Department of Burns, Third Xiangya Hospital, Central South University, Changsha, Hunan, China (mainland).
Department of Emergency Medicine, Xiangya Third Hospital, Central South University, Changsha, Hunan, China (mainland).
Med Sci Monit. 2018 May 13;24:3119-3126. doi: 10.12659/MSM.905580.
BACKGROUND This study aimed to investigate the effects of recombinant human brain natriuretic peptide (rhBNP) on IL-6, TNF-α, and IL-10 secretion in LPS-activated RAW 264.7 cells and human peripheral blood mononuclear cells (PBMCs) in vitro and to explore the related signaling pathways of the regulation mechanisms of BNP in systemic inflammatory response syndrome (SIRS). MATERIAL AND METHODS MTT assay was used to evaluate the effects of rhBNP on cell viabilities. Lipopolysaccharide (LPS) was used to induce inflammation response. The whole study was divided into 8 groups: Control, low, middle, and high concentrations of rhBNP, LPS, LPS with low, middle, and high concentrations of rhBNP. Levels of IL-6, TNF-α, and IL-10 were evaluated using the Cytometric Bead Array Kit and RT-PCR assay. Western blotting was used to test the effects of rhBNP on inflammation-related NF-kB and MAPK pathways. RESULTS Except for the concentrations ≥1.6 ng/mL, all concentrations of rhBNP showed little effect on cell viabilities of RAW264.7 cells and PBMCs after 24 h and 48 h, suggesting a weak cytotoxicity to cells. Expression of IL-6 and TNF-α significantly increased and expression of IL-10 significantly decreased at protein and mRNA levels after LPS treatment, and these effects were strongly inhibited in a dose-dependent manner by pretreatment of rhBNP. Similarly, the LPS-induced increase of NF-kB and MAPK pathway phosphorylation levels were also significantly inhibited by rhBNP. CONCLUSIONS rhBNP can regulate expression of IL-6, TNF-α, and IL-10 in LPS-activated RAW 264.7 cells and PBMCs through inhibiting NF-κB and MAPK pathways. These results may reveal potential causes of the increase of BNP in SIRS and may provide an experimental basis for treatment of SIRS.
本研究旨在探讨重组人脑利钠肽(rhBNP)对脂多糖(LPS)激活的 RAW264.7 细胞和人外周血单个核细胞(PBMC)中白细胞介素 6(IL-6)、肿瘤坏死因子-α(TNF-α)和白细胞介素 10(IL-10)分泌的影响,并探讨 BNP 调节全身炎症反应综合征(SIRS)中相关信号通路的机制。
MTT 法评估 rhBNP 对细胞活力的影响。脂多糖(LPS)用于诱导炎症反应。整个研究分为 8 组:对照组、低、中、高浓度 rhBNP 组、LPS 组、LPS 加低、中、高浓度 rhBNP 组。采用细胞因子珠阵列试剂盒和 RT-PCR 法评估 IL-6、TNF-α 和 IL-10 水平。Western blot 法检测 rhBNP 对炎症相关 NF-κB 和 MAPK 通路的影响。
除浓度≥1.6ng/mL 外,rhBNP 各浓度在 24 小时和 48 小时后对 RAW264.7 细胞和 PBMC 的细胞活力几乎没有影响,提示细胞毒性较弱。LPS 处理后,IL-6 和 TNF-α 的表达在蛋白质和 mRNA 水平上显著增加,IL-10 的表达显著降低,rhBNP 预处理呈剂量依赖性方式强烈抑制这些作用。同样,rhBNP 也显著抑制 LPS 诱导的 NF-κB 和 MAPK 通路磷酸化水平升高。
rhBNP 可通过抑制 NF-κB 和 MAPK 通路调节 LPS 激活的 RAW264.7 细胞和 PBMC 中 IL-6、TNF-α 和 IL-10 的表达。这些结果可能揭示了 SIRS 中 BNP 增加的潜在原因,并为 SIRS 的治疗提供了实验依据。