下瘀血汤对肝硬化大鼠肝组织CD68和CD163蛋白表达的影响  

Effect ofXiayuxue decoction on CD68 and CD163 protein expression in liver tissue of rats with liver cirrhosis

作　　者：刘乐平[1] 陆芩[2] 王晓斌[2] 谢洁雅[1] 丁赛丹[1] 

机构地区：[1]温州医科大学附属第一医院外科实验室,浙江温州325000 [2]温州医科大学附属第一医院神经外科,浙江温州325000

出　　处：《中国药理学与毒理学杂志》2014年第1期35-41,共7页Chinese Journal of Pharmacology and Toxicology

基　　金：温州市科技计划项目(Y20110155)~~

摘　　要：目的探讨下瘀血汤抗肝硬化的作用机制。方法 SD雄性大鼠ip给予硫代乙酰胺200 mg·kg-1每周2次,连续8周,制备大鼠肝硬化模型。造模成功后,模型大鼠于第9周首日ig给药下瘀血汤0.313,0.626和1.252 g·kg-1,每天1次,连续3周。于第11周末处死大鼠,取肝组织,用Western蛋白质印迹法和免疫组化法测定肝组织CD68和CD163蛋白表达;用激光共聚焦方法检测肝组织CD68+TUNEL+和CD163+TUNEL+双阳性细胞,观察ED1和ED2肝巨噬细胞凋亡。结果与正常对照组比较,模型组大鼠血清谷丙转氨酶(GPT)和谷草转氨酶(GOT)活性增强(P<0.01),血清总胆红素(TBil)水平升高(P<0.01),血清白蛋白(Alb)水平显著降低(P<0.01)。与模型组比较,下瘀血汤0.626和1.252 g·kg-1组大鼠血清GPT和GOT活性以及Tbil水平均显著降低(P<0.05,P<0.01),Alb水平显著升高(P<0.01)。Western蛋白质印迹和免疫组化实验结果显示,与正常对照组比较,模型组CD68表达升高(P<0.01),CD163表达明显降低(P<0.01);与模型组相比,下瘀血汤0.626和1.252 g·kg-1组CD68表达明显降低(P<0.01),CD163表达显著升高(P<0.01)。激光共聚焦实验结果表明,与正常对照组比较,模型组大鼠CD68+TUNEL+双阳性细胞显著增加(P<0.05);与模型组比较,下瘀血汤0.626和1.252 g·kg-1组CD68+TUNEL+双阳性细胞进一步增加(P<0.05,P<0.01);所有组别均未检测到CD163+TUNEL+双阳性细胞。结论下瘀血汤对肝硬化大鼠肝功能具有改善作用,可抑制肝硬化大鼠肝组织CD68表达,促进CD163表达。OBJECTIVE To investigate the mechanism of anti-liver cirrhosis of Xiayuxue decoction.METHODS Ten male SD rats were randomly chosen as the normal control group,and the remaining40 rats were ip given thioacetamide 200 mg·kg^- 1,twice each week for 8 weeks,to prepare the rat model with liver cirrhosis. At the 9th week,the model rats were randomly assigned to model,Xiayuxue decoction 0. 313,0. 626 and 1. 252 g·kg^- 1groups. The rats were ig given Xiayuxue decoction once daily for 3 weeks. At the 11th week,CD68 and CD163 protein expression in liver tissue was detected with Western blotting and immunolistochemistry method. CD68＋TUNEL＋and CD163＋TUNEL＋double positive cells in liver tissues were analyzed under a laser confocal microscope. RESULTS Compared with normal control group,the activity of serum glutamic-pyruvic transaminase（ GPT） and glutamic-oxalacetic transaminase（ GOT） was enhanced（ P 0. 01）,the level of total bilirubin（ Tbil） was elevated（ P 0. 01）,while the level of albumin（ Alb） was decreased in model group（ P 0. 01）. Compared with model group,the activity of serum GPT and GOT and the level of Tbil were decreased（ P 0. 05,P 0. 01）,while the level of Alb was increased in Xiayuxue decoction group. Western blotting and immunohistochemistry results showed that the expression of CD68 was increased（ P 0. 01） and that of CD163 was significantly reduced（ P 0. 01） in the model group. Compared with model group,the expression of CD68 was reduced（ P 0. 01） and the expression of CD163 was significantly increased（ P 0. 01） in Xiayuxue decoction 0. 626 and 1. 252 g·kg^- 1groups. Laser confocal results showed that CD68＋TUNEL＋double-positive cells were significantly increased（ P 0. 05） in model group compared with the normal group. Compared with the model group,the CD68＋TUNEL＋double-positive cells were significantly increased（ P 0. 05 or P 0. 01） in Xiayuxue decoction 0. 626 and 1. 252 g·kg^- 1groups.CD163＋TUNEL＋double-positive cells were not det

关 键 词：下瘀血汤 肝硬化 CD68 CD163 肝巨噬细胞 

分 类 号：R285[医药卫生—中药学] R975[医药卫生—中医学]