Novel in vitro model for studying hepatic ischemia-reperfusion injury using liver cubes.
Novel in vitro model for studying hepatic ischemia-reperfusion injury using liver cubes.
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DOI：
10.1016/j.surg.2012.02.012
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发表时间：
2012-08
期刊：
影响因子：
3.8
通讯作者：
Anderson, Christopher D.
中科院分区：
文献类型：
--
作者：
DuBray, Bernard J., Jr.;Conzen, Kendra D.;Upadhya, Gundumi A.;Balachandran, Parvathi;Jia, Jianluo;Knolhoff, Brett L.;Alpers, David H.;Mohanakumar, Thallachallour;Chapman, William C.;Anderson, Christopher D.
虽然流入道阻断技术使外科医生能够进行日益复杂的肝脏切除术，但缺血-再灌注（IR）损伤仍然是发病率的来源。由于缺乏足够的临床前模型，改善IR损伤的努力受到阻碍。本研究的目的是开发一种简单，有效，成本效益高的方法来研究肝脏IR损伤。从正常（C57 BL/6）小鼠获得肝立方体。肝切除术后，取4 mm打孔活检，分别置于含有肝细胞培养基的培养威尔斯孔中。实验立方体经历缺氧60分钟，而控制保持常氧。在再扩散0、6和12小时后，从各个威尔斯孔收集上清液，用于转氨酶和细胞因子测量。对单个立方体进行组织学检查。与对照组相比，在实验立方体中观察到广泛的组织学损伤，分别在6小时和24小时对活化的caspase-3和TUNEL染色更强。与缺血性损伤一致的变化发生在肝脏立方体的中心，而再扩散损伤的标记物沿着周边被赞赏。与对照组相比，实验立方体中再扩散后6小时的转氨酶显著更高，p = 0.02。在12小时再扩散时，与对照组相比，实验立方体培养基中的TNF-α和IL-1β显著更高，p分别为0.05和0.03。在体外IR立方体产生一个显着的损伤，其模式是反映肝小叶结构。这种新技术可能为解偶联IR机制开辟新途径，同时促进快速筛选潜在的治疗方法。
While inflow occlusion techniques have given surgeons the ability to carry out increasingly complex liver resections, ischemia-reperfusion (IR) injury continues to be a source of morbidity. Efforts to ameliorate IR injury have been hindered in absence of adequate pre-clinical models. The goal of the present study was to develop a simple, efficient, and cost-effective means of studying hepatic IR injury. Liver cubes were procured from normal (C57BL/6) mice. Following hepatectomy, 4 mm punch biopsies were taken for individual placement in culture wells containing hepatocyte media. Experimental cubes underwent hypoxia for 60 minutes, while controls remained normoxic. Supernatants were collected from individual wells following 0, 6 and 12 hours of rediffusion for transaminase and cytokine measurement. Histologic examination was performed on individual cubes. Extensive histologic injury was seen in the experimental cubes compared to controls with greater staining for activated caspase-3 and TUNEL at 6 and 24 hours, respectively. Changes consistent with ischemic injury occurred more centrally in liver cubes whereas markers for rediffusion injury were appreciated along the periphery. Transaminases were significantly higher at 6 hours following rediffusion in experimental cubes compared to controls, p = 0.02. TNF-α and IL-1β were significantly higher in the media of experimental cubes compared to controls at 12 hours rediffusion, p = 0.05 and 0.03 respectively. In vitro IR of cubes produces a significant injury whose pattern is reflective of hepatic lobular architecture. This novel technique may open new avenues for uncoupling the mechanisms of IR while facilitating rapid screening of potential therapies.
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影响因子：
13.5
作者：
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