌A549细胞的凋亡诱导作用

目的:研究纳米雄黄对肺癌A549细胞的凋亡诱导作用。方法:采用机械研磨法制备纳米雄黄。以肺癌A549细胞株为靶细胞,采用MTT法检测细胞的增殖活性,AnnexinV/PI双染色法检测A549细胞的凋亡,流式细胞术检测BAX蛋白和Bcl-2蛋白表达、Caspase-3活性、P53蛋白的表达。结果:纳米雄黄可显著抑制A549细胞的增殖,50μg/mL和100μg/mL的纳米雄黄处理48h后,A549细胞凋亡率显著增高为12.53%和69.19%;20μg/mL和50μg/mL的纳米雄黄作用24h,A549细胞中活性caspase-3由对照的(2.25±0.17)%增高到(3.84±0.63)%和(7.35±0.33)%;P53蛋白表达总体增高但加药48h高剂量组也有所降低;Bax蛋白表达由对照的(79.50±0.50)%增高到(92.45±0.35)%和(96.9±0.00)%;Bcl-2蛋白表达由对照的(77.85±2.15)%增高到(87.10±10.5)%和(83.1±4.6)%。结论:纳米雄黄可有效诱导肺癌A549细胞发生凋亡。     

芹菜素诱导肺癌A549细胞凋亡及其机制研究

目的 研究芹菜素诱导肺癌A549细胞凋亡效应及其机制.方法 常规培养肺癌A549细胞,分别用10、20、40、80 μnol/L芹菜素作用于A549细胞;MTT法检测A549细胞增殖率;Western blotting检测聚二磷酸腺苷核糖聚合酶(PARP)、Bcl-2的蛋白表达水平;RT-PCR检测bcl-2 mRNA的表达;Annexin V/PI双染法检测A549细胞凋亡率.结果 随着芹菜素浓度的增高,A549细胞的增殖抑制率明显增高,具有统计学意义(P＜0.05);Western blotting显示PARP的剪切带逐渐增加,Bcl-2表达呈减少趋势;RT-PCR结果显示bcl-2 mRNA转录减少;0、10、20、40、80 μmol/L芹菜素对A549细胞凋亡率分别为(2.41 ±0.072)%、(10.56±0.054)%、(15.32±0.071)%、(28.98±0.012)%、(78.24±0.064)%,差异有统计学意义(P＜0.05).结论 芹菜素通过抑制Bcl-2的表达有效地诱导肺癌A549细胞凋亡.     

真菌的中药干预研究近5年进展

近年来真菌感染率逐年上升,传统抗真菌药物易产生耐药性,而中药在防治真菌感染方面具有一定的优势。本文就近5年来中药对白念珠菌、皮肤癣菌、曲霉菌、马拉色菌、串珠镰孢菌、申克孢子丝菌、新生隐球菌及真菌生物膜的干预研究进展进行综述。     

α-Glucosidase inhibitory activity of selected Philippine plants

Ethnopharmacological relevance

Antidesma bunius Spreng. (Phyllantaceae), Averrhoa bilimbi L. (Oxalidaceae), Biophytum sensitivum (L.) DC. (Oxalidaceae), Ceriops tagal (Perr.) C.B. Rob. (Rhizophoraceae), Kyllinga monocephala Rottb. (Cyperaceae), and Rhizophora mucronata Lam. (Rhizophoraceae) are used as remedies to control diabetes. In the present study, these plants were screened for their potential α-glucosidase inhibitory activity.

Materials and methods

The 80% aqueous ethanolic extracts were screened for their α-glucosidase enzyme inhibitory activity using yeast alpha glucosidase enzyme.

Results

Except for A. bilimbi with IC₅₀ at 519.86±3.07, all manifested a significant enzyme inhibitory activity. R. mucronata manifested the highest activity with IC₅₀ at 0.08±1.82 μg mL⁻¹, followed by C. tagal with IC₅₀ at 0.85±1.46 μg mL⁻¹ and B. sensitivum with IC₅₀ at 2.24±1.58 μg mL⁻¹.

Conclusion

This is the first report on the α-glucosidase inhibitory effect of the six Philippine plants; thus, partly defining the mechanism on why these medicinal plants possess antidiabetic properties.     

Investigation of plant extracts in traditional medicine of the Brazilian Cerrado against protozoans and yeasts

Aim of the study

To investigate the activities of the 217 plant extracts in traditional medicine of the Brazilian Cerrado against protozoans and yeasts.

Materials and methods

Plant extracts were prepared by the method of maceration using solvents of different polarities. The growth inhibition of chloroquine-resistant Plasmodium falciparum strain (FcB1) was determined by measuring the radioactivity of the tritiated hypoxanthine incorporated. Activity against Leishmania (Leishmania) chagasi and Trypanosoma cruzi was measured by the MTT colorimetric assay. The antifungal tests were carried out by using the CLSI method. The active extracts were tested also by cytotoxicity assay using NIH-3T3 cells of mammalian fibroblasts.

Results

Two hundred and seventeen extracts of plants were tested against Plasmodium falciparum. The eleven active extracts, belonging to eight plant species were evaluated against L. (L.) chagasi, Trypanosoma cruzi, yeasts and in NIH-3T3 cells. The results found in these biological models are consistent with the ethnopharmacological data of these plants. The ethyl acetate extract of Diospyros hispida root showed IC₅₀ values of 1 μg/mL against Plasmodium falciparum. This extract demonstrated no toxicity against mammalian cells, resulting in a significant selectivity index (SI) of 435.8. The dichloromethane extract of Calophyllum brasiliense root wood was active against Cryptococcus gattii LMGO 01 with MIC of 1.95 μg/mL; and Candida albicans ATCC 10231 and Candida krusei LMGO 174, both with MIC of 7.81 μg/mL. The same extract was also active against Plasmodium falciparum and L. (L.) chagasi with IC₅₀ of 6.7 and 27.6 μg/mL respectively. The ethyl acetate extract of Spiranthera odoratissima leaves was active against Cryptococcus gattii LMGO 01 with MIC of 31.25 μg/mL, and against Plasmodium falciparum with IC₅₀ of 9.2 μg/mL and Trypanosoma cruzi with IC₅₀ of 56.3 μg/mL.

Conclusion

The active extracts for protozoans and human pathogenic yeasts are considered promising to continue the search for the identification and development of leading compounds.     

European medicinal polypores – A modern view on traditional uses

Ethnopharmacological relevance

In particular five polypore species, i.e. Laetiporus sulphureus, Fomes fomentarius, Fomitopsis pinicola, Piptoporus betulinus, and Laricifomes officinalis, have been widely used in central European folk medicines for the treatment of various diseases, e.g. dysmenorrhoea, haemorrhoids, bladder disorders, pyretic diseases, treatment of coughs, cancer, and rheumatism. Prehistoric artefacts going back to over 5000 years underline the long tradition of using polypores for various applications ranging from food or tinder material to medicinal–spiritual uses as witnessed by two polypore species found among items of Ötzi, the Iceman. The present paper reviews the traditional uses, phytochemistry, and biological activity of the five mentioned polypores.

Materials and methods

All available information on the selected polypore taxa used in traditional folk medicine was collected through evaluation of literature in libraries and searches in online databases using SciFinder and Web of Knowledge.

Results

Mycochemical studies report the presence of many primary (e.g. polysaccharides) and secondary metabolites (e.g. triterpenes). Crude extracts and isolated compounds show a wide spectrum of biological properties, such as anti-inflammatory, cytotoxic, and antimicrobial activities.

Conclusions

The investigated polypores possess a longstanding ethnomycological tradition in Europe. Here, we compile biological results which highlight their therapeutic value. Moreover, this work provides a solid base for further investigations on a molecular level, both compound- and target-wise.     

厚朴与凹叶厚朴群体遗传学研究

目的:对厚朴与凹叶厚朴的群体遗传学进行研究,为中药厚朴的质量控制提供分子生药学方面的依据。方法:对厚朴与凹叶厚朴15个居群应用2个叶绿体基因间序列psbA-trnH和trnL-trnF进行PCR扩增并测序,计算厚朴与凹叶厚朴单倍型频率,用程序HaploNst分析遗传多样性和遗传结构,应用TCS version 1.13软件构建单倍型网状进化树。结果:厚朴与凹叶厚朴均无特有单倍型存在,但单倍型频率存在显著差异,已开始出现遗传分化的趋势,NST略大于GST。结论:厚朴与凹叶厚朴在遗传上已出现遗传分化的趋势,但尚未完全分化成彼此独立的单系。     

Effect of feeding Mucuna pruriens on helminth parasite infestation in lambs

Ethnopharmacological relevance

Mucuna pruriens is a tropical legume anecdotally reputed to have anthelmintic properties. This study was conducted to examine the validity of such claims.

Aim of the study

The aim of this study was to determine if ingestion of Mucuna seeds reduces helminth parasite infestation in lambs.

Materials and methods

Thirty-six Dorper × Katahdin ram lambs were assigned to three treatments, a cottonseed meal based control diet, a diet in which Mucuna replaced cottonseed meal and the control diet with levamisole (7.5 mg/kg body weight) administration. All diets were isonitrogenous and isocaloric. The 12 lambs in each treatment were assigned randomly to 4 pens, each containing 3 lambs. Lambs were trickle infected three times per week by gavage with infectious Haemonchus contortus larvae (2000 larvae/lamb) for 3 weeks.

Results

Levamisole treatment decreased fecal egg counts by 87% and abomasal worm counts by 83%. Mucuna intake did not statistically affect fecal egg counts or abomasal worm counts, though numerical (P > 0.10) reductions of 7.4% and 18.1%, respectively were evident. Anemia indicators, feed intake, and lamb growth were unaffected by treatment.

Conclusions

Levamisole reduced the Haemonchus parasite burden in lambs significantly but feeding Mucuna reduced the burden by levels unlikely to eliminate the clinical effects of parasitism.     

Antiplasmodial activity of extracts from seven medicinal plants used in malaria treatment in Cameroon

Aim of the study

In a search for new plant-derived biologically active compounds against malaria parasites, we have carried out an ethnopharmacological study to evaluate the susceptibility of cultured Plasmodium falciparum to extracts and fractions from seven Cameroonian medicinal plants used in malaria treatment. We have also explored the inhibition of the Plasmodium falciparum cysteine protease Falcipain-2.

Materials and methods

Plant materials were extracted by maceration in organic solvents, and subsequently partitioned or fractionated to afford test fractions. The susceptibility of erythrocytes and the W2 strain of Plasmodium falciparum to plant extracts was evaluated in culture. In addition, the ability of annonaceous extracts to inhibit recombinant cysteine protease Falcipain-2 was also assessed.

Results and discussion

The extracts showed no toxicity against erythrocytes. The majority of plant extracts were highly active against Plasmodium falciparumin vitro, with IC₅₀ values lower than 5 μg/ml. Annonaceous extracts (acetogenin-rich fractions and interface precipitates) exhibited the highest potency. Some of these extracts exhibited modest inhibition of Falcipain-2.

Conclusion

These results support continued investigation of components of traditional medicines as potential new antimalarial agents.     

金银花类药用植物IspE和IspH基因克隆和生物信息学分析

目的:克隆金银花类药用植物4-二磷酸胞苷-2-C-甲基赤藓糖激酶(4-diphosphocytidyl-2-C-methyl-D-erythritol kinase,IspE)和4-羟基-3-甲基-2-邻苯基二磷酸还原酶(4-hydroxy-3-methylbut-2-enyl diphosphate reductase,IspH)基因,并对其基因序列、蛋白特性和转录活性进行分析、比较.方法:从忍冬Lonicera japonica转录组测序结果中分析获得了IspE,IspH基因.分别以忍冬、红白忍冬L.japonica var.chinensis、红腺忍冬L.hypoglauca和水忍冬L.dasystyla新鲜花蕾为材料,利用RT-PCR技术克隆获得了4种金银花类药用植物IspE和IspH基因的全长cDNA.运用生物信息学分析软件,预测编码蛋白的结构和功能,并通过RT-PCR检测IspE和IspH在忍冬、红白忍冬、红腺忍冬、水忍冬花蕾中的转录情况.结果:金银花类药用植物IspE基因含有1个完整的开放阅读框,长度为1 221 bp,编码406个氨基酸;IspH含有一个完整的开放阅读框,长度为1 380 bp,编码459个氨基酸.IspE和IspH均为非分泌蛋白,均定位于叶绿体中.RT-PCR分析结果表明在忍冬、红腺忍冬和水忍冬的花蕾中IspE,IspH基因的转录水平没有显著差异,但红白忍冬花蕾中IspE,IspH基因的转录水平均显著高于忍冬.结论:克隆获得忍冬、红白忍冬、红腺忍冬和水忍冬中IspE,IspH基因,并证实了其在不同金银花类药用植物中的表达,为进一步研究IspE,IspH基因对萜类化合物生物合成和花香气以及颜色的影响奠定了基础.     

中国石斛属植物文献计量研究

石斛是珍稀濒危中药材,目前正处于快速发展阶段。为全面了解我国石斛属植物研究的历史和发展现状,作者以1954～2010年"中国知网中国学术期刊网络出版总库"收录的石斛研究文献为依据,采用文献计量学的原理和方法,对我国石斛属植物研究文献从文献年代分布、期刊分布与被引频率、主题分布、研究对象分布、作者与研究机构分布等方面进行了统计与分析。结果表明,我国石斛研究明显分为起步(2个)、停滞、平稳发展、快速上升5个阶段;期刊分布存在离散性与集中性并存的现象,已形成核心期刊研究群,并以《中国中药杂志》、《中草药》和《陕西中医》为代表;研究主题广泛涉及临床与药理、组织培养与种苗繁育、成分分析等多个领域,已经形成比较稳定的研究机构和团队,但研究对象差异显著,以铁皮石斛、金钗石斛和霍山石斛最为集中。我国石斛属植物的研究已取得显著成果,但种植产业发展缓慢,供需矛盾突出,预计种苗繁育与人工种植、产品开发、化学与药理等方面是未来的研究热点,其文献报道仍将进一步上升。     

牛耳朵和黄花牛耳朵的显微和化学鉴别

目的:为苦苣苔科唇柱苣苔属植物牛耳朵和黄花牛耳朵的鉴定和分类提供解剖学和化学依据。方法:采用石蜡制片法和水合氯醛透化法对2种药用植物的根状茎和叶横切面和粉末特征进行研究,应用光学显微镜观察显微结构。采用HPLC-UV法进行化学鉴别。结果:牛耳朵和黄花牛耳朵显微特征无明显区别,但是化学特征有明显的差异。结论:化学诞生特征鉴别方法可以作为2种药用植物的鉴定方法。