右美托咪定对脂多糖诱导支气管上皮细胞炎症反应的影响

Effects of Dexmedetomidine on Inflammatory Response of Bronchial Epithelial Cells Induced by Lipopolysaccharide

目的探究右美托咪定对由脂多糖(lipopolysaccharide,LPS)诱发支气管上皮细胞活化及炎性反应的影响。方法人支气管上皮细胞系(human bronchial epithelial cell line,16HBE),经纯化鉴定后分为5组:正常对照组;LPS预处理组:将1μg/ml LPS预处理支气管上皮细胞24 h;右美托咪定低剂量组、右美托咪定中剂量组、右美托咪定高剂量组为支气管上皮细胞在1μg/ml LPS预处理24 h后分别加入5、10、20 ng/ml右美托咪定处理1 h。使用Western-blot检测c-Jun氨基末端激酶(c-Jun N-terminal kinase,JNK)和磷酸化c-Jun氨基末端激酶(phosphorylated c-Jun N-terminal kinases,p-JNK)的表达;使用逆转录-聚合酶链式反应(reverse transcription polymerase chain reaction,RT-PCR)法检测各组支气管上皮细胞培养基中的肿瘤坏死因子α(tumor necrosis factor-α,TNF-α)mRNA,白细胞介素8(interleukin-8,IL-8)mRNA和白细胞介素1β(interleukin-1β,IL-1β)mRNA的表达水平;使用酶联免疫吸附试验(enzyme linked immunosorbent assay,ELISA)测定各组支气管上皮细胞培养上清液的IL-1β、IL-8和TNF-α浓度。结果与对照组比较,LPS预处理组IL-1βmRNA、IL-8 mRNA和TNF-αmRNA表达显著上调,培养基上清液中IL-8、TNF-α、IL-1β释放显著增加,p-JNK/JNK比率明显上调,差异具有统计学意义(P<0.05);与LPS预处理组比较,右美托咪定中剂量组、右美托咪定高剂量组p-JNK/JNK比率明显下调,IL-1βmRNA、IL-8 mRNA、TNF-αmRNA表达水平及IL-8、TNF-α、IL-1β浓度明显降低,组间比较差异具有统计学意义(P<0.05),而右美托咪定低剂量组上述指标与LPS预处理组比较差异均无统计学意义(P>0.05)。结论 10 ng/ml和20 ng/ml的右美托咪定可抑制LPS诱导的支气管上皮细胞炎症反应。

Objective To explore the effects of Dexmedetomidine on lipopolysaccharide（LPS）-induced inflammatory response of human bronchial epithelial cells（16 HBE）.Methods The 16 HBE were collected and cultured invitro.After purificated and identificated,16 HBE were divided into five groups：control group,LPS group was pretreated with 1μg/ml LPS for 24 h,Dexmedetomidine low-,medium-and high-dose groups were all pretreated with LPS at 1μg/ml for24 h,and respectively added 5,10,20 ng/ml Dexmedetomidine for 1 h.The expressions of c-Jun N-terminal kinases（JNK）and phosphorylated c-Jun N-terminal kinases（p-JNK）were tested by Western blot.The expressions of tumor necrosis factor-α（TNF-α）mRNA,interleukin-8（IL-8）and interleukin-1β（IL-1β）mRNA were measured by reverse transcription polymerase chain reaction（RT-PCR）in bronchial epithelial cells of every group.The concentration of IL-8,IL-1βand TNF-αwere respectively measured by enzyme linked immunosorbent assay（ELISA）.Results Compared with control group,the expressions of TNF-αmRNA,IL-1βand IL-8 mRNA in LPS group were significantly up-regulated,meanwhile,IL-8,TNF-α,IL-1β and the ratio of p-JNK/JNK in the supernatant from medium were all increased（P〈0.05）.Compared with LPS group,the expressions and concentrations of forementioned inflammatory mediators were significantly inhibited and the ratio of p-JNK/JNK down-regulated in Dexmedetomidine medium-and high-dose groups（P〈0.05）.No significant differences were found in above mediators between Dexmedetomidine low-dose group and LPS group（P〉0.05）.Conclusion Dexmedetomidine at 10 ng/ml and 20 ng/ml can inhibit the LPS-induced inflammatory response of16 HBE.