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目的研究阻塞性胆汁淤积下肝脏中转录因子NR5A2、SP1表达是否上调,以及其表达上调与胆酸转运蛋白MRP3基因的表达是否密切相关;MRP3表达上调是否具有减轻阻塞性胆汁淤积肝脏损伤的作用。方法收集阻塞性胆汁淤积组(胆结石或肝内胆管结石引起的黄疸患者手术切除的肝脏)和正常对照组(排除肝脏疾病的活检肝组织及肝转移癌无黄疸的患者肝脏)肝脏样品各15例。采用半定量PCR和免疫荧光方法检测NR5A2、SP1基因的表达,利用独立样品t检验和线性回归分析阻塞性胆汁淤积肝组织样品中MRP3 mRNA表达上调是否与NR5A2或SP1呈正相关,以及MRP3表达上调与反映肝脏损伤的指标ALT、AST是否存在负相关性。HE染色观察胆汁淤积组肝细胞坏死程度和MRP3蛋白表达高低的关系。结果阻塞性胆汁淤积肝脏中NR5A2和SP1 mRNA表达显著上调,其中NR5A2 mRNA增高3.7倍(P<0.01),SP1 mRNA上升3.2倍(P<0.01)。免疫荧光结果表明,阻塞性胆汁淤积组NR5A2和SP1蛋白表达也显著增加,NR5A2或SP1蛋白与胆酸转运蛋白MRP3 mRNA表达呈显著正相关(分别为r2=0.47,P<0.05和r2=0.51,P<0.01);MRP3蛋白表达与肝细胞坏死程度存在密切相关,并且MRP3蛋白表达量与ALT和AST均呈显著负相关(分别为r2=0.52,P<0.01和r2=0.39,P<0.05)。结论人阻塞性胆汁淤积下NR5A2和SP1表达上调,可诱导胆酸转运蛋白MRP3的表达,而MRP3表达上调可能有减轻肝脏损伤的作用。
OBJECTIVE: To investigate whether there is an upregulation of transcription factor NR5A2 and SP1 in the liver after obstructive cholestasis and whether its expression is closely related to MRP3 gene expression. Whether upregulation of MRP3 can reduce liver injury in obstructive cholestasis. Methods The obstructive cholestasis group (liver resected in patients with jaundice caused by gallstones or intrahepatic bile duct stones) and normal control group (liver in biopsy liver patients without liver disease and those without jaundice in liver metastases) were collected. example. Semiquantitative PCR and immunofluorescence were used to detect the expression of NR5A2 and SP1 genes. The independent samples t test and linear regression were used to analyze whether the up-regulation of MRP3 mRNA in obstructive cholestatic liver tissue was positively correlated with NR5A2 or SP1 and MRP3 up-regulated The indicators of liver damage ALT, AST whether there is a negative correlation. The relationship between hepatocellular necrosis and MRP3 protein expression in cholestasis was observed by HE staining. Results The expression of NR5A2 and SP1 mRNA in obstructive cholestatic liver was significantly up-regulated. The mRNA expression of NR5A2 increased by 3.7-fold (P <0.01) and SP1 mRNA increased by 3.2-fold (P <0.01). The results of immunofluorescence showed that the expressions of NR5A2 and SP1 protein were also significantly increased in obstructive cholestasis group. The expressions of NR5A2 or SP1 protein and bile acid transporter MRP3 mRNA were positively correlated (r2 = 0.47, P <0.05 and r2 = 0.51, P <0.01). MRP3 protein expression was closely related to the degree of hepatocyte necrosis, and the expression of MRP3 protein was negatively correlated with ALT and AST (r2 = 0.52, P <0.01 and r2 = 0.39, . Conclusion The expression of NR5A2 and SP1 is upregulated in obstructive cholestasis, which can induce the expression of MRP3. However, the upregulation of MRP3 may reduce the liver injury.