TIPS术后再狭窄组织免疫组化分析

Immunohistochemical analysis of restenotic tissue after transjugular portosystemic shunt

目的探讨经颈静脉肝内门腔分流术(TIPS)后再狭窄多种组织成分的变化,为揭示TIPS分流道再狭窄的形成机制提供更多的信息.方法对6只25kg左右的家猪进行TIPS术,建成TIPS猪模型.14～21 d后处死,取出肝脏TIPS组织做病理检查,包括大体标本检查,电镜检查,病理切片分别用HE染色、免疫组化分析anti-SMC-α(抗平滑肌肌动蛋白-α)、细胞增殖核抗原(PCNA)、波形蛋白(vimentin)、肌球蛋白(myoglobulin)、eNOS(内皮型一氧化氮合酶)及iNOS(诱导型一氧化氮合酶)等多个指标表达.将TIPS再狭窄组织与通畅的支架通道组织进行上述多项指标的对比分析.结果TIPS术后2～3周处死时,6只猪有4只猪出现不同程度再狭窄,其中有2只TIPS通道完全堵塞.电镜检查,再狭窄组织含大量增殖的胶原纤维、大量平滑肌细胞与成纤维细胞;抗SMC-α、PCNA在再狭窄组织中表达为强阳性,在TIPS通道通畅的组织中,也有较强表达;波形蛋白在通畅的支架通道组织中表达为强阳性,而在支架再狭窄组织中表达明显减弱;肌球蛋白正好相反,在再狭窄组织中表达较强,而在通畅的支架组织中表达减弱;eNOS在正常肝脏组织中为阳性表达,靠近TIPS再狭窄组织,表达有减弱;而iNOS在正常肝脏组织中为表达弱阳性,但靠近TIPS再狭窄组织,表达明显增强.结论猪TIPS模型中,再狭窄中主要为抗SMC-α阳性的平滑肌细胞增殖,细胞增殖迁移能力强;正常肝脏组织中主要表达eNOS,而肝脏损伤后iNOS表达明显增强.

Objective To investigate the changes of several restenotic tissue elements after transjugular portosystemic shunt, and to provide more informations for the mechanism of TIPS restenosis. Methods TIPS was performed in 6 swine to set up TIPS animal models. 14-21 days after operation, the models were sacrificed to obtain the TIPS tissues for pathological examinations, including electric microscope, HE staining, and immunohistochemical staining of anti-SMC-actin-α, PCNA, Vementin, myoglobulin, eNOS and iNOS. Then, the results were cmparatively analized beween TIPS obstructed shunt tissues and non-obstructed shunt tissues. Results Restenosis was occurred with differnet degrees in 4 swine of the 6 TIPS models. Electric microscopic results showed that the restenosis tissues were composed of over proliferated collegen, SMCs and fibroblasts. Anti-SMC-actin-α and PCNA were strongly positive expression in restenotic tissues, and also positive in patent tissues. Vimentin expressed strongly in unstenotic tissues, on the contrary, it expressed obviouly weaker in restenotic tissues. Myoglobulin expressed more strongly in restenotic tissues and weakened in unstenotic tissues, eNOS expressed positive in normal liver tissues, and expressed weaker near TIPS restenotic tissues, iNOS showed stronger expression in restenotic tissues and could hardly expressed in normal liver tissues. Conclusions Restcnotic rate may be 67 % in TIPS swine models. Restenotic tissues may be mainly composed of proliferated SMCs positively expressed anti-SMC-actin-α with strong ability of movement, eNOS may be expressed in normal liver tissues and instead iNOS be expressed in strongly injured liver tissues.