摘要
目的 分析孤儿核受体NR4A1对氧化应激诱导的血栓形成的保护作用及机制。方法 选取健康SD大鼠30只,随机分为对照组、模型组和NR4A1组,每组10只。用FeCl_3构建颈动脉血栓大鼠模型,用NR4A1激动剂促进大鼠NR4A1的表达。称取3组小鼠的血栓重量;采用苏木精-伊红染色观察颈动脉组织切片的血栓形成情况;采用蛋白质印迹法检测核转录因子-κB(NF-κB)/环氧合酶-2(COX-2)信号轴相关蛋白表达水平;采用硝酸还原酶法检测NO水平;采用酶联免疫吸附试验(ELISA)法检测内皮素-1(ET-1)、内皮型一氧化氮合酶(eNOS)、纤溶酶原激活物抑制剂-1(PAI-1)水平;采用放射免疫分析法测定血栓素(TX)B2、6-酮-前列腺素F1α(6-keto-PGF1α)水平。结果 对照组未形成血栓,NR4A1组大鼠血栓重量明显低于模型组,3组比较,差异均有统计学意义(P<0.05)。模型组与NR4A1组大鼠颈动脉组织NR4A1蛋白表达水平均低于对照组,且模型组低于NR4A1组,差异均有统计学意义(P<0.05);模型组与NR4A1组大鼠颈动脉组织p-NF-κB p65及COX-2蛋白表达水平均高于对照组,且模型组高于NR4A1组,差异均有统计学意义(P<0.05)。模型组、NR4A1组大鼠血清NO、eNOS、6-keto-PGF1α水平及NO/ET-1均低于对照组,且模型组均低于NR4A1组,差异均有统计学意义(P<0.05);模型组、NR4A1组大鼠血清ET-1、PAI-1、TXB2水平及TXB2/6-keto-PGF1α均高于对照组,且模型组均高于NR4A1组,差异均有统计学意义(P<0.05)。结论 NR4A1对NF-κB/COX-2信号通路相关蛋白表达具有负向调控作用,进而减轻大鼠深静脉血栓模型中的血栓和炎症。
Objective To analyze the protective effect and mechanism of orphan nuclear receptor NR4A1 on oxidative stress-induced thrombosis.Methods Thirty healthy SD rats were selected and randomly divided into control,model and NR4A1 groups,with 10 rats in each group.The rat model of carotid artery thrombosis was established with FeCl 3,and NR4A1 agonist was used to promote the expression of NR4A1 in rats.The thrombi of the mice in 3 groups were weighed.Carotid artery thrombosis was observed by HE staining in pathological sections.Western blotting was adopted to detected nuclear factors-κB(NF-κB)/COX-2 signal axis related protein expression level.Nitric oxide reductase method was used to detect the level of NO.Enzyme-linked immunosorbent assay(ELISA)was applied to detect the levels of endothelin 1(ET-1),endothelial nitric oxide synthase(eNOS),and plasmin activator inhibitor-1(PAI-1).Radioimmunoassay was imployed for the determination of thromboxane B2(TXB2)and 6-keto prostaglandin F1α(6-keto-prostaglandin F1alpha,6-keto-PGF1α)level.Results The control group did not form thrombus,and the weight of thrombus in NR4A1 group was significantly lower than that in the model group,with a stastically significant difference(P<0.05).The expression level of NR4A1 protein in the carotid tissue of the model group and NR4A1 group was lower than that of the control group,and the model group was lower than the NR4A1 group,and the differences were statistically significant(P<0.05).The expression levels of p-NF-κB p65 and COX-2 protein in the carotid artery tissue of the model group and NR4A1 group were higher than those of the control group,and the model group was higher than the NR4A1 group,and the differences had statistical significance(P<0.05).Serum levels of NO,eNOS,6-keto-PGF1αand NO/ET-1 in the model group and the NR4A1 group were lower than those in the control group,and the model group was lower than the NR4A1 group,with statistically significant difference(P<0.05).Serum levels of ET-1,PAI-1,TXB2 and TXB2/6-keto-PGF1αin the model group and the NR4A1 group were higher than those in the control group,and the model group was higher than the NR4A1 group,and the differences were statistically significant(P<0.05).Conclusion NR4A1 negatively regulates the expression of NF-κB/COX-2 signaling pathway related proteins,which in turn attenuates thrombosis and inflammation in the rat model of deep vein thrombosis.
作者
钟华平
李林
谢家和
ZHONG Huaping;LI Lin;XIE Jiahe(Cardiac ICU,First Affiliated Hospital of Gannan Medical College,Ganzhou,Jiangxi 341000,China;Department of Hematology,First Affiliated Hospital of Gannan Medical College,Ganzhou,Jiangxi 341000,China;Department of Cardiology,First Affiliated Hospital of Gannan Medical College,Ganzhou,Jiangxi 341000,China)
出处
《检验医学与临床》
CAS
2023年第15期2166-2169,共4页
Laboratory Medicine and Clinic
基金
江西省卫生健康委员会科技计划项目(202130655)
江西省赣州市指导性科技计划项目(GZ2020ZSF031)。
