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目的研究氧化损伤及肝细胞凋亡在肠外营养(PN)相关肝损害机制中的作用,并探讨中药丹参减轻PN相关肝损害的有效性。方法采用生后6-8 d的新西兰种白兔,体重80-110 g,随机分为3组:①正常对照组(n=10),母乳喂养;②PN组(n=10),持续输注静脉营养液(244)ml·kg-1·d-1)10 d;③PN+丹参组(n=10),在静脉营养液中加人丹参(0．2 ml·kg-1·d-1)。持续静脉营养10d后,分别取血作肝功能生化检测,取肝组织作光镜病理、电镜检查,氧化损伤(丙二醛MDA含量测定)及凋亡细胞(TUNEL法)检测。结果PN组血直接胆红素、胆汁酸均明显高于正常对照组和PN+丹参组(均P<0．05)。肝脏病理显示:PN组可见肝细胞空泡变性、脂肪变性,有多发细胞内淤胆和胆栓形成;PN+丹参组门脉区少量炎细胞浸润,无胆管扩张和胆汁淤积表现;病理学总评分3组分别为:8,22,15,组间两两比较P<0．01。电镜肝细胞细胞器超微结构变化与光镜病理改变相符, PN组可见毛细胆管扩张、微绒毛消失,线粒体肿胀、部分可见脂肪变性,以及早期凋亡表现。MDA检测PN组明显高于对照组(2．04±0．44比1．35±0．29mmol／mgpro,P<0．05)和PN+丹参组(2．04±0．44比1．19±0．14mmol／mgpro,P<0．05)。TUNEL,法检测3组肝细胞凋亡阳性率(％)分别为: 0．92±0．85、44．59±6．68、9．27±7．28,3组两两比较均P<0．01。结论PN可导致新生兔肝细胞出现氧化损伤及凋亡,产生肝损害;而中药丹参可明显减轻PN所致肝细胞损害。
Objective To study the role of oxidative injury and hepatocyte apoptosis in the mechanism of parenteral nutrition (PN)-associated hepatic damage, and to explore the effectiveness of Salvia miltiorrhiza for reducing PN related liver damage. Methods New Zealand White rabbits aged 6-8 days after birth were weighed 80-110 g and randomly divided into 3 groups: 1 normal control group (n=10), breastfeeding; 2PN group (n=10), continuous infusion Intravenous nutrient solution (244) ml·kg-1·d-1) 10 d; 3PN+ Salvia miltiorrhiza group (n=10), and intravenous Danshen (0.2 ml·kg-1·d-1) . After 10 days of continuous intravenous nutrition, blood samples were taken for biochemical tests of liver function. Liver tissues were taken for light pathology, electron microscopy, oxidative damage (measured by malondialdehyde MDA content) and apoptotic cells (TUNEL method). Results The levels of blood bilirubin and bile acid in PN group were significantly higher than those in normal control group and PN+Salvia miltiorrhiza group (all P<0.05). Liver pathology showed that in the PN group, there were vacuolar degeneration and fatty degeneration of hepatocytes, multiple intracellular cholestasis and biliary thrombus formation; PN+ Salvia miltiorrhiza group infiltrated a small amount of inflammatory cells in the portal area, no bile duct dilatation and cholestasis; pathological score The three groups were: 8, 22, 15, and the comparison between groups was P<0.01. The ultrastructure of hepatocyte organelles in electron microscopy was consistent with the pathological changes of light microscopy. In the PN group, the bile ducts dilated, the microvilli disappeared, the mitochondria swelled, and some of them showed steatosis and early apoptosis. The detection of MDA in PN group was significantly higher than that in control group (2.04±0.44 vs 1.35±0.29 mmol/mgpro, P<0.05) and PN+Salvia miltiorrhiza group (2.04±0.44 vs 1.19) ± 0.14 mmol/mg pro, P < 0.05). TUNEL, the positive rate of hepatocyte apoptosis in 3 groups was 0.92±0.85, 44.59±6.68, and 9.27±7.28, respectively. The comparison between the three groups was P< 0.01. Conclusion PN can cause oxidative damage and apoptosis of hepatocytes in newborn rabbits, resulting in hepatic damage. Salvia miltiorrhiza can significantly reduce liver cell damage induced by PN.