摘要
目的探讨整合素αvβ6在移植肝胆管纤维化中的作用,为探索新的移植物胆管病(graft cholangiopathy,GC)防治策略提供理论基础。方法小鼠被随机分为假手术组、单纯移植组和药物干预组。假手术组不行肝移植,仅行开关腹;单纯移植组和药物干预组均采用非动脉化的小鼠原位肝移植模型,供肝冷保存时间均为12 h。术后按2、4、6周3个时相点采集血液及组织标本。采用荧光实时定量PCR法测定整合素αvβ6及基质金属蛋白酶-2(matrix metalloproteinase-2,MMP-2)、转化生长因子-1(transforming growth factor-β1,TGF-β1)、转化生长因子-2(transforming growth factor-β2,TGF-β2)和Ⅰ型前胶原(procollagenα1)等纤维化相关基因的mRNA表达,同时测定血清ALT、GGT、ALP以及肝组织羟脯氨酸(Hyp)的含量,并采用Masson三色染色法评估肝内胆管的纤维化情况。计量资料多组均数间的比较采用双因素方差分析,两总体均值的比较采用t检验,相关性分析采用直线回归分析。结果严重的冷保存再灌注损伤(cold preservation/reperfusion injury,CPRI)合并动脉血供障碍可以迅速地诱发移植肝胆管纤维化。在术后第4、6周,整合素αvβ6 mRNA表达分别上调至假手术组的13.2、32.5倍,肝组织Hyp含量较假手术组分别上升87.5%、124.4%。通过特异性抑制整合素αvβ6可以明显缓解移植肝胆管纤维化,同时在一定程度上改善肝功能。结论缺血再灌注损伤是造成移植肝胆管纤维化的重要原因之一,整合素αvβ6在这一过程中可能扮演着关键性的角色,针对整合素αvβ6的干预策略可能为GC的治疗提供新的思路。
Objective To investigate the role of integrin αvβ6 in the pathogenesis of biliary fibrosis following orthotopic liver transplantation （OLT）, and to explore the new therapeutic strategies for graft cholangiopathy （GC）. Methods A total of 45 male FVB/N mice were randomly divided into 3 groups, that is, sham-operation group （control group）, OLT group, and OLT with integrin αvβ6 antagonist EMD527040 group. The non-arterialized mouse OLT model with 12 hours＇ cold-preservation was used in the later 2 groups. The expression of αvβ6 and fibrosis-related transcriptors were determined by quantitative reverse-transcription polymerase chain reaction （ RT-PCR）, and serum levels of alanine aminotransferase （ ALT）, γ-glutamyltrans- ferase （GGT）, alkaline phosphatase （ALP） and liver hydroxyproline （Hyp） content were measured. Masson Trichrome staining were performed to determine the morphological changes of intrahepatic bile ducts. Results Cold preservation/reperfusion injury （CPRI） combined with hepatic artery occlusion induced biliary fibrosis of liver allograft in the mice. During the pathogenesis of biliary fibrosis, the mRNA expression of integrin αvβ6 was up-regulated to 13.2-fold and 32.5-fold higher than control in 4 and 6 weeks after transplantation, respec- tively. The relative hepatic Hyp content was increased by 87.5% and 124.4% at the same time points when compared with control. Inhibition of integrin αvβ6 significantly retarded biliary fibrosis progression and improved hepatic function following OLT. Conclusion Ischemia-reperfusion injury is the major cause of bile duct injury after OLT. Integrin ctv136 plays a key role in the pathogenesis of bile duct injury and repair, which might be a novel marker of biliary fibrosis and an attractive target for anti-fibrotic therapies in the treatment of GC following OLT.
出处
《第三军医大学学报》
CAS
CSCD
北大核心
2013年第7期644-648,共5页
Journal of Third Military Medical University
基金
国家自然科学基金(30901466)
重庆市自然科学基金(CSTC2009BB5331)
第三军医大学青年创新基金(2009XQN22)~~
