摘要
目的探讨甲基强的松龙(methylprednisolone, MP)对脂多糖(Lipopolysaccharide, LPS)诱导急性肺损伤(acute lung injury,ALI)中STAT3-ERK1/2的影响和作用。方法8周龄C57BL/6J雄性小鼠随机(随机数字法)分为4组(每组8只):健康对照组(Control组)、LPS造模组(LPS组)、单纯MP给药组(MP组)、MP干预组(LPS+MP组)。经肺组织湿/干质量比(W/D)测定、苏木精-伊红(HE)染色、酶联免疫吸附试验(ELISA)、实时荧光定量聚合酶链反应(qRT-PCR)和免疫印迹(Western Blotting)检测,分析肺组织病理变化,血清及肺组织中炎症因子含量及表达水平,STAT3和ERK1/2信号通路表达水平。多组比较采用单因素方差分析,均数间两两比较采用LSD-t检验。结果(1)LPS+MP组小鼠肺组织的W/D显著低于LPS组[(3.01±0.84) vs (3.87±0.17),P = 0.038);(2)LPS+MP组小鼠肺组织较LPS组炎症细胞浸润少,肺泡结构基本完整;(3)LPS+MP组血清肿瘤坏死因子-α(TNF-α)和白细胞介素-6(IL-6)水平较LPS组显著下降[分别为(3.17±1.64)pg/mL vs ( 6.61±1.27) pg/mL,P = 0.003;(1.42±0.35) pg/mL vs (3.80±1.35) pg/mL, P = 0.008);肺组织TNF-α mRNA和IL-6 mRNA较LPS组显著降低[分别为(5.10±0.81) vs (12.2±5.05),P = 0.03;(1.62±1.00) vs (11.12±6.56),P = 0.026];(4)MP干预显著抑制STAT3和ERK1/2信号通路,表现为磷酸化STAT3(p-STAT3)和磷酸化ERK1/2(p-ERK1/2)表达水平降低[(0.26 ±0.05) vs (0.86 ±0.06), P < 0.001,(0.24±0.02) vs (1.34±0.32), P < 0.001]。结论甲基强的松龙对LPS诱导的急性肺损伤具有保护作用,其机制与抑制STAT3-ERK1/2信号通路、下调TNF-α和IL-6表达有关。
Objective To investigate the effects of methylprednisolone on STAT3-ERK1/2 signaling pathway in lipopolysaccharide (LPS)-induced acute lung injury (ALI). Methods The C57BL/6J male mice (8-week-old) were randomly(random number) divided into 4 groups: control group (control), LPS-induced endotoxemia model (LPS), only methylprednisolone (MP) administration group (MP), and intervention group with 2 mg/kg MP (LPS+MP)(n= 8 per group). The wet/dry (W/D) weight ratio of lung tissue, lung pathology by hematoxylin & eosin (HE) staining, serum and mRNA levels of TNF-α and IL-6 in lungs were determined. The protein levels of p-STAT3 and p-ERK1/2 in lungs were detected by Western blot. Statistical analyses were performed using One-way analysis of variance test to compare among multiple groups. Results (1)MP treatment significantly decreased the lung W/D weight ratio compared with the LPS group[(3.01±0.84) vs(3.87±0.17), P = 0.038];(2) The histopathological lesions of the lung were improved in the LPS+MP group compared with the LPS group accompanied with reduced inflammatory cell infiltration and attenuated the alveolar wall thickening;(3) The serum levels of TNF-α and IL-6 in the LPS+MP group was significantly decreased compared with the LPS group[(3.17±1.64) pg/mL vs (6.61±1.27) pg/mL, P = 0.003;(1.42±0.35) pg/mL vs (3.80±1.35) pg/mL, P = 0.008, respectively], and the mRNA levels of TNF-α and IL-6 in the LPS+MP group were significantly lower than those of the LPS group [(5.10±0.81) vs (12.2±5.05), P = 0.03;(1.62±1.00) vs (11.12±6.56), P=0.026;respectively];(4) MP therapy significantly inhibited P-STAT3 and P-ERK1/2 protein levels [(0.26±0.05) vs (0.86±0.06), P < 0.001,(0.24±0.02) vs (1.34±0.32), P < 0.001]. Conclusions Methylprednisolone protects LPS-induced acute lung injury possibly via suppressing STAT3-ERK1/2 signaling pathway and reducing TNF-α and IL-6 expression.
作者
宋佳
王春霞
熊熙
任玉倩
张育才
Song Jia;Wang Chunxia;Xiong Xi;Ren Yuqian;Zhang Yucai(Department of Critical Care Medicine,Shanghai Children’s Hospital,Shanghai Jiao Tong University,200062 Shanghai,China)
出处
《中华急诊医学杂志》
CAS
CSCD
北大核心
2019年第10期1266-1271,共6页
Chinese Journal of Emergency Medicine
基金
上海交通大学医学院高峰高原项目(DLY201618,20171928)
上海市科委"科技创新行动计划"临床医学领域项目(18411951000).
