复方龙脉宁对急性心肌梗死模型大鼠TLR4/MyD88/NF-κB p65信号通路的影响

Effect of Compound Longmaining on TLR4/MyD88/NF-κB p65 Signaling Pathway in Rats with Acute Myocardial Infarction

目的:探讨复方龙脉宁汤对盐酸异丙肾上腺素诱导的心肌梗死大鼠模型的保护作用及其对Toll样受体4(TLR4)/髓样分化因子(MyD88)/核转录因子-κB(NF-κB)信号通路的影响。方法:取雄性SD大鼠48只,随机分成6组:正常组,模型组,复方丹参滴丸组(0.072 9 g·kg^(-1)),复方龙脉宁低、中、高剂量组(0.36,0.71,1.43 g·kg^(-1))。采用背部皮下多点注射盐酸异丙肾上腺素的方法建立急性心肌梗死动物模型。采用苏木素-伊红(HE)染色观察大鼠心肌组织病理学变化;酶联免疫吸附测定(ELISA)检测大鼠血清中白细胞介素-1β(IL-1β),白细胞介素-6(IL-6),肿瘤坏死因子-α(TNF-α),单核细胞趋化蛋白-1(MCP-1),一氧化氮(NO)的含量;免疫组化法测定大鼠心肌组织中NF-κB激酶β抑制蛋白(IKKβ),NF-κB抑制蛋白α(IκBα)的表达水平;蛋白免疫印迹法(Western blot)检测大鼠心肌组织中TLR4,MyD88,NF-κB p65蛋白的表达。结果:与正常组比较,模型组大鼠心肌损伤明显,血清中IL-1β,IL-6,TNF-α,MCP-1,NO含量明显升高(P<0.05),心肌组织中IκBα蛋白表达水平明显降低(P<0.05),心肌组织中IKKβ,TLR4,MyD88,NF-κB p65蛋白表达水平明显升高(P<0.05);与模型组比较,复方龙脉宁低、中、高剂量组能明显改善大鼠心肌损伤,明显升高IκBα蛋白的表达(P<0.05),明显降低血清中IL-1β,IL-6,TNF-α,MCP-1,NO的活性及心肌组织中IKKβ,TLR4,MyD88,NF-κB p65蛋白的表达(P<0.05)。结论:复方龙脉宁对心肌梗死的保护作用,可能与调控TLR4/MyD88/NF-κB p65信号通路相关因子的表达有关。

Objective: To investigate the protective effect of compound Longmaining isoprenaline hydrochloride-induced myocardial infarction model and its effect on Toll-like receptor 4( TLR4)/myeloid differentiation factor 88( MyD88)/nuclear transcription factor-kappa B( NF-κB) signaling pathway. Method:Forty-eight male SD rats were randomly divided into 6 groups: normal group,model group,compound salvia miltiorrhiza drop pill group( 0. 072 9 g·kg-1),and low,medium and high-dose compound Longmaining decoction groups( 0. 36,0. 71,1. 43 g·kg-1). The acute myocardial infarction model was induced through subcutaneous injection with isoproterenol. The pathological changes of myocardial tissue were examined by hematoxylin-eosin( HE) staining. The levels of interleukin-1β( IL-1β),interleukin-6( IL-6),tumor necrosis factor-α( TNF-α),monocyte chemotaxis protein-1( MCP-1) and nitrogen( NO) in serum were measured by enzyme linked immunosorbent assay( ELISA). The expression levels of inhibitors of NF-κB kinase subunit-β( IKKβ),NF-κB inhibitor α( IκBα),TLR4,MyD88 and NF-κB p65 were measured by immunohistochemical staining and Western blot. Result: Compared with normal group,the myocardial injury in model group was obvious. The levels of IL-1β,IL-6,TNF-α,MCP-1 and NO in serum increased significantly( P < 0. 05),the expression level of IκBαdecreased significantly( P < 0. 05),and the expression levels of IKKβ,TLR4,MyD88 and NF-κB p65 increased significantly in myocardial tissue( P < 0. 05). Compared with model group,low,medium and high-dose compound Longmaining groups could significantly alleviate the degree of myocardial injury in rats,significantly decrease IL-1β,IL-6,TNF-α,MCP-1 and NO levels in the serum( P < 0. 05),significantly increase the expression level of IκBα( P < 0. 05),and decreased the expression levels of IKKβ,TLR4,MyD88 and NF-κB p65( P < 0. 05).Conclusion: Compound Longmaining plays a protective effect on acute myocardial infarction by regulatingthe expressions of TLR4/MyD88/NF-κB p65 signaling pathway and relevant inflammatory factors.