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目的研究肝胆汁酸合成及转运异常在结肠炎性胆汁淤积发生中的作用及其机制。方法按照体重将Wistar雄性大鼠随机分成2组:正常组和模型组。模型组大鼠给予2,4,6-三硝基苯磺酸结肠灌注制备溃疡性结肠炎(UC)模型,正常组大鼠结肠给予0.9%Na Cl。考察动物的病变活动指数、病理组织评分及髓过氧化物酶活性;测定动物的血清碱性磷酸酶(ALP)、谷氨酰转肽酶(GGT)、谷草转氨酶(AST)和谷丙转氨酶(ALT)含量;以液质联用技术测定肝总胆汁酸、脱氧胆酸及石胆酸含量;用免疫印迹技术测定肝中胆汁酸合成限速酶胆固醇7α羟化酶(Cyp7a1)及胆汁酸外排转运体多药耐药性相关蛋白2(Mrp2)的表达。结果与正常组比较,模型组病变活动指数、病理组织评分及髓过氧化物酶活性分别为4.51±1.49,(1.36±0.69)分和(0.40±0.07)U·mg~(-1),差异均有统计学意义(均P<0.05),说明UC模型制备成功。模型组大鼠肝总胆汁酸及脱氧胆酸含量分别为(85.50±18.60)μmol·L~(-1)和(79.53±32.99)ng·g~(-1),血清ALP和GGT水平分别为(259.43±58.58),(1.50±0.68)U·L~(-1),与正常组比较差异均有统计学意义(均P<0.05),提示模型组大鼠存在肝内胆汁淤积。与正常组相比,模型组大鼠肝Cyp7a1的表达灰度值为0.72±0.07;而转运体Mrp2的表达灰度值为0.66±0.04;与正常组比较差异均有统计学意义(均P<0.05)。结论模型组大鼠存在明显的肝内胆汁淤积,其发生与肝胆汁酸合成增多及胆汁酸外排减少有关。
Objective To study the role of hepatobiliary acidosis and dysregulation in the pathogenesis of colitis cholestasis and its mechanism. Methods Wistar male rats were randomly divided into 2 groups according to body weight: normal group and model group. Rats in the model group were given ulcerative colitis (UC) model by intraperitoneal injection of 2,4,6-trinitrobenzene sulfonic acid, and normal rats were given 0.9% NaCl. The animal lesion activity index, histopathological score and myeloperoxidase activity were measured. The serum levels of ALP, GGT, AST and ALT ALT). The levels of total bile acid, deoxycholic acid and lithocholic acid were measured by LC / MS. The hepatic bile acid biosynthesis rate-limiting cholesterol 7α-hydroxylase (Cyp7a1) and bile acid Expression of multidrug resistance - associated protein 2 (Mrp2) in. Results Compared with the normal group, the index of pathological changes, pathological score and myeloperoxidase activity of the model group were 4.51 ± 1.49 and 1.40 ± 0.07 (1.40 ± 0.07) U · mg -1, respectively All were statistically significant (P <0.05), indicating UC model was successfully prepared. The levels of total bile acid and deoxycholic acid in the model group were (85.50 ± 18.60) μmol·L -1 and (79.53 ± 32.99) ng · g -1, respectively. The serum ALP and GGT levels were (259.43 ± 58.58) and (1.50 ± 0.68) U · L ~ (-1), respectively, which were significantly different from the normal group (all P <0.05), suggesting that there was intrahepatic cholestasis in the model group. Compared with the normal group, the gray value of Cyp7a1 expression in the model group was 0.72 ± 0.07, while the Mrg2 expression level was 0.66 ± 0.04. The difference was statistically significant compared with the normal group (all P < 0.05). Conclusion There is obvious intrahepatic cholestasis in the model group, which is related to the increased synthesis of hepatobiliary acid and the decrease of bile acid efflux.