粉防己碱抑制心肌细胞磷酸化减轻缺血/再灌注损伤引发的炎症反应

Tetrandrine inhibited IκB-α phosphorylation regulated the expression of TNF-α、IL-6 during rat myocardial ischemic/reperfusion

目的:探讨心肌缺血/再灌注过程中粉防己碱(Tet)对核转录因子(NF)-κB的抑制因子(IκB-α)磷酸化及肿瘤坏死因子-α(TNF-α)、白细胞介素-6(IL-6)表达的影响以及机制。方法:60只SD大鼠被随机分为3组:缺血/再灌注损伤组(I/R)、假手术组、粉防己碱治疗组(Tet)。I/R组结扎大鼠左前降支造成心肌缺血30min后再灌注24h,然后处死大鼠;假手术组只穿针不结扎,余步骤同I/R组;Tet组在缺血前20min腹腔注射Tet,余步骤同I/R组。处死大鼠24h后取心肌标本,用酶联免疫吸附法(ELISA)检测心肌组织中TNF-α、IL-6表达水平,应用免疫组织化学方法检测磷酸化IκB-α(P-IκB-α)的表达。结果:Tet组与I/R组相比TNF-α、IL-6表达水平明显减低[(208.40±25.12)pg/ml∶(306.65±17.78)pg/ml,(587.40±137.40)pg/ml∶(1003.75±138.33)pg/ml,P均<0.01],而Tet组与假手术组相比表达明显增强[(208.40±25.12)pg/ml∶(61.45±9.20)pg/ml,(587.40±137.40)pg/ml∶(229.25±90.13)pg/ml,P均<0.01]。Tet组大鼠心肌细胞胞浆中P-IκB-α的表达明显低于I/R组[(0.0817±0.09)pg/ml∶(0.1755±0.01)pg/ml,但明显高于假手术组[(0.0817±0.09)pg/ml∶(0.0513±0.03)pg/ml,P均<0.01]。结论:Tet可以抑制IκB-α磷酸化,显著减少前炎症因子TNF-α、IL-6的产生,减轻心肌缺血/再灌注损伤。

Objective：To investigate the effect and mechanism of tetrandrine（Tet） on the phosphorylation of IκB-α（P-IκB-α） and the expression of TNF-α,IL-6 during rat myocardial ischemic/reperfusion（IR） period.Methods：A total of 60 male Sprague-Dawley rats were randomly divided into 3 groups：Sham group,I/R group,Tet group.The Sprague-Dawley rat underwent 30 min of left anterior descending （LAD） coronary occlusion and 24 h reperfusion to make ischemia/reperfusion （I/R） injury model in vivo.Sham group were not subjected to occlusion of artery.Tet group were injected with tetrandrine to abdominal cavity 20 min before ischemic starting.The other procedures were same to the I/R group.Samples were collected after 24h reperfusion.The expression level of TNF-α and IL-6 protein was detected by ELISA.The expression of P-IκB-α was observed by immunohistochemical stain and image analysis.Results：In Tet group the expression of TNF-α 、IL-6 were significant lower compared with those of I/R group[（208.40±25.12）pg/ml vs.（306.65±17.78）pg/ml,（587.40±137.40）pg/ml vs.（1003.75±138.33）pg/ml,P〈0.01 all] and significant higher than shame group respectively[（208.40±25.12）pg/ml vs.（61.45±9.20）pg/ml,（587.40±137.40）pg/ml vs.（229.25±90.13）pg/ml,P〈0.01 all）.In Tet group the expression of P-IκB-α were significant lower than that of I/R group[（0.0817±0.09）pg/ml vs.（0.1755±0.01）pg/ml,and significant higher than that of shame group[（0.0817±0.09）pg/ml vs.（0.0513±0.03）,P〈0.01 all].Conclusion：Tetrandrine can attenuate myocardial ischemia/reperfusion injury,inhibit the IκB-α phosphorylation and reduce the harmful cytokine TNF-α and IL-6.