【摘要】 目的 探讨长链非编码核糖核酸核内小RNA宿主基因14(long chain non-coding ribonucleic acid small nucleolar RNA host gene 14, LncRNA SNHG14)调控微小核糖核酸223-3p(microribonucleic acid-223-3p,miR-223-3p)减轻大鼠脓毒症相关性肺损伤的作用。方法 建立脓毒症大鼠模型，将大鼠随机分为SNHG14上调组、SNHG14上调对照组、SNHG14下调组、SNHG14下调对照组、miR-223-3p上调组、miR-223-3p上调对照组、miR-223-3p下调组、miR-223-3p下调对照组、模型组，另设假手术组。均经尾静脉给药，48 h后处死，取肺组织检测LncRNA SNHG14、miR-223-3p表达；检测大鼠血清肿瘤坏死因子α(tumor necrosis factor-α,TNF-α)、白细胞介素（interleukin,IL）-1β、L-18水平、肺组织湿/干重比值（wet weight/dry weight,W/D）、肺泡液体清除率（alveolar fluid clearance rate,AFC）；观察肺组织病理改变；检测肺组织人NOD样受体家族蛋白3(NOD like receptor family protein 3,NLPR3)、半胱氨酸天冬氨酸蛋白酶1(cysteine-requiring aspartate protease 1,Caspase-1)、1-氨基环丙烷-1-羧酸合酶（1-aminocyclopropane-1-carboxylate synthase,ACS）基因和蛋白表达；双荧光素酶报告基因实验验证LncRNA SNHG14对miR-223-3p的靶向调控作用。结果 与假手术组比较，模型组肺组织LncRNA SNHG14表达、血清炎症因子水平、W/D、病理改变量化评分、肺组织NLPR3、Caspase-1、ACS表达均升高（P<0.05），而miR-223-3p表达、AFC均下降（P<0.05）。与模型组和相应对照组比较，SNHG14上调组LncRNA SNHG14表达升高（P<0.05），且SNHG14上调组与miR-223-3p下调组miR-223-3p表达、AFC均下降（P<0.05），血清炎症因子水平、W/D、病理改变量化评分及肺组织NLPR3、Caspase-1、ACS表达均升高（P<0.05）。与模型组和相应对照组比较，SNHG14下调组LncRNA SNHG14表达下降（P<0.05）,SNHG14下调组与miR-223-3p上调组miR-223-3p表达、AFC均增加（P<0.05），血清炎症因子水平、W/D、病理改变量化评分及肺组织NLPR3、Caspase-1、ACS表达均下降（P<0.05）。LncRNA SNHG14与miR-223-3p存在结合位点，且前者可负反馈靶向调控后者。结论 下调LncRNA SNHG14靶向增加了miR-223-3p表达、并抑制了NLRP3通路减轻脓毒症相关性肺损伤，与抑制炎症反应有关；而上调LncRNA SNHG14则负反馈靶向了miR-223-3p、激活了NLRP3通路加重脓毒症相关性肺损伤。更多还原

【Abstract】 Objective To investigate the role of long chain non coding ribonucleic acid(LNcRNA) small nucleolar RNA host gene 14(SNHG14) in regulating microribonucleic acid 223-3p(miR-223-3p) in alleviating sepsis associated lung injury in rats. Methods Sepsis rat models were established and the rats were randomly divided into SNHG14 upregulation group, SNHG14 upregulated control group, SNHG14 downregulation group, SNHG14 downregulation control group, miR-223-3p upregulation group, miR-223-3p upregulation control group, miR-223-3p downregulation group, miR-223-3p downregulation control group, and model group. Sham operation group was set up simutalously. All of them were administered through the tail vein. After 48 hours, the lung tissues was euthanized to detect the expressions of LncRNA SNHG14 and miR-223-3p. Tumor necrosis factor α(TNF-α), interleukin-1β(IL-1β), interleukin-18(IL-18), lung wet weight/dry weight ratio(W/D) and the alveolar fluid clearance rate(AFC) were detected. Pathological changes in lung tissues were observed. Human NOD like receptor family protein 3(NLPR3), cysteine-requiring aspartate protease 1(caspase-1), and 1-aminocyclopropane-1-carboxylate synthase(ACS) genes and proteins expressions in lung tissues were detected. The dual luciferase reporter gene experiment was used to verify the targeted regulatory effect of LncRNA SNHG14 on miR-223-3p. Results Compared with the sham operation group, the model group showed increase in lung tissue LncRNA SNHG14 expression, serum inflammatory factor levels, W/D, pathological change quantification score, and lung tissue NLPR3, caspase-1 and ACS expressions(P<0.05), decrease in miR-223-3p expression and AFC(P<0.05). Compared with the model group and corresponding control groups, the SNHG14 upregulation group showed increase in LncRNA SNHG14 expression(P<0.05), and the SNHG14 upregulation group and the miR-223-3p downregulation group showed decrease in miR-223-3p expression and AFC(P<0.05), and increase in the levels of serum inflammatory factors, W/D, quantitative scores of pathological changes, and the expressions of NLPR3, caspase-1 and ACS in lung tissues(P<0.05). Compared with the model group and corresponding control groups, the expression of LncRNA SNHG14 in the SNHG14 downregulation group decreased(P<0.05), while the expression of miR-223-3p and AFC in the SNHG14 downregulation group and miR-223-3p upregulation group increased(P<0.05), which showed decrease in the levels of serum inflammatory factors, W/D, quantitative scores of pathological changes, and the expressions of NLPR3,caspase-1 and ACS in lung tissues(P<0.05). There were binding sites between LncRNA SNHG14 and miR-223-3p, and the former could negatively feedback targeted to regulate the latter. Conclusion Downregulation of LncRNA SNHG14targets an increase in miR-223-3p expression and inhibit the NLRP3 pathway to alleviate sepsis related lung injury, which is related to the inhibition of inflammatory response, while upregulation of LncRNA SNHG14 negatively feedback targeting miR-223-3p and activated the NLRP3 pathway to exacerbate sepsis related lung injury.更多还原