80%门静脉结扎后肝脏组织中MMP-9的表达与肝再生作用的相关性研究

Correlation of Expression of MMP-9 in the Liver and Liver Regeneration in Rats after 80% of the Portal Vein Ligation

目的:探讨基质金属蛋白酶-9(MMP-9)在大鼠80%门静脉分支结扎模型中大鼠增生肝脏组织中的表达及其与肝再生作用的关系。方法:健康SD雄性大鼠48只,随机平均分成假手术对照组(Sham)和门静脉结扎实验组(PVL)。观察术后1、3、7和14d保留侧肝叶重量/体重比值;下腔静脉采血后检测血清谷丙转氨酶(ALT)、谷草转氨酶(AST)值的变化;光镜下观察保留侧肝脏组织的病理形态变化;用免疫组化法检测增殖细胞核抗原(Proliferating cell nuclear antigen,PCNA)的表达,用免疫印记法检测MMP-9的表达,并进行统计分析。结果:80%门静脉分支结扎后,结扎侧肝叶呈进行性萎缩,保留侧肝叶重量/体重比值逐渐增加,7d达"平台期";与对照组明显不同,PVL组的ALT、AST的值在1h达到高峰,7d后回到正常水平;保留侧肝脏组织中PCNA阳性细胞计数与对照组比较,3d开始表达增强(P<0.05),7d以后逐渐恢复至正常水平(P>0.05);保留侧肝叶MMP-9蛋白的表达在术后3d开始增加,术后7d达到高峰。结论:MMP-9蛋白的表达在80%门静脉结扎后大鼠肝再生过程中发挥重要作用。

To evaluate the expression of MMP-2 and MMP-9 in rats after 80% portal vein ligation. Methods： Forty-eight male SD rats were randomly divided into sham operation group and portal vein ligation group. Liver weight/total body weight ratios ofunligated lobes were measured at post operation day （POD） 1,3,7 and 14 d. The serum sample was obtained from inferior vena cava to investigate liver function including alanine amino transferase （ALT） and aspartate aminotransferase （AST）. The morphological changes of the unligated liver lobes were observed by microscope. The expressions of proliferating cell nuclear antigen （PCNA） of the unligated liver lobes was deteeted by immunohistochemistry. The expression of MMP-9 protein of unligated lobes were measured by western-blot. The results were analyzed by medical statistics methods. Results： Hepatic lobe at the ligated side diminished progressively after 80% portal vein ligation. Liver weight to total body weight ratios of unligated lobes increased slowly at ld,and got ＂the plateau stage＂ at 7d;serum levels of ALT and AST with 80% PVL differed significantly from sham-operated animals. We found that the peak concentration ld after surgery returned to control levels 7d after surgery. The PCNA protein expression reached the peak on 3d and decreased slightly on 7d after operation （P 〈 0.05 ）, then gradually return to normal lately; The expressions of MMP-9 in the unligat-ed liver lobes markedly increased on 3d, and reached the peak on 7d. Conclusion： The expression of MMP-9 plays an important role in the liver regeneration in rats after 80% of the portal vein ligation.