胆道闭锁肝纤维化中TGF-β信号通路机制及体内抑制研究

Recently, the incidence of biliary atresia (BA) increases year by year, its treatment kasai-portoenterostomy only can improve clinical symptoms. Gradual progress until cirrhosis of liver fibrosis endanger children life and health which are the problems of clinical doctors baffled. The TGF - beta 1 level in liver biopsy in children with BA progress gradually with liver fibrosis increase which domestic and foreign study found. But to the place and point of TGF - beta 1 secretion the researchers each have different or even contrary studies. By collecting a large number of literature and the preliminary experiment, declarant found TGF - beta 1 level change law is closely related to liver fibrosis development in different periods and speak out that block TGF - beta 1 / Smad2/3 promote fibrosis pathway is the key to interfere with BA liver fibrosis. Based on the previous work, declarer put forward to take TGF - beta 1 / Smad2/3 profibrogenic pathway as the research direction in this issue, detection of liver tissue in children with BA in-depth and comprehensive study with the technology of BD-Pharmingen Cytometric Bead Array and so on to explore the effect of TGF - beta 1 profibrogenic pathway to BA liver fibrosis, to establish the kind of theoretical basis of the effect of TGF - beta 1 profibrogenic pathway in BA liver fibrosis.

近来胆道闭锁（BA）发病率逐年增加，其治疗手段Kasai术仅改善临床症状，逐渐进展的肝纤维化直至肝硬化危及患儿生命健康。国内外研究发现BA患儿肝活检TGF-β1水平随肝纤维化进展增加，但研究者对TGF-β1分泌部位和作用点的研究各有差别。通过搜集大量文献和前期实验，申报者发现TGF-β1水平变化规律与肝纤维化进展不同时期密切相关，认为阻断TGF-β1/ Smad2/3促纤维化通路是干扰BA肝纤维化的关键。本课题中申报者提出以TGF-β1促纤维化通路整体为研究方向，应用流式液相多重蛋白定量等技术对BA患儿肝组织进行深入、全面研究，建立BA肝内TGF-β1促纤维化通路作用机制的理论基础。

胆道闭锁（BA）患儿Kasai术后持续进展的肝纤维化是影响患儿生命健康的关键。目前国内外其他疾病的研究已明确TGF-β信号通路在纤维化发展中起重要作用，但在BA中如何作用不明。. 本课题就以下方面着手：①BA中TGF-β1 信号通路相关蛋白的表达情况②各种激酶及调节因子对 TGF-β通路的调节作用③通路各关键蛋白在BA 肝纤维化分级中的表达④分析自体肝生存时间较长患儿肝脏特点。. 本研究采用免疫组化、PCR、Western-Bloting、流式蛋白定量等技术方法，对BA肝组织中 TGF-β通路相关蛋白进行检测，得出以下结果：制定了针对胆道闭锁的肝纤维化分级； TGF-β1和Smad2蛋白及mRNA在BA早期肝纤维化中高表达，两者协同促进BA 早期肝纤维化形成；Smad3、P-Smad3、PAI-1在肝纤维化早期大量增多，PAI-1与P-Smad3的含量及变化幅度相符；整合素αvβ8、P38、ERK1/2表达增多并可相互作用激活TGF-β1信号通路；JNK2、TIMP-1及CollagenⅢ在BA中随肝纤维化程度加重而表达升高，且对BA肝纤维化起促进作用；④TGF-β1信号通路相关蛋白在肝纤维化3级时表达量最高；⑤Kasai可延缓肝纤维化进程，手术日龄及术后胆管炎发作的频次，是影响术后肝脏病理及自体肝生存时间的重要因素；TGF-β1信号通路在肝纤维化晚期肝移植时，表达量减少，促纤维化作用减轻。. 本研究通过探讨 BA中TGF-β 促纤维化通路机制，建立阻断 TGF-β 促纤维化通路的理论依据，以期能够抑制BA患儿肝纤维化进展，提高BA患儿的自体肝生存时间。