| 摘要: |
| 目的 探讨右美托咪定（DEX）在脓毒症小鼠肺泡上皮细胞炎性反应中的作用。方法 将雄性C57BL/6小鼠随机分为盲肠结扎穿孔（CLP）组和CLP+DEX组，每组36只。CLP组小鼠在CLP术前15 min腹腔注射1 mL无菌生理盐水，CLP+DEX组小鼠在CLP术前15 min腹腔注射50 μg/kg DEX。记录小鼠CLP术后24 h内的存活率，在CLP术后0、6、12、24 h时取小鼠血清和肺泡灌洗液，用酶联免疫吸附实验（ELISA）检测血清和肺泡灌洗液中白细胞介素（IL）-6、IL-1β、肿瘤坏死因子α（TNF-α）的水平。体外培养小鼠肺泡上皮细胞MLE12，分为脂多糖（LPS）组（1 μg/mL LPS）和LPS+DEX组（1 μg/mL LPS+0.2 μg/mL DEX），处理0、6、12、24 h后用ELISA法检测细胞上清液中IL-6、IL-1β、TNF-α的水平，蛋白质印迹法检测细胞外信号调节激酶（ERK）1/2和c-Jun氨基末端激酶（JNK）的磷酸化水平。结果 CLP+DEX组小鼠CLP术后24 h内的存活率高于CLP组（P<0.05），6、12、24 h时小鼠血清和肺泡灌洗液中IL-6、IL-1β、TNF-α的水平均低于CLP组（P<0.05，P<0.01）。6、12、24 h时LPS+DEX组MLE12细胞上清液中IL-6、IL-1β、TNF-α的水平和ERK1/2磷酸化水平均低于LPS组（P<0.05，P<0.01），6、12 h时JNK的磷酸化水平也低于LPS组（P<0.05，P<0.01）。结论 DEX能减少脓毒症小鼠血清和肺泡灌洗液中炎性因子的产生，提高脓毒症小鼠存活率，其机制可能与抑制ERK1/2和JNK信号通路的激活有关。 |
| 关键词: 脓毒症 右旋美托咪定 脂多糖 肺泡上皮 小鼠 |
| DOI：10.16781/j.0258-879x.2018.04.0388 |
| 投稿时间：2017-12-04修订日期：2018-03-29 |
| 基金项目: |
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| Protective effects of dexmedetomidine on alveolar epithelial cells in sepsis mice |
| XIE Fang1,ZHAO Han-wei2,YANG Kai1,CHEN Yuan-jie1,WAN Xiao-jian1,ZHU Ke-ming1* |
(1. Intensive Care Unit, Department of Anesthesiology, Changhai Hospital, Navy Medical University(Second Military Medical University), Shanghai 200433, China;
2. Intensive Care Unit, No. 123 Hospital of PLA, Bengbu 233015, Anhui, China
*Corresponding author) |
| Abstract: |
| Objective To explore the role of dexmedetomidine (DEX) in the inflammatory response of alveolar epithelial cells in sepsis mice. Methods Male C57BL/6 mice were randomly divided into cecal ligation and puncture (CLP) group and CLP+DEX group (n=36). The mice in the CLP group were intraperitoneally treated with 1 mL sterile normal saline and the mice in the CLP+DEX group were intraperitoneally injected with DEX (50 μg/kg) at 15 min before CLP. The survival rate of mice was recorded within 24 h after CLP. The serum and bronchoalveolar lavage fluid (BALF) were collected on 0, 6, 12, 24 h after CLP, and the levels of interleukin (IL)-6, IL-1β and tumor necrosis factor α (TNF-α) were detected by enzyme-linked immunosorbent assay (ELISA). The mouse alveolar epithelial cell lines MLE12 were cultured in vitro, and were divided into lipopolysaccharide (LPS) group (1 μg/mL LPS) and LPS+DEX group (1 μg/mL LPS+0.2 μg/mL DEX). The levels of IL-6, IL-1β and TNF-α in the cell supernatants were measured by ELISA, and the phosphorylation levels of extracellular signal-regulated kinase (ERK) 1/2 and c-Jun N-terminal kinase (JNK) were determined by Western blotting on 6, 12 and 24 h of cell culture. Results Compared with the CLP group, the survival rate of mice was significantly higher in the CLP+DEX group within 24 h after CLP (P<0.05). The IL-6, IL-1β, and TNF-α levels of serum and BALF were significantly lower in the CLP+DEX group than those in the CLP group (P<0.05, P<0.01). Compared with the LPS group, the levels of IL-6, IL-1β and TNF-α were significantly lower in the MLE12 cell supernatant of the LPS+DEX group on 6, 12 and 24 h of cell culture (P<0.05, P<0.01). Western blotting results showed that the phosphorylation levels of ERK1/2 on 6, 12 and 24 h of cell culture and the phosphorylation levels of JNK on 6 and 12 h of cell culture were significantly lower in the LPS+DEX group than those in the LPS group (P<0.05, P<0.01). Conclusion DEX can reduce the production of inflammatory cytokines in the serum and BALF of sepsis mice and increase the survival rate in sepsis mice, which may be related to the inhibition effect of DEX against activation of ERK1/2 and JNK signal pathways. |
| Key words: sepsis dexmedetomidine lipopolysaccharide alveolar epithelium mouse |
