利伐沙班对脑缺血再灌注大鼠内皮功能的影响

Effects of rivaroxaban on endothelial function in cerebral ischemia-reperfusion rats

目的 探究利伐沙班对脑缺血再灌注大鼠内皮功能的影响。方法 将40只大鼠随机分为假手术组、模型组和低、高剂量实验组,每组10只。假手术组仅暴露血管,不作插线处理;其余3组用大脑中动脉线栓法建立脑缺血再灌注大鼠模型。低、高剂量实验组分别于手术前7 d灌胃给予10、30 mg·kg^(-1)利伐沙班,每天1次,第7天给药1 h后建立脑缺血再灌注模型,24 h后再分别灌胃给予10、30 mg·kg^(-1)利伐沙班,每天1次,持续3 d。模型组和假手术组均在同一时间灌胃给予等量0.9%NaCl。用酶联免疫吸附试验法检测大鼠脑组织中内皮素-1(ET-1)、白细胞介素-8(IL-8)和肿瘤坏死因子α(TNF-α)水平,用蛋白质印迹法检测大鼠脑组织中p38丝裂原激活蛋白激酶(p38MAPK)和核转录因子κB p65(NF-κB p65)蛋白的表达水平。结果 低、高剂量实验组和模型组、假手术组的ET-1水平分别为(197.53±29.63)、(155.66±23.35)、(270.16±40.53)和(123.57±18.53)ng·L^(-1),IL-8水平分别为(1.51±0.23)、(0.98±0.15)、(1.96±0.29)和(0.53±0.08)ng·L^(-1),TNF-α水平分别为(0.81±0.13)、(0.57±0.09)、(1.06±0.16)和(0.25±0.04)ng·L^(-1),p-p38MAPK/p38MAPK分别为0.75±0.08、0.57±0.07、0.92±0.09和0.45±0.06,NF-κB p65蛋白相对表达水平分别为0.87±0.09、0.73±0.08、0.99±0.09和0.61±0.06。模型组的上述指标与低、高剂量实验组和假手术组比较,差异均有统计学意义(均P<0.05)。结论 利伐沙班可能通过抑制p38MAPK/NF-κB通路活化,进而抑制炎症因子释放,从而对脑缺血再灌注大鼠内皮功能发挥保护作用。

Objective To explore the effect of rivaroxaban on endothelial function in cerebral ischemia-reperfusion rats.Methods Forty rats were randomly divided into sham-operation group,model group and experimental-L,-H groups with 10 rats per group.The sham-operation group only exposed the blood vessels without suture treatment,and the other three groups were treated with middle cerebral artery suture to establish the cerebral ischemia-reperfusion model.Experimental-L,-H groups were given 10 and 30 mg·kg^(-1) rivaroxaban by gavage on the 7th day before operation,once a day,and the cerebral ischemia-reperfusion model was established 1 hour after administration on the 7th day;24 hours later,10 and 30 mg·kg^(-1) rivaroxaban were given by gavage administration for 3 days with once a day. Model and sham - operation groups received an equal volume of 0. 9% NaCl by gavage at the same time. Thelevels of endothelin - 1( ET - 1) ,interleukin - 8 ( IL - 8) and tumor necrosis factor factor - α ( TNF - α) in rat brainwere measured by enzyme - linked immunosorbent assay method. The p38 mitogen - activated protein kinase( p38MAPK) and nuclear transcription factor κB p65 ( NF - κB p65 ) protein were detected by Western blotting.Results The levels of ET - 1 in experimental - L and experimental - H groups,model group and sham - operationgroup were ( 197. 53 ± 29. 63) ,( 155. 66 ± 23. 35) ,( 270. 16 ± 40. 53) and ( 123. 57 ± 18. 53) ng·L^(-1);the IL - 8levels were ( 1. 51 ± 0. 23) ,( 0. 98 ± 0. 15) ,( 1. 96 ± 0. 29) and ( 0. 53 ± 0. 08) ng·L^(-1);the TNF - α levels were( 0. 81 ± 0. 13) ,( 0. 57 ± 0. 09) ,( 1. 06 ± 0. 16) and ( 0. 25 ± 0. 04) ng·L^(-1),p - p38MAPK/p38MAPK were0. 75 ± 0. 08,0. 57 ± 0. 07,0. 92 ± 0. 09 and 0. 45 ± 0. 06;the relative expression levels of NF - κB p65 protein were0. 87 ± 0. 09,0. 73 ± 0. 08,0. 99 ± 0. 09 and 0. 61 ± 0. 06,respectively. The differences were statistically significantbetween the experimental - L,experimental - H and sham - operation groups and the model group ( all P < 0. 05) .Conclusion Rivaroxaban may inhibit the release of inflammatory cytokines by inhibiting the activation of p38MAPK/NF - κB pathway,which may play a protective role on endothelial function in cerebral ischemia - reperfusion rats.