低分子肝素对颈动脉球囊损伤模型大鼠AngⅡ、HB-EGF表达及内膜增生的影响  

The effects of low molecular weight heparin on the expression of AngⅡ,HB-EGF and intimal hyperplasia in rat model of carotid artery balloon injury

作　　者：赵治涛 肖珂青 ZHAO Zhitao;XIAO Keqing(Department of Anesthesiology and Pain,Shandong Provincial Third Hospital,Shandong,Ji’nan 250031,China)

机构地区：[1]山东省立第三医院麻醉疼痛科,济南市250031

出　　处：《河北医药》2021年第16期2422-2426,2431,共6页Hebei Medical Journal

摘　　要：目的探究低分子肝素(LMWH)对颈动脉球囊损伤大鼠血管紧张素Ⅱ(AngⅡ)、肝素结合性表皮生长因子(HB-EGF)表达及内膜增生的影响。方法取成年健康雄性SD大鼠,随机分为假手术组、颈动脉球囊损伤模型组、LMWH低(50 U/kg)、中(100 U/kg)、高(200 U/kg)剂量组,每组10只;除假手术组只暴露并分离颈动脉外,其余4组大鼠均用2.0 mm×12 mm球囊导管(Fogarty,2F)建立颈动脉球囊损伤大鼠模型。5组大鼠均于术后第2天开始给药,LMWH低、中、高剂量组经尾静脉注射给予相应剂量LMWH药物,假手术和模型组经尾静脉注射给予等量0.9%氯化钠溶液,5组连续给药14 d,1次/d。5组大鼠于末次给药12 h后,处死大鼠,取颈动脉血管组织,用苏木精-伊红染色(HE)检测大鼠血管组织病理损伤,并检测血管管腔面积、内膜面积、内膜/中膜面积(IA/MA);酶联免疫吸附法(ELISA)检测血管组织中炎性因子白细胞介素-6(IL-6)和肿瘤坏死因子-α(TNF-α)蛋白表达;实时定量荧光PCR(qRT-PCR)法检测血管组织中HB-EGF mRNA、AngⅡmRNA表达水平;用蛋白免疫印迹法(Western Blot)检测血管组织中HB-EGF、AngⅡ、缝隙连接蛋白43(Cx43)蛋白相对表达水平。结果与假手术组比较,模型组大鼠颈动脉内皮剥落、内膜增生、狭窄等病理损伤程度、血管内膜面积、IA/MA比值、血管组织炎性因子IL-6和TNF-α含量、HB-EGF、AngⅡmRNA及蛋白表达、Cx43蛋白表达均升高(P<0.05),管腔面积降低(P<0.05);与模型组比较,LMWH低、中、高剂量组大鼠颈动脉内皮剥落、内膜增生、狭窄等病理损伤程度、内膜面积、IA/MA比值、血管组织炎性因子IL-6和TNF-α含量、HB-EGF mRNA及蛋白表达、AngⅡmRNA及蛋白表达、Cx43蛋白表达均降低(P<0.05),血管管腔面积升高(P<0.05),且LMWH各剂量组上述指标呈剂量依赖性。结论LMWH可抑制颈动脉球囊损伤模型大鼠颈动脉血管组织AngⅡ、HB-EGF基因及蛋白表达,抑制血管内膜Objective To investigate the effects of low molecular weight heparin(LMWH)on the expression of angiotensinⅡ(AngⅡ),heparin-binding epidermal growth factor(HB-EGF)and intimal hyperplasia in rats with carotid artery balloon injury.Methods The adult healthy male SD rats were selected and randomly divided into sham-operation group,carotid artery balloon injury model group(model group),low-dose LMWH(50U/kg)group,medium-dose LMWH(100U/kg)group,high-dose LMWH(200U/kg)group,with 10 rats in each group.Except for sham-operation group in which only carotid artery was exposed and separated,the 2.0mm×12mm balloon catheter(Fogarty,2F)was used in the other groups to establish rat model with carotid artery balloon injury.The rats in the five groups were administered with LMWH on the second day after the operation.The rats in low-,medium-,and high-dose LMWH groups were given the corresponding dose of LMWH via tail vein injection,and the Sham and Model groups were given the same amount of 0.9%sodium chloride solution via tail vein injection.Each group was continuously administered for 14d,once a day.The rats in the five groups were sacrificed at 12h after the last administration,and the carotid artery tissue was taken.Hematoxylin-eosin staining(HE)was used to detect the pathological damages of vascular tissue,moreover,the vascular lumen area,intima area and intima area/media area(IA/MA)were detected.Enzyme-linked immunosorbent assay(ELISA)was used to detect the expression levels of inflammatory cytokines interleukin-6(IL-6)and tumor necrosis factor-α(TNF-α)in vascular tissue.Real-time quantitative fluorescent PCR(qRT-PCR)was used to detect the expression levels of HB-EGF and AngⅡmRNA.Western blotting was used to detect the relative protein expression levels of HB-EGF,AngⅡ,and connexin 43(Cx43).Results Compared with those in sham-operation group,carotid artery endothelial exfoliation,intimal hyperplasia,stenosis and other pathological damage,vascular intima area,IA/MA ratio,the levels of inflammatory cytokines IL-6 and TN

关 键 词：低分子肝素 颈动脉球囊损伤 血管紧张素II、肝素结合性表皮生长因子 内膜增生 

分 类 号：R541.4[医药卫生—心血管疾病]