银丹心脑通软胶囊通过抑制CaN/NFATc3信号通路改善球囊损伤所致大鼠颈动脉血管内膜增生

Yindan Xinnaotong Capsule ameliorates carotid intimal hyperplasia induced by balloon injury in rats via regulating CaN/NFATc3 signaling pathway

目的探究银丹心脑通软胶囊(Yindan Xinnaotong capsule,YDXNT)是否通过调控钙调神经磷酸酶(Calcineurin,CaN)/活化T细胞核因子c3(Nuclear factors of activated T cell c3,NFATc3)信号通路改善球囊损伤所致大鼠颈动脉血管内膜增生。方法将雄性SD大鼠随机分为假手术组(Sham)、模型组(Model)和YDXNT给药组(YDXNT),采用球囊导管术建立大鼠左侧颈动脉内膜增生模型。造模次日至第14日,YDXNT给药组灌胃1.0 g/kg YDXNT混悬液,Sham和Model组灌胃等体积双蒸水。造模2周后麻醉后取左侧颈动脉标本,HE染色观察颈动脉血管病理学形态;免疫荧光技术检测颈动脉血管新生内膜中CaN、NFATc3、白介素-1β(Interleukin-1β,IL-1β)、肿瘤坏死因子α(Tumor necrosis factor-α,TNF-α)、单核趋化蛋白-1(Monocyte chemoattractant protein-1,MCP-1)蛋白表达情况。结果与Sham组相比,Model组颈动脉血管内膜厚度明显增加,管腔狭窄,新生内膜面积(Neointimal area,NIA)、NIA/中膜面积(Media area,MA)、NIA/内弹力板围绕面积(Internal elastic lamina area,IELA)明显升高;IL-1β、TNF-α、MCP-1、CaN蛋白表达明显增多(P<0.05),NFATc3蛋白核转位明显增多(P<0.05);与Model组相比,YDXNT组颈动脉血管内膜厚度明显降低,管腔增大,NIA、NIA/MA、NIA/IELA明显降低;IL-1β、TNF-α、MCP-1、CaN蛋白表达明显降低(P<0.05),NFATc3蛋白核转位明显减少(P<0.05)。结论YDXNT至少可能通过抑制CaN/NFATc3信号通路,减轻炎症反应,改善球囊损伤所致大鼠颈动脉血管内膜增生。

Objective To study the effect of Yindan-xinnaotong capsule(YDXNT)on carotid intimal hyperplasia induced by balloon injury in rats,and explore its mechanism based on calcineurin(CaN)/nuclear factors of activated T cell c3(NFATc3)signaling pathway.Methods Male SD rats were randomly divided into sham group(Sham),model(Model),and YDXNT groups,and the carotid artery was injured by balloon catheter to establish vascular restenosis model.YDXNT(1.0 g/kg)was intragastrically administered,Sham and Model groups were given the equal volume of distilled water.All above treatments were implemented from the first day to the 14th day of surgery for 2 weeks.HE staining was used to observe the pathological morphology of carotid artery;immunofluorescence was used to detect the protein expressions of CaN,NFATc3,interleukin-1β(IL-1β),tumor necrosis factor-α(TNF-α)and monocyte chemoattractant protein-1(MCP-1)in carotid neovascularization.Results Compared with the Sham group,the carotid artery intima thickness was significantly increased and the lumen was narrowed,the neointimal area(NIA),the ratio of neointimal area to media area(NIA/MA)and the ratio of neointimal area to internal elastic lamina area(NIA/IELA)was significantly increased in the Model group;the protein expressions of IL-1β,TNF-α,MCP-1 and CaN were significantly increased,the translocation of NFATc3 protein into the nucleus was increased significantly(P<0.05).Compared with the Model group,the carotid artery intima thickness was significantly decreased and the lumen was enlarged,NIA,NIA/MA and NIA/IELA were significantly decreased in the YDXNT group;the protein expressions of IL-1β,TNF-α,MCP-1 and CaN were significantly decreased(P<0.05).Translocation of NFATc3 protein into the nucleus decreased significantly(P<0.05).Conclusion YDXNT ameliorates the carotid intimal hyperplasia caused by balloon injury in rats,and the mechanism is at least related to the down-regulating of CaN/NFATc3 signaling pathway and reducing inflammatory reaction.