论文部分内容阅读
目的:探讨Toll样受体4拮抗剂TAK-242抑制小鼠心肌缺血/再灌注损伤(ischemia/reperfusion,I/R)炎症反应的分子机制。方法:选用48只雄性C57BL/6小鼠随机分为4组:假手术组(sham)、模型组(I30min/R24h)、给药组[I/R+TAK-242(3 mg·kg-1)]、干预组[I/R+TAK-242+AG490(15 mg·kg-1)]。再灌注24 h后心脏超声检测小鼠心功能,氯化三苯基四氮唑(TTC)染色法测定心肌梗死面积,HE染色观察心肌病理改变,WB检测心肌JAK2/STAT3磷酸化水平,ELISA检测血清IL-6、TNF-α、IL-10和高迁移率族蛋白B1(HMGB1)浓度。结果:与sham组比较,I/R组小鼠左心室收缩期直径(LVIDs)延长(P<0.01),左心室射血分数(LVEF)和左心室短轴缩短分数(LVFS)显著降低(P<0.001或P<0.01),心梗面积明显增加并出现心肌炎性浸润,心肌p-JAK2/p-STAT3表达明显升高(P<0.01或P<0.05),血清IL-6、IL-10、TNF-α和HMGB1水平显著升高(P<0.001或P<0.01)。与I/R组比较,TAK-242给药组小鼠LVIDs缩短(P<0.05),LVEF和LVFS显著升高(P<0.01或P<0.05),心梗面积缩小(P<0.01),心肌炎症浸润减轻,心肌p-JAK2/p-STAT3表达降低(P<0.01或P<0.05),血清IL-6和TNF-α水平明显下降(P<0.001或P<0.01),而IL-10和HMGB1浓度进一步升高(P<0.01)。与TAK-242给药组比较,AG490干预可显著加强TAK-242治疗作用,包括心肌收缩功能增强,心梗面积缩小及炎性浸润程度减轻,心肌p-JAK2/p-STAT3表达降低(P<0.05),血清IL-6、TNF-α浓度下降而IL-10、HMGB1浓度升高(P<0.01或P<0.05)。结论:Toll样受体4拮抗剂TAK-242抑制小鼠I/R炎症反应与JAK2/STAT3信号通路失活有关。
AIM: To investigate the molecular mechanism of TAK-242, an inhibitor of toll-like receptor 4, in inhibiting myocardial ischemia / reperfusion (I / R) inflammation in mice. Methods: Forty-eight male C57BL / 6 mice were randomly divided into 4 groups: sham, model group (I30min / R24h), I / R + TAK-242 )], Intervention group [I / R + TAK-242 + AG490 (15 mg · kg-1)]. Twenty-four hours after reperfusion, cardiac function was detected by echocardiography, myocardial infarction size was measured by TTC staining, myocardial pathological changes were observed by HE staining, phosphorylation of JAK2 / STAT3 was detected by WB, Serum IL-6, TNF-α, IL-10 and high mobility group box 1 protein (HMGB1) concentrations. RESULTS: Compared with sham group, left ventricular systolic diameter (LVIDs), left ventricular ejection fraction (LVEF) and left ventricular fractional shortening fraction (LVFS) in I / R group were significantly decreased (P <0.01) (P <0.01 or P <0.01, P <0.01 or P <0.01). Myocardial infarct size was significantly increased and myocardial infiltration was found. The expression of p-JAK2 / TNF-α and HMGB1 levels were significantly increased (P <0.001 or P <0.01). Compared with the I / R group, the LVIDs of TAK-242-treated mice were shortened (P <0.05), LVEF and LVFS were significantly increased (P <0.01 or P <0.05), myocardial infarct size was reduced (P <0.01 or P <0.05), the levels of IL-6 and TNF-α in serum decreased significantly (P <0.001 or P <0.01), while the levels of IL-10 and HMGB1 concentration was further increased (P <0.01). Compared with TAK-242 administration group, AG490 intervention significantly enhanced the therapeutic effect of TAK-242, including enhanced myocardial contractile function, reduced myocardial infarct size and inflammatory infiltration, and decreased myocardial p-JAK2 / p-STAT3 expression (P < 0.05). Serum levels of IL-6 and TNF-α decreased while concentrations of IL-10 and HMGB1 increased (P <0.01 or P <0.05). Conclusion: Inhibition of Toll-like receptor 4 antagonist TAK-242 on I / R inflammation in mice is associated with inactivation of JAK2 / STAT3 signaling pathway.