瑞香狼毒提取物逆转肺癌细胞耐药的免疫组化研究

目的:探讨瑞香狼毒逆转肺癌化疗耐药的机制。方法:用瑞香狼毒水提取物处理肺癌细胞株,用药前后分别用免疫组织化学SP法检测肺癌细胞中:多药耐药相关蛋白(MDM2)、肺耐药蛋白(LRP)、热休克蛋白27(HSP27)和多药耐药基因(MDR-1)的表达状况。利用形态学图像分析,比较瑞香狼毒用药前后四种耐药基因表达的差异。结果:两个肺癌细胞株瑞香狼毒用药前后四种癌基因表达情况如下:NCI-H446细胞株用药前MDR-1、LRP和MDM2细胞阳性率和辉度均高于用药后(P<0.05),而HSP27的表达用药前后差异不显著(P>0.05)。NCI-H157细胞株用药前MDR-1、LRP和HSP27细胞阳性率和辉度均高于用药后(P<0.05),而MDM2的表达用药前后差异不显著(P>0.05)。结论:瑞香狼毒提取物对两种肺癌细胞株的某些耐药基因蛋白的表达有明显的抑制作用。     

瑞香狼毒水提取物对人肺鳞状细胞癌Survivin表达的影响

目的 研究瑞香狼毒水提取物对人肺鳞状细胞癌NCI-H520细胞株的抑制作用及对Survivin表达的影响.方法 用不同浓度的瑞香狼毒水提取物处理NCI-H520细胞,分别用MTT法检测肺癌细胞抑制率、半定量RT-PCR检测Survivin mRNA水平以及用Western-blotting检测蛋白水平的表达.结果 瑞香狼毒水提取物可显著抑制NCI-H520细胞的生长,随着药物浓度的升高抑制作用逐渐增强.瑞香狼毒作用4 h后细胞内Survivin蛋白的表达水平降低,而Survivin mRNA的表达变化不明显.结论 ①瑞香狼毒水提取物可抑制NCI-H520细胞的生长,并具有浓度和时间依赖性;②瑞香狼毒水提取物可降低NCI-H520细胞的Survivin蛋白表达水平,而对mRNA水平无明显影响.     

瑞香狼毒提取物和肿瘤坏死因子对肺癌基因蛋白表达的影响

目的探讨瑞香狼毒和肿瘤坏死因子(TNF)对肺癌细胞基因蛋白表达的影响。方法用瑞香狼毒水提取物和TNF分别及联合处理肺癌细胞株NCI-H157和NCI-H446,用免疫组织化学SP法分别检测用药前后肺癌细胞中多药耐药相关蛋白(MDM2)、肺耐药蛋白(LRP)和热休克蛋白27(HSP27)的表达状况。通过形态学图像分析,比较瑞香狼毒和TNF用药前后三种基因蛋白表达的差异。结果用药前后两种肺癌细胞株3种基因蛋白表达情况如下:①瑞香狼毒组NCI-H446细胞株用药后LRP和MDM2阳性细胞灰度值均高于用药前(P<0.05);HSP27的表达用药前后差异无显著性(P>0.05)。NCI-H157细胞株用药后LRP和HSP27阳性细胞灰度值均高于用药前(P<0.05);MDM2的表达用药前后差异无显著性(P>0.05)。②TNF组NCI-H446细胞株用药后LRP阳性细胞灰度值高于用药前(P<0.05);HSP27和MDM2的表达用药前后差异均无显著性(P>0.05)。NCI-H157细胞株用药后LRP,MDM2和HSP27阳性细胞灰度值均高于用药前(P<0.05),其中LRP具有高度显著性(P<0.01)。③联合用药组NCI-H446细胞株用药后LRP和MDM2阳性细胞灰度值均高于用药前(P<0.05),HSP27的表达用药前后差异无显著性(P>0.05)。NCI-H157细胞株用药后LRP、MDM2和HSP27阳性细胞灰度值均高于用药前(P<0.05),且LRP和HSP27差异具有高度显著性(P<0.01)。结论瑞香狼毒提取物和肿瘤坏死因子对两种肺癌细胞株的某些基因蛋白的表达具有抑制作用;两种药物联合应用对基因蛋白的表达有明显的抑制作用。     

肺癌冲剂对小鼠移植性实体瘤病理形态改变的影响

为了探究抗癌中药复方肺癌冲剂的作用机制，对肺癌冲剂对荷瘤小鼠病理形态的发言煌影响进行初步观察；结果显示肺癌冲剂使移植性实体瘤瘤块坏死面积明显增大，与对照组比较；明显降低癌细胞核分裂数，与对照组比较差异显著。超微结构观察用药组瘤细胞的线粒体和内质网均肿胀、断裂、解体或形成空泡、消失，对照组则无此类改变。证实肺癌冲剂有明显地抑制瘤细胞核分裂作用，可能是通过作用于是瘤细胞的线粒体和内质网而抑制瘤细胞生长     

瑞香狼毒提取物对人增生性瘢痕成纤维细胞抑制作用的研究

 目的观察瑞香狼毒提取物对人增生性瘢痕成纤维细胞的抑制作用,为烧伤后增生性瘢痕的治疗提供实验依据。方法体外培养烧伤后的人增生性瘢痕成纤维细胞,加入不同浓度的瑞香狼毒提取物,24h后观察细胞形态学,以乳酸脱氢酶(LDH)为指标观察细胞毒性,用MTT法检测其增殖活性。结果不同浓度的瑞香狼毒提取物均能改变成纤维细胞形态,抑制细胞增殖,浓度在500μg·mL^-1以下无明显的细胞毒作用。结论瑞香狼毒提取物对人增生性瘢痕成纤维细胞具有抑制作用,并呈剂量依赖关系,具有防治增生性瘢痕的应用前景。     

胃萎清对慢性萎缩性胃炎大鼠胃黏膜超微结构的影响

目的:通过观察慢性萎缩性胃炎(CAG)大鼠胃黏膜腺上皮主细胞、壁细胞超微结构的变化,探讨健脾清热活血中药胃萎清对CAG大鼠胃黏膜超微结构的影响,初步揭示胃萎清对CAG的作用机制。方法:将CAG大鼠随机分为6组:对照组、模型组、胃萎清高、中、低剂量组及叶酸组。以N-甲基-N'-硝基-N'-亚硝基胍(MNNG)合饥饱失常法建立CAG大鼠模型,观察各组大鼠胃黏膜超微结构的变化。结果:与对照组比较,模型组大鼠胃黏膜主细胞核固缩,胞浆中出现大量空泡样变;壁细胞染色质浓缩,线粒体嵴数量减少。与模型组比较,胃萎清高、中剂量组大鼠胃黏膜主细胞核仁结构尚均一,粗面内质网基本整齐;壁细胞核质比较规则,线粒体嵴增多。胃萎清低剂量组大鼠胃黏膜主细胞细胞核核仁形态较模型组规则;壁细胞细胞核形态改变,线粒体数量减少。叶酸组大鼠胃黏膜主细胞细胞核染色质较正常,胞浆出现空泡样变;壁细胞染色质固缩,线粒体嵴数量较模型组多。结论:胃萎清能在一定程度上改善胃黏膜腺上皮主细胞、壁细胞的超微结构,推测胃萎清能改善CAG患者不适症状的机制之一可能是改善胃黏膜超微结构。     

朝天罐提取物体外抗肿瘤活性研究

目的:研究朝天罐(Osbeckia opipara,CTG)水提取物体外抗肿瘤活性。方法:采用CCK-8法和细胞克隆形成实验,研究不同浓度CTG水提取物对人肝癌细胞7721、人鼻咽癌细胞CNE-2增殖抑制作用;通过Hoechst33342荧光染色法检测细胞核染色质的变化来评价CTG对肿瘤细胞凋亡的影响。结果:CCK8和细胞克隆形成实验显示,CTG水提取物体外显著性抑制SSMG-7721和CNE-2细胞增殖; Hoechst 33342检测发现细胞核染色质发生明显改变。结论:CTG水提取物具有一定的抗肿瘤活性。     

新疆紫草提取物对GLC-82人肺腺癌细胞的体外作用

目的:研究不同浓度的紫草提取物(SE)对体外培养的GLC-82人肺腺癌细胞(简称"GLC-82细胞")的作用。方法:荧光显微镜观察细胞形态的变化;MTT法检测不同浓度SE培养细胞后细胞的存活率;划痕试验检测不同浓度的SE对细胞迁移力的影响;流式细胞仪检测细胞周期和凋亡率。结果:SE诱导细胞凋亡程度与用药剂量呈正相关,较高剂量的药物能引起GLC-82细胞亚结构发生变化,如呈现边界不清,体积缩小,核固缩,染色质凝集、边缘化,甚至细胞发生破碎,凋亡小体形成;随着SE浓度的增高GLC-82细胞向划痕区的愈合速度不断减慢;SE的浓度≥3mg/mL时,细胞周期主要表现为G1期比例的上升伴随S期比例的下降。结论:高剂量SE能够诱导GLC-82细胞凋亡及周期的改变,为临床上用药剂量提供一定的依据。     

五倍子提取物对表皮葡萄球菌的抗菌作用及其扫描和透射电镜观察

目的观察五倍子乙醇提取物对表皮葡萄球菌的体外抗菌活性及其在电镜下菌细胞形态超微结构的变化.方法采用新的中药抑菌实验方法对五倍子乙醇提取物进行236株表皮葡萄球菌的最低抑菌浓度测定,同时用电镜观察用药后细菌细胞的形态改变.结果五倍子乙醇提取物对127株耐甲氧西林的表皮葡萄球菌(methicillin-resi stant staphylococcus epidermidi s,MRSE)和109株甲氧西林敏感的表皮葡萄球菌(methicillin-sensitive staphylococcus epidermidis,MSSE)的MIC90分别为0.575、0.288 mg/mL;在电镜下观察经五倍子处理的表皮葡萄球菌菌细胞的超微结构,扫描电镜显示菌细胞形态大小不等,排列不整齐,大多数菌细胞的细胞壁破裂;在透射电镜下,菌细胞细胞壁结构、层次不请,胞浆膜层次不分明,细胞质内含物大多消失,细胞浆内包含体和染色体模糊不清.结论五倍子乙醇提取物对236株表皮葡萄球菌具有较强的抗菌作用.     

瑞香狼毒总黄酮提取物的抗肿瘤作用

目的:　研究瑞香狼毒总黄酮提取物的抗肿瘤作用。方法:体外抗肿瘤活性研究采用MTT比色法和集落形成法测定,靶细胞采用人胃癌细胞SGC- 790 1、人肝癌细胞BEL- 74 0 2和人白血病细胞HL- 6 0 ;通过提取物对小鼠的移植性肿瘤S180和H2 2生长的影响来评价体内抗肿瘤作用。结果:瑞香狼毒总黄酮提取物对体外培养的肿瘤细胞有较强的抑制作用,体外抗肿瘤活性高于长春新碱;瑞香狼毒总黄酮提取物急性毒性较小,对小鼠移植性肿瘤S180和H2 2的生长也有显著的抑制作用,并和剂量正相关,高剂量组对小鼠移植性肿瘤S180的抑制率为4 5 . 6 4 % ,H2 2的为4 7 .5 9% ,与阳性对照组环磷酰胺的抑制率接近。结论:瑞香狼毒总黄酮提取物具有较强的体内和体外抗肿瘤作用。     

四味蒙药体外抗真菌实验研究及超微结构观察

目的观察四味蒙药水提及醇提物对红色毛癣菌、须癣毛癣菌、犬小孢子菌、白色念珠菌和糠秕马拉色菌的体外抗真菌作用,并观察大花葵水提物对白色念珠菌和糠秕马拉色菌超微结构的改变。方法采用试管内药基法测定四味蒙药水、醇提物的最小抑菌浓度(MIC),扫描电镜下观察大花葵水提物作用后白色念珠菌和糠秕马拉色菌超微结构的变化。结果四种蒙药的水、醇提物分别在不同浓度时对受试菌种有较强的抑制作用,且大花葵水、醇提物对红色毛癣菌、须癣毛癣菌、犬小孢子菌的MIC为8.3 g/L。大花葵水提物对白色念珠菌和糠秕马拉色菌的MIC分别为50 g/L、12.5 g/L。经蒙药作用后,在扫描电镜下观察白色念珠菌和糠秕马拉色菌表面出现皱缩、剥脱、裂纹等改变。结论 MIC和超微结构改变提示四味蒙药能有效抑制真菌,为其抗真菌作用机制的研究提供了线索。     

南方红豆杉水提物对人肺癌A549细胞增殖抑制作用的研究

目的:研究南方红豆杉水提物对人肺癌A549细胞增殖的抑制作用和机制。方法:采用MTT法检测南方红豆杉水提物对A549细胞增殖的抑制作用;生长曲线法观察其抑制A549细胞的时间-效应曲线;流式细胞仪检测其对A549细胞周期及凋亡的影响;倒置显微镜及电镜观察其对A549细胞形态的影响。结果:南方红豆杉水提物对A549细胞的增殖有抑制作用;其抑制作用机制为使A549细胞周期出现明显的G0/G1期阻滞,诱导细胞凋亡。结论:南方红豆杉水提物对人肺癌A549细胞的增殖有抑制作用。     

海藻提取物A1和A2抑制肝癌和肺腺癌细胞增殖作用的实验研究

目的:用体外培养法研究海藻提取物A1和A2对肝癌740 2细胞、肺腺癌GLC -82细胞和胃癌MCG -80 3细胞生长的抑制作用。方法:采用MTT法进行研究。结果:海藻提取物A1和A2分别处理肿瘤细胞48h后,A1呈剂量依赖性抑制肺腺癌GLC -82和肝癌740 2细胞增殖,测得IC50 分别为2 2 61μg/mL和3 0 0 5 μg/mL ,而对胃癌MCG -80 3细胞增殖无明显抑制作用;A2对3种肿瘤细胞增殖无明显抑制作用。结论:海藻提取物A1对肝癌740 2细胞和肺腺癌GLC -82细胞的增殖有抑制作用。     

Biological activities and metabolites from Trevoa trinervis Miers

Chilean folklore medicine uses Trevoa trinervis Miers. (Rhamnaceae), ‘trevo’, for the treatment of inflammation caused by wounds and burns. The antiinflammatory and antipyretic activities of crude methanol, aqueous and successive hexane, dichloromethane, methanol extracts of the aerial part were studied. The fractionation of the most potent extract (dichloromethane), led to the isolation of the pentacyclic triterpenoids betulinic (the major metabolite), oleanolic and ursolic acids, along with β-sitosterol. Evaluation of the acute oral toxicity of the crude methanol extract showed it to be nontoxic. The crude methanol and the aqueous extracts were assayed for their antineoplastic activity against P-388 murine leukaemia, A-549 human lung carcinoma and HT-29 colon carcinoma showing no activity. Betulinic acid was also assayed showing some activity against the neoplastic cell lines, but only residual activity against Mel-28 melanona cells. © 1997 John Wiley & Sons, Ltd.     

南方红豆杉水提物诱导人肺腺癌A549细胞凋亡与紫杉醇抑瘤机制不同的研究

目的:初步探讨南方红豆杉水提物抗人肺腺癌A549细胞(以下简称"A549细胞")的作用机制,明确其与紫杉醇抗肿瘤机制的不同。方法:以流式细胞仪检测南方红豆杉水提物对A549细胞凋亡的影响,以紫杉醇为对照;以普通光学显微镜观察南方红豆杉水提物和紫杉醇对A549细胞形态的影响;以Western blotting检测细胞凋亡相关蛋白P-jnk的表达。结果:①流式细胞检测及光镜观察发现南方红豆杉水提物处理后A549细胞呈典型的早期凋亡表现(P<0.05,P<0.01),而紫杉醇组细胞以坏死及晚期凋亡表现为主(P<0.05,P<0.01),且其比例均呈现一定的剂量-效应关系;②Western blotting检测发现南方红豆杉水提物处理后A549细胞P38蛋白表达无明显变化、P-jnk蛋白随药物浓度增加表达减少(P<0.01)。结论:南方红豆杉水提物抗A549细胞的机制与紫杉醇不同,其作用机制主要与诱导肿瘤细胞凋亡有关。     

冰片对真菌细胞超微结构的影响及治疗化脓性中耳炎的临床应用

目的 :研究冰片治疗化脓性中耳炎的药理作用 ,最佳有效浓度。方法 :取患者外耳道内分泌物中分离出的黑曲菌和实验室保留的真菌菌株 ,测定冰片最低抑真菌浓度 (MIC) ,最低杀真菌浓度 (MFC)。电子显微镜下观察冰片作用后的细胞超微结构变化。结果 :发现冰片对真菌MIC为 5 % ,MFC为 10 %。电镜下见冰片作用后的黑曲菌细胞壁变厚 ,模糊不清有空泡。细胞浆内各种物质深染凝集。呈现大量脂滴。电子密度增高 ,嵴排列紊乱。有的细胞器破坏 ,呈现大量电子空白区。有的细胞扭曲变形失去原有结构。结论 :冰片可破坏真菌细胞的结构 ,导致真菌溶解死亡 ,表现出抑菌杀菌功能。最低抑真菌浓度为 5 % ,杀菌浓度为 10 %。     

Isolation, structure, and biological activities of Fellutamides C and D from an undescribed Metulocladosporiella (Chaetothyriales) using the genome-wide Candida albicans fitness test

In a whole-cell mechanism of action (MOA)-based screening strategy for discovery of antifungal agents, Candida albicans was used, followed by testing of active extracts in the C. albicans fitness test (CaFT), which provides insight into the mechanism of action. A fermentation extract of an undescribed species of Metulocladosporiella that inhibited proteasome activity in a C. albicans fitness test was identified. The chemical genomic profile of the extract contained hypersensitivity of heterozygous deletion strains (strains that had one of the genes of the diploid genes knocked down) of genes represented by multiple subunits of the 25S proteasome. Two structurally related peptide aldehydes, named fellutamides C and D, were isolated from the extract. Fellutamides were active against C. albicans and Aspergillus fumigatus with MICs ranging from 4 to 16 μg/mL and against fungal proteasome (IC?? 0.2 μg/mL). Both compounds showed proteasome activity against human tumor cell lines, potently inhibiting the growth of PC-3 prostate carcinoma cells, but not A549 lung carcinoma cells. In PC-3 cells compound treatment produced a G2M cell cycle block and induced apoptosis. Preliminary SAR studies indicated that the aldehyde group is critical for the antifungal activity and that the two hydroxy groups are quantitatively important for potency.     

海绵提取物S1～S3抑制肝癌细胞和肺腺癌细胞增殖作用的实验研究

目的:用体外培养法研究海绵提取物S1～S3对肝癌7402细胞、肺腺癌GLC-82细胞和胃癌MCG-803细胞增殖的抑制作用.方法:采用MTT法进行研究.结果:海绵提取物S1、S2和S3分别处理肿瘤细胞48 h后,S3呈剂量依赖性抑制肺腺癌GLC-82和肝癌7402细胞增殖,测得IC50分别为2.714 μg/ml和2.598 μg/ml,而对胃癌MCG-803细胞增殖无明显抑制作用;S1和S2对3种肿瘤细胞增殖均无抑制作用.结论:海绵提取物S3对肝癌7402细胞和肺腺癌GLC-82细胞的增殖有抑制作用.     

Studies of biological properties of Uncaria tomentosa extracts on human blood mononuclear cells

Ethnopharmacological relevance

Uncaria tomentosa (Willd.) DC is a lignified climbing plant from South and Central America, which (under the name of “vilcacora” or “cat's claw”) has become highly popular in many countries due to its proven immunostimmulatory and anti-inflammatory activities and also with respect to its anticancer and antioxidative effects. There are insufficient data on the mechanism of U. tomentosa action on normal blood mononuclear cells.

Aim of the study

The aim of the study was to analyze the impact of ethanol and aqueous extracts from bark and leaves of Uncaria tomentosa on the structure and function of human mononuclear cells and to find out whether the kind of extractant used modulates biological activity of the extracts studied.

Materials and methods

Plant material consisted of four different extracts: (1) ethanol extract from leaves, (2) aqueous extract from leaves, (3) ethanol extract from bark and (4) aqueous extract from bark. The effect of these extracts on protein damage as well as on free-radical formation in human peripheral blood mononuclear cells was analyzed. Moreover, changes in viability, size, and granularity as well as apoptotic alterations in human blood mononuclear cells exposed to U. tomentosa extracts were investigated.

Results

The oxidative changes were observed in mononuclear blood cells exposed to both ethanol and aqueous extracts obtained from bark and leaves. Moreover, in the cells studied the extracts from U. tomentosa induced apoptosis and a decrease in viability of mononuclear blood cells, with the exception of aqueous extract from leaves. Additionally, no statistically significant changes in the cell size were observed both for aqueous extracts from leaves and bark. Changes in the blood mononuclear cell granularity were observed at 250 μg/mL for all extracts examined. The strongest changes were observed for the ethanol extract of the bark, which increased cell granularity at 50 μg/mL and changed cell size at 100 μg/mL.

Conclusion

The conducted research showed differences in biological activity between aqueous and ethanol extracts. It was observed that ethanol extracts exhibited stronger negative effects on mononuclear blood cells. The kind of extractant used had a significant influence of the chemical composition of the tested extracts. The ethanol extract from bark containing a high amount of polyphenols and alkaloids revealed the highest pro-apoptotic effect.     

Tecoma sambucifolia: anti-inflammatory and antinociceptive activities, and 'in vitro' toxicity of extracts of the 'huarumo' of peruvian incas

Aqueous and alcoholic extracts of pods and flowers of Tecoma sambucifolia H.B.K. (Bignoniaceae) (‘huarumo’) were analysed to determine their anti-inflammatory activity (carrageenan-induced edema test), antinociceptive activity (acetic acid writhing test) and ‘in vitro’ toxicity in Chinese hamster ovary cells, human hepatome cells and human larynx epidermal carcinoma cells. The cytotoxic effects of both extracts were evaluated by two endpoint systems: neutral red uptake assay and tetrazolium assay. The results showed that all extracts have anti-inflammatory and antinociceptive activity, but the highest potency is that of the alcoholic extracts. There were significant differences in cytotoxicity between extracts and among the response of cells to them. The highest cytotoxicity was noted with the alcoholic extract, and the human hepatome cell line was the most sensitive, especially to the alcoholic extract of flowers. The aqueous pod extract appeared to have the best pharmaco-toxicological profile, since it provided a significant reduction of both pain and inflammation together with the lowest cytotoxicity.