常山酮对大鼠心脏移植急性排斥反应的抑制作用

目的:研究常山酮(HF)对大鼠异基因心脏移植急性排斥反应的影响及相关机制。方法:SPF级近交系Wistar雌性大鼠30只,SD雌性大鼠60只,随机分为3组行颈部心脏移植手术:同系对照组15对(供、受体均为SD大鼠,术前第1天及术后均用生理盐水灌胃),排斥组15对(供体为Wistar大鼠,受体为SD大鼠,术前第1天及术后均用生理盐水灌胃)及HF治疗组15对(供、受体同排斥组,术前第1天及术后均用HF溶液灌胃)。移植后第3,5,7天每组各处死5只受体大鼠,留取移植心行组织病理学及分子生物学检测。结果:与排斥组相比,HF治疗组移植心存活时间延长,免疫排斥反应减轻,血清中白细胞介素-17(IL-17)呈低水平,供心心肌组织中相关细胞因子白细胞介素-6(IL-6)、白细胞介素-8(IL-8)及肿瘤坏死因子-α(TNF-α)呈低表达,具有统计学差异。结论:常山酮可以有效抑制大鼠心脏移植模型体内由于急性排斥反应引起的多种促炎性细胞因子的异常升高,具有抗急性移植排斥反应的潜能,其机制可能与抑制了Th17分泌IL-17减少相关。     

Development and validation of a cell culture based assay for in vitro assessment of anticryptosporidial compounds

In vitro culture of Cryptosporidium parvum oocysts in HCT-8 cells was combined with immunofluorescent labelling and digital image analysis to quantify the development of the parasite by detecting and measuring the labelled area in the respective cell cultures. The number of inoculated oocysts and the labelled area correlated reliably and significantly (R ², 0.98–0.99). The effects of various concentrations of halofuginone bromide (0.00039 to 50 μM) and monensin (0.00225 to 0.144 μM) on in vitro parasite development were determined in further trials in cultures inoculated each with 10⁵ oocysts. Monensin reduced the detected area in a dose-dependant manner. In comparison to the untreated controls, the area positive for C. parvum in the cultures treated with 0.144 to 0.009 μM monensin reached a maximum of 17%, and inhibition of 40% was observed at 0.0045 μM. Halofuginone bromide also efficiently inhibited parasite in vitro reproduction, albeit at higher concentrations. At 12.5 μM or more, inhibition was at least 90%; 0.05 μM still yielded 80% inhibition, whereas at concentrations below 0.00625 μM, labelled areas abruptly increased. Both drugs appeared efficient under in vitro conditions; the applied system is suited to screen drugs for their anti-cryptosporidial capacity.     

Halofuginone inhibits neointimal formation of cultured rat aorta in a concentration-dependent fashion in vitro

Summary Halofuginone, an anticoccidial quinoazolinone, can specifically inhibit collagen type α1 (I) synthesis and gene expression, and also inhibits cultured smooth muscle cell proliferation. The aim of this study was to investigate the effect of halofuginone on neointimal formation of rat aorta after culture in a concentration-dependent manner in vitro. Thoracic aorta of Wistar rats was removed and manipulated to damage the endothelium under sterile conditions, and culture for 15 days in halofuginone-free or halofuginone-added culture medium (n=20). Segments of cultured aorta were studied by histologic and immunohistochemical methods. Proliferation of neointimal layers consisting of loose multilayer cellular structure was observed in the halofuginone-free control group after 15 days of rat aorta culture, and neointimal formation was significantly decreased as an increasing concentration of halofuginone was added. As with precultured fresh aorta, no intimal proliferation was observed in the cultured segments of aorta with 500 ng/ml halofuginone added to culture medium. The proliferation of cell nuclear antigen index was significantly higher in the halofuginone-free control group than that in the halofuginone-added groups. The present results suggest that halofuginone can inhibit neointimal formation of rat aorta after culture in a concentration-dependent fashion in vitro.     

溴氯哌喹酮抑制角膜新生血管的实验研究

目的 观察溴氯哌喹酮(Halofuginone)对鼠角膜新生血管的抑制作用.方法 用BSS液配制500 μg/ml和1000 μg/ml的溴氯哌喹酮眼液和每公斤饲料含5mg溴氯哌喹酮(5mg/kg)的鼠颗粒饲料,制作碱性成纤维生长因子(bFGF)缓释颗粒.在7～9周龄的C57B1/6小鼠角膜板层隧道内植入bFGF缓释颗粒诱发角膜新生血管,分组滴溴氯哌喹酮眼液和自由进食溴氯哌喹酮饲料,裂隙灯下测量角膜新生血管长度和计算血管网面积,对比分析各时间点组间血管长度和面积.结果 角膜和前房无明显炎症反应.3d时从扩张增粗的角膜缘血管长出新生血管芽,5～7d长到颗粒处,血管网呈扇形.10d后新生血管开始消退,垂直走行的血管消退的最慢.食物组抑制新生血管最显著,与对照组相比差异有统计学意义,7d时血管长度和面积也明显小于眼药组,7d、10d眼药组血管长度和面积小于对照组,未显示与眼药剂量相关的抑制作用.结论 溴氯哌喹酮能抑制鼠角膜新生血管,于抑制I型胶元合成和沉积、MMP-2的表达、血管平滑肌、内皮细胞的增殖有关,口服优于滴眼液.     

Pharmacokinetic and tissue distribution of doxycycline in broiler chickens pretreated with either: Diclazuril or halofuginone

Following IV injection of doxycycline in a dose of 20 mg kg⁻¹ b.wt., its serum concentration was best fitted in two-compartment open model in chickens fed either on control or on anticoccidials-containing rations. Diclazuril and halofuginone resulted in a significant short distribution half-life (t_½α) (7.17 ± 0.39 and 11.88 ± 1.05 min, respectively) and increased total body clearance (Cl_tot) 0.37 ± 0.024 and 0.295 ± 0.034 L/kg/h, respectively. Following oral dosing the tested drug absorbed with t_½ab of 41.38 ± 1.6, 17.48 ± 0.86 and 41.83 ± 1.8 min, respectively and their C_max values (3.18 ± 0.18, 5.425 ± 0.48 and 0.986 ± 0.037 μg/ml) were attained at 2.07 ± 0.097, 1.403 ± 0.074 and 2.55 ± 0.106 h. For doxycycline alone and in presence of diclazuril and halofuginone, respectively. Systemic bioavailability was 22.64 ± 3.46, 86.74 ± 9.23 and 22.38 ± 3.09%, respectively. Following IM injection t_½ab were 9.096 ± 1.34 for doxycycline alone, 16.24 ± 2.21 and 15.6 ± 1.7 min in the presence of diclazuril and halofuginone, respectively. C_max was 3.10 ± 0.28, 4.63 ± 0.57 and 0.55 ± 0.07 μg/ml reached at 0.8 ± 0.083, 1.13 ± 0.126 and 1.21 ± 0.105 h. For the antibiotic alone, and in presence of either diclazuril and halofuginone, respectively. Systemic bioavailability was 22.41 ± 3.86, 88.97 ± 12.9 and 12.31 ± 0.99% in chickens fed on anticoccidial-free, diclazuril- and halofuginone-containing rations, respectively. Both the tested anticoccidials induced higher doxycycline tissue residues in all tested tissue samples.     

良性气道狭窄的药物治疗

气道损伤时,伤口愈合过程异常是导致肥厚性瘢痕形成和气道管腔狭窄的原因,愈合过程可分为3个阶段：炎症期、增生期及成熟期.针对不同愈合阶段,所应用的预防和治疗瘢痕形成及气道狭窄的药物是不同的.具体包括：抗生素和类固醇、丝裂霉素、紫杉醇、5-氟尿嘧啶/氟羟强的松龙结合物、卤夫酮、抗反流药物、免疫抑制剂、生长因子等.本文综述了上述药物在细胞、动物及临床试验中治疗气道狭窄的作用、安全性及应用前景.     

放射联合常山酮增强肺癌疗效及机制动物实验研究

目的 探讨放射联合常山酮对增强Lewis肺癌小鼠移植瘤疗效及对肿瘤侵袭转移的影响,并探索可能的分子机制。方法 将Lewis肺癌移植瘤小鼠随机分为空白对照组、常山酮组、放射组及联合组(9只/组),处理结束后第3天,每组随机处死4只小鼠行免疫组化及ELISA检查,剩余小鼠继续观察存活时间及肝、肺转移情况。组间差异采用方差分析。结果 处理第9天,对照组、常山酮组、放射组及联合组小鼠平均肿瘤体积分别为175.2、118.5、106.6、85.6 mm³(P=0.000)。4个组中位存活时间分别为39、61.5、78.5、84.5 d (P=0.002)。联合组肝肺转移数目显著最少。免疫组化及ELISA结果显示TGF-β₁水平放射组较对照组升高,联合组较对照组低;肿瘤平均血管密度计数联合组明显少于放射组。对照组及放射组胶原蛋白表达较高,常山酮组及联合组胶原蛋白表达较低。结论 放射联合常山酮可能会增强肿瘤治疗疗效,其潜在的机制需要进一步研究。     

Halofuginone inhibits LPS-induced attachment of monocytes to HUVECs

Atherosclerosis has become a major cause of mortality for several years, however the underlying mechanism of this disorder is still complicated. Endothelial dysfunction is a hallmark in the beginning of atherosclerosis. Lipopolysaccharides (LPS) is an important risk factor contributing to endothelial dysfunction. This study demonstrates that Halofuginone, an anti-malarial drug, possesses protective effects on human umbilical vein endothelial cells (HUVECs) against LPS-induced endothelial dysfunction. Through this study, we demonstrate that Halofuginone ameliorates LPS-induced attachment of THP-1 cells to HUVECs by inhibiting the expressions of adhesion molecules, including vascular cell adhesion molecule-1 (VCAM-1) and E-selectin. Halofuginone also suppresses the production of pro-inflammatory cytokines, including tumor necrosis factor α.(TNF-α), interleukin-1β (IL-1β), and interleukin 6 (IL-6). Furthermore, Halofuginone reduces the overproduction of reactive oxygen species (ROS) by regulating the expression of NADPH oxidase 2 (NOX-2). Mechanistically, we find the protective effects of Halofuginone depend on the expression of Kruppel-like factor 2 (KLF2), which is mediated by extracellular regulated protein kinases 5 (ERK5). Totally, our findings demonstrate that Halofuginone possesses a protective function in endothelial cells, indicating a therapeutic potential to modulate endothelial dysfunction in atherosclerosis.