特利加压素对肝大部切除术后大鼠残肝的保护作用
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摘要
目的:通过建立小体积肝脏动物模型,探讨特利加压素对肝大部切除术后大鼠小体积残肝的保护作用及其可能机制。
方法:参考Higgins法,加行肝门阻断30分钟,建立大鼠小肝综合症模型(肝大部切除(70%)+肝门阻断(30分钟))。模型分为两组:实验组(特利加压素治疗组)和对照组(生理盐水组),于肝脏血流恢复再灌注后不同时间点处死大鼠,取血送检肝功能(ALT、AST、TBil),取残肝组织观察其形态学改变,采用免疫组织化学方法检测残肝组织中A20、iNOS和ET-1的表达情况。另取实验组和对照组大鼠于给药前及肝脏血流恢复灌注后0.5小时、1小时和2小时行门静脉压检测,再取两组大鼠行7日存活率检测。
结果:实验组大鼠肝功能指标上升程度均较对照组低,以再灌注后6小时(ALT:605.82±3.58 vs 709.94±2.98;AST:905.52±3.37 VS 1159.94±3.76;TBil: 4.12±0.23 vs 5.27±0.26)与12小时(ALT:711.36±3.11 vs 854.65±3.24;AST: 1042.36±3.51 VS 129.68±3.46;TBil: 5.86±0.21 vs 7.18±0.25)明显,P<0.05;残肝组织病理形态学检查显示,实验组大鼠肝组织损伤程度较对照组轻;免疫组织化学检查显示,除再灌注后0.5小时时间点外,A20在实验组肝组织中的表达明显强于对照组,在再灌注后2小时达高峰,24小时后在两组中的表达无显著性差异, ET-1自再灌注后0.5小时两组中即可见阳性表达,与对照组比较,实验组表达较弱,以再灌注24小时明显。在再灌注后0.5小时,iNOS在两组中表达均很弱,其余时间点在对照组中的表达上调明显高于实验组,以再灌注后2小时最明显。两组大鼠给药前的门静脉压无显著性差异(10.11±1.32 vs 9.98±1.33),其余时间点实验组门静脉压力较对照组为低,以再灌注后0.5小时(13.42±1.36 vs 16.21±1.41)与1小时(11.67±1.25 vs 14.49±1.42)最为显著,P<0.05。实验组大鼠7日存活率明显高于对照组(100% vs 58.3%)(P<0.05)。
结论:“肝大部切除(70%)+肝门阻断(30分钟)”动物模型稳定可靠,适合进行小肝综合症研究。特利加压素能够有效降低肝大部切除术后早期门静脉压力,从而减轻小体积残肝再灌注后早期炎性反应及肝窦机械性损伤,维持残肝微循环稳定,减轻肝细胞坏死或凋亡,最终保护残肝功能,提高大鼠存活率。
Objective:To explore the protection and possible mechanism of Terlipressin on the rat’s remnant livers after major hepatectomy by establishing a rat model of small-for-size liver.
Method : Establish a rat model of small-for-size syndrome with 70 percent hepatectomy and PTC( portal triad clamping) for 30 minutes refer to the Higgins methed。Rats were divided into two groups:Terlipressin group and Control group.When the rats being Executed at different timepoints after portal vein reperfusion,the serum liver function (ALT、AST、TBil)were measured,the liver tissues were harvested and studied with pathological observation and the expressions of A20、iNOS、ET-1 were detected by immunohistochemictry methods.Some rats for monitoring the portal vein pressure before administratoring Terlipressin and the 0.5 hour、1 hour、and 2 hours timepoints after reperfusion,also some rats for monitoring the 7-day survival rates of the two groups.
Results: The levels of liver function were significantly decreased in the Terlipressin group when compared with the Control group At 6 hours (ALT:605.82±3.58 vs 709.94±2.98;AST:905.52±3.37 VS 1159.94±3.76;TBil: 4.12±0.23 vs 5.27±0.26) and 24 hours (ALT : 711.36±3.11 vs 854.65±3.24 ; AST: 1042.36±3.51 VS 129.68±3.46;TBil: 5.86±0.21 vs 7.18±0.25) timepoints(P<0.05),though both of them increased saliently.Microscopic observation showed that the morphological changes of remnant liver in Terlipressin group at every timepoint were markedly milder than that in Control group.Immunohistochemical staining showed that A20 expression were stronger at Terlipressin group compared to Control group at 2 hours、4 hours、6 hours timepoints except of hardly expression at 0.5 hour timepoint in the two group,and there was no marked difference of A20 expression at 24 hours after reperfusion.ET-1 were overexpressed in both groups from the 0.5 hour timepoint, it was markedly down-regulated in Terlipressin group at 24 hours timepoint compared to the Control one。From 0.5 hour timepoint, iNOS was feeble expressed in each group,and it was markedly up-regulated compared to Terlipressin group at other timepoints.The obvious discrepancy showed at the 2 hours timepoint between the two groups.The portal vein pressure of Terlipressin group were significantly lower than that of homologous Control group at different timepoints.Especilly at the 0.5 hour and 1 hour timepoints after portal vein reperfusion( P<0.05),but the elementary portal vein pressures in the two groups haven’t conspicuous discrepancy(10.11±1.32 vs 9.98±1.33) ( P>0.05).The survival rate of Terlipressin group was higher obviously than the Control one(100% vs 58.3%)( P<0.05).
Conclusion: Model with 70 percent hepatectomy and PTC( portal triad clamping) for 30 minutes was so stable and fit for the study of small-for-size syndrome .Terlipressin can lower the early portal vein pressure effectively after major hepatectomy , can ameliorate the early inflammatory response of the small-for-size liver after reperfusion,can attenuate the mechanical injury of sinu hepaticus,can sustain a stable microcirculation of the remnant liver,also it can palliate hepatocellular necrosis or apoptosis,then protects the function of the remnant liver and raises the survival rate.
方法:参考Higgins法,加行肝门阻断30分钟,建立大鼠小肝综合症模型(肝大部切除(70%)+肝门阻断(30分钟))。模型分为两组:实验组(特利加压素治疗组)和对照组(生理盐水组),于肝脏血流恢复再灌注后不同时间点处死大鼠,取血送检肝功能(ALT、AST、TBil),取残肝组织观察其形态学改变,采用免疫组织化学方法检测残肝组织中A20、iNOS和ET-1的表达情况。另取实验组和对照组大鼠于给药前及肝脏血流恢复灌注后0.5小时、1小时和2小时行门静脉压检测,再取两组大鼠行7日存活率检测。
结果:实验组大鼠肝功能指标上升程度均较对照组低,以再灌注后6小时(ALT:605.82±3.58 vs 709.94±2.98;AST:905.52±3.37 VS 1159.94±3.76;TBil: 4.12±0.23 vs 5.27±0.26)与12小时(ALT:711.36±3.11 vs 854.65±3.24;AST: 1042.36±3.51 VS 129.68±3.46;TBil: 5.86±0.21 vs 7.18±0.25)明显,P<0.05;残肝组织病理形态学检查显示,实验组大鼠肝组织损伤程度较对照组轻;免疫组织化学检查显示,除再灌注后0.5小时时间点外,A20在实验组肝组织中的表达明显强于对照组,在再灌注后2小时达高峰,24小时后在两组中的表达无显著性差异, ET-1自再灌注后0.5小时两组中即可见阳性表达,与对照组比较,实验组表达较弱,以再灌注24小时明显。在再灌注后0.5小时,iNOS在两组中表达均很弱,其余时间点在对照组中的表达上调明显高于实验组,以再灌注后2小时最明显。两组大鼠给药前的门静脉压无显著性差异(10.11±1.32 vs 9.98±1.33),其余时间点实验组门静脉压力较对照组为低,以再灌注后0.5小时(13.42±1.36 vs 16.21±1.41)与1小时(11.67±1.25 vs 14.49±1.42)最为显著,P<0.05。实验组大鼠7日存活率明显高于对照组(100% vs 58.3%)(P<0.05)。
结论:“肝大部切除(70%)+肝门阻断(30分钟)”动物模型稳定可靠,适合进行小肝综合症研究。特利加压素能够有效降低肝大部切除术后早期门静脉压力,从而减轻小体积残肝再灌注后早期炎性反应及肝窦机械性损伤,维持残肝微循环稳定,减轻肝细胞坏死或凋亡,最终保护残肝功能,提高大鼠存活率。
Objective:To explore the protection and possible mechanism of Terlipressin on the rat’s remnant livers after major hepatectomy by establishing a rat model of small-for-size liver.
Method : Establish a rat model of small-for-size syndrome with 70 percent hepatectomy and PTC( portal triad clamping) for 30 minutes refer to the Higgins methed。Rats were divided into two groups:Terlipressin group and Control group.When the rats being Executed at different timepoints after portal vein reperfusion,the serum liver function (ALT、AST、TBil)were measured,the liver tissues were harvested and studied with pathological observation and the expressions of A20、iNOS、ET-1 were detected by immunohistochemictry methods.Some rats for monitoring the portal vein pressure before administratoring Terlipressin and the 0.5 hour、1 hour、and 2 hours timepoints after reperfusion,also some rats for monitoring the 7-day survival rates of the two groups.
Results: The levels of liver function were significantly decreased in the Terlipressin group when compared with the Control group At 6 hours (ALT:605.82±3.58 vs 709.94±2.98;AST:905.52±3.37 VS 1159.94±3.76;TBil: 4.12±0.23 vs 5.27±0.26) and 24 hours (ALT : 711.36±3.11 vs 854.65±3.24 ; AST: 1042.36±3.51 VS 129.68±3.46;TBil: 5.86±0.21 vs 7.18±0.25) timepoints(P<0.05),though both of them increased saliently.Microscopic observation showed that the morphological changes of remnant liver in Terlipressin group at every timepoint were markedly milder than that in Control group.Immunohistochemical staining showed that A20 expression were stronger at Terlipressin group compared to Control group at 2 hours、4 hours、6 hours timepoints except of hardly expression at 0.5 hour timepoint in the two group,and there was no marked difference of A20 expression at 24 hours after reperfusion.ET-1 were overexpressed in both groups from the 0.5 hour timepoint, it was markedly down-regulated in Terlipressin group at 24 hours timepoint compared to the Control one。From 0.5 hour timepoint, iNOS was feeble expressed in each group,and it was markedly up-regulated compared to Terlipressin group at other timepoints.The obvious discrepancy showed at the 2 hours timepoint between the two groups.The portal vein pressure of Terlipressin group were significantly lower than that of homologous Control group at different timepoints.Especilly at the 0.5 hour and 1 hour timepoints after portal vein reperfusion( P<0.05),but the elementary portal vein pressures in the two groups haven’t conspicuous discrepancy(10.11±1.32 vs 9.98±1.33) ( P>0.05).The survival rate of Terlipressin group was higher obviously than the Control one(100% vs 58.3%)( P<0.05).
Conclusion: Model with 70 percent hepatectomy and PTC( portal triad clamping) for 30 minutes was so stable and fit for the study of small-for-size syndrome .Terlipressin can lower the early portal vein pressure effectively after major hepatectomy , can ameliorate the early inflammatory response of the small-for-size liver after reperfusion,can attenuate the mechanical injury of sinu hepaticus,can sustain a stable microcirculation of the remnant liver,also it can palliate hepatocellular necrosis or apoptosis,then protects the function of the remnant liver and raises the survival rate.
引文
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[1] Holt DR, Thiel DV, Edelstein S, et al. Hepatic resections[J]. Arch Surg, 2000,135: 1353-1358.
[2]杨连粤,黄耿文,黄建华,等. 114例大肝癌的手术治疗[J].中国实用外科杂志, 2002, 22:353-355.
[3]杨甲梅,严以群,吴孟超,等.原发性肝癌肝切除后肝脏创面的处理(附628例报告)[J].中国实用外科杂志,1997,17(4):2282-229.
[4]吴孟超主编.肝脏外科学[M].第2版.上海:上海科学技术文献出版社和上海科技教育出版社,2000.343,562,569.
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[6] Perseghin G, Mazzaferro V , Sereni L P , et al. Contribution of re2 duced insulin sensitivity and secretion to the pathogenesis of hepatogenous diabetes : effect of liver transplantation. Hepatology ,2000;31(3):694-703.
[7]吴帮富,林丽莉,胡品津。胰高糖素负荷试验的血糖P-B值用于评介肝硬化病人肝功能善状况的价值。中华肝脏病杂志,2003,11(3):186-188
[8] Caban A , Wiaderkiewicz R , Kaminski M ,et al. Arterial ketone index in assessing liver function and its detoxicative capability after ischemia-reperfusion injury. Acta Biochim Polonica, 2000;47(4):1137-1146.
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[10] Herold C , Heinz R , Raderspiel-Troger M , et al.Quantitative testing of liver function in patients with cirrhosis due to chronic hepatitis C to assess disease severity.Liver,2001;21(1):26-30
[11] Herold C ,Censlmayer M ,Ocker M,et al.Inducibility of microsomal liver function may differentiate cirrhotic. Patients with maintained compared with severely compromised liver reserve.J Gast-menterol Hepatc ,2003;18(4):445-449
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[14] Caubet MS , Laplante A , Caille J , et al.[13C] aminopyrine and [13C] caffeine breath test : influence of gender ,cigarette smoking and oral contraceptives intake. Isotopes Environ Health Stud ,2002;38(2):71-77.
[15] Shimizu S ,Kammike W,Hatanaka N ,et al. New method for measuring ICGR max with a clearance meter. World J Surg ,1995;19(1):113-118.
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[17] Poyato Gonzalez A , de la Mata Garcia M , Rufian Pena S , et al. Prognostic factors in liver transplantation. Is the lidocaine test useful ? Rev Esp Enferm Dig, 1996; 88(5):344-350.
[18] Ercolani G, Grazi GL , Calliva R , et al. The lidocaine (MEGX) test as an index of hepatic function : its clinical usefulness in liver surgery. Surgery, 2000; 127(4):464-471
[19] Ravaioli M , Grazi GL ,Principe A , et al. Operative risk by the lidocaine test (MEGX) in resected patients for HCC on cirrhosis.Hepatogastroenterology, 2003;50(53):1552-1555.
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[21] Botta F , Giannini E , Romagnoli P , et al.MELD scoring system is useful for predicting prognosis in patients with liver cirrhosis and is correlated with residual liver function:a European study.Gut,2003; 52(1):134-139.
[22] Arroyo V , Gines P ,Gerbes AL ,et al.Definition and diagnostic criteria of refractory ascites and hepatorenal syndrome in cirrhosis.Hepatology, 1996; 23(1):164-176.
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