阿司匹林触发消退素D1在大鼠动脉吻合口瘤中的作用及机制研究

Role and mechanism of aspirin-triggered resolvin D1 in rat model of anastomotic aneurysm

目的探讨阿司匹林触发消退素D1(AT-RvD1)在大鼠动脉吻合口瘤中的作用机制。方法66只雄性SD大鼠随机分为假手术组18只、模型组18只、药物组(AT-RvD1)18只、抑制剂组(AT-RvD1+MRT-67307)12只,假手术组仅显露血管,余3组采用右颈总动脉离断后吻合、左颈总动脉结扎术构建吻合口瘤模型,采用免疫荧光染色检测骨桥蛋白(OPN)和α平滑肌肌动蛋白(α-SMA)表达变化,Western blot检测磷酸化(p)-TANK结合激酶1(TBK1)、p-NF-κB,OPN表达。结果与模型组比较,药物组外径、外膜、平滑肌层、内膜厚度显著缩小[(1.70±0.19)mm vs(2.60±0.10)mm,(85.93±17.34)μm vs(189.36±9.64)μm、(50.12±5.16)μm vs(143.38±10.48)μm、(34.09±6.77)μm vs(98.41±8.97)μm,P<0.05],而管腔内径显著增大[(575.78±21.15)μm vs(541.56±36.79)μm,P<0.05]。与假手术组比较,模型组血管平滑肌细胞排列紊乱,α-SMA、p-TBK1显著下调,OPN、Ki-67、p-NF-κB及炎性因子表达显著升高(P<0.05)。药物组血管平滑肌细胞结构连续、有序,Ki-67、OPN及炎性因子表达显著低于模型组(P<0.05)。抑制剂组OPN、p-NF-κB及炎性因子表达显著高于药物组(P<0.05)。结论AT-RvD1可抑制动脉吻合口瘤形成、发展,其机制与抑制炎症及血管平滑肌细胞表型转化有关。

Objective To investigate the role and underlying mechanism of AT-RvD1 in arterial anastomotic aneurysms of rats.Methods Sixty-six male SD rats were randomly divided into 4 groups,that is,sham operation group(n=18),model group(n=18),drug group(AT-RvD1,n=18)and inhibitor group(AT-RvD1+MRT-67307,n=12).The sham group was only revealed the vessels,and the other groups were inflicted with right common carotid artery dissection followed by anastomosis and left common carotid artery ligation to construct an anastomotic aneurysm model.The corresponding agents were given to the rats of the drug group and the inhibitor group.SMC synthetic marker osteopontin(OPN)and contractile markerα-SMA were analyzed by immunofluorescence staining.The expression of p-TBK1,p-NF-κB and OPN were measured by Western blotting.Results The outer diameter and the thicknesses of adventitia,smooth muscle layer and intima were significantly decreased(1.70±0.19 mm vs 2.60±0.10 mm,85.93±17.34μm vs 189.36±9.64μm,50.12±5.16μm vs 143.38±10.48μm,34.09±6.77μm vs 98.41±8.97μm,P<0.05),while the inner diameter of the lumen was obviously larger in the drug group than the model group(575.78±21.15μm vs 541.56±36.79μm,P<0.05).The model group exhibited ovviuosly disarranged VSMC,downregulatedα-SMA and p-TBK1,increased levels of OPN,Ki-67,p-NF-κB and inflammatory factors when compared with the sham group(P<0.05).In the drug group,VSMC were well-arranged in structure,and the expression levels of Ki-67,OPN and inflammatory factors were significantly decreased when compared with those in the model group(P<0.05).The expression levels of OPN,p-NF-κB and inflammatory factors were significantly higher in the inhibitor group than the drug group(P<0.05).Conclusion AT-RvD1 inhibits the formation and progression of arterial anastomotic aneurysm,which may be associated with its inhibition on inflammation and phenotypic transformation of VSMC.