白芍总苷干预颈动脉球囊损伤模型大鼠Foxp3及骨形态发生蛋白7的表达

Expression Foxp3 and bone morphogenetic protein 7 in rat models of carotid artery balloon injury after treated by total glucosides of paeony

背景:经皮冠状动脉介入治疗后再狭窄的发生影响治疗效果。研究表明,白芍总苷能通过多种途径调节自身免疫反应,能够抑制单核巨噬细胞产生和分泌炎性细胞因子,清除氧自由基、影响细胞增殖以及具有抗炎、止痛、抗应激等作用。目的:观察大鼠颈动脉损伤后Foxp3及骨形态发生蛋白7的变化及应用白芍总苷干预后表达的变化。方法:实验方案经深圳市人民医院动物实验伦理委员会批准。将SD大鼠36只随机分为4组:对照组、球囊损伤组、白芍总苷灌胃组、白芍总苷浸泡球囊组。对照组生理盐水灌胃4d后假手术,术后生理盐水灌胃13d;球囊损伤组生理盐水灌胃4d后左侧颈总动脉进行球囊损伤,术后生理盐水灌胃13d;白芍总苷灌胃组用白芍总苷灌胃4d后行球囊损伤,术后白芍总苷灌胃13d;白芍总苷浸泡球囊组用生理盐水灌胃4d后行球囊损伤,术前将球囊浸泡于白芍总苷稀释液4h,术后生理盐水灌胃13d。术后14d取左侧颈总动脉损伤血管行苏木精-伊红染色,观察内膜变化,应用RealtimeRT-PCR检测损伤血管组织中Foxp3、骨形态发生蛋白7mRNA表达。结果与结论:①白芍总苷浸泡球囊组内膜面积及内膜面积/中膜面积,较白芍总苷灌胃组及球囊损伤组减小(P<0.05),但显著高于对照组(P<0.05);②Foxp3、骨形态发生蛋白7mRNA表达:对照组<球囊损伤组<白芍总苷灌胃组<白芍总苷浸泡球囊组,差异均有显著性意义(P<0.05);③结果说明,白芍总苷通过调节Foxp3及骨形态发生蛋白7mRNA表达,调节炎症因子的平衡,抑制损伤血管的炎症反应,减轻损伤血管的狭窄。使用白芍总苷浸泡球囊法较白芍总苷灌胃法对抑制炎症导致血管狭窄效果更佳。

BACKGROUND: Restenosis after percutaneous coronary affects the outcome of surgery. Total glucosides of paeony has been shown to regulate autoimmune response by several pathways, and can inhibit mononuclear macrophage secreting inflammatory cytokines, eliminate oxygen free radicals, affect cell proliferation and roles of anti-inflammation, analgesia and anti-stress. OBJECTIVE: To observe the changes in Foxp3 and bone morphogenetic protein 7 after carotid artery balloon injury in rats and the intervention of total glucosides of paeony. METHODS: The study was approved by the Laboratory Animal Ethical Committee of Shenzhen People’s Hospital. Thirty-six Sprague-Dawley rats were randomly divided into four groups: control group, balloon injury group, total glucosides of paeony group and total glucosides of paeony immersion group. The rats in the control group underwent sham surgery at 4 days after given normal saline via gavage, followed by 13 days of normal saline via gavage. The rats in the balloon injury group underwent left common carotid artery injured by ballooning at 4 days after given normal saline via gavage, followed by 13 days of normal saline via gavage. The rats in the total glucosides of paeony group received balloon injury at 4 days after given total glucosides of paeony via gavage, and then received 13 days of total glucosides of paeony via gavage. The rats in the total glucosides of paeony immersion group received balloon injury at 4 days after given normal saline via gavage, then the balloon was immersed in total glucosides of paeony diluent for 4 hours, and then subjected to 13 days of normal saline via gavage. At 14 days after surgery, the left common carotid artery samples were removed for hematoxylin-eosin staining to observe the changes of intima. The vessels were stained with hematoxylin-eosin. The expression levels of Foxp3 and bone morphogenetic protein 7 mRNA in injured vessels were detected by real-time RT-PCR. RESULTS AND CONCLUSION:(1) The area of intima and intima/tunica media area in the total glucosides of paeony immersion group were significantly less than those in the total glucosides of paeony group(P < 0.05), and significantly more than those in the control group(P < 0.05).(2) The order of the expression levels of Foxp3 and bone morphogenetic protein 7 mRNA was as follows: control group < balloon injury group < total glucosides of paeony group < total glucosides of paeony immersion group(P < 0.05).(3) These results suggest that total glucosides of paeony can regulate the expression of Foxp3 and bone morphogenetic protein 7 mRNA and the balance of inflammatory factors, inhibit the inflammatory reaction of injured vessels and may contribute to alleviating the restenosis of injured vessels. Local application of balloon immersed in total glucosides of paeony achieved better results than gavage in inhibiting the restenosis caused by inflammation.