乌司他丁对急性重症胆管炎所致肝屏障损伤的作用和影响

Effects of Ulinastatin on hepatic barrier in experimental acute cholangitis of severe type

目的观察乌司他丁(UTI)对急性重症胆管炎所致肝屏障损伤的作用和影响。方法建立急性重症胆管炎大鼠动物模型,将实验动物分为4组:假手术组(A组)、ACST组(B组)、ACST+小剂量乌司他丁组(C组)、ACST+大剂量乌司他丁组(D组)。检测各组肝功能和内毒素,血浆中TNF-α、IL-10、IL-6水平,汇管区肝屏障细胞的凋亡,紧密连接相关蛋白及核因子p65蛋白表达水平。结果重症胆管炎大鼠肝功能恶化,血内毒素及促炎因子TNF-α、IL-6明显升高,抗炎因子IL-10降低;汇管区肝屏障大量炎性细胞浸润,胆管上皮细胞凋亡,肝屏障紧密连接相关蛋白ZO-1、Occludin表达减少。核因子κB(nuclear factorκB,NF-κB)主要的促炎单位p65蛋白明显升高。而乌司他丁干预后重症胆管炎大鼠肝功能恶化好转,血内毒素及促炎因子TNF-α、IL-6明显下调,抗炎因子IL-10升高;汇管区肝屏障炎性细胞浸润及胆管上皮细胞凋亡好转,肝屏障紧密连接相关蛋白ZO-1、Occludin表达升高。p65蛋白表达下降,其效果随着乌司他丁剂量增加而改善明显。结论乌司他丁可明显改善急性重症胆管炎所致肝屏障损伤,且与乌司他丁剂量呈正相关,乌司他丁对肝屏障的保护机制之一可能是通过抑制NF-κB信号通路来实现的,其具体机制有待于进一步研究。

Objective To investigate the effects of different doses of Ulinastatin（UTI） action on hepatic barrier in rats with experimental acute cholangitis of severe type（ACST）. Methods Forty eight Wistar rats were randomly divided into groups of sham-operation（group A）, ACST（group B）, ACST ＋ low-dose UTI（group C）,ACST ＋ high-dose UTI（group D）. After ulinastatin administration for 24 hours, blood and liver samples were collected from the rats for morphological examination, and liver function, serum level of TNF-α, IL-10 and IL-6.The distribution and expression of the p65 and tight junction（TJ） proteins, such as Occludin, ZO-1 were examined by immunohistochemistry. Results Liver function in rats with severe cholangitis deterioated seriously, the level of endotoxin and pro-inflammatory cytokines TNF-α and IL-6 significantly increased, anti-inflammatory cytokine IL-10 reduced; inflammatory cells infiltrated into portal area, epithelial cell of bile canaliculi was apoptosis,hepatic barrier tightly, connected with protein ZO-1, Occludin decreased. Nuclear factor κB/p65 significantly increased. And after the intervention of Ulinastatin, liver function and biliary epithelial apoptosis were significantly improved, the level of blood endotoxin pro-inflammatory cytokines TNF-α and IL-6 were significantly reduced, anti-inflammatory cytokine IL-10 was significantly increased, hepatic barrier tightly connected with protein ZO-1, Occludin increased. while P65 protein decreased, the effect was positively associated with the dose of UTI. Conclusion Ulinastatin can significantly repair hepatic barrier damage induced by ACST, and the protection was positively associated with the dose of UTI, The protection mechanism of ulinastatin on hepatic barrier maybe achieve through inhibition of NF-κB signaling pathway.