肝缺血再灌注对大鼠胆管f -肌动蛋白微丝的影响

Yiming Li, Hua Li, Jidong Liu, Hong Ji
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摘要

目的探讨肝缺血再灌注(I/R)对大鼠胆管f -肌动蛋白微丝的影响。方法采用大鼠肝缺血再灌注模型,缺血时间为35 min,测定血清丙氨酸转氨酶(ALT)、天冬氨酸转氨酶(AST)、γ-谷氨酰转氨酶(GGT)活性及总胆红素(TBIL)水平。透射电镜观察胆管的变化。用FITC-Phalloidin定量分析F-actin微丝的修饰,并用激光共聚焦扫描显微镜成像分析。结果F-actin染色结果与透射电镜观察结果一致。肝细胞再灌注前F-actin染色正常,再灌注后明显降低,再灌注后小管微绒毛明显缺失,与血清GGT、TBIL异常相吻合。结论再灌注引起的是f -肌动蛋白微丝的破坏和微绒毛的丢失,而非短期缺血。这些修饰可通过破坏肝小管收缩导致胆汁分泌受损,可能是大鼠肝缺血再灌注后胆汁淤积的主要机制。
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Effect of hepatic ischemia-reperfusion on bile canalicular F-actin microfilaments in rats

Objective

To investigate the effect of hepatic ischemia-reperfusion(I/R) on bile canalicular F-actin microfilaments in rats.

Methods

A rat model of hepatic ischemia-reperfusion was employed and the ischemia time was 35 min. The activity of serum alanine aminotransferase (ALT), aspartate aminotransferase(AST), γ-glutamyl transferase(GGT) and the level of total bilirubin(TBIL) were measured. Changes in the bile canaliculi 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

Modifications of F-actin staining were consistent with the observations made 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 coincided with abnormal serum GGT and TBIL levels.

Conclusion

Reperfusion, not short-term ischemia, induced a disruption of F-actin microfilaments and a loss of microvilli. These modifications could lead to the impaired bile secretion by damaging canalicular contraction, and could be the main mechanisms of cholestasis after hepatic ischemia-reperfusion in rats.

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