亲水性胆盐对减轻大鼠移植肝肝内胆管冷保存再灌注损伤的作用

Effect of hydrophilic bile salts alleviating cold preservation-reperfusion injury of graft intrahepatic bile duct in rats

目的探讨亲水性胆盐对减轻大鼠移植肝肝内胆管冷保存再灌注损伤的作用。方法应用大鼠原位肝移植胆道外引流模型，将192只SD大鼠随机分为4组，每组供、受者各24只。各组供者在供肝切取前30min经阴茎背静脉注射不同的试剂，对照组注射1ml生理盐水；疏水性胆盐（TDc）组注射牛磺脱氧胆酸钠40μmol／kg；高、低浓度亲水性胆盐（TUDC）组分别注射牛磺熊去氧胆酸钠8（）和40μmol／kg。供肝取出后置于4℃的平衡液中保存2h再植入受者。移植肝再灌注后1、3、7和（或）14d时，采用全自动生化分析仪分别检测受者血清中碱性磷酸酶、γ-谷氨酰转移酶、胆红素总量、直接胆红素和胆汁中y谷氨酰转移酶和葡萄糖的含量。移植肝再灌注后1d时，在光镜下观察肝内胆管上皮细胞的病理学改变，在荧光显微镜下观察肝内胆管上皮细胞的凋亡情况。结果高、低浓度TUDC组各项观察指标均低于对照组和TDC组（P（0．05），且肝内胆管炎症细胞的浸润、上皮细胞的损伤以及细胞的凋亡等程度均轻于对照组和TDC组（P（0．05）；与低浓度TUDC组比较，高浓度亲水性胆盐组短期内可见各项观察指标均有所降低，但远期各指标的比较，差异无统计学意义（P〉0．05）。TDC组术后各项观察指标均高于对照组（P〈0．05）。结论在供肝获取前，供者静脉注射亲水性胆盐能减轻大鼠移植肝肝内胆管的冷保存再灌注损伤。

Objective To expore the effect of hydrophilic bile salts on preservationreperfusion injury of graft intrahepatic bile duct in rats. Methods Using the model of orthotopic liver transplantation with bili- ary extra-drainage, 192 SD rats were randomly allocated into 4 groups according to different injection Control group was injected with normal sodium （1 ml）, hydrophobic bile salt group with taurodeoxycholate （TDC, 40 μmol/kg）, high and low dose hydrophilic group with tauroursodeoxycholate （TUDC） 80 and 40 μmol/kg respectively. After 2 h of preservation in Ringer＇s （LR） solution, orthotopic implantation of the graft liver was performed. The levels of serum total bilimbin （TB）, direct bilimbin （DB）, alkaline phosphatatase （AKP）, gamma glutamyl transferase （C, GT）, and bile C, GT, glucose （Glu） were assayed at day 1, 3, 7 or 14 post-operation. The morphological changes and apoptosis of intrahepatic bile duct epithelial cells were observed under light microscopy and fluorescence microscopy respectively at day 1, Results In high and low TUDC groups, all the indexes were decreased significantly （P〈O, 05）, and morphologic observation confirmed that infiltration of inflammatory cells, apoptosis and injuries of graft intrahepatic bile duct epithelial cells wcre ameliorated significantly as compared with the control and TDC groups （P〈O, 05）, In short term, all the indexes were decreased in high dose TDC group as compared with low dose TDC group, but in long term. there was no significant difference between high and low dose TDC groups （P〉0. 05）. All the indexes were increased significantly in TDC groups as compared with control group after operation （P〈0. 05）. Conclusion Hydrophilic bile salt can protect graft intrahepatic bile duct from cold ischemia and reperfusion injury.