胆管损伤后良性狭窄形成过程中c-FLIP的表达及意义

Expression of c-FLIP During Formation of Biliary Benign Stricture after Bile Duct Injury and Its Significances

目的观察凋亡抑制因子人类细胞同源FLICE样抑制蛋白(cellular homolog FLICE-like inhibitoryprotein,c-FLIP)在胆管良性狭窄形成过程中的表达和定位情况,并探讨其意义。方法利用原位杂交法分别对15只犬(实验组)胆管损伤后2、3、4、5、6个月及其配对的15只假手术组犬的吻合口组织中c-FLIP表达和定位情况进行分析,计算每张切片的平均积分光密度值,分别比较c-FLIP在实验组及其配对的假手术组的胆道吻合口间质组织与腺体组织中表达的差异。结果实验组各时相的间质组织中c-FLIP均呈强阳性表达,以纤维细胞胞浆表达为主,而腺体组织很少表达或几乎无表达;在相应时相的间质组织中的表达明显强于在腺体组织中的表达(P<0.05);间质组织或腺体组织中c-FLIP表达在不同时相之间差异均无统计学意义(P>0.05)。假手术组各时相的间质组织和腺体组织中c-FLIP均呈弱阳性表达;在相应时相的间质组织和腺体组织中c-FLIP表达差异无统计学意义(P>0.05);间质组织或腺体组织中c-FLIP表达在不同时相之间差异均无统计学意义(P>0.05)。c-FLIP在实验组间质组织中的表达明显强于相应时相的假手术组(P<0.05);而c-FLIP在腺体组织中的表达2组间差异无统计学意义(P>0.05)。结论胆管损伤后,吻合口间质组织内的c-FLIP呈持续表达状态,由此所造成的细胞凋亡持续受阻效应可能与胆管良性狭窄的形成关系密切。

Objective To observe the expression and localization of cellular homolog FLICE-like inhibitory protein（c-FLIP）in the procedure of benign biliary stricture formation and discuss the significances.Methods The method of in situ hybridization was used in anastomotic tissues from 15 dogs（experimental group）in 2,3,4,5,6 months after bile duct injury and 15 matching sham operation dogs（sham operation group）for analyzing the expression and localization of c-FLIP and calculating the average integrated optical density of each slice.Stain cells were counted under the magnification field（×400）and at least 5 fields per slice were examined.The cells stained red in the nuclei and（or）the cytoplasm were positive cells.The signals meant：Negative for cells no stained,weak positive for the cells with nuclei and（or）cytoplasm stained pink;strong positive for the cells stained the bright red;while middle positive for the cells stained between the both.The image analysis software（Image pro plus 4.5）was applied in the gland tissue and interstitial tissue in each slice to calculate the average integrated optical density for the expression of c-FLIP.Results In the experimental group,there were all strong positive expressions of c-FLIP in the interstitial tissue at all the time points,mainly expressed in the cytoplasm of fibroblast and very little or almost no expression in the glandular tissue.Positive expression of c-FLIP in the interstitial tissue was significantly stronger than that of gland tissue（P0.05）;There were no significant differences among each time point in either the interstitial tissue or gland tissue（P0.05）.In the sham operation group,there were all weak positive expressions of c-FLIP in the interstitial tissue and gland tissue at all the time points and was no significant difference（P0.05）,no difference between each phase（P0.05）.The expression of c-FLIP in the experimental group was significantly higher than that in the sham operation group in the interstitial tissue at all the time（P0.05）,while no significant that in the gland tissue（P0.05）.Conclusion After bile duct injury,the expression of c-FLIP in anastomotic interstitial tissue is sustainable,by which the continuing obstruction effect to apoptosis may have a close relationship with the formation of biliary benign stricture.