miR-326在LPS诱导的急性肺损伤大鼠模型中的作用及对NF-κB信号通路的影响
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摘要
目的:探讨微小RNA-326(miR-326)在脂多糖(Lipopolysaccharide,LPS)诱导的急性肺损伤大鼠模型中的作用及对核转录因子κB(Nuclear factorκB,NF-κB)信号通路的影响。方法:将20只成年雌性SD大鼠根据随机数字表法分为对照组和模型组,模型组腹腔注射LPS(10mg/kg),对照组给予腹腔注射等量生理盐水,6h后麻醉处死大鼠,进行大鼠肺组织切片观察,计算大鼠肺组织湿干重比,检测肺组织中白细胞介素-1β(Interleukin-1β,IL-1β)和肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)水平,检测肺组织中miR-326、转录因子NF-E2相关因子2(nuclear factor-crythroid 2-related factor 2,Nrf2)和NF-κB mRNA的表达。结果:对照组大鼠肺组织切片未见明显异常;模型组大鼠肺组织切片见细支气管腔内有大量脱落炎细胞及渗出物,泡壁毛细血管充血严重,肺泡腔及间质内可见大量红细胞,伴有血栓形成;模型组大鼠肺组织湿干重比高于对照组,差异有统计学意义(P<0.05);模型组大鼠肺组织中IL-1β和TNF-α水平高于对照组,差异有统计学意义(P <0.05);模型组大鼠肺组织中miR-326、Nrf2和NF-κB mRNA的相对表达量均高于对照组,差异有统计学意义(P <0.05)。结论:miR-326在LPS诱导的急性肺损伤大鼠模型中表达增加,增加炎症因子的表达,其作用机制可以与激活NF-κB信号通路有关。
Objective: To investigate the role of miR-326 in lipopolysaccharide (LPS)-induced acute lung injury in rats and its effect on Nuclear factor κB (NF-κB) signaling pathway. Methods: 20 adult female SD rats were divided into control group and model group according to the random number table method. The model group received intraperitoneal injection of LPS (10 mg/kg) and the control group received intraperitoneal injection of normal saline. 6 h later,all the animals were sacrificed under anesthesia,and the pulmonary tissue section was collected for wet/dry weight ratio of lung tissue (W/D) calculation. Then the expression levels of interleukin-1β (IL-1β) and tumor necrosis factor-α (TNF-α) in lung tissues were measured by enzyme linked immunosorbent assay (ELISA),moreover,the real-time fluorescence polymerase chain reaction (RTFQ-PCR) was used to detect the expression levels of miR-326,nuclear factor-crythroid 2-related factor2 (Nrf2) and NF-κB mRNA. Results: The lung tissue sections of the control group showed no obvious abnormalities,while that of the observation group was characterized by the exfoliative inflammatory cells and exudates in the bronchiolar bronchial cavity,capillary wall congestion,red cells in the alveolar cavity and interstitial,as well as thrombus formation. The W/D of the model group was higher than that of the control group (P<0.05); The experssion levels of IL-1β and TNF-α in lung tissue of the model group were significantly higher than those of the control group (P < 0.05). The relative expression levels of miR-326,Nrf2 and NF-κB mRNA in the lung tissue of the model group were higher than those in the control group (P <0.05). Conclusion: The expression of microRNA-326 increased in LPS-induced acute lung injury rat model and increased the expression of inflammatory factors. Its mechanism may be related to the activation of NF-kappa B signaling pathway.
Objective: To investigate the role of miR-326 in lipopolysaccharide (LPS)-induced acute lung injury in rats and its effect on Nuclear factor κB (NF-κB) signaling pathway. Methods: 20 adult female SD rats were divided into control group and model group according to the random number table method. The model group received intraperitoneal injection of LPS (10 mg/kg) and the control group received intraperitoneal injection of normal saline. 6 h later,all the animals were sacrificed under anesthesia,and the pulmonary tissue section was collected for wet/dry weight ratio of lung tissue (W/D) calculation. Then the expression levels of interleukin-1β (IL-1β) and tumor necrosis factor-α (TNF-α) in lung tissues were measured by enzyme linked immunosorbent assay (ELISA),moreover,the real-time fluorescence polymerase chain reaction (RTFQ-PCR) was used to detect the expression levels of miR-326,nuclear factor-crythroid 2-related factor2 (Nrf2) and NF-κB mRNA. Results: The lung tissue sections of the control group showed no obvious abnormalities,while that of the observation group was characterized by the exfoliative inflammatory cells and exudates in the bronchiolar bronchial cavity,capillary wall congestion,red cells in the alveolar cavity and interstitial,as well as thrombus formation. The W/D of the model group was higher than that of the control group (P<0.05); The experssion levels of IL-1β and TNF-α in lung tissue of the model group were significantly higher than those of the control group (P < 0.05). The relative expression levels of miR-326,Nrf2 and NF-κB mRNA in the lung tissue of the model group were higher than those in the control group (P <0.05). Conclusion: The expression of microRNA-326 increased in LPS-induced acute lung injury rat model and increased the expression of inflammatory factors. Its mechanism may be related to the activation of NF-kappa B signaling pathway.
引文
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[2]董亮,贺宏丽,刘军,等.FLT3信号通路介导常规树突状细胞启动和调节急性肺损伤早期炎症反应的机制研究[J].中华急诊医学杂志,2016,25(11):1412~1417.
[3]Yang J,Yang Z,Lu Y,et al.1053:Imipramine improves alveolar epithelial barrier function in a murine alimodel induced by LPS[J].Critical Care Medicine,2016,44(121):339~349.
[4]金争,李冬,闫东梅,等.TRIM蛋白家族调节NF-κB信号通路的研究进展[J].中国免疫学杂志,2017,33(6):924~929.
[5]Mao L,Li J,Chen W X,et al.Exosomes decrease sensitivity of breast cancer cells to adriamycin by delivering microR-NAs[J].Tumor Biology,2016,37(4):5247~5256.
[6]温保强,刘江华,WENBao-qiang,等.LPS诱导鼠急性肺损伤模型的评价分析[J].中国医学创新,2017,12(26):137~140.
[7]Wang W,Shi L,Liu W,et al.Expression and significance of TLR-4,TNF-αand IL-1 in acute lung injury[J].Journal of Binzhou Medical University,2017.12(4):47~56.
[8]Chian C F,Chiang C H,Chuang C H,et al.SN50,a cellpermeable-inhibitor of nuclear factor-κB,attenuates ventilator-induced lung Injury in an isolated and perfused rat lung model[J].Shock,2016,46(2):194~201.
[9]Li N,Zhang X,Dong H,et al.Lithium ameliorates LPS-induced astrocytes activation partly via inhibition of toll-like receptor 4 expression[J].Cellular Physiology&Biochemistry,2016,38(2):714~725.
[10]梅颖,刘朝奇.miRNA调控肿瘤相关基因c-met的研究进展[J].中国临床药理学与治疗学,2017,22(2):204~209.
[11]Wang Q,Li D,Han Y,et al.MicroRNA-146 protects A549 and H1975 cells from LPS-induced apoptosis and inflammation injury[J].Journal of Biosciences,2017,42(4):637~645.
[12]Wu C T,Huang Y,Pei Z Y,et al.MicroRNA-326 aggravates acute lung injury in septic shock by mediating the NF-κB signaling pathway[J].The international journal of biochemistry&cell biology,2018,10(101):1~11.