信号素3F对大鼠心肌细胞氧化应激、炎症反应及白细胞跨内皮迁移的影响

Effects of semaphrins 3F on oxidative stress,inflammatory response,and cross endothelial migration of leukocytes in rat cardiomyocytes

目的探讨信号素3F对心脏骤停大鼠存活心肌细胞氧化应激、炎症反应以及白细胞跨内皮迁移的影响。方法18只雄性Sprague-Dawley大鼠构建心脏骤停心肺复苏模型后随机分为模型组和信号素3F组,另取9只大鼠作为对照组;检测各组大鼠心肌细胞内活性氧(ROS)和过氧化氢(H_(2)O_(2))含量,酶联免疫吸附法(ELISA)检测心肌细胞超氧化物歧化酶(SOD)及丙二醛(MDA)含量,实时荧光定量聚合酶链反应(qPCR)检测心肌细胞中肿瘤坏死因子-α(TNF-α)、白细胞介素-1β(IL-1β)和干扰素-γ(IFN-γ)mRNA表达量;计数腹膜灌洗中白细胞数,蛋白质免疫印迹检测血小板内皮细胞黏附分子1(PECAM-1)和神经毡蛋白2(NRP2)表达。结果与模型组比较,信号素3F组心脏骤停大鼠心肌细胞中ROS、H_(2)O_(2)、MDA和SOD含量均明显增加,差异有统计学意义(P<0.05);与模型组比较,信号素3F组心脏骤停大鼠心肌细胞中TNF-α、IL-1β和IFN-γmRNA表达量均显著增加,差异有统计学意义(P<0.05);与模型组比较,信号素3F心脏骤停大鼠腹腔灌洗中白细胞计数、PECAM-1和NRP2蛋白表达量均增加,差异有统计学意义(P<0.05)。结论信号素3F影响心脏骤停大鼠存活的心肌细胞氧化应激、炎症反应及白细胞的跨内皮迁移。

Objective To investigate the effects of semaphrins 3F on oxidative stress,inflammatory response,and cross endothelial migration of leukocyte in viable cardiomyocytes after cardiac arrest in rats.Methods Eighteen male Sprague Dawley rats were used to construct the cardiopulmonary resuscitation model of cardiac arrest,and the rats were randomly divided into model group and semaphrins 3F group,another 9 rats were used as control group.Detecting the content of reactive oxygen species(ROS)and hydrogen peroxide(H_(2)O_(2))in cardiomyocytes of all groups;enzyme-linked immunosorbent assay(ELISA)was used to detect the content of superoxide dismutase(SOD)and malondialdehyde(MDA);real time fluorescence quantitative polymerase chain reaction(qPCR)was used to detect tumor necrosis factor in cardiomyocytes-α(TNF-α),interleukin-1β(IL-1β),interferon-γ(IFN-γ)expression of mRNA.Counting the number of leukocytes in peritoneal lavage of rats;the expressions of platelet endothelial cell adhesion molecule-1(PECAM-1)and neurofelt 2(Nrp2)were detected by Western blot.Results Compared with model group,contents of ROS,H_(2)O_(2),MDA,and SOD in cardiomyocytes after cardiac arrest in semaphrins 3F group were obviously increased,differences were statistically significant(P<0.05).mRNA expression of TNF-α,IL-1β,and IFN-γin cardiomyocytes after cardiac arrest in semaphrins 3F group were obviously increased,differences were statistically significant(P<0.05);leukocytes count,the expressions of PECAM-1 and NRP2 in cardiomyocytes after cardiac arrest in semaphrins 3F group received peritoneal lavage were obviously increased,differences were statistically significant(P<0.05).Conclusion Semaphrins 3F can affect the oxidative stress,inflammatory response,and cross endothelial migration of leukocytes in cardiac arrest survival rats.