灯盏乙素对心肌缺血/再灌注损伤的保护作用及机制
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摘要
目的研究灯盏乙素对大鼠离体心脏缺血/再灌注损伤的保护作用及机制。方法 SD大鼠随机分为空白组、模型组、灯盏乙素低、中、高剂量组(0.3、3、30 mg·kg~(-1)),预给药7 d,应用Langendorff离体灌流大鼠心脏复制急性心肌缺血/再灌注损伤模型。离体灌注时,给药组用含灯盏乙素的K-H液进行灌流(0. 3、3、30 mg·L~(-1))。记录心肌收缩功能;试剂盒检测大鼠心肌组织中AST、CK、LDH的活性; ELISA检测炎性因子ICAM-1、IL-1β、IL-6、IL-18、TNF-α的含量; HE染色观察心肌组织形态学的改变; Western blot检测IL-1β、NLRP3、NF-кB相关蛋白表达。结果与空白组相比,模型组大鼠离体心脏收缩功能明显降低,心肌组织AST、CK、LDH活性明显升高,ICAM-1、IL-1β、IL-6、IL-18、TNF-α等炎性因子的含量明显升高,IL-1β、NLRP3蛋白表达水平及NF-кB核转位明显升高。灯盏乙素能明显改善上述病理改变。结论灯盏乙素对心肌缺血/再灌注损伤的保护作用可能与抑制NF-кB/NLRP3/IL-1β通路有关。
Aim To study the protective effect of scutellarin(Scu) on ischemia/reperfusion(I/R) injury in isolated rat hearts and its mechanism. Methods Acute myocardial I/R injury was induced by Langendorff perfusion in isolated rat hearts. SD rats were randomly divided into five groups: blank group, model group, low, medium and high dose group of Scu(0.3, 3, 30 mg·kg~(-1)). The drug experimental group was perfused with K-H solution containing Scu(0.3, 3, 30 mg·L~(-1)) in vitro perfusion. Myocardial contractile function was recorded; AST, CK and LDH activities in rat myocardium were detected by kit; ICAM-1, IL-1β, IL-6, IL-18 and TNF-α were detected by ELISA; the morphological changes of myocardium were observed by HE staining; and the expressions of IL-1β, NLRP3 and NF-кB were detected by Western blot. Results Compared with blank group, the contractile function of isolated rat heart in model group significantly decreased, the activities of AST, CK and LDH in myocardium significantly increased, and the contents of inflammatory factors such as ICAM-1, IL-1β, IL-6, IL-18 and TNF-α significantly increased. The expression of IL-1β and NLRP3 and the nuclear translocation of NF-кB significantly increased. Scu significantly improved the above pathological changes. Conclusion The protective effect of Scu on myocardial I/R injury may be related to the inhibition of NF-кB/NLRP3/IL-1β pathway.
Aim To study the protective effect of scutellarin(Scu) on ischemia/reperfusion(I/R) injury in isolated rat hearts and its mechanism. Methods Acute myocardial I/R injury was induced by Langendorff perfusion in isolated rat hearts. SD rats were randomly divided into five groups: blank group, model group, low, medium and high dose group of Scu(0.3, 3, 30 mg·kg~(-1)). The drug experimental group was perfused with K-H solution containing Scu(0.3, 3, 30 mg·L~(-1)) in vitro perfusion. Myocardial contractile function was recorded; AST, CK and LDH activities in rat myocardium were detected by kit; ICAM-1, IL-1β, IL-6, IL-18 and TNF-α were detected by ELISA; the morphological changes of myocardium were observed by HE staining; and the expressions of IL-1β, NLRP3 and NF-кB were detected by Western blot. Results Compared with blank group, the contractile function of isolated rat heart in model group significantly decreased, the activities of AST, CK and LDH in myocardium significantly increased, and the contents of inflammatory factors such as ICAM-1, IL-1β, IL-6, IL-18 and TNF-α significantly increased. The expression of IL-1β and NLRP3 and the nuclear translocation of NF-кB significantly increased. Scu significantly improved the above pathological changes. Conclusion The protective effect of Scu on myocardial I/R injury may be related to the inhibition of NF-кB/NLRP3/IL-1β pathway.
引文
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[9] Tang Y L,Jiang J H,Wang S,et al.TLR4/NF-κB signaling contributes to chronic unpredictable mild stress-induced atherosclerosis in ApoE mice[J].PLoS One,2015,10(4):e123685.
[10] Kim M H,Jeong H J.Zinc oxide nanoparticles suppress LPS-induced NF-κB activation by inducing A20,a negative regulator of NF-κB,in RAW 264.7 macrophages[J].J Nanosci Nanotechnol,2015,15(9):6509-15.
[11] Li P,Wang Y,Turner J H.Proinflammatory mediators alter expression of nuclear factor kappa B-regulating deubiquitinases in sinonasal epithelial cells[J].Int Forum Allergy Rhinol,2015,5(7):583-9.
[12] Li Y,Liu M,Zuo Z,et al.TLR9 regulates the NF-κB-NLRP3-IL-1β pathway negatively in Salmonella-induced NKG2D-mediated intestinal inflammation[J].J Immunol,2017,199(2):761-73.
[2] 焦会芝,魏安银.灯盏花素与多种药物配伍情况的分析[J].中国疗养医学,2012,21(4):373-4.[2] Jiao H Z,Wei A Y.Analysis of the compatibility of breviscapine with a variety of drugs[J].Chin J Conval Med,2012,21(4):373-4.
[3] 褚剑锋,郑锋.灯盏细辛注射液在心血管内科中的应用新进展[J].福建中医学院学报,2005,15(增刊):44-6.[3] Chu J F,Zheng F.New progress in application of breviscapine injection in cardiovascular medicine[J].J Fujian Coll Tradit Chin Med,2005,15(Suppl):44-6.
[4] Francis Stuart S D,De Jesus N M,Lindsey M L,Ripplinger C M.The crossroads of inflammation,fibrosis,and arrhythmia following myocardial infarction[J].J Mol Cell Cardiol,2016,91:114-22.
[5] Toldo S,Marchetti C,Mauro A G,et al.Inhibition of the NLRP3 inflammasome limits the inflammatory injury following myocardial ischemia-reperfusion in the mouse[J].Int J Cardiol,2016,209:215-20.
[6] Takahashi M.NLRP3 inflammasome as a novel player in myocardial infarction[J].Int Heart J,2014,55(2):101-5.
[7] Marchetti C,Chojnacki J,Toldo S,et al.A novel pharmacologic inhibitor of the NLRP3 inflammasome limits myocardial injury after ischemia-reperfusion in the mouse[J].J Cardiovasc Pharmacol,2014,63(4):316–22.
[8] 杨晶,张晓坚,胡长平.吴茱萸次碱通过抑制TLR4/NF-κB信号通路保护大鼠心肌缺血/再灌注损伤[J].中国药理学通报,2017,33(12):1707-12.[8] Yang J,Zhang X J,Hu C P.Evovirin protects rat myocardial ischemia/reperfusion injury by inhibiting TLR4/NF-κB signaling pathway[J].Chin Pharmacol Bull,2017,33(12):1707-12.
[9] Tang Y L,Jiang J H,Wang S,et al.TLR4/NF-κB signaling contributes to chronic unpredictable mild stress-induced atherosclerosis in ApoE mice[J].PLoS One,2015,10(4):e123685.
[10] Kim M H,Jeong H J.Zinc oxide nanoparticles suppress LPS-induced NF-κB activation by inducing A20,a negative regulator of NF-κB,in RAW 264.7 macrophages[J].J Nanosci Nanotechnol,2015,15(9):6509-15.
[11] Li P,Wang Y,Turner J H.Proinflammatory mediators alter expression of nuclear factor kappa B-regulating deubiquitinases in sinonasal epithelial cells[J].Int Forum Allergy Rhinol,2015,5(7):583-9.
[12] Li Y,Liu M,Zuo Z,et al.TLR9 regulates the NF-κB-NLRP3-IL-1β pathway negatively in Salmonella-induced NKG2D-mediated intestinal inflammation[J].J Immunol,2017,199(2):761-73.