Hepatotoxicity of cantharidin is associated with the altered bile acid metabolism

Cantharidin (CTD) is an effective antitumor agent. However, it exhibits significant hepatotoxicity, the mechanism of which remains unclear. In this study, biochemical and histopathological analyses complemented with ultra-high-performance liquid chromatography–tandem mass spectrometry (UHPLC-MS/MS)-based targeted metabolomic analysis of bile acids (BAs) were employed to investigate CTD-induced hepatotoxicity in rats. Sixteen male and female Sprague–Dawley rats were randomly divided into two groups: control and CTD (1.0 mg/kg) groups. Serum and liver samples were collected after 28 days of intervention. Biochemical, histopathological, and BA metabolomic analyses were performed for all samples. Further, the key biomarkers of CTD-induced hepatotoxicity were identified via multivariate and metabolic pathway analyses. In addition, metabolite–gene–enzyme network and Kyoto Encyclopedia of Genes and Genomes pathway analyses were used to identify the signaling pathways related to CTD-induced hepatotoxicity. The results revealed significantly increased levels of biochemical indices (alanine aminotransferase, aspartate aminotransferase, and total bile acid). Histopathological analysis revealed that the hepatocytes were damaged. Further, 20 endogenous BAs were quantitated via UHPLC-MS/MS, and multivariate and metabolic pathway analyses of BAs revealed that hyocholic acid, cholic acid, and chenodeoxycholic acid were the key biomarkers of CTD-induced hepatotoxicity. Meanwhile, primary and secondary BA biosynthesis and taurine and hypotaurine metabolism were found to be associated with the mechanism by which CTD induced hepatotoxicity in rats. This study provides useful insights for research on the mechanism of CTD-induced hepatotoxicity.     

复方斑蝥胶囊对原发性肝癌患者免疫功能及临床疗效的影响

目的探讨复方斑蝥胶囊对原发性肝癌患者临床疗效及免疫功能的影响。方法选取2017年3月—2019年3月住院治疗的中晚期原发性肝癌患者共100例,按随机数字表法分为对照组与治疗组,每组各50例。对照组给予肝动脉插管栓塞化疗(Transcatheter arterial chemoembolization,TACE)治疗,治疗组在对照组治疗基础上给予复方斑蝥胶囊治疗。观察两组患者治疗前后临床效果、肿瘤标志物甲胎蛋白(A-fetoprotein,AFP)、癌胚抗原(Carcino-embryonic antigen,CEA)、糖抗原199(carbohydrate antigen 199,CA199)含量,免疫细胞因子血管内皮生长因子(vascular endothelial growth factor,VEGF)和缺氧诱导因子-1α(hypoxia inducible factor-1α,HIF-1α)水平及生活质量变化。结果治疗后,对照组总有效率为76%;治疗组总有效率为84%。治疗组总有效率明显高于对照组,差异有统计学意义(P<0.05)。治疗后对照组患者AFP、CA199含量降低,与治疗前比较,差异有统计学意义(P<0.05),CEA值虽有降低,但差异无统计学意义(P>0.05);治疗后治疗组患者血清中AFP、CEA、CA199含量显著降低,与治疗前比较,差异有统计学意义(P<0.05);且治疗组患者治疗后AFP、CEA、CA199含量降低程度优于对照组,差异有统计学意义(P<0.05)。两组患者治疗后血清中VEGF、HIF-1α含量均明显降低,与治疗前比较,差异有统计学意义(P<0.05);且治疗组患者血清中VEGF、HIF-1α含量降低优于对照组,差异有统计学意义(P<0.05)。治疗后治疗组KPS评分升高患者24例(48%),ZPS评分升高患者25例(50%),与对照组比较差异有统计学意义(P<0.05)。结论复方斑蝥胶囊联合TACE能显著提高原发性肝癌患者的临床效果,抑制肿瘤细胞的增殖,提高生存率,改善生活质量。     

去甲斑蝥素致大鼠腹腔炎症与多脏器功能失常综合征

目的：探讨去甲斑蝥素致大鼠失控炎症在多脏器功能失常综合征（ＭＯＤＳ）发病中的作用。方法：将去甲斑蝥素（一种能刺激皮肤发泡以收集巨噬细胞的致炎物质，１５ｍｇ／ｋｇ）注入６０只Ｗｉｓｔａｒ大白鼠腹腔，动态观察其对炎性介质以及器官形态和功能的影响。结果：与对照组比较，实验组动脉血氧分压（ＰａＯ２）和血小板计数明显下降，谷丙转氨酶（ＧＰＴ）和血肌酐（ＳＣｒ）明显升高，１２０小时后上述指标均接近对照组；１２小时和２４小时大鼠死亡率分别为１５％和２０％；肿瘤坏死因子（ＴＮＦ）和磷脂酶Ａ２（ＰＬＡ２）在注射后２４小时和１２小时达最高值；肺、心、肝、肾、肠等器官出现病理改变。结论：去甲斑蝥素可能通过激活巨噬细胞、释放ＴＮＦ等炎性介质诱发全身炎症和ＭＯＤＳ。     

斑蝥及其近缘种属药用资源研究进展

作者综述了近年来斑蝥及其近缘种属药用资源的研究进展.以有代表性的论文为依据,进行分析、整理和归纳.含斑蝥素昆虫,除芫菁科7属45种外,还有同翅目蜡蝉科的鼻蜡蝉属Zanna、蜡蝉属Fulgora、翅蜡蝉属Lycorma,鞘翅目拟天牛科中Oxocopis属、Heliocis属、沃黄拟天牛属Xanthochroa,曹氏拟天牛属Oedemera等7个近缘属9种.斑蝥素药用新资源丰富,斑蝥素生物合成具有生物亲缘关系,斑蝥素形成与生态有一定关系,活体提取等斑蝥资源利用的新方法受到重视.     

叶酸修饰的斑蝥素/黄芩苷共载脂质体处方工艺优化及其评价

目的 筛选二硬脂酰基磷脂酰乙醇胺-聚乙二醇2000-叶酸（DSPE-PEG 2000 folate,DSPE-PEG_2k-FA）修饰的斑蝥素/黄芩苷共载脂质体（DSPE-PEG_2k-FA-cantharidin & baicalin-lipsomes,FA-Can&Bai-Lips）的处方工艺并对其进行表征及评价。方法 由薄膜分散超声法制备FA-Can&Bai-Lips,以药脂比、胆脂比、水化温度及水化时间为考察因素,以两药包封率、粒径、多分散指数（polydispersity index,PDI）和ζ电位为评价指标,采用单因素考察和Box-Behnken设计-响应面法（Box-Behnken design-response surface methodology,BBD-RSM）优化其处方工艺。采用电位粒径仪和透射电子显微镜（TEM）分别测定其粒径、ζ电位及形态。通过溶血性实验、细胞摄取实验、CCK-8法实验及活体成像等方法进行安全性、靶向性评价。结果 FA-Can&Bai-Lips的最佳制备工艺为药脂比1∶11、胆脂比1∶6、DSPE-PEG_2k-FA用量为10%、溶剂为50 mL、旋转蒸发及水化温度为55 ℃、水化介质为pH 6.8 PBS缓冲液、水化体积为10 mL、水化时间为94 min,超声时间为8 min。经BBD-RSM优化制得的FA-Can&Bai-Lips形态规整,为类球形粒子,稳定性及安全性好,斑蝥素与黄芩苷的包封率分别为（90.19±0.67）%、（81.24±0.72）%,粒径为（172.47±4.83）nm,PDI为0.268±0.018,ζ电位为（−1.07±0.07）mV。体外评价结果表明,FA-Can&Bai-Lips能有效抑制HepG2细胞增殖,具良好的肝肿瘤靶向性。结论FA-Can&Bai-Lips具有增效减毒作用,能高效靶向肿瘤组织,是一种极具开发潜力的抗肝肿瘤纳米递送系统。     

毛细管气相色谱法测定抗鼻咽癌口服液中斑蝥素的含量

目的:建立了抗鼻咽癌口服液中斑蝥素的含量测定方法。方法:选用DB-5弹性石英毛细管柱(30 m×0.25 mm,0.25μm);检测器为FID检测器;柱温为150℃,保持25 min;气化室温度为250℃;检测器温度为320℃;载气为氦气;柱头压力为140 kPa;分流比为30∶1。结果:斑蝥素在0.02~0.24μg范围内呈良好的线性关系,r=0.999 8(n=6);样品的平均加样回收率为97.24%,RSD=1.35%(n=6)。结论:本法简便、快速,可用于抗鼻咽癌口服液的质量控制。     

加温合并喜树碱、斑蝥素对荷Lewis肺癌小鼠的作用

本文探讨了加温合并喜树碱、斑蝥素对荷Ｌｅｗｉｓ肺癌ＮＩＨ小鼠的抗肿瘤作用以及热耐受性对其作用的影响。结果：①喜树碱１６ｍｇ／ｋｇ或斑蝥素０．４５ｍｇ／ｋｇ合并２５ｍｉｎ加温能抑制肿瘤生长，在４１～４３℃范围内随温度的升高而加强；②在加温４３℃，２５ｍｉｎ时，加温合并喜树碱１２ｍｇ／ｋｇ或斑蝥素０．４５ｍｇ／ｋｇ产生相加作用；③热耐受性在４３℃，２５ｍｉｎ加温后迅速产生，２４ｈ达高峰而后递减；热耐受性极大地影响了喜树碱和斑蝥素的抗癌效果。     

Antimikrobielle Effekte des in vitro simulierten Ketoconazol-Kantharidinblasenflüssigkeitsspiegel-Profils bei Candida albicans*,**/Antimicrobial Effects of Ketoconazole Concentration Profiles in Cantharidin Blister Fluid,Simulated in Vitro,on Candida albicans

Zusammenfassung: Candida albicans (Isolat Nr. 5125/85) wurde zum einen in Sabouraud-Glukose-Bouillon, zum anderen in einem flüssigen synthetischen Aminosäuremedium gegenüber ständig schwankenden Konzentrationen von Ketoconazol exponiert. Zugrundegelegt wurden die Profile der freien wie die der Gesamtkonzentrationen, die beim Menschen nach einmaliger oraler Gabe von 200 mg in kutaner Kantharidinblasenflüssigkeit gefunden wurden. Parallel zu jedem Versuchsansatz mit Ketoconazol wurde jeweils ein Ansatz ohne Wirkstoff mitgeführt. Bei Verwendung des ersteren, die Hefeform begünstigenden Mediums fand sich nur eine relativ geringe Wachstumshemmung. In dem die Pseudomyzelform begünstigenden synthetischen Medium war demgegenüber — konzentrationsabhängig — ein deutlicher antimikrobieller Effekt zu erkennen (Keimdichtenzunahme in Gegenwart der Gesamtkonzentration auf das 2, lfache, im Kontrollversuch auf das 15, 3fache). Die größere Wirksamkeit von Ketoconazol auf die Pseudomyzelform von Candida albicans wird wiederum bestätigt, zugleich wird die geringere Sicherheitsmarge bei der systemischen Behandlung mit Azolen gegenüber der antimikrobiellen Chemotherapie mit Antibiotika deutlich. Untersuchungen vom vorliegenden Typ — wie sie in der klinischen Bakteriologie schon öfter durchgeführt wurden — könnten auch bei der Untersuchung neuer Antimykotika zunehmend an Bedeutung gewinnen. Summary: Candida albicans was exposed to continously changing concentrations of Ketoconazole in, on the one hand, Sabouraud Dextrose Broth and, on the other hand, a liquid synthetic amino-acid medium. The profiles of the free and total concentrations found in cutaneous cantha-ridin blister-fluid in humans after a single oral dose of 200 mg were used as basic information. For every experiment with ketoconazole, an identical experiment without addition of an antimycotic was carried out Using the first, yeast-promoting medium, only a relatively small growth-inhibition was evident. Using the synthetic amino-acid medium, which promotes the pseudomycelial form, however, a marked, concentration-dependent, antimicrobial effect was observed. (The colonial density of Candida albicans increased 2.1 times in the presence of the total ketoconazole concentrations, compared with 153 times in the control experiment.) The greater effectiveness of ketoconazole against the pseudomycelial form of Candida albicans is again confirmed. At the same time, the smaller safety margin associated with systemic treatment with azoles as compared with anti-microbial therapy with antibiotics, is apparent. Investigations of the present type, which have often been carried out in clinical bacteriology, may also gain significance in the investigation of new antimycotics.     

复方斑蝥注射液联合放化疗治疗局部晚期鼻咽癌的疗效及不良反应

目的:探讨复方斑蝥注射液联合放化疗在局部晚期鼻咽癌治疗中的应用价值。方法:以我中心2015年7月至2017年4月收治的157例初治局部晚期鼻咽癌患者为研究对象,将其随机分为试验组79例与对照组78例。试验组患者采用复方斑蝥注射液联合同步放化疗,对照组患者行常规同步放化疗治疗。收集临床资料,比较两组患者治疗期间相关不良反应发生率、机体免疫功能变化和近期疗效情况。结果:试验组与对照组的CR、PR、SD、PD差异无统计学意义（P=0.269＞0.05）,近期有效率（CR+PR）差异存在统计学意义（P=0.001）,试验组较优于对照组;进一步分析两组患者的4级不良反应发生率存在统计学差异（P＜0.05）,两组患者治疗前后CD3+、CD4+、CD4+/CD8+、CD8+、CD19+淋巴细胞计数下降,试验组较对照组下降趋势缓和。结论:局部晚期鼻咽癌在规范抗肿瘤治疗过程中给予复方斑蝥注射液治疗可降低患者治疗期间严重不良反应的发生率,对机体免疫功能可能有一定的保护作用,并在一定程度上可辅助提高疗效,需开展多中心、大样本临床研究进一步证实。     

基因芯片技术分析斑蝥素对肝癌细胞细胞毒作用的分子机制

目的：研究抗癌药斑蝥素对肝癌细胞细胞毒作用的分子机制。方法：应用基因芯片技术检测12．5μmol／L斑蝥素作用于肝癌QGY7703细胞24h后基因表达谱的变化。结果：斑蝥素抑制细胞表达参与细胞周期进程基因(如p27、ref—1、DNA、polymerase delta、XRCC9等)、能量代谢基因(如malate dehydrogenase,ADP/ATP translocase等)、致瘤活性基因(如c-myc,tre等)以及肿瘤特异表达基因(如bladder cancer related protein等)。相反，斑蝥素促进了多种细胞生长抑制基因(如BCRA2,BTG2,dual-specificity protein phosphatase等)以及凋亡相关基因(如ATL-derived PMA-responsive peptide等)的表达。结论：斑蝥素改变调控细胞周期进程、细胞增殖、能量代谢以及凋亡级联反应的基因表达可能是其细胞毒作用的机制。