肝脏靶向表达大鼠白介素-10基因对猪血清诱导的大鼠肝纤维化的抑制  

Targeting expression of rat interleukin-10 in liver suppresses pig serum-induced hepatic fibrosis in rats

作　　者：黄月红[1] 陈运新[1] 郑伟达[2] 张莉娟[1] 陈治新[1] 王小众[1] 

机构地区：[1]福建医科大学附属协和医院消化内科,福州350001 [2]福建医科大学附属协和医院急诊科,福州350001

出　　处：《第三军医大学学报》2011年第13期1353-1357,共5页Journal of Third Military Medical University

基　　金：福建省自然科学基金(2009J05065;2010J05062;2010J01165);福建卫生厅青年科研项目(2010-1-10)~~

摘　　要：目的探讨肝脏靶向表达rIL-10基因对猪血清诱导的大鼠肝纤维化的抑制作用。方法 rIL-10基因通过尾静脉快速大容量注射法转移至大鼠体内,RT-PCR法检测大鼠肝、肾、脾和肺组织rIL-10 mRNA的表达及S-P免疫组化法检测rIL-10蛋白在肝脏中表达。30只体质量100～120 g清洁级雄性SD大鼠按随机数字表法分为正常对照组(n=6)、纤维化模型对照组(n=8)、rIL-10基因干预组(n=8)和空质粒对照组(n=8)。正常对照组与其余3组大鼠腹腔分别注射生理盐水和猪血清,0.5 ml/只,每周2次。第5周开始造模同时,rIL-10基因干预组和空质粒对照组大鼠分别从尾静脉注射pcDNA3-rIL-10和pcDNA3空质粒,每周1次。第8周末处死全部大鼠收集组织和血清备用。HE、天狼星红染色分别检测各组肝组织病理形态学和Ⅰ、Ⅲ胶原的改变,生化检测血清ALT和AST含量。结果 RT-PCR和免疫组化证实rIL-10基因经尾静脉快速大容量注射转染到大鼠体内后主要在肝组织中转录及表达;纤维化模型和空质粒对照组:HE染色显示大鼠肝细胞脂肪变性,炎性细胞浸润,大量胶原沉积形成粗细不等的纤维隔,部分假小叶形成;天狼星红染色显示肝组织中大量Ⅰ、Ⅲ型胶原纤维沉积;血清中ALT和AST表达水平较正常对照组明显升高(P<0.01)。与对照组比较,rIL-10基因干预组大鼠HE染色显示肝细胞变性坏死程度减轻、炎细胞的浸润减少,胶原沉积明显减少,天狼星红染色显示Ⅰ、Ⅲ型胶沉积面积显著下降(P<0.01),血清中ALT和AST表达水平显著降低(P<0.01)。结论 尾静脉快速大容量注射法可使rIL-10基因在肝组织靶向表达,靶向表达的rIL-10有效抑制猪血清诱导的大鼠肝纤维化形成。Objective To determine the effect of the targeting expression of rat interleukin-10（rIL-10） gene in the liver on the progress of porcine serum-induced hepatic fibrosis in rats.Methods pcDNA3-rIL-10 was transferred to the rat liver by hydrodynamic-based transfection.RT-PCR and SP-immunohistochemistry analysis were used to detect the expression and distribution of rIL-10 mRNA and protein in the kidneys,lungs,spleen,liver and heart in 24 h after transfection.A total of 30 clean-grade male SD rats were randomly divided into normal control group（n=6）,liver fibrotic model group（n=8）,rIL-10 treatment group（n=8） and null-plasmid control group（n=8）.Rat liver fibrotic model was established by intraperitoneal injection of pig serum,0.5 ml/d,twice a week for 8 weeks,and the rats of normal control group received an injection of saline.From the 5th week after pig serum injection,the rats of the later 3 groups underwent an intraperitoneal injection of Ringer＇s solution containing 1 μg/g pcDNA3-rIL-10 or pcDNA3 through tail vein once a week.At the end of the 8th week,all rats were put to death and then collected blood and liver sample.HE staining was used to detect the changes of liver tissue sections pathomorpholohy.The change of ALT and AST in serum and collagen type Ⅰ,Ⅲ in the liver were measured by biochemistry analysis and Picrosirius staining respectively.Results RT-PCR and immunohistochemistry confirmed that the presence of rIL-10 transcription and expression mainly in the liver.HE staining showed that rat hepatic fibrosis model induced by pig serum was established successfully.Compared with the null-plasmid control group,the liver in the fibrotic model group showed hepatocytic fatty degeneration,inflammatory cells invasion,massive collagen deposition to form fiber partition in difference thickness,and even false hepatic lobules.Compared with liver fibrotic model and null-plasmid control group,rIL-10 treatment group showed significant therapeutic effects： decreased necrosis in hepatocytes and

关 键 词：大鼠白介素-10 裸质粒DNA 尾静脉注射 基因干预 肝纤维化 

分 类 号：R575.2[医药卫生—消化系统] R394.2[医药卫生—内科学]