【摘要】 目的:研究不同白酒引起的酒精性肝纤维化、肝硬化合并肝癌大鼠肝组织中P53基因突变及其相关ASPP蛋白家族的表达。方法:运用A、B(分为低剂量D和高剂量G)2种白酒和黄曲霉素B1(AFB1)对大鼠进行处理,制备酒精性肝纤维化、肝硬化合并肝癌大鼠模型,用PCR及DNA测序技术检测大鼠肝脏组织中P53基因第5、6、7、8外显子的突变情况;用免疫组织化学Envision法检测大鼠肝脏组织中P53、P53凋亡刺激蛋白1(ASPP1)、ASPP2、ASPP家族抑制因子(iASPP)的表达。结果:成功制备了酒精性肝纤维化、肝硬化合并肝癌大鼠模型,不同白酒诱导大鼠肝纤维化、肝硬化合并肝癌的程度不同。免疫组化检测结果发现,P53及iASPP表达的阳性率在白酒B组(BD组和BG组)明显高于白酒A组(AD组和AG组),差异有统计学意义(P<0.05);ASPP1、ASPP2的阳性率在白酒B组(BD组和BG组)明显低于白酒A组(AD组和AG组),差异有统计学意义(P<0.05);iASPP表达量高的酒精性肝病和肝癌中,ASPP1、ASPP2表达量降低,差异有统计学意义(P<0.05);P53基因的突变检测显示P53的突变方式以碱基插入为主,其次为碱基突变及缺失;AD组和AG组突变率为42.3%,BD组和BG组突变率为78.3%,差异具有统计学意义(P<0.05)。结论:不同白酒对大鼠肝脏肝纤维化、肝硬化和肝硬化并肝癌的诱发程度不一样,可能与其对P53基因突变及P53相关蛋白(ASPP1、ASPP2、iASPP)表达影响不同有关。更多还原

【Abstract】 Objective: To explore of the expression of P53 gene and ASPP protein family in liver tissue of rats with hepatic fibrosis,hepatocirrhosis and hepatocirrhosis with hepatoma. Methods: The hepatic fibrosis,hepatocirrhosis combined with hepatoma rat model was established by using aflatoxin B1( AFB1) and 2 kind of liquor( liquor A and B with low and high content) respectirely. Using immunohistochemistry to detect rat liver tissue P53,ASPP1,ASPP2,iASPP expression. Application of PCR technology to test rat liver tissue P53 gene exon 5,6,7,8 expression. Results: The positive reaction rates of P53 and iASPP in group B were higher than those of group A,difference had statistical significance( P < 0. 05). The positive reaction rates of ASPP1 and ASPP2 in group B were lower than those of group A,difference had statistical significance( P < 0. 05). In hepatic fibrosis,hepatocirrhosis and hepatocirrhosis with hepatoma,iASPP highly expressed,and ASPP1,ASPP2 lowly expressed,difference had statistical significance( P < 0. 05). P53 gene mutation detection showed that the main mutation way of P53 gene exon 5,6,7,8 was bases insert,followed by bases mutations and missing. group A mutation rate was 42. 3%,group B mutation rate was 78. 3%,difference had statistical significance( P <0. 05). Conclusions: Different liqueur has diverse inducing effects on hepatic fibrosis,hepatocirrhosis and hepatocirrhosis with hepatoma,which may relate to their different influence on P53 gene mutation and P53 associated protein expression.更多还原