摘要
目的:观察以eNOS基因导入肝硬化门静脉高压症大鼠后,肝内eNOS表达和肝窦容积的变化;探讨eNOS基因转染对肝窦容积的影响。方法:①建立CC14肝内型肝硬化大鼠模型,分别取10只肝硬化大鼠和10只正常大鼠肝组织,用免疫组化、图像分析系统测定eNOS含量和肝窦容积;②取30只肝硬化大鼠,随机分成3组:内eNOS治疗组(n=10)、Tris对照组(n=10)和空质粒对照组(n=10)。分别经门静脉注射脂质体-eNOS质粒、Tris缓冲液脂质体-空质粒5d后,用RT-PCR和免疫组化方法检测3组大鼠肝内eNOSmRNA、eNOS含量和肝窦容积;结果:①硬化肝内eNOS含量和肝窦容积分别为(3.81±1.09)%和(3.51±0.72)%,均显著低于正常对照组的(14.78±3.06)%和(9.32±1.05)%(P<0.01);两组大鼠肝内eNOS含量与肝窦容积之间均呈显著正相关,相关系数r分别为0.676(P<0.05)和0.703(P<0.05)。②经门静脉注射各组之规定内容5d后,eNOS基因治疗组大鼠肝内eNOSmRNA、eNOS含量和肝窦容积分别为(1.22±0.14)U、(17.38±4.62)%和(5.77±1.49)%,均显著高于Tris处理组或空质粒处理组(P<0.01);3组的eNOS含量与肝窦容积之间均呈显著正相关,r分别为0.799(P<0.01)、0.767(P<0.01)和0.748(P<0.05)。结论:肝硬化时,因肝窦内皮细胞受损,肝内eNOS含量减少,肝内NO产量减少。
Objective:To evaluate the effect of the expression of endothelial nitric oxide synthase(eNOS)on the capacity of the hepapic sinusoids by lipid-mediated eNOS gene transfer in cirrhotic rats.Methods:Ten of130male Sprague-Dawley rats were taken as normal controll group and the residual rats were induced to hepatic cirrhosis by intraperitoneal injection of carbon tetrachloride(CCl 4 ).Fourty cirrhotic rats were divided into four groups randomly:untreated group(n=10),eNOS-treated group(n=10),control plasmid-treated group(n=10)and Tris buffer-treated group(n=10).Liposome-eNOS was tansduced into rats of eNOS-treated group through the injection to portal vein;Tris buffer-treated group and control plasmid-treated group were received Tris buffer or control plasmid respectively.Five days later,the animals were sacrificed.Reverse Transcription-Polymerase Chain Reaction(RT-PCR)was performed to detect the expression of eNOS mRNA.Immunohistochemistry and image analysis were performed to detect the expression of eNOS and the capacity of liver sinusoid.The untreated cirrhotic group and normal controll group rats were killed,and the expression of eNOS and capacity of hepatic sinusoid were detected by the same method as described above.Results:①The content of eNOS and capacity of liver sinusoid in untreated cirrhotic group were(3.81±1.09)%and(3.51±0.72)%respectively,which were much lower than that of the normal group[(14.78±3.06)%and(9.32±1.05)%respectively](P<0.01).The content of eNOS and the capacity of liver sinusoid were positively correlated in the both two groups,r=0.676(P<0.05)and r=0.703(P<0.05)respectively.②Five days after eNOS gene transfer,the expression of eNOS mRNA,content of eNOS and capacity of liver sinusoid were(1.22±0.14)U,(19.43±7.37)%and(5.77±1.49)%respectively,which were much higher than that in the control groups(P<0.01).The expression of eNOS and the capacity of liver sinusoids were positively correlated in the all the cirrhotic rats treated with the3methods,r=0.799(P<0.01),r=0.767(P<0.01)and
出处
《外科理论与实践》
2003年第4期329-332,共4页
Journal of Surgery Concepts & Practice
基金
国家自然基金(30070740)
上海市科委基金(99XD14004)资助项目
关键词
肝硬化
ENOS基因
基因治疗
肝窦容积
大鼠
门静脉高压症
Portal vein Hypertension Endothelial nitric oxide synthase Capacity of liver sinusoids Gene therapy
