Sun Jing, Shi Shaolan, Wang Qun, Yu Kezhou, Wang Rong
Department of Nephrology, Shandong Provincial Hospital, Shandong University Jinan 250021, China.
Int J Clin Exp Pathol. 2015 Feb 1;8(2):1555-64. eCollection 2015.
In the present study, we investigated whether CVVH can reduce HMGB1, TLR4, NF-κB and other serum cytokine levels, preventing organ injury in a dog sepsis model. A total of 10 dogs were injected with LPS and treated with either CVVH group (n = 5) or nothing (Control, n = 5) for 24 h. EILSA was used for examining the concentration of TNF-α, IL-6, HMGB 1 and TLR4. The histological change of lung, liver and kidney tissues was determined. The mRNA expression of HMGB1, TLR4 and NF-κB was examined by RT-PCR. The protein of HMGB1 and phosphated NF-κB was examined by Western-blot. The levels of serum HMGB1 came to the peak at 8 h, 16 h and then declined. The LPS-induced increase in HMGB1 level was suppressed by CVVH compared with Control. Likewise, serum TNF-α and IL-6 levels decreased with CVVH along with a significant improvement in the function of main organs. Histologic examination revealed significant reduction in inflammation in lung; liver and kidney tissues harvested 24 h after CVVH compared with Control. The mRNA of HMGB1, TLR4 and NF-κB in the kidney was expressed at high level after LPS administration, which was significantly decreased by CVVH. The increased protein expression of HMGB1 and phosphated NF-κB was reduced after CVVH compared with control. CVVH by reducing the level of HMGB1, TLR4, NF-κB and other cytokines could weaken the cascade of cytokines and restore the immune system, and reduce the damage of important organs in sepsis.
在本研究中,我们调查了连续性静脉-静脉血液滤过(CVVH)是否能降低高迁移率族蛋白B1(HMGB1)、Toll样受体4(TLR4)、核因子κB(NF-κB)及其他血清细胞因子水平,从而在犬脓毒症模型中预防器官损伤。总共10只犬注射脂多糖(LPS),并分为CVVH组(n = 5)或不做处理(对照组,n = 5),处理24小时。采用酶联免疫吸附测定法(ELISA)检测肿瘤坏死因子-α(TNF-α)、白细胞介素-6(IL-6)、HMGB 1和TLR4的浓度。测定肺、肝和肾组织的组织学变化。采用逆转录-聚合酶链反应(RT-PCR)检测HMGB1、TLR4和NF-κB的mRNA表达。采用蛋白质免疫印迹法检测HMGB1和磷酸化NF-κB的蛋白表达。血清HMGB1水平在8小时、16小时达到峰值,随后下降。与对照组相比,CVVH抑制了LPS诱导的HMGB1水平升高。同样,CVVH使血清TNF-α和IL-6水平降低,主要器官功能显著改善。组织学检查显示,与对照组相比,CVVH后24小时采集的肺、肝和肾组织炎症明显减轻。LPS给药后,肾中HMGB1、TLR4和NF-κB的mRNA高水平表达,CVVH使其显著降低。与对照组相比,CVVH后HMGB1和磷酸化NF-κB的蛋白表达增加减少。CVVH通过降低HMGB1、TLR4、NF-κB及其他细胞因子水平,可减弱细胞因子级联反应,恢复免疫系统,并减轻脓毒症中重要器官的损伤。