大鼠肝脏缺血再灌注对胆小管纤维形肌动蛋白微丝的影响_临床医学论文.docVIP

大鼠肝脏缺血再灌注对胆小管纤维形肌动蛋白微丝的影响_临床医学论文 大鼠肝脏缺血再灌注对胆小管纤维形肌动蛋白微丝的影响_临床医学论文 作者：黎一鸣，刘继东，吉鸿，李顺乐 【摘要】 目的 探讨大鼠肝缺血再灌注(I/R)对胆小管纤维形肌动蛋白(F微丝的影响。方法 采用肝缺血35min再灌注模型，检测血清中ALT、AST、GGT和TBIL的含量；电镜观察胆小管的变化；异硫氢胍荧光素标记的鬼笔环肽(FITC显示胆小管F微丝的分布变化，激光共聚焦显微镜采集图像并定量分析。结果 F微丝荧光变化和电镜超微结构的改变相符合，Factin的荧光染色再灌注前正常，而再灌注后明显减弱；胆小管微绒毛再灌注前没有消失，而再灌注后却大量脱落。这与血清GGT和TBIL的异常改变一致。结论 再灌注而非缺血可造成胆小管F微丝破坏、微绒毛丧失，导致胆小管收缩减弱，胆汁排泄功能受损。这可能是大鼠肝I/R后胆汁淤积发生的主要机制。 【关键词】 肝缺血再灌注；胆汁淤积；纤维形肌动蛋白F ABSTRACT: Objective To investigate the effect of hepatic ischemia(I/R)on bile canalicular Factin microfilaments in rat. Methods A model of rat hepatic ischemiareperfusion was adopted and the ischemia time was 35min. The levels of serum ALT, AST, GGT, and TBIL were measured. The changes of bile canaliculus were observed by transmission electron microscope. The modification of Factin microfilaments was quantified by using FITCPhalloidin and analyzed by confocal laser scanning microscopy imaging. Results This modification of Factin staining was consistent with the observation by transmission electron microscopy. The staining of Factin was normal in hepatocytes before reperfusion, but decreased significantly after reperfusion, and there was a marked loss of canalicular microvilli after reperfusion, which were in coincidence with abnormality of serum GGT and TBIL. Conclusion Reperfusion not ischemia induced disruption of Factin microfilaments and a loss of microvilli, and these modifications could lead to impaired bile excretion by damaging canalicular contraction, which maybe the main mechanism of cholestasis after rat hepatic ischemiareperfusion. KEY WORDS: hepatic ischemia reperfusion; cholestasis; Factin 肝脏缺血再灌注后常有胆汁淤积的发生[1，其机制尚未完全了解。F微丝沿质膜分布，尤其在胆小管区域，它对胆小管收缩引起胆汁排泄起重要作用[4]。目前，国内尚无对肝缺血再灌注时胆小管F微丝变化的研究报道。本实验通过异硫氢胍荧光素标记的鬼笔环肽显示大鼠肝脏胆小管F微丝在经历缺血再灌注后的变化，用激光共聚焦显微镜采集图像并定量分析，以期探讨上述胆汁淤积发生的机制。 1 材料与方法 1.1 材料 雄性SD大鼠36只，体重200～250g，由西安交通大学医学院实验动物中心提供。异硫氢胍荧光素标记的鬼笔环肽(FITC和抗荧光衰减封片剂均购自Alexis公司。日立HITACHI公司H透射式电子显微镜由西安交通大学医学院电子显微镜中心提供。激光共聚焦显微镜(型号：TCS Sp2)