摘要
目的探讨3,3'-二吲哚甲烷(DIM)对脂多糖(LPS)诱导的新生大鼠心肌细胞炎症损伤的作用及其机制。方法提取分离并培养新生大鼠心肌细胞,分为4组:对照组、LPS(10 mg·L^-1)组、DIM(20μmol·L^-1)组及LPS+DIM组。CCK8实验检测DIM对细胞活性的影响;免疫荧光染色观察各组心肌细胞形态改变;用实时-荧光定量聚合酶链免疫反应检测12,24 h促炎介质mRNA表达水平;同时用Western blotting法分析DIM对MAPKs信号通路的影响。结果与LPS组比较,LPS+DIM组早期炎症因子,包括白细胞介素6(IL-6)和肿瘤坏死因子α(TNF-α)以及晚期促炎细胞因子高迁移率族蛋白1(HMGB1)mRNA水平显著降低(P<0.05)。与LPS组比较,LPS+DIM组Toll样受体-4(TLR-4)、p-ERK1/2、p-P38和p-JNK等蛋白水平显著降低。结论DIM通过抑制TLR-4/MAPKs信号通路,调节炎症因子IL-6、TNF-α和HMGB1的过度产生,从而改善LPS诱导的新生大鼠心肌细胞的炎症损伤。
Objective This study aimed to evaluate the effects of 3,3′-diindolylmethane(DIM)on LPS-induced inflammatory injury in neonatal rat cardiomyocytes and explore its molecular mechanisms.Methods The neonatal rat cardiomyocytes were isolated and cultured,which was divided into 4 groups:control group,LPS(10 mg·L^-1)group,DIM(20μmol·L^-1)group and LPS+DIM group.Cell viability was measured by CCK8,and the morphology of cultured neonatal rat cardiomyocytes was observed with immunofluorescence staining.The mRNA expression levels of pro-inflammatory mediators were measured with real-time PCR and the protein expression levels were examined with Western blotting analysis.Results The mRNA levels of early inflammatory factors in the LPS+DIM group,including interleukin-6(IL-6)and tumor necrosis factor alpha(TNF-α)and late pro-inflammatory cytokine high mobility group box 1(HMGB1),were significantly lower than those of the LPS group(P<0.05).Moreover,in the LPS+DIM group,the elevated levels of proteins such as Toll like receptor-4(TLR-4)and phosphorylated extracellular signal-regulated kinases 1/2(ERK1/2),phosphorylated P38 and phosphorylated c-Jun NH2-terminal kinase(JNK)induced by LPS were significantly lower than those of the LPS group.Conclusion DIM had a protective effect on LPS-induced inflammatory injury in neonatal rat cardiomyocytes,and modulated the excessive production of pro-inflammatory cytokines IL-6,TNF-αand HMGB1 via suppressing the TLR-4/MAPKs signaling pathways.
作者
杨文健
焦蓉
刘月
孙晓东
桑明
YANG Wenjian;JIAO Rong;LIU Yue;SUN Xiaodong;SANG Ming(Department of Pediatrics,the First People's Hospital of Xiangyang City,Affiliated Hospital of Hubei University of Medicine,Xiangyang 441000,China)
出处
《医药导报》
CAS
北大核心
2019年第9期1146-1151,共6页
Herald of Medicine
