血必净注射液对缺氧/复氧大鼠心肌TLR4--NF-κВ--IL-1β通路的影响

Effect of Xuebijing injection on TLR4--NF-κВ--IL-1βpathway of myocardial hypoxia/reoxygenation in rats

目的:探讨血必净注射液(XBJI)对缺氧/复氧大鼠心肌TLR4--NF-κВ--IL-1β通路的抑制作用。方法:健康雄性SD大鼠36只,体重(280±30)g,随机分为6组(n=6):正常组(N组)、平衡灌注组(BP组)、模型组(M组)、小剂量XBJI组(XBJIL组)、中剂量XBJI组(XBJIM组)、大剂量XBJI组(XBJIH组)。通过langendorff离体心脏灌流装置,建立缺氧/复氧大鼠心肌炎性反应模型。用ELISA方法检测心肌组织中白介素1β(IL-1β)的浓度;Western blot检测心肌组织中核因子-κВp65(NF-κВp65)蛋白、Toll样受体4(TLR4)蛋白的表达;RT-PCR检测心肌组织中NF-κВp65 mRNA及TLR4 mRNA的表达;光镜观察其心肌光学显微结构的改变。结果:与M组比较,XBJIL组、XBJIM组、XBJIH组心肌IL-1β的浓度、NF-κВp65及TLR4蛋白、NF-κВp65及TLR4 mRNA表达量有不同程度的下降,均以XBJIM组下降最为明显(P<0.05,P<0.01);M组心肌结构损伤严重,XBJI处理后上述变化均得到改善,以XBJIM最为明显。结论:不同剂量XBJI可通过抑制TLR4--NF-κВ--IL-1β信号转导通路减轻缺氧/复氧大鼠心肌的炎性反应,以4 ml/100ml的XBJI疗效最佳。

Objective： To investigate the role of Xuebijing injection（XBJI, traditional Chinese medicine）, in inhibiting TLR4--NF IL-1/3 pathway of myocardial hypoxia/reoxygenation in rats. Methods： Thirty six male SD rats （280 _＋ 30）g were randomly divided into six groups（ n = 6） ： normal group （ N group）, balanced peffusion group （ BP group）, model group （ M group）, low dose XBJI group（ XBJIL group）, middle dose XBJI group（XBJIM group）, high dose XBJI group （XBJIH group）. By langendorff isolated heart perfusion device to establish the model of myocardial hypoxia/reoxygenation in rats. ELISA was used to detect the concentration of interleukin-1β （IL-1β）; Western blot was used to detect the expression of nuclear factor kappa B p65 （ NF-xB 1365 ）protein and toll like receptor 4（TLR4 ） protein; and RT-PCR to deter- mine the expression of NF-xB p65 mRNA and TLR4 mRNA;To observe microstrueture changes of hypoxia/reoxygenation myocardial by light microscopy. Results： Compared with M group,the IL-l^3 eoneentration,NF-tcB 1365 and TLR4 protein,NF-xB p65 and TLR4 mRNA of XBJIL group,XBJ/M group, XBJIH group expression decreased in varying degrees,and decreased most obviously all in XBJIM group（ P 〈 0.05, P 〈 0. 01 ）; Myocardical structural damage was serious in M group,and improved after treatment XBJI,the most obvious was the XBJIM. Conclusion： Different dose of XBJI can inhibit TLR4--NF-xB--IL- 1β signal transduction pathway and reduce several inflammatory reaction after myocardial hypoxia/reoxygenation iniury,the 4 ml/100 rnl of XBJI is the best.