METHODS: HSCs were derived from the livers of adul male Sprague-Dawley rats. IL-6 expression was evalu ated using real-time quantitative polymerase chain reaction and enzyme linked immunosorbent

assay. The phosphorylation activity of p38 mitogen activated pro tein kinases (MAPK) and extracellular regulated pro tein kinases (ERK) 1/2 upon induction by IL-17A and suppression by IL-17RA shRNA were examined using Western blotting.RESULTS: IL-6 expression 哪里 induced by IL-17A was significantly increased compared to control in HSCs (P < 0.01 in a dose-dependent manner). Suppression of IL17RA using lentiviral-mediated shRNA inhibited IL-6 expression induced by IL-17A compared to group with only IL-17A treatment (1.44 ± 0.17 vs 4.07 ± 0.43, P < 0.01). IL-17A induced rapid phosphorylation of p38 MAPK and ERK1/2 after 5 min exposure, and showed the strongest levels of phosphorylation of p38 MAPK and ERK1/2 at 15 min in IL-17A-treated HSCs. IL-6 mRNA expression induced by IL-17A (100 ng/mL) for 3 h

exposure was inhibited by preincubation with specific inhibitors of p38 MAPK (SB-203580) and ERK1/2 (PD-98059) compared to groups without inhibitors preincubation (1.67 selleck化学 ± 0.24, 2.01 ± 0.10 vs 4.08 ± 0.59, P < 0.01). Moreover, lentiviral-mediated IL-17RA shRNA 1 inhibited IL-17A-induced IL-6 mRNA expression compared to random shRNA in HSCs (1.44 ± 0.17 vs 3.98 ± 0.68, P < 0.01). Lentiviral-mediated IL17RA shRNA 1 inhibited phosphorylation of p38 MAPK and ERK1/2 induced by 15 min IL-17A (100 ng/mL) exposure. CONCLUSION: Down-regulation of the IL-17RA receptor by shRNA decreased IL-6 expression induced by IL-17A via p38 MAPK and ERK1/2 phosphorylation in HSCs. Suppression of IL-17RA expression may be a strategy to reduce the inflammatory response induced by IL-17A in the liver.
目的探讨丝裂原活化蛋白激酶(mitogen-activated protein kinase,MAPK)活化与脓毒症大鼠肺组织和血浆肿瘤坏死因子-α(TNF-α)表达及组织炎症反应的关系。方法采用盲肠结扎穿孔术(CLP)模型制造大鼠腹腔感染。随机将56只大鼠分为正常对照组、CLP组和CLP+MAPK抑制剂(SB203580)处理组,各组又分不同时间点亚组。采用反转录多聚酶链式反应和酶联免疫吸附试验检测试剂盒分别测定血浆和肺TNF-αmRNA和蛋白水平;同时测定肺髓过氧化酶(MPO)活性、HE染色病理切片检测肺组织炎症反应程度。结果与正常大鼠相比,CLP后各时间点肺血浆和肺组织TNF-αmRNA、蛋白含量均升高明显;同时肺MPO和组织病理学评分升高明显。MAPK抑制剂处理后,与CLP组相应时间点相比,MPO和病理学评分均显著下降;TNF-αmRNA表达和蛋白含量同时显著降低。结论抑制MAPK活化可缓和脓毒症所致大鼠肺组织炎症反应,减轻肺组织损伤。
目的:研究NF-κB信号传导通路在EAE发病中的作用,观察中药复方益肾达络饮干预实验性自身免疫性脑脊髓炎(EAE)的疗效,探讨益肾达络饮治疗EAE的作用机制。方法:采用免疫蛋白质印迹法、免疫组化法测定EAE小鼠中枢神经组织中NF-κBp65、IκB-α蛋白表达的变化。结果:中药复方益肾达络饮能降低EAE的复发率(P<0.05),改善EAE模型小鼠神经功能评分(P<0.05)。与模型组相比,激素组、中药组、中药加激素组NF-κBp65表达水平明显降低,IκB-α表达水平明显升高(P<0.05)。结论:在EAE中,NF-κB介导的炎性级联反应是EAE小鼠CNS炎性浸润、轴索损伤的重要原因;中药复方益肾达络饮能降低EAE的复发率,有效改善EAE小鼠神经功能损伤;中药益肾达络饮干预EAE的作用机制与对NF-κB信号通路的调节密切相关。
为了探索丙二醛(malonaldehyde,MDA)抑制间充质干细胞(mesenchymal

那个 stem cells,MSCs)成骨分化的作用机制,用不同浓度的丙二醛孵育MSCs,进行成骨诱导培养,检测碱性磷酸酶活性和钙结节形成;并检测p38和JNK表达水平和磷酸化程度,用这两种信号分子的特异性阻断剂进行验证.结果发现,丙二醛浓度依赖性地降低MSCs碱性磷酸酶的活性,抑制钙结节的形成;并引起p38和JNK的表达上调和磷酸化增强,诱导JNK由胞浆向胞核转位;p38和JNK阻断剂对丙二醛的上述效应有拮抗作用.结果表明,丙二醛可抑制MSCs的成骨诱导分化,其作用机制涉及p38和JNK信号通路的参与.