正常及肝纤维化小鼠大范围肝切除后的肝再生研究

The research of liver regeneration between normal mice and liver fibrosis mice after a wide range of hepatectomy

目的比较正常及肝纤维化小鼠行肝脏大范围切除后的肝再生情况,研究肝纤维化对肝切除后肝脏损伤和肝再生的影响。方法选择120只C57/BL6小鼠,用随机数字表法将其分为对照组和实验组,每组各60只。实验组背部皮下注射40%四氯化碳(CCl4)玉米油溶液诱导肝纤维化模型,对照组给予等量玉米油。6周后肝纤维化成模,两组行肝左、中叶切除。分别于术后3小时、6小时、12小时、24小时、48小时和72小时处死小鼠,采集血液标本与残存肝脏组织,检测血清丙氨酸转氨酶(ALT)、天冬氨酸转氨酶(AST)水平,肝组织病理,肝重体重比值和增殖细胞核抗原(PCNA),观察肝脏损伤及再生差异。结果肝切除后,实验组各时间点ALT、AST呈先升高后降低趋势,均在12小时达到峰值,且与对照组相比具有统计学差异〔ALT(U/L):923.11±41.26比869.55±33.65;AST(U/L):976.69±48.65比744.77±21.42,均P<0.05〕;肝脏病理显示,实验组正常肝小叶破坏,有假小叶形成,肝细胞索排列紊乱,肝细胞出现变性坏死,损伤较重,而对照组肝小叶及细胞形态正常,损伤较轻;肝体重比及PCNA检测显示,相对于对照组,实验组肝再生延迟或缓慢,增殖细胞核抗原(PCNA)阳性表达细胞数在48小时达到高峰,且PCNA阳性细胞数显著降低〔PCNA阳性细胞数(个):92.33±10.68比176.33±14.74,P<0.05〕。结论肝纤维化病变可使小鼠大范围肝切除后肝脏损伤加重,肝再生减缓。

Objective To compare the liver regeneration after a wide range of hepatectomy between normal mice and liver fibrosis mice, studying the influences of liver fibrosis on liver damage and liver regeneration after hepatectomy. Methods A total number of 120 C57/BL6 mice were divided into control group and experimental group using random number table, fibrosis was induced in the experimental group by administration of 40% carbon tetrachoride （CCl4） corn oil solution, while the control group were given corn oil only. Two groups were subjected to the left and middle hepatic lobe liver resection after 6 weeks. The blood samples and remaining liver tissue were collected at 3 hours, 6 hours, 12 hours, 24 hours, 48 hours and 72 hours after hepatectomy to detect the serum alanine aminotransferase （ALT）, aspartate transaminase （AST） levels, liver tissue pathology, the liver weight ratio and proliferating cell nuclear antigen （PCNA）, the differences of damage and regeneration were observed. Results ALT and AST were significantly higher in the experimental group as compared to the controls after hepateetomy. They reached peak value at 12 hours and had significant differences compared with control group [ALT （U/L） ： 923.11 ±41.26 vs. 869.55±33.65 ; AST （U/L） ：976.69±48.65 vs. 744.77±21.42, all P 〈 0.05]. Pathology results showed that experimental group were damaged more seriously than controls. Liver regeneration was slow in the experimental group as compared to the controls in the liver weight ratio and PCNA staining. In 48 hours, it reached peak value and the number of PCNA positive cells were significantly decreased compared with control group （number of PCNA positive cells ：92.33± 10.68 vs. 176.33±14.74, P 〈 0.05）. Conclusion Liver fibrosis lesions may aggravate liver injury and slow down the liver regeneration after a wide range of hepateetomy in mice.