Fluconazole induces genotoxicity in cultured human peripheral blood mononuclear cells via immunomodulation of TNF-α, IL-6, and IL-10: new challenges for safe therapeutic regimens

Context: Fluconazole (FNZ) is a drug used in antifungal therapy. However, the minimum FNZ dose to interfering with immune responses or inducing DNA damage is still unknown.

Objective: This study investigated the toxicological profile of FNZ on cultured human peripheral blood mononuclear cells (PBMCs) treated with different concentrations of this azole.

Materials and methods: Cultured PBMCs were exposed to FNZ (6, 12, 30, 60 and 120?μg/mL) and the toxicological profile was assessed by the following parameters: cytotoxic and nuclear division index (necrotic, apoptotic and viable cells), DNA damage (alkaline comet test), mutagenic potential (micronucleus test), cytokine modulation (IL-1, IL-6, IL-10, TNF-α, IFN-γ), and predictive toxicity (Osiris^® and LAZAR^® programs).

Results: Our results demonstrated that FNZ induced cellular DNA damage and mutagenicity at concentrations above the plasma peak (>30?μg/mL) and 6?μg/mL, respectively, which was associated with increased TNF-α, and decrease IL-6 and IL-10 concentrations. These effects may be related to increased apoptosis and cytotoxic nuclear division index in the cultured PBMCs. In silico results indicated potential mutagenic, tumorigenic, irritant, and carcinogenic effects, which were partially confirmed by the above assays.

Discussion and conclusions: Together, these findings suggest the need to rationalize the use of FNZ, especially if it is used for long periods or with concomitant pathologies requiring azole therapy that may increase FNZ's plasma concentration.     

Use of a high-throughput umu-microplate test system for rapid detection of genotoxicity produced by mutagenic carcinogens and airborne particulate matter

In the present study, we developed a rapid umu-microplate test system that uses the nitroreductase- and O-acetyltransferase-overproducing Salmonella typhimurium strain NM3009 and the O-acetyltransferase-overproducing S. typhimurium strain NM2009 to detect genotoxic activity in small volume samples. The assay was used to test the genotoxicity of several standard mutagens and environmental samples. Exponentially growing cultures of NM3009, NM2009, and the parental strain TA1535/pSK1002 were incubated in 96-well microplates with test chemicals both in the presence and in the absence of rat liver S9. The relative beta-galactosidase activities were then determined colorimetrically using either chlorophenol red-beta-D-galactopyranoside (CPRG) or O-nitrophenyl-beta-D-galactopyranoside (ONPG) as a measure of umuC gene induction activity. The sensitivities of NM3009 without S9 mix and NM2009 with S9 mix to nitroarenes and aromatic amines were up to 24- to 75-fold higher than those of the parent strain. Induction of umuC gene expression was detected more readily with CPRG than ONPG. The umu-microplate assay also detected genotoxicity in organic extracts of particulate matter from air samples collected in Osaka City, Japan. The pattern of the responses suggested that the genotoxic activity of the particulate extract was due primarily to nitrated polycyclic aromatic hydrocarbons. Our results indicate that the umu-microplate assay may be a useful way of carrying out rapid screens for genotoxicity in small-volume environmental samples.     

藿香正气片增强镇静催眠药的镇静与催眠作用的研究

目的研究藿香正气片增强镇静催眠药对小鼠的镇静催眠作用.为临床增添了一个中西药结合的镇静催眠合剂提供药理依据.方法应用抖笼换能器法观察藿香正气片对小鼠的自发活动的影响.由此观察藿香正气片与镇静催眠药的协同作用.测定藿香正气片与镇静催眠药对小鼠的入睡时间及睡眠持续时间的作用.两个实验均分成四组,生理水组、藿香正气片组、安定组、安定+藿香正气片组.结果1.对小鼠自发活动的影响.(1)藿香正气片组与生理盐水组比较,具有极显著性差异(P＜0.01).(2)安定组与安定+藿香正气片组比较具有显著性差异(P＜0.05).2.藿香正气片增强镇静催眠药对小鼠的睡眠作用.(1)藿香正气片组与生理盐水组比较具有显著性差异(P＜0.05).(2)藿香正气片+安定组与安定组比较具有极显著性差异(P＜0.01).结论藿香正气片与安定合用对镇静及催眠确有协同作用.     

阿斯匹林对胃癌细胞株SGC-7901的细胞毒作用及体外增效作用

目的：观察阿斯匹林（ASA）对胃癌细胞株SGC－7901的细胞毒作用以及联用其它抗肿瘤药的增效作用。方法：应用流式细胞仪（FCM）测定细胞周期,噻唑蓝（MTT）法测定药物细胞的抑制率。结果：MTT显示体外ASA对SGC－7901有细胞毒作用,与ASA的浓度和作用时间有显著的相关性。同时可使SGC－7901细胞周期中S期及G2／M期比例升高,G1期比例下降,呈一定的剂量效应关系。低浓度ASA与5－氟脲噻啶（5－Fu)、阿霉素（DOX）和丝列霉素（MMC）合用,可增强它们对SGC－7901的杀伤作用,与各药物有相加或协同作用。结论：ASA体外对SGC－7901有细胞毒作用,可明显阻断SGC－7901细胞在S期和G2／M期;增强上述药物对SGC－7901的杀伤作用。鉴于所试药物对细胞周期的影响不同,提示ASA的体外增效作用可能与此相关。     

姜黄素与羟基脲对K562细胞的体外联合作用

目的 了解姜黄素与羟基脲合用对人白血病Ｋ５６２细胞的体外作用。方法 采用ＭＴＴ法测定药物的体外杀伤作用,应用ＡＯ／ＥＢ荧光染料染色观察药物诱导细胞凋亡,金氏公式进行联合用药分析。结果 姜黄素５,１０ｕｇ／ｍｌ,羟基脲２５,５０ｕｇ／ｍｌ同时给药,对Ｋ５６２细胞可产生单纯相加的协同杀伤效果。姜黄素与羟基脲同时给药诱导Ｋ５６２细胞凋亡率高于羟基脲单独用药,低于姜黄素单独用药。结论 姜黄素与羟基脲同时联     

工程化纳米囊泡结合化疗用于增强肿瘤免疫治疗

目的：结合免疫治疗与化疗,以改善肿瘤的治疗效果,为安全有效的肿瘤免疫治疗策略的发展提供更多见解。方法：使用工程化纳米囊泡,囊泡膜上表达PD-1受体,可以靶向肿瘤细胞表面的PD-L1,通过破坏PD-1/PD-L1免疫抑制通路增强抗肿瘤反应。同时,囊泡包裹的化疗药物阿霉素可以进入肿瘤细胞核,抑制DNA与RNA的合成,诱导肿瘤细胞死亡。结果：实验证实,制备的PD1-阿霉素材料具备良好的稳定性、安全性,能准确靶向肿瘤部位,阿霉素在细胞核部位起作用,能有效地进行肿瘤杀伤。结论：本研究首次将PD-1免疫检查点抑制与化疗药物阿霉素相结合,利用PD-1囊泡安全性高、长循环的特点作为包裹化疗药物阿霉素的载体,这种方法可以进行肿瘤细胞的有效靶向与治疗,实现肿瘤的有效清除。     

Developmental toxicity and genotoxicity studies of 1,1,1,3,3,3-hexachloropropane (HCC-230fa) in rats.

The potential developmental toxicity and the in vitro and in vivo genotoxicity of HCC-230fa were assessed. In the developmental toxicity study, groups of 25 mated Crl:CD(R)(SD)BR rats were exposed (whole body) by inhalation to HCC-230fa over days 7-21 of gestation; the day of confirmed mating was designated as gestation day 1 (GD1). Exposures were 6 h per day at concentrations of 0, 0.5, 2.5, or 25 ppm. Body weight, food consumption, and clinical observation data were collected during the study. On day 22 of gestation, the dams were euthanized and examined grossly. The fetuses were removed and subsequently weighed, sexed, and examined for external, visceral, head, and skeletal alterations. Evidence of maternal and developmental toxicity was observed at 25 ppm and was noted as significant, compound-related reductions in mean maternal body weight, weight change, and food consumption. Significant fetal effects also were observed at 25 ppm as compound-related reductions in mean fetal weight and increased fetal malformations (filamentous tail, situs inversus, absent vertebrae) and variations (rudimentary cervical ribs, delayed sternebral ossification). There was no evidence of either maternal or developmental toxicity at 0.5 or 2.5 ppm. The genotoxicity of HCC-230fa was examined in a bacterial reversion assay and in erythrocyte micronucleus studies in two species by different routes of administration. No increases in the number of revertants were observed in the bacterial reversion assay. In one micronucleus study, HCC-230fa was administered by inhalation to rats as part of a 90-day study at doses indicated above. For the second study, ICR mice were given a single ip dose at 0, 166, 330, or 660 mg/kg. In both micronucleus studies, a significant increase in micronucleated erythrocytes was observed. The results of these studies suggest that HCC-230fa affects rapidly dividing cells and may have long-term consequences for occupational exposures.     

硝酸羟胺亚慢性染毒对大鼠骨髓嗜多染红细胞微核率的影响

背景与目的:研究硝酸羟胺亚慢性染毒对大鼠骨髓嗜多染红细胞微核率的影响.材料与方法:设置硝酸羟胺6.97、13.93、27.86 mg/kg 3个不同剂量组和生理盐水阴性对照组,间日腹腔注射大鼠,连续染毒90d后脱颈椎处死2/3实验动物;剩余1/3实验动物停止染毒再饲养30 d后同法处死;观察并统计大鼠骨髓嗜多染红细胞的微核率.结果:硝酸羟胺27.86 mg/kg组诱导的大鼠骨髓嗜多染红细胞微核率与阴性对照组、6.97mg/kg组相比差异均具有显著性;6.97和13.93 mg/kg组与阴性对照组比较差异无显著性;而恢复期各染毒组与阴性对照组相比差异均无显著性.结论:硝酸羟胺亚慢性染毒对大鼠具有一定的遗传毒性,在一定的时间内该毒性可能具有可逆性但无延迟性.     

骆驼蓬总碱抗肿瘤及其协同抗瘤作用

用小鼠移植性肿瘤和裸鼠异体移植人肿瘤模型研究骆驼蓬总碱（TAH）抗肿瘤作用及其协同抗瘤作用。结果表明：TAH对小鼠S-180、LⅡ、肝癌以及裸鼠移植人鼻咽癌（CNE2）和人（BEL-7402）肝癌均有抗肿瘤作用，并发现TAH与顺铂（DDP）或阿霉素（ADM）合用，具有协同抗肿瘤作用。     

金属离子对果胶酶的协同作用研究

以香蕉皮为原料,在固定pH值、温度、加热时间的实验条件下,采用酸醇沉淀的实验工艺,探讨不同金属离子(铜、铅、镁)及同种离子不同浓度对果胶提取率的影响,从而确定金属离子对果胶酶的协同作用。