两面针碱、花椒棚碱对肺腺癌A549细胞的抑制作用研究

目的观察两面针碱以及花椒棚碱对抗肺腺癌A549细胞的抑制作用。方法采用MTT比色法观察检测,两面针碱及花椒棚碱对A549细胞抑制效应,并计算Ic50。结果两面针碱、花椒棚碱对A549细胞的Ic50分别为0．68μg．mL-1、173．4lμg．mL-1。两面针碱在1．015μg．mL-1。浓度下能够诱导66．O％的A549细胞凋亡,在显微镜下显示为细胞的皱缩、核裂解、核固缩等现象。结论两面针碱体外对A549细胞有杀伤作用,能显著抑制A549细胞的生长,而花椒棚碱高浓度溶液虽然能显示出抑制A549细胞的生长,但体外对A549细胞无杀伤作用。     

人硫氧还蛋白1在大肠杆菌中的重组表达及其对高糖导致的人脐静脉内皮细胞损伤的保护作用

目的 克隆人硫氧还蛋白1(hTRX1)基因并构建其原核表达载体,获得重组人硫氧还蛋白1(rhTRX1),评价rhTRX1对高糖导致的人脐静脉内皮细胞(HUVEC)损伤的保护作用.方法 采用逆转录PCR方法扩增hTRX1基因片段,插入pET22b(+)质粒并转化大肠杆菌Rosetta-gami(2),获得工程菌Rosetta-gami(2)-pET22ab(+)/hTRX1.用SDS-PAGE电泳和Western blot鉴定重组蛋白的正确性,用镍亲和层析纯化重组蛋白.采用高糖制备HUVEC损伤模型,MTT比色法检测HUVEC的存活率,生化方法测定HUVEC中乳酸脱氢酶(LDH)的外漏率以及细胞上清液中一氧化氮(NO)的水平.结果 构建的工程菌及其产生的重组蛋白rhTRX1正确.与正常对照组比较,高糖损伤组HUVEC的存活率降低、LDH外漏率明显升高,NO的水平明显地降低.不同剂量rhTRX1处理组HUVEC的存活率升高、LDH的外漏率明显降低,NO的水平明显地升高.结论 成功克隆并在大肠杆菌中表达rhTRX1、获得结构正确的rhTRX1,rhTRX1对高糖诱导的HUVEC损伤具有保护作用.     

粉防己碱对人肺癌A549细胞血管生成拟态的影响

目的:研究粉防己碱对人肺癌A549细胞血管生成拟态的影响。方法:采用细胞黏附试验观察粉防己碱对A549细胞黏附作用的影响;通过细胞迁移试验检测粉防己碱对A549细胞迁移能力的影响;通过血管生成拟态试验观察粉防己碱对A549细胞体外类血管生成的作用;实时定量聚合酶链反应(RT-PCR)法检测粉防己碱对A549细胞血管生成拟态相关基因表达的影响。结果:5、10、20μmol/L粉防己碱作用A549细胞30 min后,细胞的黏附作用受到抑制,并与浓度呈正相关;与对照比较,5、10、20μmol/L粉防己碱处理A549细胞12、24 h后,A549细胞迁移数减少,A549细胞管腔形成数明显减少,管腔长度明显缩短;A549细胞以粉防己碱处理后,血管生成拟态相关基因VE-cadherin、VEGF的表达减弱。结论:粉防己碱可抑制A549细胞黏附、细胞迁移和血管生成拟态,其机制可能与抑制血管生成拟态相关基因VE-cadherin、VEGF的表达有关。     

Sansanmycin A对铜绿假单胞菌体内外抗菌活性的初步研究

目的评价sansanmycin A对临床分离铜绿假单胞菌的体内、外抗菌活性。方法体外试验以微量肉汤稀释法测定sansanmycin A同时对照10种药物对150株铜绿假单胞菌的最低抑菌浓度(MIC),以平皿计数法测定最低杀菌浓度(MBC);半体内试验以微量稀释法测定sansanmycin A的血清抑菌活性(SBS)和血清杀菌活性(SBA)。结果 Sansanmycin A对铜绿假单胞菌体外抗菌活性与哌拉西林相似,其MIC50和MBC50分别为8和16μg/mL,MIC范围为2~>512μg/mL,MBC范围为2~>512μg/mL,头孢曲松和替卡西林对铜绿假单胞菌的MIC50和MBC50较sansanmycin A高2~4倍。Sansanmycin A对铜绿假单胞菌显示浓度依赖性杀菌作用。Sansanmycin A对铜绿假单胞菌峰时SBS和SBA中位数分别为1:256和1:128。结论 Sansanmycin A对铜绿假单胞菌显示较强的体外和体内杀菌活性,有进一步研究和开发的价值。     

藤茶双氢杨梅树皮素诱导人肺癌A549细胞凋亡及其机制研究

目的：探讨藤茶双氢杨梅树皮素（ampelopsin,APS）抗人肺癌A549细胞的作用。方法：以MTT法和集落形成法观察APS对人肺癌A549细胞的体外抑制作用;采用RT-PCR技术测定APS对人肺癌细胞株A549抑瘤基因p53表达的影响。结果：APS能明显抑制体外培养的A549细胞的生长,MTT法中IC50为9.8μg·mL^-1;细胞生长曲线法提示其对A549细胞的生长也有明显的抑制作用;集落形成试验中,当药物浓度为14.84,9.90,6.60,4.40μg·mL^-1时,抑制率分别为100%,91.6%,64.5%和23.4%;当浓度为25μg·mL^-1时,APS能显著增强p53基因的表达水平。结论：APS对体外培养的人肺癌A549细胞的生长有明显的抑制作用,对抑癌基因p53表达的影响是其抗肿瘤的作用机制之一。     

海洋放线菌N331生物碱活性组分的研究

目的 对海洋放线菌N331发酵液中的抗菌活性成分进行研究.方法 采用大孔树脂吸附、硅胶柱层析分离纯化抗菌活性成分,并对其理化性质和生物活性进行初步研究.结果 从海洋放线菌N331的发酵液中分离到一种结晶状活性组分,鉴定为生物碱类物质,由几种极性相近的化合物组成;其对金黄色葡萄球菌敏感茼及其耐药菌的最小押菌浓度均为4μg·mL-1,对口腔上皮癌KB细胞和肺癌A549细胞的半数抑制浓度(IC50)分别为2.0,3.0μg·mL-1,与顺铂的抗肿瘤效价相当.结论 海洋放线菌N331能产生抗茸及细胞毒性较强的生物碱类活性物质.     

杨梅中一个新化合物的抗肿瘤活性的初步研究

目的对从植物杨梅中分离得到的一个新化合物一杨梅新醇进行抗肿瘤活性研究。方法通过SRB法检测细胞的存活率,Hoechst33258核染色法、琼脂糖凝胶电泳法和流式细胞术来检测人肺癌细胞A549细胞的凋亡情况,来初步评价杨梅新醇的抗肿瘤活性。结果杨梅新醇的浓度依赖性地抑制肿瘤细胞体外生长活性,浓度为32μg／mL、16μg／mL、8μg／mL、1．9μg／mL的杨梅新醇对人肺癌细胞A549的杀伤率分别为：90．99％、36．48％、21．53％、19．46％。结论杨梅新醇具有一定的抗肿瘤活性,有望成为一种新型天然抗肿瘤活性化合物。     

腺病毒Ad-PTEN对肺腺癌细胞的体外抑制作用

目的探讨含PTEN基因的重组腺病毒载体(Ad-PTEN)对肺腺癌细胞株A549的体外抑制作用。方法采用细胞内同源重组方法构建PTEN基因重组腺病毒载体,293细胞扩增病毒,纯化后转染至人肺腺癌A549细胞株。观察Ad-PTEN的细胞生长抑制作用、细胞凋亡及对细胞超微结构的影响。结果 RT-PCR鉴定证实重组腺病毒载体Ad-PTEN构建成功。体外转染后能在A549细胞内转录和表达,并诱导A549细胞凋亡,抑制其生长和增殖。结论成功构建了含PTEN基因的重组腺病毒载体Ad-PTEN。Ad-PTEN引起肺腺癌细胞株A549明显的细胞形态学改变,生长增殖抑制,诱导A549细胞凋亡,是PTEN基因抑制肺腺癌细胞增殖的主要作用机制之一。     

重组人5型腺病毒在体外对A549细胞的影响

目的探讨重组人5型腺病毒对A549细胞增殖、细胞周期及凋亡的影响。方法采用MTT法、细胞迁徙试验、流式细胞术、TUNEL法等方法检测重组人5型腺病毒对A549细胞生物学特性的影响。结果重组人5型腺病毒在体外对A549细胞有显著的生长抑制作用,且具有时间和浓度依赖性;一定质量浓度的重组人5型腺病能显著降低A549细胞迁徙活性,诱导细胞凋亡,阻滞A549细胞于G0/S期。结论重组人5型腺病在肿瘤的临床治疗中有着重要的作用和广阔的前景。     

基于糖酵解的金钗石斛乙醇提取物抗肺癌的药理机制研究

目的 研究金钗石斛乙醇提取物(DNEE)治疗肺癌的药理作用机制。方法 采用人肺癌A549细胞进行体外实验,MTT法检测50～800μg·mL^-1DNEE对A549细胞的抑制率,比色法观察细胞的葡萄糖消耗量和乳酸生成量。以小鼠肺癌LLC细胞皮下移植荷瘤小鼠为疾病模型评估DNEE 200和600 mg·kg^-1的体内抗肿瘤作用,测量和绘制荷瘤小鼠体重变化曲线和肿瘤生长曲线。Western blot法检测A549细胞和肿瘤组织中缺氧诱导因子(HIF)-1α、葡萄糖转运蛋白(GLUT)4和己糖激酶(HK)2蛋白表达水平,氯化钴(CoCl2)诱导HIF-1α高表达分析HIF-1α对GLUT4和HK2蛋白表达的影响,分子对接技术探讨DNEE抗肿瘤主要活性成分与HIF-1α、GLUT4和HK2的相互作用。结果 与空白对照组相比,50～800μg·mL^-1 DNEE组A549细胞抑制率呈浓度依赖性增加(P <0.05),200、400μg·mL^-1 DNEE组A549细胞中葡萄糖消耗量和乳酸生成量显著降低(P...     

辽东葱木皂苷体内外抗肿瘤作用的实验研究

目的通过体内外抗肿瘤实验观察辽东葱木皂苷的抗肿瘤作用.方法:在小鼠异体移植性S180肿瘤模型上观察辽东葱木皂苷的抗肿瘤作用,同时用MTT比色法检测辽东葱木皂苷对腺癌A549和喉癌Hep2细胞株的体外细胞毒作用.结果:辽东葱木皂苷对荷S180小鼠肿瘤生长有明显抑制作用,高、中、低剂量组抑瘤率分别是70.31%,47.23%,41.31%;而且能明显抑制体外培养的肿瘤细胞的生长,对两种细胞有明显的细胞毒效应,其IC50分别是0.0155 mg/ml和0.0246mg/ml;结论:辽东葱木皂苷具有一定的体内外抗肿瘤活性.     

肿瘤相关巨噬细胞对非小细胞肺癌MCP-1、MIP-1的影响

目的 观察肿瘤相关巨噬细胞(TAMs)U937和非小细胞肺癌细胞(NSCLC)A549在体外不同的共培养条件下,对单核细胞趋化蛋白-1(MCP-1)、巨噬细胞炎性蛋白-1(MIP-1)表达的影响.方法 采用Jeremy培养方法,分为三组:1组用不同浓度的U937细胞与A549细胞共培养24h;2组同浓度的U937细胞与A549细胞共培养不同时间;3组用A549在不同时间段单独培养,采用原位杂交方法分别检测三组A549细胞中MCP-1、MIP-1mRNA表达情况.结果 随着U937细胞浓度逐渐增高以及共培养时间的逐渐延长,癌细胞A549中MCP-1、MIP-1mRNA表达的阳性系数均值也相应地增高,明显高于U937细胞浓度为0×105/ml的共培养组以及各时段对照组(均P<0.05).结论 TAMs细胞U937和NSCLC细胞A549的相互作用,与A549细胞MCP-1,MIP-1mRNA的阳性表达密切相关,并存在着浓度和时间的依从性.     

应用A549细胞单层模型研究蛋白多肽类药物肺部吸收的特性应用A549细胞单层模型研究蛋白多肽类药物肺部吸收的特性

药物通过呼吸进行吸收的路径依次为气管→支气管→肺泡,其中通过肺泡上皮细胞吸收占90％以上。由于肺泡上皮细胞使大分子的蛋白多肽类药物不易通过,是它们渗透的主要屏障,因此对肺泡膜屏障特性、药物透过过程及渗透特性的研究有助于更     

Differential effects of single-walled carbon nanotubes on cell viability of human lung and pharynx carcinoma cell lines

Carbon nanotubes (CNTs) are attracting significant attention as a novel material for future innovations. Many in vitro studies have assessed the cytotoxicity of CNTs, but the effects of CNTs differ depending on the cell lines and the synthetic method adopted for fabricating CNTs. In the present study, the differential effects of single-walled CNTs (SWCNTs) on the cell viability of A549 cells from human lung carcinomas and FaDu cells from human head and neck carcinomas were investigated. The SWCNTs used in the present study were synthesized with nickel and yttrium (SO-SWCNTs), and iron (FH-P-SWCNTs) as catalysts. Cell viability was evaluated on the basis of cell-membrane biomass, adenosine triphosphate (ATP) content, and intracellular metabolic capacity. After 24-hr exposure of A549 and FaDu cells to 1.0 mg/ml SO-SWCNTs, the cell-membrane biomass of A549 cells decreased to 43% as compared to the control cells, whereas that of FaDu cells remained over 90%. After 24-hr exposure of A549 and FaDu cells to 1.0 mg/ml SO-SWCNT, the intracellular metabolic capacity decreased to 24% and 37%, respectively, and the ATP content decreased to 40% and 54%, respectively. SWCNTs had a greater impact on the viability values of A549 cells than on those of FaDu cells. In addition, cells exposed to FH-P-SWCNTs exhibited a higher viability than those exposed to SO-SWCNTs. Caspase 3/7 activity was not increased at maximum concentration of 1.0 mg/ml SO-SWCNTs. It was surmised that sensitivity to SWCNTs differs among the 2 cell lines; additionally, SWCNT characteristics may produce different effects on these cell lines.     

Antitumor and antiangiogenic effects of GA-13315, a gibberellin derivative

This study showed that 13-chlorine-3,15-dioxy-gibberellic acid methyl ester (GA-13315), a gibberellin derivative, possessed high antitumor and antiangiogenic activity in vitro and in vivo. Cytotoxicity assays showed that GA-13315 was a potential and efficient antitumor compound, with inhibitory concentration 50 (IC₅₀) values ranging from 0.13 to 30.28 μg/ml in 12 human tumor cell lines, and it showed moderate toxicity to peripheral blood mononuclear cells with an IC₅₀ value of 14.2 μg/ml. Administration of 0.5 or 2.5 mg/kg GA-13315 for 23 days significantly inhibited tumor growth of human non-small cell lung tumor (A549) xenografts, with relative growth rates ranging from 29.91% to 35.05%. Acute toxicity was determined in ICR mice, and the lethal dose 50 (LD₅₀) was 4.19 g/kg after intragastric administration. The high antitumor potency of GA-13315 occurred in parallel with its antiangiogenic activity. In vitro, GA-13315 inhibited recombinant human epithelial growth factor-induced chemotactic motility and capillary-like tube formation of primary cultured human endothelial cells. Furthermore, GA-13315 decreased the factor VIII⁺ microvessel density and vascular endothelial growth factor expression in A549 tumors, indicating its antiangiogenic efficacy in vivo. These results indicate that the antiangiogenic activity of GA-13315 contributes to its anticancer properties. Further studies are needed to investigate the use of GA-13315 as an anticancer drug.     

Biological activity of beta-dolabrin, gamma-thujaplicin, and 4-acetyltropolone, hinokitiol-related compounds

Beta-dolabrin, gamma-thujaplicin, and 4-acetyltropolone, the components of Aomori Hiba (Thujopsis dolabrata SIEB. et ZUCC. var. hondai MAKINO), showed antifungal activity on seven kinds of plant-pathogenic fungi, antibacterial activity against two kinds of Legionella sp., and in vitro cytotoxic effect on murine P388 lymphocytic leukemia cell line. Firstly, beta-dolabrin, gamma-thujaplicin and 4-acetyltropolone had clear antifungal activity against seven kinds of plant-pathogenic fungi tested. In particular, beta-dolabrin and 4-acetyltropolone showed strong antifungal activity against Pythium aphanidermatum IFO 32440, with minimum inhibitory concentration (MIC) values of 6.0 microg/ml. Secondly, beta-dolabrin, gamma-thujaplicin and 4-acetyltropolone had obvious growth-inhibitory effect on two kinds of Legionella sp. 4-Acetyltropolone especially had strong antibacterial activity toward Legionella pneumophila SG 1, and its MIC value was 3.1 microg/ml. These three compounds showed cytotoxic effects against murine P388 lymphocytic leukemia cell line in vitro. The cytotoxic effect of three compounds in the murine P388 lymphocytic leukemia cell line were clear when cell growth was measured using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) method. At 48 h after treatment, gamma-thujaplicin and 4-acetyltropolone at 0.63 microg/ml inhibited cell growth of murine P388 lymphocytic leukemia by 85% and 65%, respectively. At the same time after treatment, the growth of the murine P388 lymphocytic leukemia cell line was completely suppressed by the three compounds at concentrations higher than 5.0 microg/ml. Among these three compounds, gamma-thujaplicin had the strongest cytotoxic activity on the growth of this tumor cell line in vitro.     

CCK-8法检测AT#9对肿瘤细胞的生长抑制作用

目的:研究复方制剂AT#9 的体外抗肿瘤作用.方法:采用细胞毒性检测CCK-8 法检测AT#9 对小鼠乳腺癌细胞EMT6、人肺癌细胞A549 和人前列腺癌细胞PC3M 的体外抑制生长作用.结果:AT#9 对三种肿瘤细胞的半数抑制浓度IC50 分别为1824μg/ml、3219μg/ml 和5854μg/ml.结论:AT#9 在体外对三种不同的肿瘤细胞有不同程度的生长抑制作用,且呈浓度依赖关系.     

In vitro antimicrobial activity of a chitooligosaccharide mixture against Actinobacillus actinomycetemcomitans and Streptococcus mutans.

The purpose of this study was to evaluate the in vitro antibacterial activity of a chitooligosaccharide mixture (MW 2000–30 000 Da) with a deacetylation degree of 91.5% against two representative oral pathogens, Actinobacillus actinomycetemcomitans and Streptococcus mutans. A 0.1% concentration of the chitooligosaccharides (derived from the exoskeletons of marine crustaceans) was used to estimate antibacterial activity. Approximately 2 log colony forming units (CFU)/ml of A. actinomycetemcomitans were inactivated by 0.1% chitosan after 30 min, while 120 min exposure inactivated about 4.5 log CFU/ml of this organism. In contrast, the level of inactivation against S. mutans was less than 0.5 log CFU/ml after an exposure of up to 120 min. Electron microscopy showed that the exposure of A. actinomycetemcomitans to the chitooligosaccharides resulted in the disruption of cell membranes and that it could be considered for the treatment of periodontal diseases associated with A. actinomycetemcomitans.     

Influenza A virus replication is inhibited in IFN-λ2 and IFN-λ3 transfected or stimulated cells

Interferons lambda (IFN-λ) are the most recently defined members of the class III cytokine family. To investigate whether IFN-λ2 and IFN-λ3 displayed antiviral activity against influenza A virus (IAV), a number of cell lines induced with IFNs - as well as two established cell lines (A549-IFN-λ2 and A549-IFN-λ3) - were infected with IAV. Our results indicate that IFN-λ2 has statistically significant antiviral activity in A549-IFN-λ2 (P=0.0028) although less so than IFN-λ3, which reduced viral titer to 10% (P<0.0001). The reverse was observed for cells treated with IFNs, with IFN-λ2-treated A549 cells inhibiting IAV infection more efficiently than IFN-λ3-treated A549 cells. The antiviral effect on IFN-stimulated cells was most apparent on Vero cells (compared with MDCK and HeLa). Both IFNs significantly inhibited IAV replication and inhibition was observed in a dose-dependent manner, with an optimal IFN concentration of 20 ng/ml. IFN-λ2 was more potent than IFN-λ3 on Vero cells while IFN-λ3 appeared more efficient than IFN-λ2 on MDCK and HeLa cells.